MX2008005125A - Antimicrobial collagenous constructs - Google Patents

Abstract

Bioengineered collagen constructs with antimicrobial properties are provided. The bioengineered collagen constructs comprise a sheet-like layer of purified collagenous tissue matrix derived from a tissue source, such as the tunica submucosa of small intestine or a processed intestinal collagen layer derived from the tunica submucosa of small intestine, treated with an antimicrobial agent. The constructs arc biocompatible. The present invention has a variety of applications, including wound dressing and surgical repair devices. Methods for treating a damaged or diseased soft tissue are provided. Methods for treating a wound in need of care and treatment are also disclosed.

Description

ANTIMICROBIAL COLLAGEN CONSTRUCTIONS Field of the Invention This invention is in the field of regenerative medicine and tissue building engineering. The invention is directed to bioconstructed constructions prepared from processed tissue material or matrix, derived from animal sources. The bioconstructed constructions of the invention are prepared using methods that preserve the biocompatibility, cell compatibility, strength and capacity for bio-remodeling of the processed tissue matrix. Antimicrobial properties are imparted to bioconstructed constructions, which are used for grafts, implants, tissue repair, wound repair and remodeling, or other use in a mammalian host. Brief Description of the Background of the Invention The field of regenerative medicine and engineering or tissue technique combines the methods of technique or construction with the principles of life science to understand the structural and functional relationships in normal and pathological mammalian tissues. The purpose of regenerative medicine and tissue building is the development and final application of biological substitutes to rebuild, maintain and improve tissue functions. Collagen is the main structural protein in the body and constitutes approximately one third of the total body protein. It comprises most of the organic matter of the skin, tendons, bones and teeth, and occurs as fibrous inclusions in most other body structures. Some of the collagen properties include high tensile strength; low antigenicity, due in part to the masking of potential antigenic determinants by the helical structure; and low extensibility, semipermeability, and solubility. In addition, collagen is a natural substance for cell adhesion. These properties and others make collagen an adequate material for tissue construction and the production of biocompatible implantable substitutes and bio-removable prostheses. Methods for obtaining collagen tissue and tissue structures from explanted mammalian tissues and processes to construct prostheses from these tissues and tissue structures have been extensively investigated to heal wounds, repair surgeries and to replace tissues or organs. It is a continuous purpose of search engines to develop bioconstructed constructions that can be easily used to raise the standard of medical care that patients receive. Brief Description of the Invention Biologically derived collagen materials such as the intestinal submucosa are used in the repair or replacement of tissues and continue to be developed and improved. New bioconstructed and biorremodelables constructions are now imparted with antimicrobial properties to improve their performance characteristics in regenerative medicine, including wound healing and tissue repair and replacement. Methods for mechanical and chemical processing of the proximal porcine jejunum are described to generate an acellular, simple layer of intestinal collagen (ICL) derived from the intestinal submucosa that can be used to form laminates with antimicrobial properties to heal, repair and replace. Processing removes cells and cell debris while maintaining the native collagen tissue matrix structure. The resulting sheet from the processed fabric matrix is used to prepare a single layer and a multilayer laminate, crosslinked constructions with desired specifications. The efficacy of single-layer wound dressing products, multi-layered laminated patches for soft tissue repair as well as the use of tubular constructions as a vascular graft has been investigated. This processed tissue material derived from the intestine provides the necessary physical support, while generating minimal adhesions, and is capable of integrating into the surrounding native tissue and becomes infiltrated with host cells. Bioremodelation in vivo does not compromise the mechanical integrity of these constructions. The intrinsic properties and functional characteristics of the implant, such as the modulus of elasticity, suture retention and ultimate tensile strength are important parameters that can be manipulated for specific requirements by varying the number of layers and the crosslinking conditions. Antimicrobial qualities are now imparted for these constructions to control or decrease the microbial activity in the treatment site where these constructions are used. It is an object of the invention to provide a bioconstructed collagen construct with antimicrobial properties comprising a laminated tissue layer layer of purified collagen tissue derived from a tissue source, such as the tunica submucosa of the small intestine or a layer of collagen from the intestine. processed derivative of the tunica submucosa of the small intestine, treated with an antimicrobial agent. When the bioconstructed collagen construct of the invention is used as a bandage or wound dressing for treating the wound of a mammalian patient, the construction is applied to the wound base to substantially cover the wound in such a way that the tissue of the wound itself The patient is provided with a moist environment to facilitate the regeneration of the skin tissue while the antimicrobial agent in the construction controls or decreases the activity in the wound. The construction is biocompatible, meaning that the construction is non-cytotoxic, does not cause dermal desensitization and does not cause primary skin irritation. In one embodiment, the wound dressing comprises a sheet of processed intestinal collagen derived from the tunica submucosa of the small intestine having a thickness between about 0.05 to about 0.07 mm and an antimicrobial agent. Given the sheet-like geometry of the purified tissue matrix, it can be layered and then the layers chemically bonded together to provide a multilayer construction. Therefore, another embodiment is a construct comprising two, or more, layers of purified tissue matrix that have been joined together and treated with an antimicrobial agent. The constructions of the invention can be meshed, perforated or fenestrated either for better conformation to a wound base, better drainage of wound exudates, or both. It is a further object in this aspect of the invention to treat a wound that needs care and treatment, particularly one that needs antimicrobial intervention and protection, where the wound comprises any of one of the following types of wounds: partial and thickness wounds Total, pressure ulcers, diabetic ulcers, chronic vascular ulcers, tunnel / subsurface wound injuries, surgical wounds, donor site wounds for autografts, post-moh surgery wounds, wounds of post-laser surgery, wound dehiscence, trauma wounds, abrasions, lacerations, second degree burns, lacrimal or drained skin wounds. It is another object of the invention to provide a surgical repair device such as a patch or mesh, for the treatment and repair of soft tissues and organs, comprising two or more layers, such as between two and ten potatoes, of processed intestinal collagen derived of the tunica submucosa of the small intestine that are bonded and cross-linked together to form a multi-layered construct that is biocompatible and bio-removalable that, when implanted in damaged or diseased soft tissue, undergoes controlled biodegradation that occurs with cell replacement living adequate in such a way that the original implanted prosthesis is remodeled by the living cells of the patient. It is a further object in this aspect of the invention to provide a method for treating a damaged or diseased soft tissue in need of antimicrobial repair and intervention, comprising an implant of a prosthesis comprising two or more chemically bonded, superimposed layers of intestinal collagen. processed submucosa derived from the tunica small intestine, connected with an antimicrobial agent, which, when implanted in damaged or diseased soft tissue, undergoes controlled biodegradation that occurs with the replacement of adequate living cell in such a way that the original implanted prosthesis is remodeled by the living cells of the patient. For example, damaged or diseased soft tissue that needs repair are wounds, abdominal and thoracic wall defects, muscle flap reinforcement, rectal and vaginal prolapse, pelvic floor reconstruction, hernias, suture line reinforcement and reconstructive procedures. Detailed Description of the Invention The invention is directed to a bioconstructed collagen construct (e.g., prosthesis, graft) comprising a laminated tissue layer layer of purified collagen tissue, a processed tissue material, derived from native tissues, such as the processed intestinal collagen layer derived from the tunica submucosa of the small intestine, treated with an antimicrobial agent. The bioconstructed collagen construct can be either a single layer of processed tissue matrix or it can be a variety of bonded, superimposed layers of the processed tissue matrix. When the bioconstructed collagen construct of the invention is used to treat a wound of a mammalian subject, it is applied to the wound bed to substantially cover the wound such that the skin tissue of the subject itself is provided as well as an environment. moist to facilitate regeneration of skin tissue and an antimicrobial composition to control or decrease activity microbial in the wound and in the periphery of the wound. When the bioconstructed collagen construct of the invention is used as a surgical device, it is implanted to an implant site in a mammalian subject to serve as a repair of functioning, augmentation or replacement of the body part or tissue structure. The antimicrobial agent controls or decreases microbial activity in the wound or implant site by preventing the adhesion and proliferation of bacteria in the construct. The antimicrobial constructions of the invention are biocompatible, meaning that the construct is non-cytotoxic, does not cause dermal desensitization and does not cause primary skin irritation. The prostheses of the invention are also "bio-remodelables", which means that they will undergo controlled biodegradation that occurs concomitantly with remodeling and replacement with new endogenous matrix provided by the host or patient cells to create new tissue. In this way, a prosthesis of this invention when imparted with an antimicrobial agent and when used as a replacement tissue, has multiple properties. First, it functions as a body part that replaces or covers the wound. Second, while still functioning as a substitute body part, it functions as a tempering of remodeling for the inward growth of host cells. Third, it provides activity local antimicrobial to the treatment site. The prosthetic material of this invention is a processed tissue matrix of purified collagen developed from collagen tissue derived from the mammal that can bind to itself or another purified tissue matrix, processed and imparted with antimicrobial properties to form the prosthesis for grafting or implanting a site on a subject's body that requires treatment. The invention includes methods for making prostheses constructed of tissue from processed tissue material where the methods do not require adhesives, sutures, or staples to join the layers together while maintaining the bio-removability of the prostheses. The terms "processed tissue matrix" and "processed tissue material" mean a native, normally cellular tissue that has been procured from an animal source, preferably a mammal, and mechanically clean and concomitant tissues and chemically clear of cells, cellular debris and provided substantially free of extracellular matrix components without collagen. The processed tissue matrix, while substantially free and purified from components without collagen, maintains much of its native matrix structure, organization, strength, and shape. Compositions of processed tissue material for preparing the bioconstructed grafts of the invention are derived from animal tissues comprising collagen with such collagen tissue sources which include, but are not limited to: intestine, dermis, fascia lata, pericardium, dura mater, placenta, and other flat and planar structures comprising a collagen tissue matrix. The structure and geometry of these tissue matrices makes them capable of being easily cleaned, manipulated and assembled into a form for preparing the bioconstructed grafts of the invention. Other suitable tissue sources with a similar planar structure, geometry and matrix composition can be identified, procured and processed by the skilled artisan in other animal sources according to the invention. One of such tissue matrix composition processed to prepare the bioconstructed grafts of the invention is a layer of intestinal collagen derived from the tunica submucosa of the small intestine. Suitable sources for the small intestine are mammalian organisms such as human, cow, pig, sheep, dog, goat or horse while the pig small intestine is an easily available source. The prostheses of the invention can be prepared from the processed intestinal collagen layer (sometimes called the "intestinal collagen layer" or "ICL") which is a processed tissue material derived from the tunica submucosa of the porcine small intestine. In a method to obtain this layer of intestinal collagen, the small intestine is harvested of a mammal and concomitant mesenteric tissues are excessively severed from the intestine. The tunica submucosa is separated, or delaminated, from the other layers of the small intestine by mechanically compressing the intestinal raw material such as between opposite rollers similar to those of a machine that covers sausages to eliminate the muscular layers (tunica muscularis) and the mucosa (tunica) mucous membrane). When the tunica submucosa of the small intestine is harder and more rigid than the surrounding tissue, the rollers compress the softer components of the submucosa, resulting in a mechanically clean tissue matrix. In the examples that follow, the porcine small intestine was mechanically cleaned using an intestine cleaning machine and then chemically purified or cleaned in a series of solutions to provide a processed tissue matrix. This mechanically and chemically cleansed intestinal collagen layer derived from the tunica submucosa of the small intestine is herein referred to as "i CL" and is a type of matrix or processed tissue material from which the antimicrobial constructions of the invention are prepared. .

In the composition, the processed ICL tissue material is a Type I cellular telopeptide collagen, approximately 93% by dry weight, with less than about 5% dry weight of glycoproteins, glycosaminoglycans, proteoglycans, lipids, proteins without collagen and acids nucleics such as DNA and RNA and is substantially free of cells and cell debris. The processed ICL fabric material retains much of its matrix structure and strength. Importantly, the biocompatibility and bio-removability of the tissue matrix is preserved in part by the cleaning process as it is free of bound detergent residues that would adversely affect the bio-removability of the collagen. Additionally, the collagen molecules have retained their telopeptide regions when the tissue has not undergone treatment with enzymes during the cleaning process. To obtain the processed tissue matrix, an appropriate animal and a tissue source is determined. The tissue is processed both mechanically and chemically to remove concomitant tissues and to remove collagen-free components from the tissue to result in a matrix of processed tissue. By way of example, ICL is a type of processed tissue matrix used in the production of the bioconstructed graft prostheses of the invention. The methods described below are following the process tissue to provide a processed tissue matrix and to fabricate bioconstructed graft prostheses comprising ICL and an antimicrobial agent. To obtain porcine ICL, the tunica submucosa of the porcine small intestine is used as a starting material for the bioconstructed graft prostheses of the invention. He A pig's small intestine is harvested, the concomitant tissues are removed and then the intestine is cleansed mechanically using an intestinal cleansing machine that forcibly removes fat, muscle layers and mucosal from the tunica submucosa using a combination of mechanical action and washing using water The mechanical action can be described as a series of rollers that compress and stretch out the successive layers of the tunica submucosa when the intact intestine runs between them. The submucosa tunica of the small intestine is harder and more rigid compared to the surrounding tissue, and the rollers compress the softer components of the submucosa. Other means of mechanical cleaning in the art can be determined by the skilled artisan to include other physical manipulation such as scraping, squeezing, compressing and rubbing. The result of mechanical cleaning is such that the submucosal layer of the intestine remains only, a mechanically cleansed intestine. After mechanical cleaning, a chemical cleaning treatment is used to remove mechanically cleaned cell and matrix components from the intestine, preferably performed under aseptic conditions at room temperature. The mechanically cleansed intestine is the descending longitudinal bowel of the lumen and then cut into sections of approximately 15 cm to 50 cm in length. HE material weight and it was placed in containers in a ratio of approximately 100: 1 v / v of solution to the intestinal material. In most of the preferred chemical cleaning treatment, such as the method described in U.S. Patent Nos. 5,993,844 and 6,599,690 to Abraham, the descriptions of which are incorporated herein, the collagen tissue is contacted with an effective amount of chelating agent, such as tetrasodium salt of ethylenediaminetetraacetic acid (EDTA) under alkaline conditions, such as by the addition of sodium hydroxide (NaOH); followed by contact with an effective amount of acid where the acid contains a salt, such as hydrochloric acid (HCl) containing sodium chloride (NaCl); followed by contact with an effective amount of buffered saline such as 1 M sodium chloride (NaCl) / phosphate buffered saline (PBS); finally followed by a rinse stage using water. Each treatment step is preferably carried out using a rotation or stirring platform to improve the actions of the chemical and rinsing solutions. The result of the cleansing processes is a layer of processed intestinal collagen, or ICL, a matrix of mechanically processed tissue and cleaned chemical derived from the tunica submucosa of the small intestine. After rinsing, the ICL is then removed from the cleaning containers and gently compressed or dried to remove the excess Water. At this point, the ICL can be stored frozen at -80 ° C, at 4 ° C in sterile phosphate buffer, or dried until it is fabricated into a prosthesis. If stored dry, the ICL sheets are flattened on a surface such as a flat plate, preferably a porous plate or membrane, such as a polycarbonate membrane, and any marks on the abluminal side of the material are removed using a scalpel, and The ICL sheets can be left to dry in a laminar flow hood at room temperature and humidity. The ICL is a flat sheet structure that can be used as a single layer material or to manufacture various types of constructions that are used as prostheses in the form of prostheses that ultimately depend on their intended use. To form multilayer prostheses of the invention, the ICL sheets are laminated using a method that continues to preserve the biocompatibility and bio-removability of the processed matrix material but also that is able to maintain its strength and structural characteristics for its realization as a replacement fabric. The processed tissue-derived tissue matrix retains the structural integrity of the native tissue matrix, that is, the collagen matrix structure of the original tissue remains substantially intact and maintains physical properties so that it would exhibit many intrinsic and functional properties when implanted . When they prepare laminated with several layers of ICL, the ICL sheets are covered to contact with other sheets. The contact area is a link region where the layers contact each other, any of the layers are directly superimposed on each other, or partially in contact or overlap for the formation of more complex structures. In completed constructions, the binding region must be able to withstand suturing and tension while being manipulated in the clinic, during the implant and during the initial healing phase while functioning as a replacement body part. The binding region must also maintain sufficient resistance until the population of the patient's cells and subsequently the bio-remodeling of the prosthesis forms a new tissue. The processed tissue matrix is used as a single-layer prosthesis or is formed in a joined prosthesis, of flat, tubular or complex-shaped layers. When the prostheses of the invention comprise two or more layers or matrix of processed tissue, the layers are joined by chemically crosslinking the layers together using a crosslinking agent. While chemical crosslinking is used to join multiple layers of processed tissue matrix together, the degree of chemical crosslinking can be varied to modulate bio-remodeling rates throughout the prosthesis, which is the speeds at which a prosthesis is both reabsorbed as replaced by host and tissue cells. In other words, the higher degree of crosslinking imparted to the prostheses of the invention, the decrease in the rate of bio-remodeling of the prostheses will suffer, adversely, the lower degree of cross-linking, the faster the rate of bio-remodeling. Surgical indications dictate the degree and / or speed of bio-remodeling required by the prosthesis. For example, when a single layer construction is used as a wound dressing, the prosthesis may or may not be chemically crosslinked. For example, as a surgical repair patch, or mesh, the prosthesis is a multi-layered construction that has a low degree of crosslinking so that the prosthesis would be bio-remodeled at a faster rate. For example, as a bladder sling to support a hypermovable bladder to prevent urinary incontinence, the prosthesis is a multi-layered construction that has a high degree of crosslinking so that the prosthesis is not bio-remodeled so fast, that is, it persists in substantially the same conformation in which it is implanted over a longer period of time. The collagen matrix or construction, when in laminated form, generally has two surfaces of large, opposite area. To treat the collagen matrix processed with an antimicrobial agent, the antimicrobial agent is applied by contacting it either laterally of the Collagen matrix processed or they can be bound on both sides. Alternatively, the fibers, absorbent qualities of the processed collagen matrix may be effective to apply the antimicrobial agent to the interior of the processed collagen material, such as in the interstices of the fibrous processed tissue matrix, such as by immersing the collagen matrix in a solution that contains the antimicrobial agent and that allows the solution to penetrate the matrix through absorption. Another method of providing the antimicrobial agent within a multilayer construction is to treat single layers of the processed fabric matrix and then laminate and bond the layers together. For multilayer configurations, the methods include conducting the manufacturing steps of treating the matrix sheets with an antimicrobial agent, layered the sheets of the matrix to form multiple layers and crosslinking the construction with a crosslinking agent in any order , including the following: treating the matrix sheets with an antimicrobial agent, layering the sheets of the matrix to form multiple layers, then crosslinking with a crosslinking agent; treat the sheets of the matrix with an antimicrobial agent, crosslink with a crosslinking agent, then layered the sheets of the matrix to form multiple layers; crosslink with a crosslinking agent, treat the sheets of the matrix with an antimicrobial agent, then put layered the sheets of the matrix to form multiple layers; crosslinking with a crosslinking agent, layering the sheets of the matrix to form multiple layers, then treating the matrix sheets with an antimicrobial agent; layered the sheets of the matrix to form multiple layers, crosslink with a crosslinking agent, then treat the sheets of the matrix with an antimicrobial agent; or, layered the sheets of the matrix to form multiple layers, treat the sheets of the matrix with an antimicrobial agent, then crosslink with a crosslinking agent. In some cases, the cross-linking of the matrix may not be practical after coating with an antimicrobial agent, as part of the antimicrobial agent may be washed out by the cross-linking agent. Untreated and treated layers can be layered in different arrangements to provide a prosthesis having a localized antimicrobial agent. Methods for forming such multilayer configurations including processed tissue matrix sheets in layers that have been treated with an antimicrobial agent and untreated matrix sheets together to form a multilayer construction and crosslink the layers together. For example, at least two matrix sheets can be layered, crosslinked, and antimicrobially treated to form a treated matrix construction, and then one or more untreated matrix sheets can be layered either or both of the upper and lower surfaces of the treated matrix construction, and then the resulting construction can be crosslinked to form a combination construction. Alternatively, at least two untreated matrix sheets can be layered and crosslinked to form an untreated matrix construction, and then one or more treated matrix sheets can be layered on either or both of the upper and lower surfaces of the matrix. untreated matrix construction, and then the resulting construction can be cross-linked to form a combined matrix construction. In another example, treated and untreated matrix sheets can be alternately layered and then crosslinked to form a combination construct. In yet a separate example, treated matrix sheets that have been treated with two different antimicrobial agents may be arranged in an alternating or other order to provide a combination construct. In still yet another example, the orientation of the treated and untreated sheets together, or for matrix constructions treated or untreated in a combination matrix construction can be configured by taking advantage of the quality without many sides of the ICL material. To treat only selected parts or areas of a prosthesis, portions of the surface of the material can be treated with an antimicrobial agent by masking portions of the surfaces that are treated in such a way that the mask obstructs the antimicrobial agent from contacting the material while allowing other areas of the surface to be treated. Another way of locating an antimicrobial agent in a collagen matrix is to partially submerge the collagen material in a bath or container such that only a portion of the collagen matrix contacts the antimicrobial agent and other portions remain free of contact with the collagen matrix. antimicrobial treatment. Yet another way to locate the antimicrobial agent on the surface of the material is to spray, or otherwise propel, the antimicrobial agent onto a surface of the material while leaving the opposite surface untreated. Single-layer or multi-layer constructions are treated with an antimicrobial agent to impart antimicrobial properties to the construction. At least one antimicrobial agent is applied to the constructions of the invention by contacting all or only part of the construction to the antimicrobial agent. Preferred antimicrobial agents include silver-based antimicrobial agents and chemical-based antimicrobial agents. An antibiotic agent can also be included in the composition. A combination of agents can be used to treat the collagen material to provide a broad spectrum of antimicrobial activity, for example, an antimicrobial agent based in silver and an antimicrobial agent based on chemistry; an antimicrobial agent based on chemistry and an antibiotic agent; an antimicrobial agent based on silver and an antibiotic agent; or a combination of the three types of agents. Antimicrobial agents based on silver can be selected to impart antimicrobial properties to prostheses comprising a processed tissue matrix. Silver can be applied to collagen constructions in various forms. Antimicrobial agents based on silver include silver or compounds containing silver that have some degree of antimicrobial activity and are compatible with both the collagen construct and the patient. Pure silver, also referred to as elemental or noble silver, is relatively chemically inactive and is not reacted with water or oxygen at normal temperatures and is not soluble in dilute acids and bases. Ionic silver can also be used. It is believed that silver in ionic form has not yet produced microbial resistance and it seems less likely to do otherwise antimicrobial agents. While it is desired to be bound by the theory, silver in this ionic form is effective against bacteria, yeast and fungi and extracellular viruses directly affecting cellular and cell wall respiration and transport as well as reproduction. Silver in others forms can also be used, including but not limited to: silver oxide; silver nitrate; sulfazidine silver (silver sulfadiazine (I) comprises an insoluble polymer compound that releases silver ions slowly in its role as an antimicrobial and antifungal agent and can be used topically in the treatment of severe burns to prevent bacterial infection); silver imidazoate; arglaes (AgKaP04); Colloidal Silver; Silver crystals, such as silver nanocrystals, also called "nanocrystalline silver" is another way of providing and prolonged release of silver cation and its radicals for a rolled or implant site. Nanocrystalline silver compositions ("nanoplata") are made according to different different methods. One method, a pulsatile plasma method, produces nanocrystals with an organized crystal lattice structure that lacks atomic disorder but exhibits tight particle size distribution and morphology with a high degree of fidelity. The pulsatile plasma process uses two rods of conductive feedstock. The rods are fed into the reaction chamber filled with controlled gas, most likely an inert gas such as argon, at atmospheric pressure. The rods are connected to a powerful, pulsating discharge power supply. The nanomaterial is synthesized by rapidly discharging power from the supply of Pulsatile power through the feed material rods. The high-power discharge eliminates the raw material to create a high-pressure, high-temperature metal plasma. Because using single gas dynamics, the plasma rapidly expands into the surrounding gas to create a homogeneous gas-phase suspension of nanoparticles. The nanoparticles produced are continuously collected using a closed mesh system. A blower recirculates the controlled gases transporting the particles to the collection system. This method produces nanocrystalline silver particles from 10 nm to 100 nm in diameter depending on the parameters of the process. Generally, particle sizes are selected from 15 nm to 40 nm, or from 20 nm to 25 nm for use in the invention. Other methods for making nanosilver compositions include those described in U.S. Patent No. 6,719,987 to Burrell. These methods create crystals that have a crystal lattice characterized by atomic disorder. In the Burrell process, the material that is deposited is generated in the vapor phase, for example by evaporation or bubbling, and is transported in a large volume in which the temperature is controlled. Atoms of the colloid material with atoms of the atmosphere of gas processing, energy lost and are quickly condensed from the vapor phase on a substrate cold, such as a cooled index of liquid nitrogen. Atomic disorder is created by conditions which limit diffusion in such a way that sufficient atomic disorder is retained in the material. The deposition is conducted at low substrate temperatures for silver, from -10 ° to 100 ° C; the elaboration of gas pressures at higher than the normal values are used; angles of incidence lower than approximately 30 °; and higher deposition rates that create a higher than the normal atom flux. The atomic disorder can also be achieved by the presence of different atoms or molecules in the metal matrix by a process called "doping" or by the incorporation of reactive gases (ie, oxygen) into the chamber. According to this method, oxygen is a constituent in the process gas. Chemical-based antimicrobial agents can be applied to collagen constructs to impart antimicrobial properties to constructions. While there is no exhaustive list, chemical antimicrobial agents can be selected from the following: poly (hexamethylene biguanide) hydrochloride (PHMB); Chlorhexadine gluconate; bis-amido polybiguanides such as those described in U.S. Patent No. 6,316,669, the disclosure of which is incorporated herein; honey; benzalkonium chloride, triclosan (2,4,4'-trichloro-2'-hydroxydiphenylether); and salt of quaternary ammonium silyl (octadecyl-demethyl chloride) trimethoxysilyl propyl ammonium). In addition to an antimicrobial agent, the collagen construct may additionally comprise an antibiotic agent. An antibiotic agent is one that is produced by microorganisms to destroy or inhibit the growth of other microorganisms. Generally, antibiotics are molecules of low molecular weight (not protein) produced as secondary metabolites, mainly by microorganisms that live in the earth. Most of these microorganisms form some type of spore or other latent cell, and some are considered to be related between antibiotic production and sporulation processes. Among the molds, notable antibiotic producers are Penicillium and Cephalosporium, which are the main source of beta-lactam antibiotics that include penicillin and related compounds. In bacteria, Actinomycetes, notably Streptomyces species, produce a variety of types of antibiotics including aminoglycosides, for example, streptomycin, macrolides, e.g., erythromycin, and tetracyclines. Bacillus species that form the endospore produce polypeptide antibiotics such as polymyxin and bacitracin. Antibiotics can have a cidal effect, which is a "destructive" effect, or a static effect, which means an "inhibitory" effect, on a range of microbes. The spectrum of action of an antibiotic agent is the range of bacteria or other microorganisms are affected by it. Effective antibiotics against prokaryotes that destroy or inhibit a wide range of Gram-positive and Gram-negative bacteria are called "broad spectrum"; those effective mainly against Gram-positive or Gram-negative bacteria are "narrow spectrum"; and those effective against an organism or simple disease are "limited spectrum". A preferred antibiotic compound that is used is a broad spectrum antibiotic. Antibiotic compounds can be provided with the collagen construct in combination, such as a combination of a narrow spectrum Gram-positive compound and a narrow spectrum Gram-negative compound; however, any combination of broad, narrow and narrow spectrum antibiotic range can be used. Antibiotics for use in the invention include: beta-lactams (penicillins and cephalosporins), such as penicillin G, cephalothin; semi-synthetic penicillin (which may also include clavulanic acid), such as ampicillin, amoxicillin and methicillin; monobactams, such as aztreonam; carboxypenems, such as imipenema; aminoglycosides, such as streptomycin; gentamicin; glycopeptides, such as vancomycin; lincomycins, such as clindamycin; macrolides, such as erythromycin; polypeptides, such as polymyxin; bacitracin; polyenes, such as amphotericin; nystatin; rifamycins, such as rifampicin; tetracyclines, such as tetracycline; semi-synthetic tetracycline, such as doxycycline; and chloramphenicol. The processed tissue matrix can be used as a single layer alone in a single layer prosthesis or used to fabricate prostheses having two or more layers. If used as a single layer, the antimicrobial agent is contacted with the processed tissue matrix to impart antimicrobial properties to the matrix. Either before or after contact with an antimicrobial agent, the processed tissue matrix can be cross-linked to control the rate of bio-remodeling and biodegradation of the material. In other words, the single-layer antimicrobial constructions are: single layer and treated with an antimicrobial agent; simple layer crosslinked with a crosslinking agent is then treated with an antimicrobial agent; or the single layer treated with an antimicrobial agent then crosslinked with a crosslinking agent. Methods for manufacturing multilayer prostheses comprising two or more layers of processed tissue matrix are described using ICL. One embodiment of the invention is directed to flat sheet prostheses, and methods for making and using flat sheet prostheses, comprising two or more ICL layers bonded and crosslinked for use as an implantable biomaterial capable of being bio-remodeled by cells. of the patient. Due to the ICL flat blade structure, the prosthesis is easily made to comprise any number of layers, preferably between 2 and 10 layers, more preferably between 2 and 6 layers, with the number of layers depending on the strength and volume required for the final intended use of the construction. The ICL has structural matrix fibers that run in the same general direction. When in layers, the orientations of the layer can be varied for the effectiveness of the general orientations of tissue fiber in the layers of processed tissue. The sheets can be layered so that their fiber orientations are in parallel or at different angles. The layers can also be superimposed to form a construction with continuous layers through the area of the prosthesis. Alternatively, since the final size of an overlying arrangement is limited by the circumference of the intestine, the layers may be staggered, in mounting arrangement to form a sheet construction with a surface area larger than the dimensions of the starting material but without continuous layers through the area of the prosthesis. Complex features such as a conduit or network of conduit or channels running between the layers or traversing the layers, for example, may be introduced. In the manufacture of a multilayer construction comprising ICL, an aseptic environment and sterile tools are preferably employed to maintain construction sterility when starting with the sterile ICL material.

To form a multilayer ICL construction, a first rigid sterile support member, such as a rigid polycarbonate sheet, is placed in the sterile field of a laminar flow cabinet or cabinet. If the ICL sheets are not yet in a hydrated state from the mechanical and chemical cleaning processes, they are hydrated in aqueous solution, such as water or phosphate buffered saline. The ICL sheets are dried with sterile absorbent cloths to absorb excess water from the material. If they are not yet done, the ICL material is adjusted from any of the lymphatic markers on the serosal surface, on the abluminal side. A first adjusted ICL sheet is placed on the polycarbonate sheet and is manually smoothed to the polycarbonate sheet to remove any air bubbles, bends and creases. A second adjusted ICL sheet is placed on top of the first sheet, again manually removing any bubbles, folds and creases. This is repeated until the desired number of layers is obtained for a specific application, preferably between 2 and 10 layers. ICL has a less lateral quality of a native tubular state: an internal mucosal surface that is coated from the intestinal lumen in the native state and an opposite exterior serosal surface that is coated with ablumen. It has been found that these surfaces have characteristics that can affect the post-operative realization of the prosthesis but can be effective for the realization of the improved device. Currently with the use of synthetic devices, the adhesion formation may need the need for re-operation to release the adhesions from the surrounding tissue. In the formation of a pericardial patch or hernia repair prosthesis having two ICL layers, it is preferred that the binding region of the two layers is between the serosal surfaces as the mucosal surfaces have been shown to have a capacity to resist the formation of post-operative adhesion after implantation. In other embodiments, it is preferred that one surface of the ICL patch prosthesis be non-adhesive, non-adherent and the other surface have an affinity for adhering to the host tissue. In this case, the prosthesis would have a mucosal surface and the other serosal surface. In yet another embodiment, it is preferred that the opposing surfaces be capable of creating adhesions to develop tissues together that come into contact on either side, so that the prosthesis would have serosal surfaces on both sides of the construction. Because only the two outer sheets are potentially contacted, other body structures when implanted, the orientation of the inner layers, if the construction is comprised of more than two, is of minor importance as it will probably be without contributing to the formation of post-operative adhesion.

After the formation of layers of the desired number of ICL sheets, the sheets are then joined by dehydrating them together in their binding regions, that is, when the sheets are in contact. While not wishing to be bound by theory, dehydration causes the collagen fibers of the ICL layers to come together when the water is removed from between the fibers of the ICL matrix. The layers can be dehydrated either open faced on the first support member or, between the first support member and a second support member, such as a second polycarbonate sheet, placed before drying on the upper layer of ICL and secured to the first support member to keep all the layers in a smooth planar arrangement together with or without a small amount of pressure. To facilitate dehydration, the support member may be porous to allow air and moisture to pass through the dewatering layers. The layers may be dried in air, in a vacuum or by chemical means such as by acetone or an alcohol such as ethyl alcohol or isopropyl alcohol. The dehydration can be completed at ambient humidity, between about 10% Rh to about 20% Rh, or less; or from about 10% to about 20% w / w moisture, or less. Dehydration can easily be performed at the angle of the frame fastener the polycarbonate sheet and the ICL layers up to the face of the air flow next to the laminar flow cabinet for at least about 1 hour to 24 hours at room temperature, about 20 ° C and at room humidity. While not necessary, in one embodiment, the dehydrated layers are rehydrated before crosslinking. The dehydrated ICL layers are detached from the porous support member together and rehydrated in an aqueous rehydration agent, preferably water, by transferring them to a vessel containing the aqueous rehydration agent for at least about 10 to about 15 minutes at a temperature between about 4 ° C to about 20 ° C to rehydrate the layers without separating or delaminating them. The dehydrated, or dehydrated and rehydrated bound layers are then cross-linked together to the binding region by contacting the ICL in layers with a cross-linking agent, preferably a chemical cross-linking agent, preferably a chemical cross-linking agent that preserves the bio -Removable material of ICL. As mentioned above, dehydration leads to the collagen fibers in the matrices of the adjacent ICL layers together and crosslinking those layers together forms chemical bonds between the components to join the layers. The cross-linking of the attached prosthetic device also provides strength and durability to the device to improve the properties of handling. Various types of crosslinking agents are known in the art and can be used such as ribose and other sugars, oxidizing agents and dehydrothermal methods (DHT). A preferred crosslinking agent is 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). In another preferred method, sulfo-N-hydroxysuccinimide is added to the EDC crosslinking agent as described by Staros, J.V., Biochem. 21, 3950-3955, 1982. In addition to chemical crosslinking agents, the layers can be joined together with fibrin-based gums or medical grade adhesives such as polyurethane, vinyl acetate or polyepoxy. In the most preferred method, EDC is solubilized in water at a concentration of preferably between about 0.1 mM to about 100 mM, more preferably between about 1.0 mM to about 10 mM, most preferably at about 1.0 mM. In addition to water, phosphate buffered saline or buffer (2- [N-morpholino] ethanesulfonic acid) (MES) can be used to dissolve the EDC. Other agents can be added to the solution, such as acetone or an alcohol, up to 99% v / v in water, typically 50%, to make crosslinking more uniform and efficient. These agents remove water from the layers to bring the matrix fibers together to promote crosslinking between those fibers. The ratio of these agents to the water in the crosslinking agent can be used to regulate the crosslinking. The EDC crosslinking solution is prepared immediately before using as EDC would lose its activity over time. To contact the crosslinking agent with the ICL, the bound, hydrated ICL layers are transferred to a container such as a shallow pan and the crosslinking agent gently decant to the pan ensuring that the ICL layers are both covered and floatable free and that air bubbles are not present under or within the layers of the ICL constructions. The container is covered and the ICL layers are allowed to crosslink between about 4 to about 24 hours, more preferably between 8 to about 16 hours at a temperature between about 4 ° C to about 20 ° C. The crosslinking can be regulated with temperature: at lower temperatures, the crosslinking is more effective as the reaction is decreased; at higher temperatures, the crosslinking is less effective as the EDC is less stable. After crosslinking, the crosslinking agent is decanted and disposed of and the constructions are rinsed in the pan by contacting them with a rinse agent to remove the residual crosslinking agent. A preferred rinse agent is water or another aqueous solution. Preferably, sufficient rinsing is achieved by contacting the chemically bonded construction three times with equal volumes of sterile water for approximately five minutes for each rinse. As described herein, antimicrobial properties are imparted to the constructions by contacting them with an antimicrobial agent either by contacting, or treating, each layer of tissue matrix individually processed or by contacting, or treating, an intermediate structure of several layers. The method of treating the tissue matrix processed in simple or multilayered form with an antimicrobial agent will vary with the type of antimicrobial agent used but must be one that preserves the bio-remodelable, biomechanical and biocompatible properties of the processed tissue matrix. When nanocrystalline silver is selected as the antimicrobial agent, it can be applied to the collagen matrix material by contacting the collagen matrix material to the nanocrystalline plate. The antimicrobial agent can be applied by coating the processed matrix material by coating it upon suspending the agent in solution. The nanocrystalline silver is dispersed in water, aqueous solution, or in a solvent to form a dispersed nanocrystalline silver solution. The ICL is immersed in a tray with a volume of nanocrystalline silver solution such that, when the ICL is immersed, the nanocrystalline silver adheres to the ICL. The solution may also be agitated or agitated during the immersion. The ICL is then removed from the solution and placed in an environment that allows the water or solvent to evaporate from the ICL with silver attached to result in an ICL construction with a nanocrystalline silver coating. Other methods for coating ICL with a dispersion containing nanocrystalline silver include spraying and allow the solvent to evaporate from the ICL. When PHMB is selected as the antimicrobial agent, the PHMB is added to the solution using 0.09% - 0.5% in v / v water in which the processed tissue matrix is immersed in such a way that the PHMB solution saturates the matrix. After a sufficient time of saturation of the solution in the processed tissue matrix, the matrix is removed from the solution and allowed to dry in such a way that the PHMB remains in the matrix when the solvent evaporates. The processed tissue matrices and the constructions can be treated or modified, either physically or chemically, before or after the manufacture of a multi-layer, attached graft prosthesis. Physical modifications such as training, conditioning by stretching and relaxation, or drilling, meshing or fenestration of clean tissue matrices and constructions can be performed. The conditioning decreases the total strain of the material while the perforation, meshing or fenestration provides for either the best conformation for a wound bed or better passage and drainage of exudates, or both. Chemical modifications such as growth factors of link, selected extracellular matrix components, genetic material, and other agents that would affect the bio-remodeling and repair of the body part that is treated, repaired, or replaced can also be performed. Methods for physically modifying the tissue matrix and constructions of the invention can be determined by one of skill in the art when considering the performance characteristics required of the constructions. The constructions may be provided with a design of perforations communicating across opposite sides of the construction using a pressure with a mold having needles, blades, or pins arranged in the pattern on the face of the mold. The construction is placed on a platform and the mold is pressed in such a way that the needles, blades or pins are pressed through the construction to the platform while the construction is punctured. A method for providing cuts or a mesh design uses a skin meshing apparatus similar to those currently used in skin grafting procedures derived from itself. An appliance is a mesh-former of Zimmer's skin. The constructions can be laser-drilled to create miera-sized pores through the completed prosthesis to aid in cell inward growth using an excimer laser (e.g. at KrF or ArF wavelengths). The constructions can be perforated, fenestrate or laser pierced, but is preferably done before decontamination or terminal sterilization. For some indications it is preferred that the perforations or holes drilled with laser communicate through all the layers of the prosthesis to assist in passage of the cell or drainage of fluid. For other indications, it is preferred that it does not pass all the way through the layers so that the holes provide cellular access to the interior of a multilayer construction or to aid in the neovascularization of the construction. Other physical modifications for the construction of simple layer are perforations or fenestrations that communicate between both sides of the construction. Another physical modification for the simple layer construction is the construction meshing, which is, to provide an arranged design of cuts for the construction so that it is compared to a mesh. The cuts are made in a design with a mesh ratio similar to that provided by a skin mesh former such as those made by Zimmer. The mesh ratios can be set from 1: 1.5 to 2: 1. To make a single layer ICL construction with antimicrobial properties, the ICL is spread on the mucosal downward side in a smooth polycarbonate sheet, ensuring the elimination of folds, air bubbles and visual lymphatic markings. The scattering of the ICL on the sheet of Polycarbonate is made to optimize the dimensions. The material is optionally dried suitably on its entire surface. At least one antimicrobial agent is then applied to the material. The material is optionally modified in a physical way by meshing or perforation and then cut to size and packaged and finally sterilized by sterilization specifications. In an alternate embodiment, the antimicrobial treatment is applied to the material after the material has been physically modified with a mesh, perforations or fenestrations. After the processed tissue matrix is prepared and covered with an antimicrobial agent, the constructions are adjusted to the desired size. For illustration, a usable size is about 15.2 cm x 15.2 cm (about 6 square inches) but any size can be prepared and used to graft a patient. The antimicrobial treated collagen matrix is then packaged in a container that is sealed for final sterilization, storage and distribution. The antimicrobial treated collagen matrix can be packaged in the container in a dry or wet state. Preferred packaging materials are compatible with the antimicrobial treatment, the collagen matrix and, if packaged in a wet state, any agents that maintain the moisture of the product. For constructions treated with sensitive antimicrobial agents to light, such as silver-based antimicrobial agents, packaging materials that prevent or filter the passage of light are used to package the product to prevent the reduction of antimicrobial activity and discoloration of the constructions. The constructions are then finally sterilized using means known in the medical device sterilization art. A preferred method for sterilization is by contacting the constructs with sterile 0.1% neutralized peracetic acid (PA) treatment with a sufficient amount of 10 N sodium hydroxide (NaOH), according to U.S. Patent No. 5,460,962 , the description of which is incorporated herein. Decontamination is performed in a container on a shaker platform, such as 1 L Nalge containers, for approximately 18 ± 2 hours. The constructions are then rinsed by contacting them with three volumes of sterile water for 10 minutes each rinse. In a more preferred method, the ICL constructs are sterilized using gamma irradiation between 25-37 kGy. Gamma irradiation significantly, but not detrimentally, decreases Young's modulus, ultimate tensile strength, and contraction temperature. The mechanical properties after gamma irradiation are still sufficient for use in a range of applications and gamma is a means preferred for sterilization as widely used in the field of implantable medical devices. Dosimetry indicators are included with each run or sterilization test to verify that the dose is within the specific range. The constructions are packaged using a packaging material and designated to be compatible with the composition of the construction and ensure sterility during storage. A preferred packaging means is a double-layer peelable package where the main package is a hermetically sealed blister package comprised of a tray modified with polyethylene terephthalate glycol (PETG) with a thin-film top of paper surface which is enclosed in a secondary heat seal bag comprised of a polyethylene / polyethylene terephthalate (PET) laminate. Together, both the primary and secondary packaging and the ICL construction contained herein are sterilized using gamma radiation. The prostheses of this invention can be flat, tubular or of complex geometry. The conformation of the formed prosthesis would be decided by its intended use. Thus, when bonding layers of the prosthesis of this invention are formed, the mold or support member of the plate can be made to accommodate the desired shape. Flat multi-layer prostheses can implanted to repair, augment or replace damaged or diseased organs, such as the abdominal wall, pericardium, hernias and various other organs and structures including, but not limited to, bone, periosteum, perichondrium, intervertebral disc, articular cartilage, dermis, intestine , ligaments, and tendons. In addition, flat multi-layer prostheses can be used as a vascular or intra-cardiac patch, or as a replacement heart valve. Flat sheet prostheses, whether in single or multiple layers, are used in wound healing to cover a subject's wound to form a wet barrier, to provide comfort and comfort to the wound site and to provide antimicrobial activity as The site of the wound is stabilized to initiate healing. Flat blades for supporting the organ can also be used, for example, to support prolapsed or hypermobile organs using the blade as a sling for the organs, such as bladder or uterus. Tubular prostheses can be used, for example, for replacement through sections of tubular organs such as vasculature, esophagus, trachea, intestine, and fallopian tubes. These organs have a basic tubular shape with an external surface and an internal luminal surface. In addition, flat sheets and tubular structures can be formed together to form a complex structure to replace or augment the cardiac or venous valves.

The ICL material used in the manufacture of the antimicrobial constructions of the invention are biocompatible. The biocompatibility test has been performed on prostheses made of ICL according to both Tripartita and ISO-10993 guidelines for biological evaluation of medical devices. "Biocompatible" means that the prostheses are non-cytotoxic, hemocompatible, non-pyrogenic, endotoxin-free, non-genotoxic, non-antigenic, and without provoking a dermal sensitization response, without eliciting a primary skin irritation response, without case of toxicity acute systemic, and without causing subchronic toxicity. These biocompatible qualities are discussed in more detail below. Test articles of ICL-prepared constructs shown without biological reactivity (Grade 0) or cytotoxicity observed in L929 cells after the exposure period test article when using the test entitled "L929 Agar Cover Test for In Vitro Cytotoxicity" (L929 Agar Overlay Test for Cytotoxicity In Vitro). The cellular response observed for the positive control article (Grade 3) and the negative control article (Grade 0) confirm the validity of the test system. Tests and assessments were conducted in accordance with USP guidelines. The prostheses of the invention are considered non-cytotoxic and meet the requirements of the Cover Test with Agar L929 for Cytotoxicity ln Vitro. Hemocompatibility test (in vitro hemolysis, using the modified ASTM extraction method test, in vitro) of the prosthesis of the invention was conducted according to the modified ASTM extraction method. Under the conditions of the study, the mean hemolytic index for the device extract was 0% while positive and negative controls were performed as anticipated. The results of the study indicate that the prostheses of the invention are nonhemolytic and hemocompatible. The prostheses of the invention were subjected to a pyrogenicity test following the current USP protocol for the pyrogen test in rabbits. Under study conditions, the total increase in rabbit temperatures during the observation period was within acceptable USP limits. The results confirmed that the prostheses of the invention are non-pyrogenic. The prostheses of the invention are free of endotoxin, preferably at a level of < 0.06 EU / ml (per cm2 of product). Endotoxin refers to a particular pyrogen that is part of the cell wall of gram-negative bacteria, which is poured by bacteria and contaminating materials. The prostheses of the invention do not elicit a dermal sensitization response. There are no reports in the literature that would indicate that the chemicals used to clean porcine intestinal collagen cause a sensitization response, or I would modify the collagen to elicit a response. The sensitization test results in the prostheses of the invention formed from chemically cleaned ICL indicate that the prostheses do not elicit a sensitization response. The prostheses of the invention do not elicit a primary skin irritation response. The results of the chemically cleaned ICL irritation test indicate that the prostheses of the invention formed from chemically cleaned ICL do not elicit a primary skin irritation response. Acute systemic toxicity test and intracutaneous toxicity were performed on chemically cleaned ICL used to prepare prostheses of the invention, the results of which demonstrate a lack of toxicity among the prostheses tested. Additionally, in animal implant studies there was no evidence that porcine intestinal collagen chemically cleaned caused acute systemic toxicity. The subchronic toxicity test of the prostheses of the invention containing porcine intestinal collagen confirmed lack of subchronic toxicity of the device. There were no reports in the literature that would indicate that the chemicals used to clean the porcine intestinal collagen would affect the potential for genotoxicity, or modify the collagen to cause such a response. The test of genotoxicity of the prostheses of the invention containing porcine intestinal collagen confirmed lack of genotoxicity of the device. The purpose of the chemical cleaning process for porcine intestinal collagen used to prepare prostheses of the invention is to minimize antigenicity by elimination of cells, cell remnants, and matrix components without collagen and non-elastin. The prostheses of the invention containing porcine intestinal collagen confirm a lack of antigenicity of the device, as confirmed by implant studies conducted with porcine intestinal collagen chemically cleaned. The ICL constructs of the invention are preferably interpreted to be virally inactivated. In the manufacturing process, the effectiveness of two chemical cleaning procedures, the alkaline chelating solution of NaOH / EDTA (pH 11-12) and the HC / NaCl acidic saline solution (pH 0-1), was tested to inactivate four relevant viruses and models. The model viruses were chosen based on the source of porcine material, and to represent a wide range of physical-chemical properties (DNA, RNA, hidden and non-hidden viruses). The viruses included are pseudo-rabies virus, bovine viral diarrhea virus, reovirus-3 and porcine parvovirus. The studies were conducted based on FDA and ICH guidance documents, including: CBER / FDA "Points to Consider in the Characterization of Cell Lines Used to Produce Biologicals (1993)"; ICH "Note for Guidance in Quality of Biotechnological Products: Evaluation of Viral Safety of Biotechnological Products Derived from Cell Lines of Human or Animal Origin" (CPMP / ICH / 295/95); and, Part of CPMP Biotechnology Development "Note for Guidance in Virus Validation Studies: The Design, Contribution and Interpretation of Studies that Validate the Inactivation and Elimination of Viruses" (CPMP / BWP / 268/95). The results of the study show that the cumulative viral inactivation of the two stages of chemical cleaning is an evacuation of more than 106 for the four model viruses. The data indicate that chemical cleaning procedures are a strong and effective process that maintains the potential for inactivation of a wide variety of viral agents. The prostheses of the invention are bio-remodelables. While functioning as a substitute body part, the prostheses of the invention also function as a bio-removable matrix scaffold for the inward growth of host cells. "Bio-remodeling" is used herein to mean the production of endogenous structural collagen, vascularization, and cellular repopulation by growing inwardly from host cells at a rate approximately equal to the rate of biodegradation, reformation and replacement of the matrix components of the implanted prosthesis by host cells and enzymes. Graft prostheses retain their structural characteristics while being remodeled by the subjects in whom they are implanted in all, or substantially all, host tissue, and as such, are functional as an analogous repair or replacement tissue. In addition to these bio-remodelables qualities, the prostheses of the invention made of two or more layers of processed tissue matrix are prepared to incorporate desirable biomechanical properties. Young's modulus (MPa) is defined as the linear proportional constant between stress and strain. The Final Drive Resistance (N / mm) is a measurement of the resistance through the prosthesis. Both of these properties are a function of the number of ICL layers in the prosthesis. When used as a carrying load or support device, it must be able to withstand the rigors of physical activity during the initial healing phase and remodeling throughout. The rolling resistance of the bonding regions is measured using an adhesion test. Immediately after the surgical implant, it is important that the layers are not delaminated under physical stress. In animal studies, unexplained materials showed any evidence of delamination. Before implantation, the adhesion force between two opposing layers is approximately 8.1 ± 2.1 N / mm by a multilayer construction crosslinked by 1 mM EDC. Shrinkage Temperature (° C) is an indicator of the degree of crosslinking of the matrix. The higher the contraction temperature, the more crosslinked matrix is in the material. An ICL irradiated with gamma, not reticulated has a contraction temperature of approximately 60.5 ± 1.0 ° C. In the preferred embodiment, cross-linked EDC prostheses would preferably have a shrinkage temperature between about 64.0 ± 0.2 ° C to about 72.5 ± 1.1 ° C for devices that are crosslinked in 1 mM EDC to about 100 mM EDC in 50% of acetone, respectively. The mechanical properties include mechanical integrity such that the prosthesis resists creep during bio-remodeling, and is additionally flexible and suturable. The term "flexible" means good handling properties for easy clinical use. The term "suturable" means that the mechanical properties of the layer include suture retention that allows needles and suture materials to pass through the prosthesis material at the time of suturing the prosthesis to sections of native tissue. During suturing, such prostheses should not tear as a result of the tension forces applied to them by the suture, they should not tear when the suture is knotted The ability to suture the prosthesis, that is, the ability of the prosthesis to resist tearing while being sutured, is related to the intrinsic mechanical strength of the prosthesis material, the thickness of the graft, the tension applied to the suture, and the speed to which the knot is terminated forced. The suture retention of a flat, highly crosslinked 6-layer prosthesis is crosslinked in 100 mM EDC and 50% acetone is approximately 6.7 ± 1.6 N. The suture retention for a 2-layer prosthesis cross-linked in 1 mM EDC in water is approximately 3.7 N ± 0.5 N. The preferred lower suture retention strength is approximately 2N for a flat 2-layer, cross-linked prosthesis as a surgeon force in suturing is approximately 1.8 N. As used herein , the term "without dragging" means that the biomechanical properties of the prosthesis imparts durability so that the prosthesis is not stretched, distended, or expanded beyond normal limits after implantation. As described below, the total stretch of the implanted prosthesis of this invention is within acceptable limits. The prosthesis of this invention acquires a resistance to stretching as a function of post-implant bio-remodeling by replacing structural collagen with host cells at a faster rate than the loss of resistance mechanics of implanted materials due to biodegradation and remodeling. The processed woven material of the present invention is "semi-permeable", although it has been layered and bonded. The semi-permeability allows the inward growth of host cells to remodel or for deposition of agents and components that would affect bio-remodelability, cell-cell growth, adhesion or promotion prevention, or blood flow. The "non-porous" quality of the prosthesis prevents the passage of fluids destined to be retained by the implant of the prosthesis. Adversely, pores, perforations, fenestrations, cuts or a mesh can be formed in the prosthesis if pore quality or perforated is required for an application of the prosthesis. The mechanical integrity of the prosthesis of this invention is also in its ability to be draped or bent, as well as the ability to cut or adjust the prosthesis obtaining a clean edge without delaminating or tearing the edges of the construction. A sheet of processed intestinal collagen derived from the tunica submucosa of the small intestine normally has a thickness between about 0.05 to about 0.07 mm. Since the geometry as lamina of the purified tissue matrix can be layered and then chemically bonded together to provide a multilayer construction with greater thickness. The processed tissue matrix layers of the bonded, multi-layer prosthetic device of the invention may be of the same collagen material, such as two or more layers of ICL, or of different collagen materials, such as one or more ICL layers. and one or more layers of fascia lata (thigh apneurosis). Constructs with antimicrobial properties can be used for the management or control of wounds including: partial and total thickness wounds, pressure ulcers, venous ulcers, diabetic ulcers, chronic vascular ulcers, tunnel / weakened wounds, surgical wounds (such as donor site wounds for autografts, post-Moh's surgery wounds, post-laser surgery wounds, wound dehiscence), traumatic wounds (such as abrasions, lacerations, second-degree burns, and skin cuts) and drainage wounds The wound dressing is a single layer sheet or a multi-layer construction of porcine intestinal collagen mechanically and chemically cleaned, at least one antimicrobial agent, each sheet of processed tissue matrix is approximately 0.05 to approximately 0.07 mm in thickness, containing Fenestrations that communicate between both sides of the sheets. The product comprises mainly porcine type I collagen (approximately> 95%) in its native form, with less than about 0.7% lipids and undetectable levels of glycosaminoglycans (approximately <0.6%) and DNA (approximately <0.1 ng / μ?). Porcine intestinal collagen is substantially free of cells and cell remnants. The wound dressing of the invention may or may not be crosslinked, but cross-linked, cross-linked to a degree to regulate and control biodegradation, bio-remodeling, or replacement of the bandage by patient cells. Physical modifications can be provided to the wound advantage to allow the passage of fluids. The wound dressing provides a moist healing environment, a barrier to control and minimize bacterial contamination, and pain relief to a patient by covering the sensory nerve terminals. An antimicrobial agent provides effective protection against microbial contamination in and around the wound site.

The wound dressing is applied to a patient with a wound that needs treatment. Before application, the wound area can be cleansed and debrided from scar tissue using techniques that standard wound dressing. Preferably, debridement is to a degree where the edges of the wound contain viable tissue. To apply the bandage of the invention, it is cut to the contour of the injured area. If the wound is larger than a simple bandage piece, multiple pieces may be used and overlap to provide full wound cover. If the bandage is in a dry state, it can be rehydrated using sterile saline or other isotonic solution. The edge of the bandage should be in contact with the intact tissue then smoothed instead of ensuring that the bandage is in contact with the bed of the fundamental wound. If excess exudate is collected under the sheet, small openings can be cut into the sheet to allow the exudate to drain. The antimicrobial wound dressing of the invention can be used as an interface between the wound and the conventional or secondary wound dressing. Alternatively, the antimicrobial wound dressing of the invention can be used as a cover for a skin replacement, such as an autograft, allograft or cultured skin construct. When used as a cover, the wound dressing provides a moist wound environment such that cells from either skin replacement can populate the wound bed to enclose the wound faster. After application, a secondary, non-adherent dressing suitable for maintaining a moist wound environment is preferably applied. The optimal secondary dressing is determined by the location of the wound, size, depth and preference of the user. The secondary bandage can be changed as needed to maintain a clean, moist wound arch. The frequency of bandage changes will vary depending on the type, size and depth of the wound which is treated, the volume of exudates produced and the type of bandage used. When healing occurs, the wound dressing may need to be replaced in which case additional applications of wound dressing can be performed until the healing is completed. Other embodiments of the invention are directed to multilayer constructions imparted with antimicrobial properties. The prosthetic device of this invention has two or more overlapping collagen layers that are bonded together. As used herein, "collagen bonded layers" means a composite of two or more layers of the same or different collagen material treated in such a way that the layers are superimposed on each other and sufficiently held together by semi-lamination and cross-linking. chemistry. More specifically, the prosthetic device is a surgical mesh or graft intended to be used for implant to reinforce soft tissue including, but not limited to: abdominal and thoracic wall defects, muscle flap reinforcement, rectal and vaginal prolapse, reconstruction of the pelvic floor, hernias, bridge-shaped openings in fascial defects, suture line reinforcement and reconstructive procedures. A prosthetic mesh or graft of the invention comprises a five layer lamina of porcine ICL and an antimicrobial agent, of approximately 0.20 mm a approximately 0.25 mm in thickness. The product consists mainly of Type I porcine collagen (approximately> 95%) in its native form, with less than about 0.7% lipids and undetectable levels of glycosaminoglycans (approximately <0.6%) and DNA (approximately <0.1 ng). / μ?). Porcine intestinal collagen is substantially free of cells and cell remnants. The prostheses are supplied sterile in sheet form in sizes ranging from 5 x 5 cm to 12 x 36 cm in double layer peelable packaging. The prosthesis has a denaturing temperature of approximately 58 ± 5 ° C; a tensile strength of more than 15N; a suture retention force of more than 2 N using a braided silk suture; and, an endotoxin level of 5.0.06 EU / ml (per cm2 of product). In particular, the prosthesis is a flat sheet construction consisting of five ICL layers and an antimicrobial agent, bound and crosslinked with 1 mM with 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) in water. To form this construction, a first ICL sheet is of mucosal downward side spread on a smooth polycarbonate sheet; ensure the elimination of folds, air bubbles and visual lymphatic markings. The spreading of the ICL was done to optimize dimensions. Three sheets of ICL (mucosal descending side) are formed in layers on top of the first, ensuring the elimination of creases, bubbles air and visual lymphatic markings when each sheet is formed in layers. The fifth sheet should be layered with the mucosal side surface, ensuring the elimination of folds and air bubbles. The visual lymphatic marks are eliminated before the formation of layers of this fifth sheet. The layers are dried together for 24 + 8 hours. The layers are now dried together and then crosslinked in 1 mM EDC in water for 18 ± 2 hours in 500 mL of crosslinking solution per 30 cm of the five layer sheet. Each product is rinsed with sterile water and then cut with final size specifications while hydrating. In an alternate modality, the prosthetic device is a surgical sling with at least one antimicrobial agent that is intended for implantation to reinforce and support soft tissues where weakness exists including but not limited to the following procedures: pubourethral support, prolapse repair (urethral) , vaginal, rectal and colon), reconstruction of the pelvic floor, support of the bladder, sacrocolposuspension, reconstructive procedures and tissue repair. In another more preferred embodiment, the prosthetic device is a surgical sling comprised of three to five layers of crosslinked ICL, attached, treated with at least one antimicrobial agent. To fabricate a five-layer device, ICL is a mucosal downward side spread on a smooth polycarbonate sheet; ensuring the elimination of folds, air bubbles and visual lymphatic markings. The spreading of the ICL was done to optimize dimensions. A second, third and fourth lamina of ICL (mucosal descending side) are layered n the top of the first, ensuring the elimination of folds, air bubbles and visual lymphatic markings when each sheet is formed into layers. The fifth sheet is formed in layers with the upper surface of the mucosal side, ensuring the elimination of folds and air bubbles. Visual lymphatic markers must be eliminated before the formation of layers of this fifth sheet. (A three layer construction is made by a first ICL sheet of mucosal downward side spread on a smooth polycarbonate sheet, ensuring the elimination of folds, air bubbles and visual lymphatic markings, a second sheet of ICL (mucosal descending side) in layers in the upper part of the first, and a third layer in layers in the upper part of the second sheet with the upper surface of the mucosal side). The layers are dried for 24 ± 8 hours and dried once, crosslinked in 10 mM EDC in 90% acetone for 18 ± 2 hours in 500 mL of crosslinking solution per 30 cm of five layer sheet. Each joined, reticulated construction is rinsed with sterile water and cut to final size specifications while hydrating. Providing pubourethral support, the sling can be used for the treatment of urinary incontinence resulting from urethral hypermobility or intrinsic sphincter deficiency. The surgical sling consists of a laminated sheet of five layers of porcine intestinal collagen, approximately 0.20 mm to approximately 0.25 mm in thickness, and an antimicrobial agent. The device is cross-linked with 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). The device consists mainly of porcine type I collagen (approximately> 95%) in its native form, with less than about 0.7% lipids and undetectable levels of glycosaminoglycans (approximately <0.6%) and DNA (approximately <0.1 Ng). / μ?). Porcine intestinal collagen is free of cells and cell remnants. The denaturation temperature (DSC) of the prosthesis is greater than about 63 ° C; its tensile strength is greater than about 15N; its suture retention strength is greater than approximately 2N using a 2-0 braided silk suture; and the final endotoxin level is < _0.06 EU / ml (per cm2 of product). While the bio-remodelable aspects of the sling may be varied and actuated, the sling prosthesis of the invention is not a replacement body part, but an organ support device implanted as an assistive structure. It is preferred that the ICL layers of the sling be more highly cross-linked to reduce the bio-removability of the sling. The sling prosthesis is a structure of highly biocompatible, flexible collagen, which, when implanted, maintains required structural support and strength while functioning as an organ support device. In still another alternate modality, the prosthetic device is a hard repair patch that is intended for implantation to repair the dura mater, a strong membrane that protects the central nervous system. The hard repair device of the invention comprises four layers of cross-linked ICL, attached, and an antimicrobial agent. To fabricate a four-layer device, ICL is a mucosal downward side spread on a smooth polycarbonate sheet, ensuring elimination of folds, air bubbles and visual lymph marks. The spreading of the ICL was done to optimize dimensions. A second and third lamina of ICL (mucosal descending side) is formed in layers in the upper part of the first, ensuring elimination of folds, air bubbles and visual lymphatic markings when each sheet is layered. The fourth sheet is layered with the upper surface of the mucosal side, ensuring elimination of folds and air bubbles. The visual lymphatic marks must be eliminated before the formation of layers of this fourth sheet. The layers are dried for 24 ± 8 hours and dried once, cross-linked at about 0.1 mM to about 1 mM EDC in buffer (2- [N-] acid. morpholino] ethanesulfonic acid (MES) for 18 ± 2 hours in 500 measure crosslinking solution per 30 cm of four layer sheet. Each bonded, reticulated structure is rinsed with sterile water and cut to final size specifications while hydrating. The hard repair device consists of a laminated sheet of four layers of porcine intestinal collagen, from about 0.14 mm to about 0.21 mm thick. The device is cross-linked with 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). The device consists mainly of porcine type I collagen (approximately> 95%) in its native form, with less than about 0.7% lipids and undetectable levels of glycosaminoglycans (approximately <0.6%) and DNA (approximately <0.1 Ng). / μ?). Porcine intestinal collagen is free of cells and cell remnants. The denaturation temperature (DSC) of the prosthesis is greater than about 63 ° C; its tensile strength is greater than about 15N; its suture retention strength is greater than about 2N using a 2-0 braided silk suture; and the final endotoxin level is < 0.06 EU / ml (per cm2 of product). The hard repair device is biocompatible and bio-remodelable in such a way that, when implanted in a patient in need of hard repair, its functions as a hard replacement while overtime, is bio-remodeled by host cells that degrade both and they replace the device in such a way that a new host tissue replaces the overtime device. For example, a multilayer construction of ICL is used to repair body wall structures. It can also be used as, for example, a pericardial patch, a myocardial patch, a vascular patch, a bladder wall patch, or a hernia repair device (such as a tension-free patch or a plug) or used as a sling to support hypermobile or prolapsed organs (rectocele, prolapse vault, cystocele). The construction of several layers is useful for treating connective tissue such as a rotator cuff or capsule repair. The construction of several layers is useful for hard repair to repair skull defects after craniotomy procedures or to repair hard canal along with the spinal cord. The material is useful in annular repair when annular fibrosis is herniated (i.e., slipped disc) and used as a plug in the gap created by the slipped disc or as a hollow cover, or both. The material is useful in plastic surgery procedures such as mastopexy, abdominal surgery, and in facial plastic surgery (cheek and forehead stretches). Both simple and multilayer ICL materials can be used as a wound cover or bandage to assist in wound repair. In addition, it can also be implanted flat, rolled or bent to increase the volume of tissue and accretion. A number of ICL layers can be incorporated into the construction to increase volume or resistance indications. Prior to implantation, the layers may be further treated or coated with collagen or other components of extracellular matrix, hyaluronic acid, heparin, growth factors, peptides, or cultured cells. The following examples are provided for a better explanation of the practice of the present invention and should not be construed in any way to limit the scope of the present invention. It will be appreciated that the device designed in its composition, conformation, and thickness is to be selected depending on the final indication for construction. Those skilled in the art will recognize that various modifications may be made to the methods described herein while not departing from the spirit and scope of the present invention. EXAMPLES Example 1: Chemical Cleaning of the Mechanically Cleaned Porcine Thin Bowel The pig's small intestine was harvested and mechanically washed, using a Bitterling gut cleaning machine (Nottingham, UK) that forcibly removes the fat, muscle and mucosal layers from The tunic submucosa using a combination of mechanical action and washing with water. The Mechanical action can be described as a series of rollers that compress and pull away the successive layers of the tunica submucosa when the intact intestine is tested between them. The submucosa tunica of the small intestine is comparatively harder and more rigid than the surrounding tissue, and the rollers squeeze the softer components of the submucosa. The result of the cleaning machine was such that only the submucosal layer of the intestine remained. The remainder of the process, chemical cleaning according to International PCT Application No. WO 98/49969 of Abraham, et al., The description of which is incorporated herein by reference, is made under aseptic conditions and at room temperature. The chemical solutions were all used at room temperature. The intestine was then cut longitudinally down the lumen and then cut into sections of 15 cm. The material weight was placed in containers at a ratio of approximately 100: 1 v / v of solution to intestinal material. To each container containing intestine was added approximately 1 L of 0.22 μ? T solution? (miera) sterilized filter 100 mM tetrasodium salt of ethylenediaminetetraacetic acid (EDTA) / 10 mM sodium hydroxide solution (NaOH). The containers were then placed on a shaker table for approximately 18 hours at approximately 200 rpm. After stirring, the EDTA / NaOH solution was removed from each bottle. To each vessel was added approximately 1 L of 0.22 μ? T solution. filter sterilized 1 M hydrochloric acid (HCI) / 1 M sodium chloride solution (NaCl). The containers were then placed on a shaker table for between about 6 to 8 hours at approximately 200 rpm. After stirring, the HCI / NaCl solution was removed from each container. To each vessel was then added about 1 L of 0.22 pm solution of sterilized 1 M sodium chloride (NaCl) / 10 mM phosphate buffered saline (PBS). The containers were then placed on a shaker table for approximately 18 hours at approximately 200 rpm. After stirring, the NaCl / PBS solution was removed from each container. To each vessel was added approximately 1 L of 0.22 μ? T solution. filter sterilized PBS chloride 10 mM. The containers were then placed on a shaker table for approximately two hours at approximately 200 rpm. After stirring, phosphate-buffered saline was then removed from each container. Finally, approximately 1 L of 0.22 pm of sterilized water filter was added to each vessel. The containers were then placed on a stirring board for approximately one hour at approximately 200 rpm. After stirring, the water was then removed from each container. Processed ICL samples were cut and fixed for histological analysis. Hemotoxilin and cosine (H & E) and Masson's trichrome dye solution was performed on both cross section and longitudinal section samples of both control and treated tissues. The tissue samples of ICL processed cell-free and cell waste while the untreated control samples appear normally and expectedly very cellular. This single layer ICL material can be used as a single layer or used to form bonded multilayer constructions, or constructions with tubular complex and planar geometrical aspects. Example 2: Method for Fabricating a Multilayer ICL Construction ICL processed according to the method of Example 1 was used to form a multilayer construction having 2 ICL layers. A sterile porous polycarbonate sheet (pore size, manufacturer) was placed down in the sterile field of a laminar flow cabinet. ICL was stained with sterile TEXWIPES (LYM-TECH Scientific, Chicopee, MA) to absorb excess water from the material. The ICL material was adjusted from its lymphatic markings on the abluminal side and then in pieces of approximately 15.2 cm in length (approximately 6 inches). A first adjusted ICL sheet was placed on the polycarbonate sheet, mucosal descending side, which manually removes any air bubbles, folds and folds. A second adjusted ICL sheet was placed on the upper part of the face, or abluminal side, of the first sheet with the abluminal side of the second sheet contacting the abluminal side of the first sheet, which manually removes again any bubbles of air, folds, and creases. The polycarbonate sheet with the ICL layers was angled up to the ICL layers that cover the near air flow of the laminar flow cabinet. The layers were allowed to dry for about 18 ± 2 hours in the cabinet at room temperature, approximately 20 ° C. The dried ICL layers were detached from the polycarbonate sheet together without being separated or delaminated and transferred to a room temperature water bath for about 15 minutes to hydrate the layers. The chemical crosslinking solution of 100 mM EDC / 50% Acetone was prepared immediately before crosslinking as EDC would lose its activity over time. The hydrated layers were then transferred to a shallow pan and the crosslinking agent was gently decanted into the pan ensuring that the layers were both covered and floating free and that no air bubbles were present under or inside the constructions. The pan was covered and allowed to settle for approximately 18 ± 2 hours in a ventilation hood. The crosslinking solution was decanted and placed. The constructions were rinsed in the pan three times with sterile water for approximately five minutes for each rinse. Using a scalpel and a ruler, the constructions were adjusted to the desired size. The constructions were decontaminated with 0.1% sterile peracetic acid (PA) treatment neutralized with sodium hydroxide and 10N NaOH according to U.S. Patent No. 5,460,962, the disclosure of which is incorporated herein. The constructions were decontaminated in 1 L Nalge containers on a shaker platform for approximately 18 ± 2 hours. The constructions were then rinsed with three volumes of sterile water for 10 minutes each rinse and the PA activity was monitored by the Minncare strip test to ensure their removal from the constructions. The constructions were then packaged in plastic bags using a vacuum sealer which in turn was placed in hermetic bags for gamma irradiation between 25.0 and 35.0 kGy. Example 3: Inhibition Test Zone The purpose of this Example is to illustrate how to test for an effective amount of an antimicrobial substance at single layer constructs prepared from the processed tissue material derived from porcine intestinal submucosa (ICL) prepared for the method described in Example 1 which would provide a microbial barrier as well as a microbial destructor. Various antimicrobial substances were applied to ICL by either solubilization or suspension of the substances in solution, as follows: 1. 0.5% silver nitrate solution 2. 1% silver nitrate solution 3. 1-2% solution Nanocrystalline Silver in Alcohol Isopropyl ether (IPA) 4. 1-2% silver nanocrystalline solution in PBS 1X 5. 1-2% polyhexamethylene biguanide (PHMB) in phosphate buffered saline 6. 1-2% polyhexamethylene biguanide (PHMB) in purified water. A chemically cleaned ICL was cut into pieces of 4 cm x 6 cm and each one was placed, in both hydrated and dehydrated states, in approximately 100 mL of each solution and allowed to settle overnight. An ICL treated was then removed from the solutions and dried. To serve as a control condition, untreated ICL samples were used. Circular pieces of each sample piece were cut. Agar plates were disrupted with bacteria. The samples were plated and allowed to incubate for 24 hours for observation. The solutions of nanocrystalline silver (N.S) made in IPA completely dispersed and settle or hold in the solution, while the N.S. in PBS 1X it does not hold in the solution and the agitation required before adding the solution to the ICL. The IPA evaporated during the night and appears still and uniform in both hydrated or dehydrated pieces of ICL, the dehydrated piece maintains a definite shape and is easier to manipulate. Both pieces turned a charcoal black color. The ICL side of the descending surface on the tray had a smaller side although it does not seem to affect the coating of the ICL or the effectiveness of the material in creating a large zone of inhibition (ZOI). The products of N.S. created from the best ZOI and also created from ZOI that do not allow the growth of bacteria in the ZOI 24 hours after 4 days as something from other samples produced. Although the silver nitrate samples showed ZOI that were smaller than the N.S. but larger than others, the reported toxicity of silver nitrate does not necessarily make it the ideal additive for ICL. The known antimicrobial properties of silver nitro makes it useful for comparing N.S. against silver nitrate, and N.S. exceeds the effectiveness of silver nitrate. The PHMB solutions seem both the same, while the ICL after wetting does not. The PBS solutions leave the ICL with what appears to be a different salt residue while the WFI solutions leave the ICL without apparent change. They both gave similar ZOI. They were smaller and did not create them in such a way as with the ZOI of N.S. Both nanocrystalline silver and PHMB were found to be effective antimicrobial agents that can be applied to ICL constructs as evidenced by their bactericidal effect in this assay. Example 4: Treatment of 2-Layer Collagen Constructions Containing Antimicrobial Nanocrystalline Silver or PHMB ICL constructions of 2 laminated layers (both laminated and cross-linked) were prepared and then treated with an antimicrobial agent to produce two-layer constructions with antimicrobial qualities . Twelve ICL constructions of 2 layers, each approximately 9 cm x 9 cm in size, were prepared according to Example 2. In their preparation, these ICL constructs were crosslinked using a buffer of 10 mM EDC / 0.1 M MES [2- (N-morpholino) ethanesulfonic acid] (Pierce, Rockford, IL) for 16-20 hours and were rinsed three times in sterile filtered water for 30 minutes. The constructions were then treated with an antimicrobial agent. Antimicrobial agents were prepared either as solutions or as dispersions. Five were prepared dispersions of nanocrystalline silver by mixing 10, 1.0, 0.1, 0.01, 0.001 grams of nanocrystalline silver (Nanotechnologies (Ag-20) or equivalent, Austin, TX) in 1 L of dispersing agent, isopropyl alcohol (sterile filtered water is also a dispersing agent acceptable). A PHMB solution of 0.2%, 0.1%, 0.02%, 0.002% was prepared by mixing Cosmocil CQ (20% PHBM solution, ArchChemicals, Inc., Norwalk, CT) with RDOI / WFI. To coat the laminated and crosslinked constructions with an antimicrobial agent, the hydrated constructions were placed in trays of 9.5 cm x 9.5 cm. Each agent was decanted 50 meters in the trays, and the trays were placed on a stirring board. The coating times were 10 seconds, 1 hour, 3 hours and overnight (approximately 18 hours). With the completion of the established coating times, the samples were allowed to dry on polycarbonate sheets in the sterile air flow of a safe biological laminar flow cabinet until dried at 10-20% Rh. The resulting constructions were 2-layer ICL constructions that have been laminated, cross-linked and treated with an antimicrobial agent to impart antimicrobial qualities to the constructions. Example 5: Treatment of Single Layer Collagen Matrix with Antimicrobial Nanocrystalline Silver or PHMB Used to Fabricate 2-Layer Antimicrobial Constructions Single-layered, cross-linked ICL constructions were prepared, treated with an antimicrobial agent and then laminated to form two-layered constructions to produce two-layer processed tissue matrix constructions with antimicrobial qualities. A more antimicrobial agent was incorporated between the ICL processed tissue matrix layers by laminating the constructions after treatment with the antimicrobial agent. The ICL was prepared according to Example 1, spread on polycarbonate sheets and lymphatic markers were removed. Pieces were cut into sizes of approximately 10 cm x 9 cm. Each piece of ICL was then crosslinked in a buffer of 10 mM EDC / 0.1M MES [2- (N-morpholino) ethanesulfonic acid] (Pierce, Rockford, IL), 2 liters per 26 pieces and stirred on an established shaker board a, 4, for 16-20 hours and then rinsed three times at a minimum of 2 L in sterile filtered water for a minimum of 20 minutes per rinse. Antimicrobial agents were prepared either as solutions or as dispersions. Three nanocrystalline silver dispersions were prepared by mixing 1.0, 0.1, 0.01 grams of a nanocrystalline silver composition (Nanotechnologies (Ag-20) or equivalent, Austin, TX) in 1 L of dispersing agent, such as isopropyl alcohol (sterile filtered water). it is also an acceptable dispersing agent). HE prepared PHMB solutions of 0.2%, 0.1%, 0.02% by mixing Cosmocil CQ (20% solution of PHMB, ArchChemicals, Inc., Norwalk, CT) with RODI / WFI. To cover ICL with an antimicrobial agent, pieces were placed in sterile containers of 125 square meters (Nalgene) with four pieces per container. 100 ml of solution or dispersion containing the antimicrobial agent was decanted in each container to submerge the constructions. The ICL remained submerged for 3-6 hours while the vessels were agitated on a rotating shaker platform. The treated antimicrobial ICL was then layered to form two-layer constructions by placing a treated piece of ICL flat against a polycarbonate sheet. A second treated piece of ICL was then placed directly on top and spread over the first such that no air bubbles appeared between the layers. The polycarbonate sheets with the constructions were then placed in the sterile air flow of a laminar flow biological safety cabinet until dried at 10-20% Rh for a minimum of 12 hours. The resulting constructions were 2-layer ICL constructions that have been cross-linked and treated with an antimicrobial agent in such a way that it imparts antimicrobial qualities to the constructions, and then laminates of such way that they incorporate an antimicrobial agent not just on the exterior surfaces of the constructions but also between the layers of the constructions. Example 6: Treatment of 2-Layer Collagen Constructions Containing Antimicrobial Nanocrystalline Silver ICL constructions of 2 laminated layers (both laminated and cross-linked) were prepared and then treated with an antimicrobial agent to produce two-layer constructions with antimicrobial properties. Twelve ICL constructions of 2 layers, each approximately 35-40 cm x 9 cm in size, were prepared according to Example 2. During their preparation, these ICL constructs were cross-linked using a 10 mM EDC buffer / MES 0.1 M [2- (N-morpholino) ethanesulfonic acid] (Pierce, Rockford, IL) in water for 16-20 hours and rinsed three times in sterile filtered water for 30 minutes. The prepared constructs were then treated with an antimicrobial agent. Antimicrobial agents were prepared as dispersions.

Three dispersions of nanocrystalline silver were prepared by mixing 10., 5.0, 1.0 grams of nanocrystalline silver (Nanotechnologies (Ag-20) or equivalent, Austin, TX) in 1 L of isopropyl alcohol as a dispersing agent for nanocrystalline silver (sterile filtered water as well). he is an agent acceptable dispersant). To cover the constructions with an antimicrobial agent, the constructions were placed in 1 L square containers (Nalgene) with four constructions per container. 200 ml of solution or dispersion containing an antimicrobial agent was decanted in each container to submerge the constructions. The constructions remained submerged for 3-6 hours while the vessels were agitated on their side on a rotating stirring platform. Constructions that have been contacted with nanocrystalline silver dispersions once in 1 L of sterile filtered water were rinsed. Antimicrobial treated constructions were then placed flat on polycarbonate sheets in the sterile air flow of a laminar flow biological safety cabinet until dried at 10-20% Rh for a minimum of 12 hours. The resulting constructions were 2-layer ICL constructions that have been laminated, cross-linked and treated with an antimicrobial agent to impart antimicrobial qualities to the constructions. Example 7: Treatment of a Single-Layer Collagen Matrix with Antimicrobial Nanocrystalline Silver Used to Fabricate 2-Layer Antimicrobial Constructions Single layer ICL constructions were prepared reticulated, cross-linked, treated with an antimicrobial agent and then laminated to form two-layer constructions to produce two-layer constructions with antimicrobial qualities. By laminating the constructions after treatment with the antimicrobial agent, it is possible to incorporate more antimicrobial agent into the constructions by coating the outer surfaces and it occurs between the layers of the construction. ICL was prepared according to Example 1. It was then spread onto polycarbonate sheets 35-40 cm long x 9 cm wide and lymphatic markers were removed. Each piece of ICL was then crosslinked in a buffer of 10 mM EDC / 0.1 M MES [2- (N-morpholino) ethanesulfonic acid] (Pierce, Rockford, IL), 3 liters per 30 pieces, stirred on a shaker table set at 4, for 16-20 hours and then rinsed three times in 3-5 L of sterile filtered water for 30 minutes per rinse. Antimicrobial agents were prepared as dispersions. Four dispersions of nanocrystalline silver were prepared by mixing 1.0, 5., 1.0 grams of nanocrystalline silver (Nanotechnologies (Ag-20) or equivalent, Austin, TX) in 1 L of a dispersing agent such as isopropyl alcohol. A nanocrystalline silver of 5.0 g in 1 L of RODI was also prepared. To cover ICL with an antimicrobial agent, pieces of ICL were placed in sterile 250 mL containers (Nalgene) with four pieces per container. 200 ml of solution or dispersion containing an antimicrobial agent was decanted in each container to submerge the constructions. The ICL remained submerged for 3-6 hours while the containers were agitated on a rotating shaker platform. The ICL that has been contacted with the nanocrystalline silver dispersion were rinsed once in 1 L of sterile filtered water. Each treated antimicrobial ICL was then layered to form two-layer constructions by placing a treated piece of ICL flat against a polycarbonate sheet. A second treated piece of ICL was then placed directly on top and spread over the first such that no air bubbles were present between the layers. The polycarbonate sheets with the constructions were then placed in the air flow of a laminar flow biological safety cabinet until dried at 10-20% Rh for a minimum of 12 hours. The resulting constructions were 2-layer ICL constructions that have been cross-linked and treated with an antimicrobial agent to impart antimicrobial qualities to the constructions, and then laminated to incorporate more antimicrobial agent between the layers of the construction. Example 8: Post-crosslinking and pre-lamination coating Single-layer ICL constructions were prepared reticulated and reticulated, then treated with an antimicrobial agent and then laminated to form two-layered constructions to produce two-layered constructions with antimicrobial qualities. Laminating the constructs after treatment with the antimicrobial agent, more antimicrobial agent was incorporated between the ICL layers. ICL was prepared according to Example 1. ICL pieces were then spread on polycarbonate sheets and lymphatic markers were removed and were approximately 35-40 cm long x 9 cm wide. Each piece of ICL was then crosslinked in 10 mM EDC buffer / 0.1 M MES [2- (N-morpholino) ethanesulfonic acid] (Pierce, Rockford, IL), 3 liters per 30 pieces, stirred on a shaker table set to 4 , for 16-20 hours and then rinsed three times in 3-5 L of sterile filtered water for 30 minutes per rinse. Antimicrobial agents were prepared either as solutions or as dispersions. A nanocrystalline silver dispersion was prepared by mixing 5.0 grams of nanocrystalline silver (Nanotechnologies (Ag-20) or equivalent, Austin, TX) in 1 L of dispersing agent, RODI. A 0.1% PHMB solution was prepared by mixing 5.0 mL of Cosmocil CQ (20% of PHMB solution) per 1000 mL of RODI / WFI. To coat ICL with the PHMB agent, 28-30 pieces of ICL were placed in a clean 5L Pyrex glass bottle. 3000 mL of PHMB solution was added to the 0. 1%. The ICL remained submerged for 3-6 hours while the containers were agitated on a rotating shaker platform. To coat ICL with the nanocrystalline silver dispersion, 200 mL were added in sterile 250 mL containers (Nalgene). Four pieces of ICL were placed in each container and shaken for 3-6 hours. The ICL that has been contacted with the nanocrystalline silver dispersion was rinsed once in 250 mL of RODI for 15 minutes and once in 500 mL of RODI for a minimum of 5 minutes until it was laminated. The ICL that has been contacted with the PHMB solution was not rinsed. The treated antimicrobial ICL was then layered to form two-layer constructions by placing a treated piece of ICL flat against a polycarbonate sheet. A second treated piece of ICL was then placed directly on top and spread over the first such that no air bubbles were present between the layers. The polycarbonate sheets with the constructions were then placed in the sterile air flow of a laminar flow biological safety cabinet until dried at 10-20% Rh for a minimum of 12 hours. The resulting constructions were 2-layer ICL constructs that have been cross-linked and treated with an antimicrobial agent to impart antimicrobial qualities to the constructions, and then laminated to incorporate more antimicrobial agent between the layers of the construction. Example 9: Evaluation of Three Antimicrobial Dressings in Proliferation of Methicillin Resistant Staphylococcus aureus (MRSA) in a Partial Thick Wound Model The antibacterial activity of three antimicrobial dressings in partial thickness wounds that were colonized with resistant Staphylococcus aureus was examined to Methicillin (MRSA). A pig was used as the experimental search animal since its skin is morphologically similar to human skin. The pig weighed approximately 25-30 kg and remained in housing for two weeks before starting the experiment. This pig was fed a basal diet ad libitum and was housed only in the animal facility (complying with USDA) with controlled temperature (19-21 ° C) and lights (12h / 12h LD). The pig was anesthetized with Telazol (5 mg / kg), Xylazine (2 mg / kg), Atropine (0.05 mg / kg) I.M., and inhalation of a combination of isoflurane and oxygen. The hair in the back of the pig was cut with a standard animal cutting machine. The skin on both sides of the back of the pig was prepared by washing with a non-antibiotic soap (Neutrogena®) and sterile water. The animal was dried from the spot with a sterile gauze. Thirty six were made (36) partial thickness wounds (10x7x0.3mm) in the dorsal skin using a specialized electrokeratome. The wounds were then inoculated with Methicillin-resistant Staphylococcus aureus of ATCC 33593. To prepare the bacterial inoculation, a fresh culture of pathogenic isolation obtained directly from the American Type Culture Collection (ATCC), Rockville, Maryland, was used. The inoculum was Methicillin-resistant Staphylococcus aureus from ATCC 33593. The culture of lyophilized bacteria was recovered by the standard ATCC recovery protocol. All inocula suspensions were made by scraping overnight growth from a culture dish in 5 ml of normal saline until the turbidity of the suspension is equivalent to that of a MacFarland # 8 Standard Turbidity. This would result in a suspension concentration of about 108 colonies that form units / ml (CFU / ml). The suspension of 108 was serially diluted to make a suspension inoculated with a concentration of approximately 108 CFU / ml. A small amount of the inoculated suspension was placed in a culture medium to quantify the exact concentration of viable organisms. The inocula suspension was used directly to inoculate each site of the wound. An aliquot of 0.025 ml (25 μm) of the suspension would be deposited in a sterile glass cylinder (22 mm diameter) in the center of each wound. The suspension is purged lightly at each test site for ten seconds using a sterile Teflon spatula and allowed to dry for 3 minutes. In the course of 10 minutes of inoculation, all the inoculated wounds were converted with a bandage of polyurethane film for 24 hours before the initiation of the treatment to give the time of bacteria to colonize the wounds. Three additional wounds were created and inoculated to obtain a CFU / ml baseline before the initiation of treatments: Inoculum: Log 4.24 CFU / mL; Baseline Accounts: Log 5.38 CFU / mL. Six wounds were assigned to several treatment groups as seen by the experimental design below. The animal subject is monitored daily for any observable signs of pain or discomfort. To help minimize possible discomfort, an analgesic (Duragesic - fentanyl transdermal system eluting at 25 pg / hr) was used during the entire experiment. 1) Bandage A was a construction treated with PHMB from 2 layers made according to Example 8 2) Bandage B was a construction treated with 2-layer nanocrystalline silver made in accordance with the Example 8 3) Bandage C was a 1-layer nanocrystalline silver treated construction made in accordance with Example 8 4) Control (oil gauze or polyurethane film only) 5) Positive control (antimicrobial competitor bandage or Bactroban ointment) 6) Control treatment groups All exposed to air, untreated except untreated were covered with polyurethane film ( as discussed with sponsor). Treatment groups were randomly assigned to any 1, 2, 3, 4, 5 or 6 area. A total of 3 injuries per treatment group per day were assessed. Three wounds were cultured from each treatment group at 24 and 72 hours post-treatment. At each sampling time, sites were cultivated quantitatively. Each site was cultivated only once. The area was covered by a sterile glass cylinder (22 mm outside diameter) held in place by two handles. A 1 ml depuration solution was pipetted into the glass cylinder and the site was cleaned with a sterile Teflon spatula for 30 seconds. Serial dilutions were made and depuration solutions were quantified using the Spiral Silversystem, which deposits a defined small amount (40 μ?) Of suspension on the surface of a rotating agar plate. MRSA was growing on selective media prepared with Mueller Hinton agar, 4% NaCl and 6 micrograms of oxacillin / ml. Oxacillin was used instead of methicillin because it is more stable. After the inoculation period (24 hours), colonies were counted in the plates and the units forming the colony were calculated per ml_ (CFU / ml). The results from this study are presented in Table 1. The results show that the 2-layer PHMB, 2-layer nanocrystalline silver and 1-layer nanocrystalline silver were effective in bacteria that eradicate wounds.

Example 10: Mechanical Testing Techniques and Properties of Multilayer ICL Prostheses Preferred embodiments of multilayer ICL patch constructions were tested. The constructions of 2, 4 and 6 layers of ICL crosslinked with 100 mM EDC in 50% Acetone (100/50) and constructions of 6 layers crosslinked with 7 mM EDC / 90% acetone v / v in water (7 / 90) and 1 mM EDC in water (1/0) were evaluated throughout a variety of measurements. The results are summarized in Table 2. A flexible failure test was performed using a servo-hydraulic MTS test system with TestStar-SX software. Tapes of 1.25 cm in width were extracted to fail in uniaxial tension at a constant tension speed of 0.013 s "1. The slope of the linear region (EY) and the final tensile strength (UTS) were calculated from the curves of Strengths and elongations The adhesion strength between the layers was tested using a standard protocol for adhesive testing (ASTM D1876-95) The adhesion strength is the average force required to detach two layers of laminated ICL apart from one another. constant velocity of 0.5 cm / sec. Differential scanning calorimetry was used to measure the thermal flow to and of a sample under thermally controlled conditions. defined as the start temperature of the denaturation peak in the temperature energy graph. Suture retention was not performed on 2 or 4 layer reticulated constructions in 100 mM EDC and 50% acetone since the suture retention (3.7N ± 0.5N) for a 2-layer construction cross-linked in 1 mM EDC and without acetone (much less crosslinked) was well above the minimum specification of 2 N. The rolling resistance between layers of ICL and shrinkage temperature are dependent on the concentration of crosslinking and the addition of acetone rather than the number of layers in a building.

Example 11: Method for Treating a Wound with an Antimicrobial Treated Construction Any of a single sheet ICL layer of the Example 1 or an integrated ICL multilayer sheet construction formed by the method of Example 2 is used to treat a full density skin wound. The sheet is formed in mesh or is fenestrated to create small openings to allow infiltration of wound exudate. Skin wounds including second degree burns, lacerations, tears and abrasions; excision wounds or surgical removal of donor site cancerous development or autograft sites; and skin ulcers such as venous, diabetic, pressure (decubitus ulcers), and other chronic ulcers are managed using ICL in simple or multi-layered form. The ICL matrix of collagen protects the wound bed while maintaining moisture and allows drainage of the wound. The ICL was applied before the wound, the wound bed was prepared for its application. Patients with burn injuries requiring grafting were selected. ICL was placed either directly on the excised wound bed or on the expanded or unexpanded mesh autograft at a ratio of 2: 1 or more. Test sites (ICL) and control sites (autograft), when used, are of the same mesh ratio. Burn wound sites that are grafted are prepared, such as debridement, before treatment in accordance with standard practice so that the area of burned skin is He excised completely. Excised pearls appear clean and clinically uninfected. Patients suffering from surgical excision are anesthetized locally. The pre-operative area is cleaned with an anti-microbial / antiseptic skin cleanser (Hibiclens®) and rinsed with normal saline. Dense partial deep wounds are made on the skin and the skin is grafted elsewhere unless it is cancerous. ICL was applied to the wound bed and sterile bandages were applied. In any case of injury, appropriate post-operative care is provided to the patient in examining, cleaning, changing bandages, etc., of the treated wounds. A complete record of the condition of the treated sites is kept to document all the procedures, necessary medications, frequency of bandage changes and any observations made. The wound beds remain protected from the external environment and moisture to aid in wound management and healing. The bandage of the wound is tested in an animal model. The wound dressing construction of the invention is either a single or multi-layered construction made of ICL formed by the methods of Examples 1 and 2. A rat full density wound healing model (a commonly used model) used for wound dressing products) was used to assess the performance of a construction of wound dressing made of a simple layer material of ICL. A total of 20 animals, four per evaluation time point, had two (2) 2 cm x 2 cm of full density excision wounds created on their back. The test and control articles were cut slightly longer than the periphery of the wound and applied dry to any wound after a randomized application scheme. The bandages were rehydrated by the wound fluid and saline as needed. Secondary bandages of petrolatum gauze were applied to each test and control article and changed weekly or at each evaluation time point. The wounds were evaluated at 3, 7, 14, 28 and 42 days post-treatment. Evaluations include speed and percentage of wound closure (based on wound traces), erythema, exudate, and histology of explanted wound sites. According to the results of the analysis of the percentage and speed of wound closure, the sites treated for the construction of the wound dressing showed slightly faster, although not statistically significant, wound closure than the control sites. The time analysis to complete the wound closure did not find a difference between the test and the treated control sites. The results of erythema, exudate and histology analysis were equivalent for the two products. Histology showed that the construction of the wound dressing made of layer ICL simple, presented healing characteristics required over time, re-epithelialization of the wound, and resorption of collagen materials. There was no evidence of an adverse reaction to the construction by the test subjects. Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it would be obvious to one of skill in the art that certain changes and modifications may be practiced within the scope of the appended claims.

Claims (36)

1. CLAIMS 1. A bioconstructed collagen construct, characterized in that it comprises: a layer of purified collagen tissue matrix derived from the tunica submucosa of a small intestine, the purified collagen tissue matrix has been treated with an antimicrobial agent, wherein the Construction is biocompatible and facilitates tissue growth when applied to a wound bed.
2. 2. The bioconstructed collagen construction according to claim 1, characterized in that the construction is mesh-shaped, fenestrated or perforated.
3. 3. The bioconstructed collagen construction according to claim 2, characterized in that the meshing ratio of the mesh is 1: 1.5. The bioconstructed collagen construction according to claim 1, characterized in that the antimicrobial agent is selected from the group consisting of nanocrystalline silver and PHMB. The bioconstructed collagen construction according to claim 1, characterized in that the construction is chemically linked with a crosslinking agent of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride. 6. The construction of bioconstructed collagen from according to claim 1, characterized in that the construction has a thickness between about 0.05 to about 0.07 mm. 7. The bioconstructed collagen construction according to claim 1, characterized in that it is bio-remobable. A bioconstructed collagen construct, characterized in that it comprises: two or more chemically bonded layers of processed tissue material treated with an antimicrobial agent, wherein when the construction is implanted in a wound bed of an animal patient, the construction undergoes biodeg controlled radation that occurs with adequate living cell replacement in such a way that the construction is remodeled by the living cells of the patient. 9. The bioconstructed collagen construction according to claim 9, characterized in that the construction is mesh, fenestrated or perforated. 10. The bioconstructed collagen construction according to claim 9, characterized in that the meshing ratio of the mesh is 1: 1.5. 11. The bioconstructed collagen construct according to claim 8, characterized in that the antimicrobial agent is selected from the group consisting of nanocrystalline silver and PHMB. 12. The bioconstructed collagen construction according to claim 8, characterized in that the construction is chemically linked with a cross-linking agent of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride. 13. The bioconstructed collagen construction according to claim 8, characterized in that the construction is biocompatible. 14. The bioconstructed collagen construction according to claim 8, characterized in that the processed tissue material is tunica submucosa of small intestine. 15. A method for preparing an antimicrobial bio-removable prosthesis having two or more joined layers, its matrix of processed tissue matrix, characterized in that it comprises the steps of: (a) layered separation of two or more sheets of matrix layers of processed fabric, hydrated; (b) dehydrating the fabric layers to adhere the layers together; (c) crosslinking the layers of fabric with a crosslinking agent to bond the layers together; (d) rinsing the layers to remove the crosslinking agent; and (e) treating the bonded layers with an antimicrobial agent; wherein the prosthesis, when implanted in a mammalian patient undergoes controlled biodegradation that occurs with the replacement of the appropriate living cell such that the prosthesis is remodeled by the living cells of the patient. 16. A method for preparing an antimicrobial bio-removable prosthesis having two or more joined, superimposed layers of processed tissue matrix derived from the tunica submucosa of the small intestine, characterized in that it comprises the steps of: (a) layered separation of two or more layered layers of processed tissue matrix, hydrated; (b) dehydrating the fabric layers to adhere the layers together; (c) crosslinking the layers of fabric with a crosslinking agent to bond the layers together; (d) rinsing the layers to remove the crosslinking agent; and (e) treating the bonded layers with an antimicrobial agent; wherein the prosthesis, when implanted in a mammalian patient undergoes controlled biodegradation that occurs with the replacement of the appropriate living cell such that the prosthesis is remodeled by the living cells of the patient. 17. A method for preparing an antimicrobial wound dressing having a single layer of processed tissue matrix derived from the tunica submucosa of the intestine thin, characterized in that it comprises the steps of: (a) purifying a tissue matrix of the tunica submucosa; (b) treating the tissue matrix with an antimicrobial agent, wherein the wound dressing, when applied to a wound, maintains a moisture environment in the wound to facilitate regeneration of the skin tissue in the wound. 18. The method according to claim 15, 16 or 17, characterized in that the wound dressing is mesh-shaped, fenestrated or perforated. 19. A method for treating a wound, characterized in that it comprises the steps of: (a) cleaning the wound and surrounding area of the wound; (b) applying an antimicrobial wound dressing comprising a tissue matrix of purified collagen derived from the tunica submucosa of the small intestine treated with an antimicrobial agent. The method according to claim 21, characterized in that the antimicrobial wound dressing is mesh-shaped, perforated or fenestrated. 21. A method for treating a wound, characterized in that it comprises the steps of: (a) cleaning the wound and surrounding area of the wound; (b) apply a skin replacement selected from the group consisting of an autograft, an allograft, or a construction of cultured skin to the wound; and (c) applying an antimicrobial wound dressing comprising a tissue matrix of purified collagen derived from the tunica submucosa of the small intestine treated with an antimicrobial agent to the replacement skin. 22. The method according to claim 21, characterized in that the replacement of the skin is mesh-shaped, perforated or fenestrated. 23. The method according to claim 21, characterized in that the antimicrobial wound dressing is in the form of a mesh before application to the wound. The method according to claim 21, characterized in that the wound is selected from the group consisting of full density wounds, pressure ulcers, venous ulcers, diabetic ulcers, chronic vascular ulcers, tunnel wounds, undetermined wounds. , surgical wounds, donor site wounds for autografts, post-moh surgery wounds, post-laser surgery wounds, wound dehiscence, trauma wounds, abrasions, lacerations, second degree burns, lacrimal or drained skin wounds. 25. A method for repairing or replacing a damaged tissue, characterized in that it comprises the steps of: implanting a prosthesis in a patient, the method comprising two or more joined layers, superimposed on material of collagen treated with an antimicrobial agent, wherein the prosthesis, when implanted in a mammalian patient, undergoes controlled biodegradation which occurs with the replacement of the appropriate living cell such that the implanted prosthesis is remodeled by the living cells of the patient . 26. The method according to claim 25, characterized in that the prosthesis comprises two to five sheets of processed intestinal collagen derived from the tunica submucosa of the small intestine, the sheets are bonded and cross-linked together with 1-ethyl-3- hydrochloride ( 3-dimethylaminopropyl) carbodiimide at a concentration between 0.1 to 100 mM. 27. The method according to claim 25, characterized in that the prosthesis is a hernia repair patch, a femoral hernia repair plug, a pericardial patch, a bladder sling, a uterine sling, intra-cardiac patch, replacement heart valve, a vascular patch, an annular fibrosis repair plug, an annular fibrosis repair patch, a rotator cuff repair prosthesis, a duramater repair patch, a cystocele repair device, a repair device of retrocele, a sling of vaginal vault prolapse repair, an implant by plastic surgery. 28. The method according to claim 21, characterized in that damaged or diseased soft tissue in need of repair is used to treat abdominal and thoracic wall defects, muscle flap reinforcement, rectal and vaginal prolapse, pelvic floor reconstruction, hernias, suture line reinforcement and reconstructive procedures. 29. A modified intestinal collagen layer, characterized in that it comprises a layer of intestinal collagen derived from the tunica submucosa of the small intestine, the layer has been treated with an antimicrobial agent. 30. The modified intestinal collagen layer according to claim 29, characterized in that the antimicrobial agent is nanocrystalline silver. 31. The modified intestinal collagen layer according to claim 30, characterized in that the nanocrystalline silver lacks atomic disorder. 32. The modified intestinal collagen layer according to claim 29, characterized in that the antimicrobial agent is PHMB. 33. A processed tissue matrix comprising a mammalian cell tissue substantially free of collagen-free components, characterized in that the processed tissue matrix has been treated with an antimicrobial agent. 34. The processed tissue matrix according to claim 33, characterized in that the agent Antimicrobial is nanocrystalline silver. 35. The processed tissue matrix according to claim 34, characterized in that the nanocrystalline silver lacks atomic disorder. 36. The processed tissue matrix according to claim 33, characterized in that the antimicrobial agent is PHMB.