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WO2010036009A2 - Support poreux pour la régénération tissulaire guidée et procédé de préparation de ce support - Google Patents

Support poreux pour la régénération tissulaire guidée et procédé de préparation de ce support Download PDF

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Publication number
WO2010036009A2
WO2010036009A2 PCT/KR2009/005397 KR2009005397W WO2010036009A2 WO 2010036009 A2 WO2010036009 A2 WO 2010036009A2 KR 2009005397 W KR2009005397 W KR 2009005397W WO 2010036009 A2 WO2010036009 A2 WO 2010036009A2
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WIPO (PCT)
Prior art keywords
layer
collagen
silk
group
bone
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Ceased
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English (en)
Korean (ko)
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WO2010036009A3 (fr
Inventor
박정극
서영권
한미정
윤희훈
송계용
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Industry Academic Cooperation Foundation of Dongguk University
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Industry Academic Cooperation Foundation of Dongguk University
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Publication of WO2010036009A2 publication Critical patent/WO2010036009A2/fr
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Anticipated expiration legal-status Critical
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/28—Bones
    • A61F2/2846—Support means for bone substitute or for bone graft implants, e.g. membranes or plates for covering bone defects
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2310/00—Prostheses classified in A61F2/28 or A61F2/30 - A61F2/44 being constructed from or coated with a particular material
    • A61F2310/00005—The prosthesis being constructed from a particular material
    • A61F2310/00365—Proteins; Polypeptides; Degradation products thereof

Definitions

  • the present disclosure relates to a biodegradable porous support for inducing guided tissue regeneration, in particular regeneration of periodontal tissue, and a method of making the same.
  • Periodontal tissue supporting teeth is composed of alveolar bone, periodontal ligament tissue and connective tissue.
  • the loss of alveolar bone due to the progression of periodontitis is accompanied by loss of the periodontal ligament tissue, the normal recovery of the alveolar bone and periodontal ligament tissue is impossible due to the overgrowth of connective tissue in the tissue area lost after periodontal treatment.
  • the periodontal ligament tissue may not be normally differentiated, which may lead to tooth loss.
  • chitosan and synthetic polymers which have been conventionally used in biodegradable shielding membranes, have a problem of causing inflammatory reactions in the body compared to natural polymers in terms of biocompatibility, and natural polymers have advantages such as high biocompatibility and low antigenicity. It is weak and quickly decomposed in the inflammatory area has the disadvantage of poor treatment efficiency.
  • step 1) forming a silk layer using woven silk from which sericin is removed; 2) applying a collagen solution to the silk layer of step 1) and then lyophilized to form a collagen sponge layer; And 3) crosslinking the resulting structure of step 2), a method for producing a porous support for induced tissue regeneration.
  • the porous support according to the present disclosure is biodegradable and does not require additional removal surgery, is excellent in tissue regeneration and mechanical properties, and hardly causes an immune response after transplantation compared to a synthetic polymer support or a natural-synthetic polymer support. Therefore, it is excellent in biocompatibility, and the woven silk is used to suture and improve the physical properties of the support, so that it can be usefully used as a substrate for promoting bone regeneration of the orthopedic region as well as alveolar bone or periodontal ligament.
  • FIG. 1 and 2 are schematic cross-sectional and photographic views of a porous support, respectively, according to one embodiment of the present disclosure.
  • FIG. 3 is a photograph of pulp tissue obtained from an extracted wisdom tooth and pulp cells separated and cultured therefrom.
  • FIG. 5 is a result of MTT analysis showing cell proliferation 10 days after inoculation of pulp cells on a porous support obtained by mixing hyaluronic acid (HA) with collagen by concentration and applying it to a silk layer.
  • HA hyaluronic acid
  • FIG. 7 is a photo of the porous support of FIG. 5 cultured in differentiation medium for 4 weeks, pocket transplanted to the back of nude mice, and biopsied at 8 weeks:
  • A hyaluronic acid-free group
  • B hyaluronic acid 0.4 ml addition group
  • FIG. 8 shows the results of H / E staining and Vonkossa staining (200-fold) of the biopsy tissue of FIG. 7 :
  • FIG. 11 is a photomicrograph of the porous support of FIG. 9 cultured in differentiation medium for 4 weeks and pocket transplanted to the back of nude mice followed by biopsy at week 8.
  • FIG. 12 is the result of H / E staining and Vonkossa staining (200-fold) of the biopsy tissue of FIG. 11 :
  • Figure 13 is the result of inoculating periodontal cells to the porous support to which the bone morphogenetic protein (BMP-2) is applied and confirmed the proliferation of the cells through MTT analysis after 3 days of culture.
  • BMP-2 bone morphogenetic protein
  • FIG. 15 is a photomicrograph of the porous support of FIG. 13 cultured in differentiation medium for 4 weeks, pocket transplanted to the back of nude mice, and biopsy at 8 weeks:
  • A, C BMP-2 untreated group
  • B, D BMP-2 treated group
  • FIG. 16 is the result of H / E staining and Vonkossa staining (200-fold) of the biopsy tissue of FIG. 15 :
  • FIG. 18 is a photograph of the porous support of FIG. 17 cultured in differentiation medium for 4 weeks, followed by pocket transplantation on the back of nude mice followed by biopsy at week 8.
  • B nanohydroxyapatite and BMP-2 treatment group.
  • FIG. 19 is the result of H / E staining and Vonkossa staining (200-fold) of the biopsy tissue of FIG. 18 :
  • FIG. 20 is a view of pocketing porous supports including a PGA layer and a silk fiber layer on the back of a nude mouse instead of the porous support prepared in Example 3 and the sericin-free woven silk layer according to the present disclosure.
  • the next two weeks are micrographs of inflammation.
  • Example 21 is a result of measuring the tensile strength of the porous support prepared in Example 6 and the porous support without the silk layer as a comparative example according to the present disclosure:
  • the porous support of the present disclosure may be usefully used as a support for tissue regeneration because of excellent biocompatibility and physical properties of the support.
  • the subject refers to a subject in need of tissue transplantation, and the subject includes a mammal including a human.
  • the silk layer is formed of woven silk from which sericin causing an inflammatory reaction is removed.
  • Woven silk without sericin can be sutured to improve the properties of the scaffold, and it is well compatible with surrounding tissues without causing an inflammatory reaction even after implantation in the body.
  • silk fibers 300-500 ⁇ m in diameter can be woven into a cotton substrate using a weaving machine and then treated with alkali salts such as soda ash (Na 2 CO 3 ) and sodium hydroxide (NaOH) to remove sericin. have.
  • the thickness of the silk layer may be 1 to 2 mm, but is not limited thereto.
  • the silk layer may be coated with nanohydroxyapatite and / or bone morphogenic protein.
  • nanohydroxyapatite is a major component constituting the bone matrix along with collagen to promote the activity of bone cells and to combine with collagen to promote osteoblast differentiation of stem cells.
  • the nanohydroxyapatite may have a particle diameter of 1 to 1000 nm.
  • Bone morphogeneic protein (BMP) is used to promote bone formation, BMP-2 or BMP-12 may be used.
  • the collagen sponge layer is porous and promotes cell adhesion, migration and proliferation.
  • the collagen sponge layer may be formed using collagen alone or a mixture of collagen and hyaluronic acid and / or glycosaminoglycans.
  • the thickness of the collagen sponge layer is not limited thereto, but may be formed to about 1 to 4 mm.
  • Collagen together with nanohydroxyapatite, is a major component of the bone matrix, promoting bone cell activity, and binding to nanohydroxyapatite to promote bone cell differentiation.
  • Collagen may be used both insoluble collagen and soluble collagen, preferably from a mammal such as cattle or from marine life such as the skin of tibia fish. More preferably fish with colorless skin, most preferably flounders, for example fishes such as Seodae, Munchi flounder, Turbot, Brill can be used.
  • Hyaluronic acid is known to promote bone differentiation, but is not limited thereto, hyaluronic acid having a molecular weight of 1.8 to 2 million may be used.
  • Glycosaminoglycans play a role of crosslinking between collagen, and chondroitin, chondroitin sulfate, heparan, heparan sulfate, dermatan sulfate and the like can be used.
  • the porous support may further include a shielding layer on one surface of the collagen sponge layer or the silk layer.
  • the shielding layer may be formed in contact with the silk layer.
  • the shielding layer serves to inhibit the invasion of other soft tissue cells, but is not limited thereto.
  • other soft tissue cells amnion, small intestinal submucosa, collagen membrane, gelatin membrane, and the like may be used, and generally 0.5 to 1 mm. It may have a thickness of.
  • the material constituting the shielding layer is not particularly limited as long as it can suppress the invasion of other soft tissue cells.
  • FIGS. 1 and 2 Schematic cross-sections and photographs of a porous support according to one embodiment of the present disclosure are shown in FIGS. 1 and 2 , respectively.
  • Porous support for induced tissue regeneration according to the present disclosure described above 1) forming a silk layer using woven silk from which sericin is removed; 2) applying collagen to the silk layer of step 1) and then lyophilized to form a collagen sponge layer; And 3) crosslinking the resulting structure via step 2).
  • the description of the above porous support is applied as is.
  • step 1) sericin is removed from the woven silk and washed with a detergent to form a silk layer.
  • the silk layer may have a thickness of about 1 to 2 mm, but is not limited thereto.
  • Removal of sericin may use an alkali salt such as soda ash, sodium hydroxide, for example, 0.02 to 0.1 M, preferably 0.02 to 0.05M soda ash can be used, to sufficiently remove the sericin causing the inflammatory reaction For 8 hours at 100 ° C.
  • an alkali salt such as soda ash, sodium hydroxide, for example, 0.02 to 0.1 M, preferably 0.02 to 0.05M soda ash can be used, to sufficiently remove the sericin causing the inflammatory reaction For 8 hours at 100 ° C.
  • the silk layer may be coated with nanohydroxyapatite and / or bone morphogenic protein.
  • Nanohydroxyapatite may have a particle diameter of 1 to 1000 nm, but is not limited thereto, but in a concentration of 0.01 to 0.3 g / ml, preferably 0.1 to 0.2 g / ml, more preferably 0.15 g / ml Can be used.
  • the osteogenic protein can be coated at a concentration of 100 to 500 ng / cm 2 , preferably 200 ng / cm 2 , on the silk layer, for example by dissolving 1 ⁇ g / ml of the osteogenic protein in a carbodiimide crosslinking reagent. .
  • the osteogenic protein can be BMP-2 or BMP-12.
  • the nanohydroxyapatite is first coated in order to minimize the loss of the bone morphogenic protein and then at 4 to 20 ° C, preferably at 4 ° C, for 24 to 48 hours, Preferably it is dried for 48 hours and then coated with the osteogenic protein and dried under the same conditions as above.
  • step 2) a collagen solution is applied to the silk layer obtained in step 1) or the silk layer coated with nanohydroxyapatite and / or bone forming protein, and then lyophilized to form a porous collagen sponge layer.
  • the collagen sponge layer is porous and promotes cell adhesion, migration and proliferation.
  • Collagen may be used alone or in the form of a mixture of collagen and hyaluronic acid and / or glycosaminoglycans.
  • the collagen is dissolved in an acidic solution at a concentration of less than 2% (w / v), preferably 1 to 1.5% (w / v) to prepare a creamy solution, followed by a solution of hyaluronic acid and glycosaminoglyco.
  • hyaluronic acid may be used at a concentration of 5 to 15% (v / v), preferably 10% (v / v), and glycosaminoglycan is 0.5 to 5% (v / v), preferably Can be used at a concentration of 1% (v / v).
  • acetic acid or hydrochloric acid may be used as the acid solution, and the concentration of the acid solution may be 0.001 to 0.01 M, but is not limited thereto.
  • Lyophilization is not limited thereto, but may be performed at -30 to -100 ° C, preferably at -50 to -80 ° C.
  • the crosslinking can be carried out physically or chemically.
  • heat drying eg, 72 hours at 105 ° C.
  • UV irradiation at a temperature of 4 to 25 ° C. and a power condition of 5 to 20 W (For example, irradiation with 254 nm of UV for 4 hours at conditions of 4 ° C. and 10 W).
  • chemical crosslinking for example, it may be performed by treating diphenylphosphoryl azide, glutaraldehyde, hexamethylene isocyanate, succinimide or carbodiimide at 4 to 25 ° C.
  • Such crosslinking can be carried out for 12 to 24 hours.
  • the shielding layer may be further formed on one surface of the collagen sponge layer or the silk layer, and preferably, the shielding layer may be formed to contact the silk layer.
  • the shielding layer performs a function of inhibiting the invasion of other soft tissue cells, and amnion, small intestinal submucosal membrane, collagen membrane, gelatin membrane, and the like may be used, and the thickness may be 0.5 to 1 mm. If it can suppress the invasion of other soft tissue cells is not particularly limited.
  • Periodontal wisdom teeth were used for the experiment with the patient's consent.
  • the extracted tooth was transferred to a laboratory in a serum-free medium (DMEM, Gibco, USA) in a refrigerated state (4-8 ° C.), and then the tooth was washed in 70% ethanol for 30 seconds and an antibiotic-antimycotic agent: 100 unit / ml penicillin G sodium, 100 unit / ml streptomycin sulfate, 0.25 ⁇ g / ml amphotericine B; WellGen, Korea) was washed thoroughly for 3 minutes.
  • DMEM serum-free medium
  • an antibiotic-antimycotic agent 100 unit / ml penicillin G sodium, 100 unit / ml streptomycin sulfate, 0.25 ⁇ g / ml amphotericine B; WellGen, Korea
  • the isolated pulp cells were primaryly incubated at 37 ° C. for 7 days in ⁇ -MEM medium containing 10% fetal bovine serum, and cultured under the same culture conditions, replacing the same medium once every two days.
  • FIG. 3 shows pulp tissue and primary cultured pulp cells obtained from extracted wisdom teeth
  • FIG. 4 shows FACS (fluorescence-activated cell sorting) results showing that pulp cells have stem cell characteristics.
  • Silk (Won Corporation, Korea) was woven into a mesh type using a weaving machine, and then washed in a 0.02 M Na 2 CO 3 solution followed by a 0.3% ivory cleaner to remove sericin. After washing for 3 days with distilled water and dried at 4 °C to form a silk layer.
  • the collagen solution thus prepared was applied onto the silk layer obtained in Example 2 and lyophilized at -80 ° C.
  • the collagen-coated silk layer was first subjected to 20 ml of 40% (v / v) ethanol containing 50 mM 2-morpholineoethane sulfonic acid (MES, Fluka Chemic AG, pH 5.5) for crosslinking. 30 minutes with 50 mM morphoethanesulfonic acid (MES, pH 5.5), 24 mM ethyldimethylaminopropyl carbodiimide (1-ethyl-3- (3-dimethyl aminopropyl) carbodiimide, Fluka Chemic AG) and Treatment with 40% (v / v) ethanol containing 5 mM hydroxysuccinimide (Fluka Chemic AG) for 12 hours.
  • 40% (v / v) ethanol containing 50 mM 2-morpholineoethane sulfonic acid MES, Fluka Chemic AG, pH 5.5
  • MES 2-morpholineoethanesulfonic acid
  • MES 2-morpholineoe
  • the pulp cells obtained in Example 1 were inoculated at a concentration of 5 ⁇ 10 5 cells on the previously prepared support sample. After inoculation, the support samples were grown with 10% fetal bovine serum (FBS; BioWhittaker TM , CAMBREX) and antibiotics (100 units / ml penicillin, 100 ⁇ g / ml streptomycin, 1 ⁇ g / ml amphotericin, Invitrogen).
  • FBS fetal bovine serum
  • CAMBREX fetal bovine serum
  • antibiotics 100 units / ml penicillin, 100 ⁇ g / ml streptomycin, 1 ⁇ g / ml amphotericin, Invitrogen.
  • the support sample was placed in a 12-well plate and incubated for 2 hours by adding 1 ml of MTT solution (0.33 mg / ml DMEM) to each well. Thereafter, the MTT solution was removed and 1 ml of isopropanol acidified with 0.04 N hydrochloric acid was added for 20 minutes at room temperature. The extracted solution was measured for absorbance at 540 nm using a spectrophotometer (Amersham Pharmarcia). As a result, as shown in Figure 5 , it can be seen that the pulp cells proliferate well in all the supports, in particular, the pulp cells proliferate more well in the support containing hyaluronic acid.
  • RNA isolation was performed using Trizol (Sigma). The total RNA obtained was quantified using a spectrophotometer (Amersham Pharmarcia), and then the same amount of total RNA was synthesized using hexamer primers (Promega, USA) and superscript II reverse transcriptase (Gibco BRL). It was.
  • the support sample was pocket-grafted to the back of nude mice.
  • lumpoon Yuhan Corp., Korea
  • ketamine Byer Korea Ltd., Korea
  • anesthetized by intraperitoneal injection of about 300 ⁇ l and mice were sterilized with povidone.
  • the support was pocket-grafted on the fascia, and the surgical site was sutured in six places with a suture.
  • H / E staining and immunochemical staining (Vonkossa) staining and immunochemical staining (Vonkossa)
  • the surgical site was incised to remove the scaffold and the 4% formaldehyde solution. After fixing with alcohol, dehydration using alcohol, clear with xylene, and embedded with paraffin were cut. The sections obtained by cutting were subjected to histology, stained with H / E and Boncosa, and observed under an optical microscope. The biopsy was performed with reference to histology (National Association of Clinical Pathology Professors, Korea Medical University 1992) and the pathological tissue staining guide (Kim Ju- ho, Shinkwang Publishing Co., 1993).
  • Nanohydroxyapatite was mixed with 0.001N HCl at a concentration of 0 g / ml, 0.03 g / ml, 0.15 g / ml, 0.3 g / ml to prepare a suspension, and the silk layer prepared in Example 2 was dipped therein. It was taken out and dried in air. The coating of the nanohydroxyapatite on the silk layer and drying was repeated two to three times, followed by applying 1% atelo collagen and freeze-drying at -80 ° C.
  • the collagen-coated silk layer was first treated with 20 ml of 40% (v / v) ethanol containing 50 mM morphoethaneethanesulfonic acid (MES, pH 5.5) for 30 minutes, and 50 mM morpholine Treatment was performed for 12 hours with 40% (v / v) ethanol containing ethanesulfonic acid (MES, pH 5.5), 24 mM ethyldimethylaminopropyl carbodiimide and 5 mM hydroxysuccinimide.
  • 40% (v / v) ethanol containing 50 mM morphoethaneethanesulfonic acid (MES, pH 5.5) for 30 minutes
  • 50 mM morpholine Treatment was performed for 12 hours with 40% (v / v) ethanol containing ethanesulfonic acid (MES, pH 5.5), 24 mM ethyldimethylaminopropyl carbodiimide and 5 mM hydroxy
  • a sample of a constant size was prepared using a 6 mm diameter punch, and then sterilized by gamma irradiation ( ⁇ -irradiation) of 10 KGy and stored at -20 ° C until use.
  • Example 1 The pulp cells prepared in Example 1 were inoculated to the thus prepared support sample at a concentration of 2.8 ⁇ 10 6 cells, and cell proliferation and differentiation were induced in the same manner as in Example 3.
  • MTT assay and RT-PCR were performed on the support sample in the same manner as in Example 3 except that the MTT assay was performed 3 days and 3 weeks after the culture.
  • Figs . 9 and 10 it was shown that the cells proliferate well in all the supports, and type 3 collagen, fibronectin, osteocalcin, and osteonectin, which are extracellular matrix expressed in osteocytes. It can be seen that osteopontin, osteoprotegerin, and BMP-2 are well expressed.
  • 0.15 g / ml nanohydroxyapatite-coated scaffolds showed better cell proliferation and more extracellular matrix expression.
  • the resulting structure was then treated for 30 minutes with 20 ml of 40% (v / v) ethanol containing 50 mM morphoethaneethanesulfonic acid (MES, pH 5.5) for crosslinking, and 50 mM morphoethanesulfonic acid ( MES, pH 5.5), 40% (v / v) ethanol with 24 mM ethyldimethylaminopropylcarbodiimide and 5 mM hydroxysuccinimide for 12 hours. After washing with 0.1 M sodium hydrogen phosphate (Na 2 HPO 4, pH 9.0) for 12 hours, 6 hours with 1 M sodium chloride, 2 days with 2 M sodium chloride, and 2 days with tertiary distilled water.
  • sodium hydrogen phosphate Na 2 HPO 4, pH 9.0
  • a support sample was prepared with a punch of 6 mm in diameter, sterilized by gamma irradiation ( ⁇ -irradiation) of 10 KGy, and stored at ⁇ 20 ° C. until use.
  • Example 1 The pulp cells prepared in Example 1 were inoculated at a concentration of 2 ⁇ 10 6 cells on the previously prepared support sample, and cell proliferation and differentiation were induced in the same manner as in Example 3.
  • MTT assay and RT-PCR were performed on the support sample in the same manner as in Example 3 except that the MTT assay was performed 3 days after inoculation of pulp cells.
  • the type 3 collagen, osteocalcin and the like which are extracellular matrices that proliferate well and are expressed in osteoblasts, are well expressed in all supports, and in particular, BMP- It can be seen that the cells proliferate better and the extracellular matrix is expressed even more well on the 2 coated support.
  • the support sample was pocket-transplanted into the back of a nude mouse in the same manner as in Example 3, and biopsy was performed at 8 weeks, followed by Boncosa staining and osteonectin. Staining and alcian blue staining were performed.
  • the calcium deposition on all the support is well formed, the expression of Boncosa, osteonectin and alkian blue was found to be well, especially in the BMP-2 coated support The better the deposition, the better the expression of Boncosa, osteonectin and alcian blue, and thus, it can be seen that it is effective in bone differentiation.
  • Crosslinking was carried out in the following manner. That is, the construct was treated with 20 ml of 40% (v / v) ethanol containing 50 mM morphoethane ethanesulfonic acid (MES, pH 5.5) for 30 minutes and then 50 mM morphoethane ethanesulfonic acid (MES, pH 5.5). ), 40% (v / v) ethanol with 24 mM ethyldimethylaminopropyl carbodiimide and 5 mM hydroxysuccinimide for 12 hours.
  • 40% (v / v) ethanol containing 50 mM morphoethane ethanesulfonic acid (MES, pH 5.5) for 30 minutes and then 50 mM morphoethane esulfonic acid (MES, pH 5.5).
  • lyophilization was again performed at -80 ° C, and a support sample was prepared using a 6 mm diameter punch, and then sterilized by 10 KGy gamma irradiation ( ⁇ -irradiation) and stored at -20 ° C until use.
  • the support sample was subjected to RT-PCR in the same manner as in Example 3.
  • the extracellular matrix, type 1 collagen, type 3 collagen, osteonectin, osteoprotegerin, osteopontin, osteocalcin and the like expressed in osteoblasts It can be seen that it is well expressed, in particular nanohydroxyapatite and BMP-2 can be seen that even more well expressed on the applied support.
  • a PGA layer and silk fibers formed of a surgical suture and PGA used as various biomaterials
  • a porous support was prepared in the same manner as in Example 3 except that a silk fiber layer formed of Korea) and 0.4 mL of hyaluronic acid were used.
  • porous support including the PGA layer prepared above and the porous support including the silk fiber layer and the porous support prepared using 0.4 ml of hyaluronic acid in Example 3 according to the present disclosure were subjected to gamma irradiation ( ⁇ -irradiation) of 10 KGy. Sterilized.
  • the histology was performed 2 weeks after pocket transplantation into the back of the nude mouse in the same manner as described in Example 3.
  • the porous support including the sericin-free silk layer according to the present disclosure can be seen that the inflammatory response is significantly reduced even when implanted in the body.
  • a porous support was prepared in the same manner as in Example 6 without forming a silk layer and without coating with nanohydroxyapatite and BMP-2.
  • a porous support prepared in Example 6 according to the present disclosure without a coating using nanohydroxyapatite and BMP-2 was prepared in a size of 10 mm ⁇ 5 mm, and then a tensile strength meter (H5KT, HOUNSFIELD, UK) was used. Tensile strength was measured at a rate of 2 mm / min (10% strain rate).
  • the tensile strength of the porous support prepared according to the present disclosure is 40 ⁇ 8 N, which is about 20 times stronger than the porous support (2 ⁇ 0.5 N) prepared as a comparative example Appeared to have.

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Abstract

La présente invention concerne un support poreux pour la régénération tissulaire guidée et un procédé de préparation de ce dernier. Bien que le support poreux selon la présente invention ne nécessite pas d'opération d'ablation additionnelle, il permet la biodégradation avec d'excellentes propriétés de régénération tissulaire et mécaniques et il est réalisé uniquement en polymère naturel qui élicite une très faible réponse immunitaire et offre une biocompatibilité supérieure qui joue un rôle sur la greffe. De plus, on utilise de la soie tissée pour assurer les sutures de sorte que les propriétés physiques du support sont améliorées. Par conséquent, cette invention est utilisée en pratique pour la régénération de tissus tels que les os alvéolaires et les ligaments parodontaux et en tant que substrat pour stimuler la régénération osseuse en chirurgie orthopédique.
PCT/KR2009/005397 2008-09-23 2009-09-22 Support poreux pour la régénération tissulaire guidée et procédé de préparation de ce support Ceased WO2010036009A2 (fr)

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KR1020080093339A KR100937736B1 (ko) 2008-09-23 2008-09-23 유도 조직 재생을 위한 다공성 지지체 및 그의 제조방법
KR10-2008-0093339 2008-09-23

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CZ307053B6 (cs) * 2013-12-30 2017-12-20 Student Science, s. r. o. 3 D kolagenové porézní kompozitní nosiče pro akcelerovanou regeneraci kostí
US20210052771A1 (en) * 2018-04-06 2021-02-25 The Regents Of The University Of California Nanoparticulate mineralized collagen glycosaminoglycan scaffold with an anti-resorption factor

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KR101106018B1 (ko) * 2010-02-09 2012-01-17 차의과학대학교 산학협력단 열적 안정성을 갖는 생분해성 고분자를 이용한 조직재생용 지지체의 제조방법
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