TWI424861B - And a method for producing the regenerated film and the method for producing the regenerated film - Google Patents

Description

Regeneration film applied to tissue repair engineering and preparation method of the same

The present invention relates to the field of tissue repair and regeneration of medicine, and more specifically, to a membrane-like substance which assists and promotes tissue repair engineering, the membrane-like substance has permeability of unequal inflow and outflow rates, and biological tissue Degradability, and can be a drug carrier. The present invention relates to the construction of the film-like substance and a method of developing the film-like substance.

Tissue repair refers to the process by which local tissues or cells are damaged or killed by certain pathogenic factors and repaired by regeneration of adjacent healthy cells to restore tissue integrity. The speed and completeness of the repair process are affected by many factors, including the damage type, nutrition, blood supply, infection, and tissue defect rate. Tissue regeneration refers to the process of cell division and proliferation after tissue damage to complete the repair. In addition to autologous repair and regeneration processes, the use of excipients at the wound site can promote tissue repair regeneration.

The regenerative membrane is one of the above-mentioned auxiliary materials, and the regenerative membrane is placed on the surface of the tissue damage, and the invading damage surface such as unnecessary cells, fibrous tissue or epithelial tissue is isolated in the area covered thereby, and the blood can be maintained. The drug or drug is passed and the waste generated in the tissue repair and regeneration project is excluded. The placement of the regenerative membrane allows for the fabrication and organization of a tissue-textured repair regenerative space.

Taking the repair of bone as an example, the regenerated film can be divided into two types: non-absorbable regenerative film and absorbable regenerative film.

Common materials that are not absorbable are Gore-TEXR, MilliporeR, BiobarrierR. It contains special biochemical materials such as e-polytetrafluoroethylene (e-PTFE), which is a significant aid to the osseointegration of dental implants. In addition to inhibiting the invasion of epithelial and fibroblasts, It also promotes bone cell growth. It also has the following properties: cell occlusivity, space making, tissue integration, biocompatibility, good stability and reliability. Sex.

Absorbable regenerative membranes have been developed since 1990 and can be divided into three categories depending on the composition of the materials.

The first type is collagen regeneration membrane: cross-linking treatment can improve its physical properties and absorption time, so that its absorption time can be extended to more than 8 weeks, which is beneficial to bone regeneration applications, such as Bio-GageR, Bio. -MendR, AtrisorbR.

The second type is the polymer regeneration film: mainly composed of polylactic acid and polyglycolic acid. The functional integrity time is extended to 8-10 weeks, and the complete absorption takes 6-7 months to provide complete tissue isolation. . Such as Resolut XtR.

The third category is the general term for other recycled membranes other than the first two materials.

At present, there are many kinds of regenerative membranes used for tissue damage regeneration. Most of the materials used in the past are synthetic materials. The advantages of the materials are good operability and good biocompatibility. For example, bone damage can be combined with titanium-containing materials or other materials. Fill the material to achieve a more stable and larger space for bone growth. Since such a regenerative membrane is a non-absorbable material, it is necessary to re-examine the regenerated membrane about 4 to 8 weeks after the operation, and if it is exposed, it may cause infection and affect bone growth. Nowadays, the regenerative membrane has developed many kinds of absorbable materials, which can be slowly absorbed by the human body after being placed. The advantage is that the second operation can be dispensed with. There are many reports that the clinical effects of absorbable and non-absorbable regenerative membranes are much the same.

The above-mentioned absorbable or non-absorbable regenerative membranes have comparable porosity and maintain permeability, and the permeability is determined by the outflow rate and the inflow rate on both sides of the membrane, and the outflow rate and the inflow rate. The rate is almost equal, we call it peer-to-peer. However, peer-to-peer permeability will dilute the contribution of the regenerative membrane to tissue repair and regeneration. Because the initial, middle and end of tissue damage have different reaction changes, in the initial stage, the lesion is full of hematoma, blood clots and even inflammatory reactions, and a large number of necrotic cells and tissue fluids should be accelerated to the extramembranous diffusion to eliminate inflammation. In the middle and late stages, growth factors begin to grow and function. At this time, the regenerative membrane must prevent epithelial cells and connective tissue from permeating through the membrane and invading the injured area. When the regenerated membrane is equivalent to permeation, it will result in an equal rate of diffusion to the membrane and a rate of penetration into the membrane, resulting in a decrease in the rate of elimination of the unfavorable substance and inability to effectively control the invasion of the unfavorable substance.

From this, it can be seen that the inflow and outflow permeability of the regenerated membrane has an important influence on tissue repair and regeneration. However, most of the research on recycled membranes has focused on the selection and combination of materials, and has not studied and developed the inflow and outflow permeability.

The purpose of the present invention is to provide a regenerative membrane for use in tissue repair engineering, which has the characteristics of asymmetry of inflow and outflow permeability, and is a regenerative membrane with a high unidirectional permeability. The high unidirectional permeability can be high inflow or high outflow, and it is determined by the demand of the damaged part to determine the application mode of the high outflow or high inflow to the damaged area. In the application mode of high outflow, the regenerative model can accelerate the elimination of a large amount of waste cells such as necrotic cells and tissue fluid generated in the tissue damage, reduce the inflammatory reaction, and insulate the invasive lesions such as epithelial cells and connective tissue, thereby assisting and promoting Organize the restoration project. The regenerative membrane can be a drug carrier, and in a high inflow application mode, the beneficial substance or drug for repairing and regenerating tissue is introduced into the damaged tissue, thereby accelerating the tissue repair regeneration function.

The purpose of the present invention is to provide a regenerative membrane for use in tissue repair engineering which is biodegradable and which does not require a second surgical removal after being implanted into the injury site during tissue defect repair surgery.

The object of the present invention is to provide a method for producing the above-described regenerated film which has a large pore layer and a small pore layer, the large pore layer having a large pore surface having a small pore surface. The method of the invention can further control the difference in the ratio of the pore size and the number of the large pore layer and the small pore layer. The greater the difference, the higher the asymmetry of the permeability and the higher the high unidirectional permeability.

The purpose of the present invention is to provide a method for producing the above-described regenerated film which can further control the difference in the ratio of the macroporous layer to the small pore layer, thereby adjusting the high unidirectional permeability.

The object of the present invention is to provide a method for producing the above-mentioned regenerated film which does not have a conventional cortical effect and thus maintains a desired high unidirectional permeability.

The disadvantage of the cortical effect of the asymmetric regenerated film of the present invention is improved by the hydrophilic gel layer process.

A regenerative film applied to a tissue repairing process, the regenerative film comprising: a first void layer and a second pore layer adjacent to each other; the first pore layer and the second pore layer are distributed with a plurality of connected to the external environment a pore, and a pore size of the first pore layer is larger than a pore of the second pore layer.

A method for preparing a regeneration film applied to a tissue repairing process, comprising: step 1: mixing a polymer material with a first solvent in a predetermined ratio to form a polymer solution; and step 2, attaching the polymer solution to a mold surface And forming a regenerated film semi-finished product; in step 3, the regenerated film semi-finished product is subjected to a pore-forming process by a second solvent, and the above-mentioned pores are formed on the film wall of the regenerated film semi-finished product.

For the convenience of the description, the central idea expressed in the column of the above summary of the present invention is expressed by a specific embodiment. Various items in the embodiments are depicted in terms of ratios, dimensions, amounts of deformation, or displacements that are suitable for illustration, and are not drawn to the proportions of actual elements, as set forth above. In the following description, like elements are denoted by the same reference numerals.

As shown in the first figure, the cross section of the regenerated film of the present invention is depicted by an industrial drawing method. The regenerated film 10 of the present invention is made of a biodegradable polymer, and one side of the regenerated film 10 is a first The pore layer 11, the other side of the regeneration film 10 is a second pore layer 12, each of the first pore layer 11 and the second pore layer 12 has a plurality of pores 111, 121 communicating with the external environment, and the first pore layer 11 The pores 111 are larger in size than the pores 121 of the second pore layer 12. The surface of one side of the regenerated film 10 is a first surface 13 , and the surface of the other side of the regenerated film 10 is a second surface 14 having a hole 111 of the first pore layer 11 , the second The surface 14 has an aperture 121 of the second aperture layer 12, the aperture 111 of the first surface 13 being sized larger than the aperture 121 of the second surface 14. As shown in the second figure, the surface states of the first surface 13 and the second surface 14 are respectively depicted by an industrial drawing method. As shown in the third figure, the asymmetric permeability of the regenerated film of the present invention is depicted by an industrial drawing method. The permeability of the regenerated film 10 from the first pore layer 11 and the first surface 13 to the second pore layer 12 and the second surface 14 is higher than that from the second pore layer 12 and the second surface 14 based on the difference in pore size. The permeability of the first pore layer 11 and the first surface 13. In other words, the permeability of both sides of the film of the regenerated film 10 is asymmetrical. Moreover, the greater the difference in the ratio of the size and the number of the above pores, the higher the asymmetry of the permeability and the higher the high unidirectional permeability. In the legend of the third figure, a high unidirectional permeability from the first pore layer 11 to the second pore layer 12 is described.

Based on the demand for tissue degradability, the regenerated film of the present invention is made of a polymer material, and is an example of the polymer material of the present invention, including but not limited to polylactic acid glycol copolymer (poly(DL-lactci acid-co-glycolic acid). ), hereinafter referred to as PLGA), poly ε-caprolactone (PCL), polylactide acid (PLA), chitosan, and the like.

The method for preparing a regenerated film by using the above materials mainly comprises the following steps: Step 1: mixing a polymer material with a first solvent in a predetermined ratio to prepare a polymer solution; and Step 2, attaching the polymer solution to a mold surface Forming a regenerated film semi-finished product; in step 3, performing a slit hole forming process on the regenerated film semi-finished product with a second solvent, and forming a first pore layer and a second pore layer on the two sides of the regenerated film semi-finished product; , cleaning, drying, stripping, and completing a regenerated film with asymmetric pores.

The first solvent which can be used in the present invention is not particularly limited as long as it can dissolve the polymer material therein. When the polymer material to be used is a polylactic acid glycolic acid copolymer, polycaprolactone, or polylactic acid, examples of the first solvent include, but are not limited to, 1,4-dioxane, Chloroform (chloroform) and hexafluoroisopropanol (HFIP). When the polymer material to be used is chitosan, an example of the first solvent includes an aqueous acetic acid solution or the like, but is not limited thereto. The concentration of the polymer solution is not particularly limited in the present invention, but is preferably 10% (w/w) in order to make it easy to handle and form a film.

The mold used in the second step is preferably a flat mold, and the shape of the mold can be prepared according to the desired shape of the film. The material of the flat plate mold is not particularly limited in the present invention, and any one which does not react with the polymer material and/or the first solvent and which can facilitate the release of the film can be applied to the present invention. Examples which may be mentioned include, but are not limited to, metals, glass, quartz, and the like.

The method of adhering the polymer solution to the surface of the mold in the foregoing step 2 is not particularly limited in the present invention, and any conventional polymer solution can be applied to the surface of the mold, and can be applied to the present invention. Including, but not limited to, conventional methods such as dipping-casting, pouring, and the like.

The pore size of the outer layer of the regenerated membrane semi-finished membrane wall can be adjusted by the polarity of the second solvent in the third step. When the concentration of the second solvent is lower (the higher the polarity), the pore size is larger; The higher the concentration of the second solvent (the smaller the polarity), the smaller the pore size. For example, when the aqueous ethanol solution is used as the second solvent, the concentration of the aqueous ethanol solution is preferably from 10 to 60% (v/v), more preferably from 20 to 40% (v/v). The greater the difference in pore size between the outer layer of the membrane wall and the inner layer of the membrane wall, the higher the asymmetry of the permeability of the regeneration membrane, that is, the asymmetric unilateral high flow rate of the material permeating into and out of the membrane wall.

The conventional immersion-casting method forms a cortical effect on the surface of the regenerated film, and the regenerated film 10 of the present invention is taken as an example to explain the phenomenon of the cortex effect. The ideal regenerated film 10 should be as described in the foregoing embodiments, with the first pore layer 11 and the first surface 13 having a larger pore size than the second pore layer 12 and the second surface 14. However, when the cortical effect is produced, the pore size of the first surface 13 is actually smaller than that of the second surface 14, which has a severe effect on the originally expected high unidirectional permeability. In order to solve the problem, the present invention pre-coats a hydrophilic gel layer on the surface of the mold before the step 2 is performed. The hydrophilic gel layer is a substance that is insoluble or poorly soluble in the first solvent, including but not Limited to bio-polyvinyl alcohol (PVA), gelatin or F127 gel. The hydrophilic gel can block the second solvent from invading between the polymer material and the flat membrane when the regenerated membrane semi-finished product is immersed in the second solvent to form a film, and the hydrophilicity of the hydrogel is higher than that of the polymer. a solution, so that the hydrophilic gel layer absorbs most of the second solvent, allowing the inner side of the regenerated film to contact the second solvent with a minimum, thereby maintaining the pore size of the first pore layer 11 and the first surface 13 larger than the second The ideal condition of the void layer 12 and the second surface 14.

The following experiment uses PLGA polymer material dissolved in 1-4 dioxan (first solvent), and adheres it to a glass plate mold, and then uses 95% and 40% ethanol (second solvent) to perform the impregnation-precipitation reaction. .

Please refer to the fourth and fifth figures for the scanning electron micrograph (SEM) of the regenerated film after the 95% ethanol immersion-precipitation method, wherein the fourth picture is the cross section of the regenerated film, and the fifth picture is the regenerated film. Upper and lower surfaces. The sixth and seventh figures are electron micrographs (SEM) of the regenerated film after 40% ethanol impregnation-precipitation method, wherein the sixth picture is the cross section of the regenerated film, and the seventh picture is the first surface of the film and the first Two surfaces. The regenerated film of the 95% ethanol impregnation-precipitation method was used as a control group. Under the observation of the electron microscope, the cross section showed a symmetric small pore structure layer 30, and both the upper surface and the lower surface of the regenerated film were small pore surfaces. Layers 31, 32. The sixth and seventh figures are the regenerated film 10 of the present invention, and the electron micrograph (SEM) shows that the cross section has the first pore layer 11 and the second pore layer 12, and the first pore layer 11 and the second pore layer Each of the 12 has a plurality of pores 111, 121 in communication with the external environment, and the pores 111 of the first pore layer 11 are larger in size than the pores 121 of the second pore layer 12. The first aperture layer 11 has a first surface 13 and the second aperture layer 12 has a second surface 14 having a pore size greater than a pore size of the second surface 14. From the SEM results, it was revealed that the cortical effect caused by the above-described conventional stain-precipitation method did not occur in the present invention.

The regenerated film of the present invention has an asymmetric permeability and can be proved by the following experiment.

The permeability test was carried out by using 10% glucose solution and placed in four glass tubes respectively. The port of the first glass tube was sealed by the regeneration membrane of the present invention (40% ethanol in the control group), and the first surface was sealed. The ports of the two glass tubes were sealed by the regenerative membrane of the present case (40% ethanol in the control group), and the third and fourth glass tubes were respectively regenerated by the control group (95% ethanol impregnation). The upper surface and the lower surface of the film are adhered to the seal. Next, the glass tubes containing the solutions were each immersed in 5 ml of a phosphate buffer solution loaded in a bottle. Thereafter, after 1, 2, and 3 days, 20 μl was taken out from the phosphate buffer solution in the bottle, and the glucose concentration was separately analyzed. Here, the glucose concentration measurement was carried out by an ultraviolet spectrophotometer (Hitachi Co., U-2000) at a wavelength of 500 nm.

Please refer to the eighth figure for the analysis of the results of the two-way permeability of the glucose permeable membrane wall. As can be seen from the eighth figure, the control group (40% regenerated film) showed a remarkable high unidirectional permeability (<0.001), and the first pore layer 11 (large pore layer) to the second pore layer 12 (small) The permeability of the pore layer is greater than the permeability of the second pore layer 12 (small pore layer) to the first pore layer 11 (large pore layer). However, since the control group was all of the small pore layers 31, 32, the permeability of both sides was also observed in the above manner, and the results showed that the film had a low bilateral flow rate.

In this case, the animal experiment is further carried out. Please refer to Annex I. Under the aseptic operation, two round bone wounds with a diameter of 5 mm and a depth of 0.5 mm can be made from the outer grinding head of the eight-week white rat head bone cover, and the case is covered on the case. There are two ways of regenerating the film. The first type is that the first surface 13 of the regenerated film of the present invention covers the surface of the bone damage portion 40 (as shown in FIG. 9); the second type is the second surface 14 of the regenerated film of the present invention. Covers the surface of the bone lesion 40 (as in the tenth image). The first type of coverage constitutes an environment of high outflow and low inflow permeability (HOLI), and the second type of coverage constitutes high inflow and low outflow (High inflow and low outflow). Permeability, HILO) environment. In addition, it is compared with a low external permeability group (low permeability, LP) and a non-regeneration membrane control group.

At 4, 8, and 12 weeks, the skull was removed for X-ray photography, and the bone defect regeneration percentage (BDRp) was quantified. For the results, please refer to the eleventh. Fig., the results of this experiment can be observed at various time points, especially 8 and 12 weeks. The results of using HOLI group are faster than those of other groups. It can be proved that in the environment of high outflow, the regenerative mold can accelerate the elimination of a large amount of waste such as necrotic cells and tissue fluid generated by the tissue damage, reduce the inflammatory reaction, and insulate the harmful substances such as connective tissue such as epithelial cells from invading the damage. Thereby assisting and promoting the organization of the restoration project.

In addition, the rats were further scanned by computer tomography and reconstructed by bone reconstruction. The results are shown in Annex II. The bone damage of the HOLI group can be seen in the computerized tomographic images of the fourth and the 12th week. The repair has been completed (white arrow), but the repair efficiency of his group of recycled membranes is poor.

After the X-ray photography experiment, the H&E staining method was used for tissue section staining. The results of the eighth week are shown in Annex III. The third part is the slice result of HOLI in the eighth week, and the fourth group is the LP group of the case. NB is the regenerative bone, OB is the old bone, the black arrow is the bone island, and the new bone is darker in color than the immature new bone. The result shows that the asymmetric bone regeneration membrane of this creation is for the new bone formation of bone damage in the early growth. Faster, the distribution of bone islands is more dense, which means that the maturity of new bones is higher. As can be seen from Annex IV, the distribution of bone islands is less and the color is darker, which means that the group is still undergoing bone in the eighth week. Chemical action, which is the accumulation of surface collagen, and the maturity of new bone is poor.

In summary, the asymmetric regenerative membrane of the present invention has the following advantages: the material used for the regenerated membrane is a highly biocompatible polymer, which is biodegradable and does not require a second surgical removal during tissue defect repair surgery. Recycled film.

The regenerated film of the present invention has an asymmetric pore structure, that is, a pore layer having a size. The asymmetric structure provides a reproducible membrane with an asymmetric permeability.

The asymmetric membrane permeability of the regenerated membrane of the present invention can be adjusted by the concentration of the solvent during the manufacturing process.

The disadvantage of the cortical effect of the asymmetric regenerated film of the present invention is improved by the hydrophilic gel layer process.

The asymmetric regeneration membrane of the present invention has a high unidirectional permeability, and can form an environment of high outflow and low inflow permeability (HOLI) at the tissue damage, and can discharge a large amount of inflammatory metabolites caused by damage. And block the effective epithelial cells from invading the defect to affect growth.

The asymmetric regeneration membrane of the present invention has a high unidirectional permeability, which can constitute a high outflow and low inflow permeability (HOLI) environment for tissue damage, or a high internal flow and low external flow permeability ( High inflow and low outflow permeability, HILO). The one-way flow rate needs to select a high outflow environment or a high inflow environment by the demand of the damaged part.

The regenerated film of the present invention may be a drug carrier. In addition to the tricalcium phosphate disclosed in the present invention, various growth factors such as PDGF (platelet derived growth factor) and TGF-β (transforming growth) may be cross-linked by physical or chemical methods. Factors), VEGF (vascular endothelial growth factor), EGF (epithelial growth factor), IGF (insulin-like growth factor); and peptides that help tissue regeneration such as RGD; or platelet-rich plasma with platelet content Referred to as PRP), the most commonly used material in the medical world today. When the regenerative membrane is a drug carrier, the manner of covering the tissue damage site should preferably form a high influent environment.

The regenerated membrane created in this case is applied to the repair of hard bone defects to illustrate the advantages of the membrane. However, the application of the regenerative membrane to the repair of human tissue is not only the repair of hard bone defects, but also the large amount of inflammatory substances and the invasion of other tissues when the tissue is damaged. Asymmetric regenerative membrane with one-way high permeability.

Although this case is illustrated by a preferred embodiment, it is true that those skilled in the art can Various forms of change are made in the spirit and scope of the case. The above embodiments are only used to illustrate the present case and are not intended to limit the scope of the present invention. All kinds of modifications or changes that are not in violation of the spirit of the case are the scope of patent application in this case.

10‧‧‧Regeneration film

11‧‧‧First pore layer

111‧‧‧ pores

12‧‧‧Second pore layer

121‧‧‧ pores

13‧‧‧ first surface

14‧‧‧ second surface

30‧‧‧Small pore structure

31‧‧‧Small pore surface layer

32‧‧‧Small pore surface layer

40‧‧‧Bone injury

In the first figure, the cross section of the regenerated film of the present invention is depicted by an industrial drawing method.

In the second figure, the surface features of the first surface and the second surface of the regenerated film of the present invention are depicted by an industrial drawing method.

In the third figure, the asymmetric permeability of the regenerated film of the present invention is depicted by an industrial drawing method.

The fourth figure is an electron microscope (SEM) cross-sectional view of the regenerated film after the 95% ethanol impregnation-precipitation method.

The fifth figure is the upper surface and the lower surface of the regenerated film of the fourth figure.

Fig. 6 is an electron microscope (SEM) cross-sectional view of the regenerated film after immersion-precipitation using a 40% ethanol.

In the seventh figure, the first surface and the second surface of the film are reproduced in the sixth figure.

Figure 8 is a graph showing the results of the two-way permeability of the glucose permeable membrane wall.

In the ninth figure, the first surface of the regenerated film of the present invention covers the surface of the bone damage surface, and a schematic diagram of a high outflow low internal flow permeability environment is formed.

In the tenth figure, the second surface of the regenerated film of the present invention covers the surface of the bone damage surface, and forms a high internal flow and low external flow permeability environment.

In the eleventh figure, X-ray photography was performed by removing the head cover of the mouse, and the result of the bone defect regeneration percentage (BDRp) was quantified.

Attachment 1, under the aseptic operation, the eight-week white rat skull bones are made of two external straight surgical rods. The circular bone with a diameter of 5 mm and a depth of 0.5 mm is damaged and covered with the regenerated film of the present invention.

In Annex II, the experimental rats were further scanned by computed tomography and reconstructed using image reconstruction to observe the repair of bone injury.

Annex III, the results of tissue section staining by H&E staining of damaged tissue in high outflow and low inflow permeability (HOLI) environment.

Annex IV, the results of tissue section staining by H&E staining of damaged tissue in a low permeability (LP) environment.

10‧‧‧Regeneration film

11‧‧‧First pore layer

111‧‧‧ pores

12‧‧‧Second pore layer

121‧‧‧ pores

13‧‧‧ first surface

14‧‧‧ second surface

Claims (10)

A regenerated film applied to a tissue repairing process, characterized in that the regenerated film is made of a biodegradable polymer, one side of which is a first pore layer, and the regenerated film The other side is a second pore layer; the first pore layer and the second pore layer are each distributed with a plurality of pores communicating with the external environment, and the pore size of the first pore layer is larger than the pore of the second pore layer size. The regenerated film applied to the tissue repair engineering according to the first aspect of the invention, wherein the surface of one side of the regenerated film is a first surface, and the surface of the other side of the regenerated film is a second surface, the first surface An aperture having the first aperture layer, the second surface having an aperture of the second aperture layer, the first surface having a pore size greater than a pore size of the second surface. A method for preparing a regenerated film for use in a tissue repair engineering according to the first aspect of the patent application, comprising: step 1: mixing a polymer material with a first solvent in a predetermined ratio to prepare a polymer solution; and step 2 The polymer solution is attached to a surface of a mold to form a regenerated film semi-finished product; in step 3, the regenerated film semi-finished product is subjected to a pore-forming process by a second solvent, and the pores are formed on the film wall of the regenerated film semi-finished product. The method for preparing a regenerated film for tissue repair engineering according to the third aspect of the invention, wherein the polymer material is selected from the group consisting of biodegradable polylactic acid glycol copolymer (poly(DL-lactci acid- One or a combination of co-glycolic acid (PLGA), polycaprolactone (PCL), and polylactide acid (PLA). The method for preparing a regenerated film for tissue repair engineering according to the fourth aspect of the invention, wherein the first solvent comprises 1,4-dioxane, chloroform (chloroform) and hexafluoroisopropanol (HFIP). The method for preparing a regenerated film for use in a tissue repair engineering according to the third aspect of the invention, wherein the second solvent is an aqueous ethanol solution. The method for preparing a regenerated film for use in a tissue repair engineering according to claim 6, wherein the concentration of the second solvent is 10 to 60% (v/v). The method for preparing a regenerated film for use in a tissue repair engineering according to claim 6, wherein the concentration of the second solvent is 20 to 40% (v/v). The method for preparing a regenerated film for tissue repair engineering according to claim 3, wherein the pore size of the outer layer of the regenerated film semi-finished film is adjusted by the polarity of the second solvent in the third step. The method for preparing a regenerated film for use in a tissue repairing process as described in claim 3, wherein, before performing step two, a surface of the mold is precoated with a layer of a second solvent to invade the polymer material and the mold. Hydrophilic gel layer.