WO2010081408A1 - Bioactive tissue regeneration film and preparation method thereof - Google Patents

Bioactive tissue regeneration film and preparation method thereof Download PDF

Info

Publication number
WO2010081408A1
WO2010081408A1 PCT/CN2010/070133 CN2010070133W WO2010081408A1 WO 2010081408 A1 WO2010081408 A1 WO 2010081408A1 CN 2010070133 W CN2010070133 W CN 2010070133W WO 2010081408 A1 WO2010081408 A1 WO 2010081408A1
Authority
WO
WIPO (PCT)
Prior art keywords
silk fibroin
bioactive
tissue
tissue regeneration
factor
Prior art date
Application number
PCT/CN2010/070133
Other languages
French (fr)
Chinese (zh)
Inventor
张玉峰
刘朝阳
马兆成
Original Assignee
武汉本药康华生物科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 武汉本药康华生物科技有限公司 filed Critical 武汉本药康华生物科技有限公司
Publication of WO2010081408A1 publication Critical patent/WO2010081408A1/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/005—Ingredients of undetermined constitution or reaction products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/16—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices

Definitions

  • the invention relates to a biomedical engineering material and a preparation method thereof, in particular to a three-dimensional tissue repairing material capable of inducing tissue regeneration and a preparation method thereof, and belongs to the technical field of biomedical materials.
  • Membrane materials can be mainly divided into absorbable and non-absorbable films.
  • Non-absorbable non-degradable film material represented by polytetrafluoroethylene (PTFE).
  • PTFE polytetrafluoroethylene
  • the film has the characteristics of stable physical and chemical properties, low friction coefficient, no stickiness, hydrophobicity, heat resistance, corrosion resistance, bio-aging resistance, and easy sterilization. At the same time, it has good biocompatibility and does not affect the regeneration process. Therefore, it has been widely used and has been regarded as the "gold standard". However, its clinical application is time-consuming and difficult. After 4 to 6 weeks of placement, it requires secondary surgery to remove the tissue that has been repaired. The removal time is not easy to determine. At the same time, it causes secondary trauma to the patient, and it has been used less frequently.
  • biodegradable biofilms There are two main types of biodegradable biofilms reported in the literature: one is collagen membrane made of animal collagen or collagen fiber, and the other is membrane made of polylactic acid and medical plasticizer or collagen fiber. Most of these biodegradable biofilms are composed of collagen and polylactic acid. However, when they are implanted into the body, they are prone to acid production during the degradation process, which may lead to an inflammatory reaction in the implanted area, and there is a risk of rejection in the body.
  • the object of the present invention is to overcome the deficiencies of the prior art and to provide a bioactive tissue regeneration membrane which is biocompatible, degradable in vivo and has a barrier function, and also has tissue-induced regenerability and a preparation method thereof.
  • the bioactive tissue regeneration membrane is a two-layer composite structure, and the smooth dense layer is a cell closure layer, which can prevent unwanted soft tissue mucosal cells from invading into the transplant defect area; the proximal transplantation area is a loose multi-space structure, which can bind organisms. Active factor, which can promote tissue regeneration.
  • a bioactive tissue regeneration membrane which is a two-layer composite structure, characterized in that: one layer is a tissue isolation layer composed of silk fibroin, and its appearance The surface is smooth and dense, the inner surface is a scratched rough surface; the other layer is a tissue inducing layer composed of a mixture of silk fibroin or silk fibroin and biologically active factors attached to the rough surface of the tissue separation layer.
  • the outer surface is loose and long and empty.
  • the mixture of the silk fibroin and the biologically active factor has a bioactive factor content of 3-10 mg/ml, and the bioactive factor is bone morphogenetic protein, platelet-derived growth factor, insulin-like growth factor, fibroblast growth factor or transformation. Growth factor.
  • the present invention also provides a method for preparing the above bioactive tissue regeneration membrane, which comprises the steps of:
  • the scratch roughening treatment is performed by sharpening a side of the tissue separation layer composed of silk fibroin to form a staggered scratch having a depth of 1 to 10 ⁇ m.
  • the 10 ⁇ 30% silk fibroin solution mixed with the bioactive factor has a bioactive factor content of 3-10 mg/ml, and the bioactive factors are bone morphogenetic protein, platelet-derived growth factor, insulin-like growth factor, and One or more of fibroblast growth factor or transforming growth factor.
  • the smooth layer of the bioactive tissue regeneration membrane provided by the present invention is a cell closure layer, which can prevent unwanted soft tissue mucosal cells from invading into the transplant defect area, and can also serve as a platform for supporting tissue repair to assist wound recovery.
  • the bioactive tissue regeneration membrane allows blood vessels to grow inward into the protein fiber space, thereby promoting blood vessel density at the transplant site.
  • the rough surface of the bioactive tissue regeneration membrane is composed of a large number of nano-scale protein bundles, with a large space between the protein bundles, which can fix the tissue to heal early blood clots and support new tissue regeneration. Tissue growth is better promoted by the addition of growth factors.
  • the biologically active tissue is re
  • the biofilm can be digested into natural amino acids. Unlike films made from polylactic acid and polyglycolide, a large amount of acid is produced during the degradation process, thereby damaging the repaired tissue.
  • the bioactive bone-inducing regenerative membrane provided by the invention has wide application range and can be widely used in orthopedics, cranial and maxillofacial surgery, ophthalmology, general, extrathoracic, dental implant, periodontal surgery and reconstructive surgery. A number of clinical disciplines such as neurosurgery.
  • the preparation method provided by the invention is simple, the process is mature and easy to control.
  • Figure 1 is a scanning electron micrograph of the bioactive tissue regeneration membrane section
  • Figure 2 is a scanning electron micrograph of cells cultured on the surface of bioactive tissue regeneration membrane.
  • Example 1 Bioactive tissue regeneration membrane for periodontal tissue regeneration
  • the silk fibroin was dissolved in deionized water to prepare a 10% silk fibroin solution, and then the solution was uniformly coated on the scratched film surface which was formed, and then placed in a -80 ° C refrigerator. After freezing for 10 hours, it was dried in a vacuum freeze dryer and released into a mold. According to the clinical requirements, it is cut into a suitable size and packaged, and then sterilized by ethylene oxide for use.
  • the silk fibroin film may be first cross-linked and then scratched before the scratch-roughening treatment with the probe.
  • Example 2 Bioactive tissue regeneration membrane for bone tissue regeneration (1)
  • the clean and impurity-removed silk was selected in a 0.5% Na 2 CO 3 aqueous solution, and the gel was degummed twice at 100 ° C for 1 hour each time, and the bath ratio was 1:50.
  • the degummed silk fibroin was dried to dryness in a drying oven at 80 °C.
  • 10 g of the silk fibroin fiber after degumming was dissolved in 100 ml of a calcium chloride I ethanol I water ternary solution (molar ratio of 1:2:8), and dissolved by magnetic stirring at 70 ° C under constant temperature.
  • the silk fibroin solution obtained above is subjected to dialysis, desalting and purification to obtain a water-soluble silk fibroin solution having a concentration of 2.5 to 3.5%, and concentrated and dried to obtain silk fibroin.
  • the silk fibroin is dissolved in deionized water to prepare a silk fibroin solution having a concentration of 30%, and then electrospinning is performed, and the spun film surface is sprayed according to a conventional condition, that is, A nanostructure-like staggered silk protein bundle was formed on the film surface, and then it was frozen in a refrigerator at -70 ° C for 10 hours, and then vacuum-dried in a vacuum freeze dryer to release the mold. According to the clinical requirements, it is cut into a suitable size and packaged, and then sterilized by ethylene oxide for use.
  • Example 3 Growth factor composite bioactive tissue regeneration membrane for bone tissue regeneration
  • the flat plate can be selected from non-stick plates such as glass plates and polytetrafluoroethylene plates, and the specifications can be based on clinical needs. As a result, there is no effect on the effects of the present invention.

Abstract

Provided is a double-deck composite structured bioactive tissue regeneration film which has a tissue isolation layer consisting of silk fibroins, and a tissue induction layer adhered to the tissue isolation layer, which consists of silk fibroin beams or silk fibroin beams mixed with biological active factors. The film is smooth and dense on one side, with loose porous structure on the other side. A method for preparing the bioactive tissue regeneration film is also provided. The method comprises the following steps: the silk fibroin solution is swollen, stirred, deaired and casted into film; one side of the film is scratched to be coarsened; the silk fibroin solution or the silk fibroin solution mixed with biological active factors is coated on the scratched side; the film is obtained after freezing, vacuum drying and demoulding. The film is nontoxic and harmless, has perfect biocompatibility, promote the adherence and growth of cells, and can be used for substituting collagen in clinical medicine.

Description

说 明 书  Description
一种生物活性组织再生膜及其制备方法 技术领域  Bioactive tissue regeneration membrane and preparation method thereof
本发明涉及一种生物医学工程材料及其制备方法， 特别涉及一种可诱导组织 再生的三维组织修复材料及其制备方法， 属生物医用材料技术领域。  The invention relates to a biomedical engineering material and a preparation method thereof, in particular to a three-dimensional tissue repairing material capable of inducing tissue regeneration and a preparation method thereof, and belongs to the technical field of biomedical materials.
背景技术 Background technique
引导组织再生术是目前临床上对组织缺损修复的最佳方法， 其原理是利用生 物膜的物理屏障功能将骨缺损区与周围组织隔离，创造一个相对封闭的组织环境， 从而使骨组织的再生功能得到最大程度的发挥。 它的应用为牙周病的治疗， 牙种 植区骨量不足及其它骨缺损的修复、 骨折的愈合提供了一个新的有效途径。  Guided tissue regeneration is currently the best clinical method for tissue defect repair. Its principle is to use the physical barrier function of biofilm to isolate the bone defect area from the surrounding tissue, creating a relatively closed tissue environment, thus regenerating bone tissue. The function is maximized. Its application provides a new and effective way for the treatment of periodontal disease, the lack of bone mass in the dental implant area and the repair of other bone defects and the healing of fractures.
膜材料主要可分为可吸收性和不可吸收性膜两种。 ①不可吸收性非降解膜材 料， 以聚四氟乙烯 (PTFE) 为代表。这种膜具有理化性能稳定、摩擦系数小、无黏 性、 疏水、 耐热、 耐腐蚀、 耐生物老化、 消毒杀菌容易等特点。 同时生物相容性 好， 不影响再生过程， 因此应用广泛， 曾被奉为 "金标准"。 但其临床应用操作费 时而且困难， 放置 4〜6 周后需二次手术摘除， 易损坏已经修复好的组织， 取出 时间也不易确定， 同时给患者造成二次创伤， 现在已较少应用。 ②可吸收性降解 膜材料。 由于这些材料植入体内后不需取出， 减少了患者的痛苦和手术难度， 因 此得到较广泛的应用。 目前文献报道的可降解生物膜主要有两种： 一种为动物胶 原蛋白或胶原纤维制成的胶原膜， 另一种为以聚乳酸和医用增塑剂或胶原纤维制 成的膜。 这些可降解生物膜大多由胶原及聚乳酸等构成， 但将它们植入体内后， 在降解过程中容易产酸从而可能导致植入区发生炎症反应， 同时存在着体内排斥 反应的风险。  Membrane materials can be mainly divided into absorbable and non-absorbable films. 1 Non-absorbable non-degradable film material, represented by polytetrafluoroethylene (PTFE). The film has the characteristics of stable physical and chemical properties, low friction coefficient, no stickiness, hydrophobicity, heat resistance, corrosion resistance, bio-aging resistance, and easy sterilization. At the same time, it has good biocompatibility and does not affect the regeneration process. Therefore, it has been widely used and has been regarded as the "gold standard". However, its clinical application is time-consuming and difficult. After 4 to 6 weeks of placement, it requires secondary surgery to remove the tissue that has been repaired. The removal time is not easy to determine. At the same time, it causes secondary trauma to the patient, and it has been used less frequently. 2 absorbable degradation of membrane materials. Since these materials do not need to be taken out after being implanted in the body, the pain of the patient and the difficulty of surgery are reduced, and thus it is widely used. There are two main types of biodegradable biofilms reported in the literature: one is collagen membrane made of animal collagen or collagen fiber, and the other is membrane made of polylactic acid and medical plasticizer or collagen fiber. Most of these biodegradable biofilms are composed of collagen and polylactic acid. However, when they are implanted into the body, they are prone to acid production during the degradation process, which may lead to an inflammatory reaction in the implanted area, and there is a risk of rejection in the body.
发明内容 Summary of the invention
本发明的目的在于克服现有技术存在的不足， 提供一种生物相容性好、 能在 体内降解和具有分隔屏障作用， 同时还具有组织诱导再生性的生物活性组织再生 膜及其制备方法。 该种生物活性组织再生膜为双层复合结构， 其平滑的致密层是 细胞闭合层， 可以阻止不需要的软组织粘膜细胞侵袭到移植缺陷区； 近移植区则 为疏松多空结构， 可结合生物活性因子， 能够很好的促进组织再生。  SUMMARY OF THE INVENTION The object of the present invention is to overcome the deficiencies of the prior art and to provide a bioactive tissue regeneration membrane which is biocompatible, degradable in vivo and has a barrier function, and also has tissue-induced regenerability and a preparation method thereof. The bioactive tissue regeneration membrane is a two-layer composite structure, and the smooth dense layer is a cell closure layer, which can prevent unwanted soft tissue mucosal cells from invading into the transplant defect area; the proximal transplantation area is a loose multi-space structure, which can bind organisms. Active factor, which can promote tissue regeneration.
为实现上述目的， 本发明采用的技术方案是： 一种生物活性组织再生膜， 该 膜为双层复合结构， 其特征在于： 一层为由丝素蛋白构成的组织隔离层， 其外表 面呈光滑致密状， 内表面为有划痕的粗糙面； 另一层为附着在组织隔离层粗糙面 上的由丝素蛋白或丝素蛋白与生物活性因子的混合物构成的组织诱导层， 其外表 面呈疏松多空结构。 In order to achieve the above object, the technical solution adopted by the present invention is: a bioactive tissue regeneration membrane, which is a two-layer composite structure, characterized in that: one layer is a tissue isolation layer composed of silk fibroin, and its appearance The surface is smooth and dense, the inner surface is a scratched rough surface; the other layer is a tissue inducing layer composed of a mixture of silk fibroin or silk fibroin and biologically active factors attached to the rough surface of the tissue separation layer. The outer surface is loose and long and empty.
所述丝素蛋白与生物活性因子的混合物中， 生物活性因子的含量为 3-10mg/ml, 生物活性因子为骨形成蛋白、血小板衍生生长因子、胰岛素样生长因 子、 成纤维细胞生长因子或转化生长因子。  The mixture of the silk fibroin and the biologically active factor has a bioactive factor content of 3-10 mg/ml, and the bioactive factor is bone morphogenetic protein, platelet-derived growth factor, insulin-like growth factor, fibroblast growth factor or transformation. Growth factor.
本发明还提供了上述生物活性组织再生膜的制备方法， 其特征在于包括以下 步骤：  The present invention also provides a method for preparing the above bioactive tissue regeneration membrane, which comprises the steps of:
(1)将丝素蛋白溶于去离子水配成浓度范围为 10~30%的丝素蛋白溶液备用； (1) Dissolving silk fibroin in deionized water to prepare a silk fibroin solution having a concentration ranging from 10 to 30%;
(2)将浓度为 10~30%的丝素蛋白溶液在室温下充分溶胀后， 搅拌至丝素蛋白 完全溶解、 脱泡， 然后倒入平板模上流延成膜， 再室温挥发、 于 50~70°C真空干 燥制成由丝素蛋白构成的组织隔离层； (2) After the silk fibroin solution having a concentration of 10-30% is fully swelled at room temperature, stir until the silk fibroin is completely dissolved and defoamed, then poured into a flat mold and cast into a film, and then volatilized at room temperature, at 50~ Vacuum drying at 70 ° C to form a tissue isolation layer composed of silk fibroin;
(3)在已制成的组织隔离层一面进行划痕粗化处理， 再将 10~30%的丝素蛋白 溶液或者混有生物活性因子的 10~30%的丝素蛋白溶液， 均匀涂布或者采用静电 纺丝方法喷涂在有划痕的粗糙面上；  (3) Scratch roughening treatment on one side of the prepared tissue isolation layer, and uniformly coating 10 to 30% of the silk fibroin solution or 10 to 30% of the silk fibroin solution mixed with the bioactive factor Or spraying on a rough surface with scratches by electrospinning;
(4)在 -40~-80°C条件下冷冻 8~12小时， 然后在真空冷冻干燥器中真空干燥， 脱模即可。  (4) Freeze at -40 to -80 °C for 8 to 12 hours, then vacuum dry in a vacuum freeze dryer to release the mold.
所述的划痕粗化处理， 是在由丝素蛋白构成的组织隔离层的一面上用尖状物 刻划， 使表面形成深度为 1~10 微米的交错状划痕。 所述混有生物活性因子的 10~30%的丝素蛋白溶液中， 生物活性因子的含量为 3-10mg/ml， 生物活性因子为 骨形成蛋白、 血小板衍生生长因子、 ***、 成纤维细胞生长因子或 转化生长因子的一种或几种。  The scratch roughening treatment is performed by sharpening a side of the tissue separation layer composed of silk fibroin to form a staggered scratch having a depth of 1 to 10 μm. The 10~30% silk fibroin solution mixed with the bioactive factor has a bioactive factor content of 3-10 mg/ml, and the bioactive factors are bone morphogenetic protein, platelet-derived growth factor, insulin-like growth factor, and One or more of fibroblast growth factor or transforming growth factor.
本发明具有以下优点：  The invention has the following advantages:
1. 本发明提供的生物活性组织再生膜平滑的致密层是细胞闭合层，可以阻止 不需要的软组织粘膜细胞侵袭到移植缺陷区， 也可作为一个支持组织修复的平台 辅助创面恢复。 当这些软组织细胞生长至闭合时， 生物活性组织再生膜允许血管 向内生长到蛋白纤维空间中， 所以能促进移植点的血管密集。  1. The smooth layer of the bioactive tissue regeneration membrane provided by the present invention is a cell closure layer, which can prevent unwanted soft tissue mucosal cells from invading into the transplant defect area, and can also serve as a platform for supporting tissue repair to assist wound recovery. When these soft tissue cells are grown to be closed, the bioactive tissue regeneration membrane allows blood vessels to grow inward into the protein fiber space, thereby promoting blood vessel density at the transplant site.
生物活性组织再生膜的粗糙面是由大量的纳米级别的蛋白束构成， 在蛋白束 之间具有大量的空间， 可以固定组织愈合早期的血凝块以及支持新的组织再生。 加入生长因子后更能很好的促进组织再生。 在组织再生完全后， 生物活性组织再 生膜可以被酶解为天然的氨基酸。 不像聚乳酸和聚乙交酯制成的膜在降解过程中 会产生大量的酸， 从而伤害修复的组织。 The rough surface of the bioactive tissue regeneration membrane is composed of a large number of nano-scale protein bundles, with a large space between the protein bundles, which can fix the tissue to heal early blood clots and support new tissue regeneration. Tissue growth is better promoted by the addition of growth factors. After the tissue regeneration is complete, the biologically active tissue is re The biofilm can be digested into natural amino acids. Unlike films made from polylactic acid and polyglycolide, a large amount of acid is produced during the degradation process, thereby damaging the repaired tissue.
2. 本发明提供的这种生物活性骨诱导再生膜适用范围广， 可广泛用于骨科、 颅、 颌面外科、 眼科、 普外、 胸外、 牙种植科、 牙周外科和修复重建外科及神经 外科等多个临床学科。  2. The bioactive bone-inducing regenerative membrane provided by the invention has wide application range and can be widely used in orthopedics, cranial and maxillofacial surgery, ophthalmology, general, extrathoracic, dental implant, periodontal surgery and reconstructive surgery. A number of clinical disciplines such as neurosurgery.
3. 本发明提供的制备方法简单， 工艺成熟， 易于控制。  3. The preparation method provided by the invention is simple, the process is mature and easy to control.
附图说明 DRAWINGS
图 1为生物活性组织再生膜切面的扫描电镜图  Figure 1 is a scanning electron micrograph of the bioactive tissue regeneration membrane section
图 2为生物活性组织再生膜表面培养细胞后的扫描电镜图  Figure 2 is a scanning electron micrograph of cells cultured on the surface of bioactive tissue regeneration membrane.
具体实施方式 detailed description
下面通过具体实施例对本发明的技术方案做进一步说明， 其目的在于帮助更 好的理解本发明的内容， 但这些具体实施方案不以任何方式限制本发明的保护范 围。 本实施方案中所用的原料可在市场上购得， 或可用本领域已知的方法得到。 本实施方案中所说的浓度均为质量百分比浓度。  The technical solutions of the present invention are further described in the following by means of specific embodiments, which are intended to facilitate a better understanding of the present invention, but the specific embodiments do not limit the scope of protection of the present invention in any way. The starting materials used in this embodiment are commercially available or can be obtained by methods known in the art. The concentrations referred to in this embodiment are all mass percentage concentrations.
实施例 1 : 用于牙周组织再生的生物活性组织再生膜  Example 1 : Bioactive tissue regeneration membrane for periodontal tissue regeneration
(1) 选取干净除杂的蚕丝于 0.5% Na2CO3水溶液中， 100°C脱胶两次， 每次 1 小时， 浴比 1 : 50。 脱胶后的丝素置于 80°C的干燥箱中干燥至干。 取脱胶后的丝 素纤维 10克溶于 100ml的氯化钙 I乙醇 I水三元溶液 （摩尔比为 1 : 2 : 8) 中， 在 70°C恒温磁力搅拌溶解。 将上述获得的丝素溶液进行透析、 脱盐、 纯化， 得到 浓度为 2.5~3.5%的水溶性丝素溶液， 浓缩干燥即可得丝素蛋白。 (1) Select clean and impurity-removed silk in 0.5% Na 2 CO 3 aqueous solution, degumming twice at 100 ° C for 1 hour each time, bath ratio 1: 50. The degummed silk fibroin was dried to dryness in a drying oven at 80 °C. 10 g of the silk fibroin fiber after degumming was dissolved in 100 ml of a calcium chloride I ethanol I water ternary solution (molar ratio of 1:2:8), and dissolved by magnetic stirring at 70 ° C under constant temperature. The silk fibroin solution obtained above is subjected to dialysis, desalting and purification to obtain a water-soluble silk fibroin solution having a concentration of 2.5 to 3.5%, and concentrated and dried to obtain silk fibroin.
(2)用天平称取 6g丝素蛋白， 加入到 20ml的去离子水中， 搅拌溶解 1~3小 时至丝素蛋白完全溶解，脱泡 1~3小时至溶液中无气泡后，倒入 5x5cm的平板模， 本实施例选用的是平板玻璃模上平淌流延成膜，室温下挥发 1~2天后于 70°C下真 空干燥 8小时至膜干燥成型。 此时用探针进行划痕粗化处理， 使该膜的一面上形 成深度为 1~10微米的交错状划痕。 将丝素蛋白溶解到去离子水中， 配成 10%的 丝素蛋白溶液， 然后将该溶液均匀涂布在已制成的有划痕的膜面上， 然后将其放 入 -80°C冰箱冷冻 10小时后， 放入真空冷冻干燥器中干燥， 脱模即成。 根据临床 要求裁剪成适当大小的块状后封装， 经环氧乙烷消毒备用。 在用探针进行划痕粗 化处理之前， 还可对丝素蛋白膜先进行交联处理， 再划痕。  (2) Weigh 6g of silk fibroin with a balance, add to 20ml of deionized water, stir and dissolve for 1~3 hours until the silk fibroin is completely dissolved, defoam for 1~3 hours until no bubbles in the solution, pour into 5x5cm For the flat plate mold, the flat glass mold is cast on a flat glass mold, and the film is volatilized at room temperature for 1 to 2 days, and then vacuum dried at 70 ° C for 8 hours to form a film. At this time, the probe was subjected to scratch roughening treatment to form a staggered scratch having a depth of 1 to 10 μm on one side of the film. The silk fibroin was dissolved in deionized water to prepare a 10% silk fibroin solution, and then the solution was uniformly coated on the scratched film surface which was formed, and then placed in a -80 ° C refrigerator. After freezing for 10 hours, it was dried in a vacuum freeze dryer and released into a mold. According to the clinical requirements, it is cut into a suitable size and packaged, and then sterilized by ethylene oxide for use. The silk fibroin film may be first cross-linked and then scratched before the scratch-roughening treatment with the probe.
实施例 2: 用于骨组织再生的生物活性组织再生膜 (1) 选取干净除杂的蚕丝于 0.5%Na2CO3水溶液中， 100°C脱胶两次， 每次 1 小时， 浴比 1 : 50。 脱胶后的丝素置于 80°C的干燥箱中干燥至干。 取脱胶后的丝 素纤维 10g溶于 100ml的氯化钙 I乙醇 I水三元溶液（摩尔比为 1 : 2 : 8)中， 在 70°C恒温磁力搅拌溶解。 将上述获得的丝素溶液进行透析、 脱盐、 纯化， 得到浓 度为 2.5~3.5%的水溶性丝素溶液， 浓缩干燥即可得丝素蛋白。 Example 2: Bioactive tissue regeneration membrane for bone tissue regeneration (1) The clean and impurity-removed silk was selected in a 0.5% Na 2 CO 3 aqueous solution, and the gel was degummed twice at 100 ° C for 1 hour each time, and the bath ratio was 1:50. The degummed silk fibroin was dried to dryness in a drying oven at 80 °C. 10 g of the silk fibroin fiber after degumming was dissolved in 100 ml of a calcium chloride I ethanol I water ternary solution (molar ratio of 1:2:8), and dissolved by magnetic stirring at 70 ° C under constant temperature. The silk fibroin solution obtained above is subjected to dialysis, desalting and purification to obtain a water-soluble silk fibroin solution having a concentration of 2.5 to 3.5%, and concentrated and dried to obtain silk fibroin.
(2) 用天平称取 6g丝素蛋白， 加入到 30ml的去离子水中， 搅拌溶解 1~3小 时至丝素蛋白完全溶解，脱泡 1~3小时至溶液中无气泡后，倒入 5x5cm的平板模， 本实施例选用的是平板玻璃模上平淌流延成膜， 挥发 2天后于 50°C下真空干燥 7 小时至膜干燥成型。此时用探针划痕粗化处理， 使该膜的一面上形成深度为 1~10 微米的交错状划痕。 将丝素蛋白溶解在去离子水中， 配成浓度为 30%的丝素蛋白 溶液，然后采用静电纺丝法，按照常规条件在已制成的有划痕的膜面上进行喷纺， 即在膜面上形成纳米结构样的交错状蚕丝蛋白束，然后将其放入冰箱中在 -70°C下 冷冻 10小时后， 放入真空冷冻干燥器中真空干燥， 脱模即成。根据临床要求裁剪 成适当大小的块状后封装， 经环氧乙烷消毒备用。  (2) Weigh 6g of silk fibroin with a balance, add to 30ml of deionized water, stir and dissolve for 1~3 hours until the silk fibroin is completely dissolved, defoam for 1~3 hours until no bubbles in the solution, pour into 5x5cm For the flat plate mold, the flat glass mold was cast on a flat glass mold, and after volatilization for 2 days, it was vacuum dried at 50 ° C for 7 hours until the film was dried. At this time, the probe is subjected to roughening treatment to form a staggered scratch having a depth of 1 to 10 μm on one side of the film. The silk fibroin is dissolved in deionized water to prepare a silk fibroin solution having a concentration of 30%, and then electrospinning is performed, and the spun film surface is sprayed according to a conventional condition, that is, A nanostructure-like staggered silk protein bundle was formed on the film surface, and then it was frozen in a refrigerator at -70 ° C for 10 hours, and then vacuum-dried in a vacuum freeze dryer to release the mold. According to the clinical requirements, it is cut into a suitable size and packaged, and then sterilized by ethylene oxide for use.
实施例 3 : 用于骨组织再生的生长因子复合生物活性组织再生膜  Example 3: Growth factor composite bioactive tissue regeneration membrane for bone tissue regeneration
(1) 选取干净除杂的蚕丝于 0.5% Na2CO3水溶液中， 100°C脱胶两次， 每次 1 小时， 浴比 1 : 50。 脱胶后的丝素置于 80°C的干燥箱中干燥至干。 取 10g脱胶后 的丝素纤维溶于 100ml的氯化钙 I乙醇 I水三元溶液（摩尔比为 1： 2： 8)中， 在 70°C恒温磁力搅拌溶解。 将上述获得的丝素溶液进行透析、 脱盐、 纯化， 得到浓 度为 2.5~3.5%的水溶性丝素溶液， 浓缩干燥即可得丝素蛋白。 (1) Select clean and impurity-removed silk in 0.5% Na 2 CO 3 aqueous solution, degumming twice at 100 ° C for 1 hour each time, bath ratio 1: 50. The degummed silk fibroin was dried to dryness in a drying oven at 80 °C. 10 g of degummed silk fibroin fiber was dissolved in 100 ml of calcium chloride I ethanol I water ternary solution (molar ratio 1:2:8), and dissolved by magnetic stirring at 70 ° C under constant temperature. The silk fibroin solution obtained above is subjected to dialysis, desalting and purification to obtain a water-soluble silk fibroin solution having a concentration of 2.5 to 3.5%, and concentrated and dried to obtain silk fibroin.
(2) 用天平称取 6g丝素蛋白， 加入到 40ml的去离子水中， 搅拌溶解 1~3小 时至丝素蛋白完全溶解，脱泡 1~3小时至溶液中无气泡后，倒入 5x5cm的平板模， 本实施例选用的是平板玻璃模上平淌流延成膜， 挥发 1天后于 60°C下真空干燥 8 小时。 此时用探针划痕粗化处理， 使膜面上形成深度为 1~10微米的交错状划痕。  (2) Weigh 6g silk fibroin with a balance, add to 40ml of deionized water, stir and dissolve for 1-3 hours until the silk fibroin is completely dissolved, defoam for 1-3 hours until there is no bubble in the solution, pour into 5x5cm For the flat plate mold, the flat glass mold was cast on a flat glass mold to form a film, and after volatilization for 1 day, it was vacuum dried at 60 ° C for 8 hours. At this time, the probe is subjected to roughening treatment to form a staggered scratch having a depth of 1 to 10 μm on the film surface.
(3) 将 0.5g生物活性因子重组人基因骨形成蛋白的冻干粉， 加入到用丝素蛋 白与去离子水配成的 30%丝素蛋白液中混合均匀， 再采用静电纺丝法， 按照常规 条件进行喷纺， 即在膜面上形成携带生长因子的蛋白束， 然后将其放入冰箱中在 -50°C下冷冻 12小时后， 放入真空冷冻干燥器中真空干燥， 脱模即成。 根据临床 要求裁剪成适当大小的块状后封装， 经环氧乙烷消毒备用。  (3) 0.5 g of biologically active factor recombinant human bone-forming protein lyophilized powder, added to a 30% silk fibroin solution prepared by using silk fibroin and deionized water, and uniformly mixed, and then electrospinning method, Spray spinning according to conventional conditions, that is, forming a protein bundle carrying a growth factor on the film surface, and then freezing it in a refrigerator at -50 ° C for 12 hours, and then vacuum-drying in a vacuum freeze dryer to release the mold. Serve. According to the clinical requirements, it is cut into a suitable size and packaged, and then sterilized by ethylene oxide for use.
其中， 平板模可选用玻璃板、 聚四氟乙烯板等不粘板， 其规格可根据临床需 要而定， 对本发明的效果没有影响 Among them, the flat plate can be selected from non-stick plates such as glass plates and polytetrafluoroethylene plates, and the specifications can be based on clinical needs. As a result, there is no effect on the effects of the present invention.

Claims

权 利 要 求 书 Claim
1. 一种生物活性组织再生膜， 该膜为双层复合结构， 其特征在于： 一层为由丝 素蛋白构成的组织隔离层， 其表面呈光滑致密状； 另一层为附着在组织隔离 层粗糙面上的由丝素蛋白束或丝素蛋白与生物活性因子的混合物构成的组织 诱导层， 其表面呈疏松多空结构。  A bioactive tissue regeneration membrane, the membrane being a two-layer composite structure, characterized in that: one layer is a tissue separation layer composed of silk fibroin, the surface of which is smooth and dense; the other layer is attached to the tissue isolation A tissue-inducing layer composed of a silk fibroin bundle or a mixture of silk fibroin and a biologically active factor on a rough surface of the layer, the surface of which is loose and long-spaced.
2. 根据权利要求 1所述的生物活性组织再生膜， 其特征在于： 再生组织面为粗 糙多孔。  The bioactive tissue regeneration membrane according to claim 1, wherein the regenerated tissue surface is rough and porous.
3. 根据权利要求 1所述的生物活性组织再生膜， 其特征在于： 所述丝素蛋白与 生物活性因子的混合物中， 生物活性因子的含量为 3-10mg/ml。  The bioactive tissue regeneration membrane according to claim 1, wherein the content of the bioactive factor in the mixture of the silk fibroin and the biologically active factor is 3-10 mg/ml.
4. 根据权利要求 1所述的生物活性组织再生膜， 其特征在于： 所述生物活性因 子为骨形成蛋白、 血小板衍生生长因子、 ***、 成纤维细胞生 长因子或转化生长因子中的一种或几种。  The bioactive tissue regeneration membrane according to claim 1, wherein the bioactive factor is a bone morphogenetic protein, a platelet-derived growth factor, an insulin-like growth factor, a fibroblast growth factor or a transforming growth factor. One or several.
5. 一种权利要求 1所述的生物活性组织再生膜的制备方法， 其特征在于包括以 下步骤：  A method of producing a bioactive tissue regeneration membrane according to claim 1, comprising the steps of:
(5)将丝素蛋白溶于去离子水配成浓度范围为 10~30%的丝素蛋白溶液备用； (5) dissolving the silk fibroin in deionized water to prepare a silk fibroin solution having a concentration ranging from 10 to 30%;
(6)将浓度为 10~30%的丝素蛋白溶液在室温下溶胀后， 搅拌至丝素蛋白完全 溶解、 脱泡， 然后倒入平板模上流延成膜， 再室温挥发、 于 50~70°C真空干燥制 成由丝素蛋白构成的组织隔离层； (6) After the silk fibroin solution having a concentration of 10-30% is swollen at room temperature, stir until the silk fibroin is completely dissolved and defoamed, then poured into a flat plate and cast into a film, and then volatilized at room temperature, at 50-70. °C vacuum drying to form a tissue isolation layer composed of silk fibroin;
(7)在已制成的组织隔离层一面进行划痕粗化处理， 再将 10~30%的丝素蛋白 溶液或者混有生物活性因子的 10~30%的丝素蛋白溶液， 均匀涂布或者采用静电 纺丝方法喷涂在有划痕的粗糙面上；  (7) Scratch roughening treatment on the prepared tissue isolation layer, and uniformly coating 10 to 30% of the silk fibroin solution or 10 to 30% of the silk fibroin solution mixed with the bioactive factor Or spraying on a rough surface with scratches by electrospinning;
(8)在 -40~-80°C条件下冷冻 8~12小时， 然后在真空冷冻干燥器中真空干燥， 脱模即可。  (8) Freeze at -40 to -80 °C for 8 to 12 hours, then vacuum dry in a vacuum freeze dryer to release the mold.
6. 根据权利要求 5所述的生物活性组织再生膜的制备方法， 其特征在于： 所述 划痕粗化处理， 是在由丝素蛋白构成的组织隔离层的一面上用尖状物刻划， 使表面形成深度为 1~10微米的交错状划痕。  The method for producing a bioactive tissue regeneration membrane according to claim 5, wherein the scratch roughening treatment is performed by a pointed object on one side of a tissue isolation layer composed of silk fibroin , to form a staggered scratch with a depth of 1 to 10 microns.
7. 根据权利要求 5所述的生物活性组织再生膜的制备方法， 其特征在于： 采用 静电纺丝方法将丝素蛋白喷涂在有划痕的粗糙面上。  The method for producing a bioactive tissue regeneration membrane according to claim 5, wherein the silk fibroin is sprayed on the rough surface having scratches by an electrospinning method.
8. 根据权利要求 5所述的生物活性组织再生膜的制备方法， 其特征在于： 所述 混有生物活性因子的 10~30%的丝素蛋白溶液中， 生物活性因子的含量为 3- 10mg/ml o The method for preparing a bioactive tissue regeneration membrane according to claim 5, wherein: the content of the bioactive factor in the 10 to 30% silk fibroin solution mixed with the bioactive factor is 3- 10mg/ml o
9. 根据权利要求 5所述的生物活性组织再生膜的制备方法， 其特征在于： 所述 生物活性因子为骨形成蛋白、 血小板衍生生长因子、 ***、 成 纤维细胞生长因子或转化生长因子中的一种或几种。  The method for preparing a bioactive tissue regeneration membrane according to claim 5, wherein the bioactive factor is bone morphogenetic protein, platelet-derived growth factor, insulin-like growth factor, fibroblast growth factor or transforming growth. One or several of the factors.
PCT/CN2010/070133 2009-01-13 2010-01-12 Bioactive tissue regeneration film and preparation method thereof WO2010081408A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN200910060497.5 2009-01-13
CN 200910060497 CN101474430B (en) 2009-01-13 2009-01-13 Tissue regeneration membrane with bioactivity and preparation method thereof

Publications (1)

Publication Number Publication Date
WO2010081408A1 true WO2010081408A1 (en) 2010-07-22

Family

ID=40835257

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2010/070133 WO2010081408A1 (en) 2009-01-13 2010-01-12 Bioactive tissue regeneration film and preparation method thereof

Country Status (2)

Country Link
CN (1) CN101474430B (en)
WO (1) WO2010081408A1 (en)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120148639A1 (en) * 2009-08-21 2012-06-14 Kyoto University Substrate for feeding cells and/or tissues, cell/tissue-feeder and method for the production of the same, method for the regeneration of tissues, and method for the production of porous bodies
CN113041392A (en) * 2021-04-01 2021-06-29 浙江理工大学 Preparation method of silk fibroin-gelatin bioactive composite hydrogel loaded with insulin particles
CN113398338A (en) * 2021-06-30 2021-09-17 华东理工大学 Double-layer repairing film for guiding tissue regeneration and preparation method thereof
CN114028619A (en) * 2021-11-02 2022-02-11 武汉亚洲生物材料有限公司 Double-layer artificial periosteum and preparation method and application thereof
CN114028620A (en) * 2021-11-02 2022-02-11 武汉亚洲生物材料有限公司 Mineralized artificial periosteum and preparation method and application thereof
CN114028611A (en) * 2021-11-02 2022-02-11 武汉亚洲生物材料有限公司 Absorbable double-layer artificial periosteum and preparation method and application thereof

Families Citing this family (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101474430B (en) * 2009-01-13 2013-01-09 武汉大学 Tissue regeneration membrane with bioactivity and preparation method thereof
CN101829025A (en) * 2010-04-27 2010-09-15 马兆成 Bioactive facial mask and preparation method therefore
TWI424861B (en) * 2010-07-30 2014-02-01 Univ Central Taiwan Sci & Tech And a method for producing the regenerated film and the method for producing the regenerated film
GB201118000D0 (en) * 2011-10-19 2011-11-30 Orthox Ltd An implantable repair device
CN102847194A (en) * 2012-09-17 2013-01-02 浙江星月生物科技股份有限公司 Stent type silk fibroin film insoluble in water, and preparation and application of stent type silk fibroin film
CN103007364B (en) * 2012-12-20 2014-05-14 北京市意华健科贸有限责任公司 Aliphatic polyester double-layered asymmetric guided tissue regeneration membrane and preparation method thereof
CN103223193B (en) * 2013-04-10 2014-12-10 浙江大学 High strength silk fibroin bone repair scaffold material preparation method
KR101573838B1 (en) * 2014-06-13 2015-12-07 대한민국 Artificial biomembrane using Cocoon and Method for manufacturing thereof
CN104474589B (en) * 2014-12-23 2019-03-12 山东国际生物科技园发展有限公司 A kind of guide tissue regeneration film and the preparation method and application thereof
CN104667349B (en) * 2015-02-06 2017-01-25 福州大学 Growth factor-loading silk fibroin/collagen bracket material and preparation method thereof
CN104707180B (en) * 2015-02-06 2017-01-25 福州大学 BMP loaded silk fibroin/collagen scaffold material and preparation method thereof
CN106552294B (en) * 2015-09-25 2021-06-01 上海市东方医院 Biological patch material for heart repair
CN105327401B (en) * 2015-11-17 2018-07-17 上海纳米技术及应用国家工程研究中心有限公司 Fibroin albumen bilayer imitates the preparation method of periosteum material
CN105524472A (en) * 2015-12-22 2016-04-27 傅泽红 Keratin/silk fibroin blended membrane and preparation method thereof
CN106236326B (en) * 2016-08-26 2018-08-10 天津市赛宁生物工程技术有限公司 Big stage Bone Defect Repari system
CN106361468B (en) * 2016-08-26 2019-01-15 天津市赛宁生物工程技术有限公司 A kind of Bone Defect Repari system of big stage bone defect
CN106668940A (en) * 2016-12-27 2017-05-17 上海纳米技术及应用国家工程研究中心有限公司 Double-layer silk fibroin film, preparation method and application
CN107812248A (en) * 2017-10-26 2018-03-20 锡山区东港玉英家电经营部 A kind of W model production methods for three layers of tip composite membrane for improving union
CN107551324B (en) * 2017-10-27 2020-02-18 北京华信佳音医疗科技发展有限责任公司 Preparation and application of suturable hard membrane repairing material
CN108904885B (en) * 2018-09-25 2021-02-09 清华大学深圳研究生院 Controllable degradable biological water condensation film and manufacturing method and application thereof
CN109291550B (en) * 2018-10-15 2020-05-01 南通纺织丝绸产业技术研究院 Composite barrier film and preparation method thereof
CN109876193B (en) * 2019-04-01 2020-05-19 西安交通大学 Double-layer composite periodontal defect repair material based on photo-crosslinked hydrogel and preparation method thereof
CN111298198B (en) * 2019-04-18 2022-03-01 上海交通大学医学院附属第九人民医院 Double-layer absorbable bionic barrier film and preparation method and application thereof
CN111617320B (en) * 2020-04-20 2022-03-08 广州迈普再生医学科技股份有限公司 Bone induction regeneration membrane and preparation method and application thereof
CN112190771A (en) * 2020-10-23 2021-01-08 扬子江药业集团广州海瑞药业有限公司 Bone tissue regeneration guide membrane and preparation method thereof
CN112402703A (en) * 2020-10-27 2021-02-26 北京幸福益生高新技术有限公司 Double-layer porous composite biological membrane containing bioactive mineral material and preparation method thereof

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004000915A2 (en) * 2002-06-24 2003-12-31 Tufts University Silk biomaterials and methods of use thereof
US20060095137A1 (en) * 2004-10-29 2006-05-04 Seoul National University Industry Foundation Nanofibrous nonwoven membrane of silk fibroin for guided bone tissue regeneration and manufacturing method thereof
CN101036802A (en) * 2006-12-30 2007-09-19 苏州大学 Antheraea pernyi silk fibrion biology medicine material and the preparation method
WO2008103017A1 (en) * 2007-02-23 2008-08-28 Pusan National University Industry-University Cooperation Foundation Biodegradable porous composite and hybrid composite of biopolymers and bioceramics
CN101474430A (en) * 2009-01-13 2009-07-08 武汉大学 Tissue regeneration membrane with bioactivity and preparation method thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004000915A2 (en) * 2002-06-24 2003-12-31 Tufts University Silk biomaterials and methods of use thereof
US20060095137A1 (en) * 2004-10-29 2006-05-04 Seoul National University Industry Foundation Nanofibrous nonwoven membrane of silk fibroin for guided bone tissue regeneration and manufacturing method thereof
US20080292667A1 (en) * 2004-10-29 2008-11-27 Seoul National University Industry Foundation Nonwoven Nanofibrous Membranes of Silk Fibroin for Guided Bone Tissue Regeneration and Their Preparation Method
CN101036802A (en) * 2006-12-30 2007-09-19 苏州大学 Antheraea pernyi silk fibrion biology medicine material and the preparation method
WO2008103017A1 (en) * 2007-02-23 2008-08-28 Pusan National University Industry-University Cooperation Foundation Biodegradable porous composite and hybrid composite of biopolymers and bioceramics
CN101474430A (en) * 2009-01-13 2009-07-08 武汉大学 Tissue regeneration membrane with bioactivity and preparation method thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
KIRKER-HEAD, C. ET AL.: "BMP-silk composite matrices heal critically sized femoral defects", BONE, vol. 41, no. 6, August 2007 (2007-08-01), pages 247 - 255 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120148639A1 (en) * 2009-08-21 2012-06-14 Kyoto University Substrate for feeding cells and/or tissues, cell/tissue-feeder and method for the production of the same, method for the regeneration of tissues, and method for the production of porous bodies
US9440005B2 (en) * 2009-08-21 2016-09-13 National Institute Of Agrobiological Sciences Substrate for feeding cells and/or tissues, cell/tissue-feeder and method for the production of the same, method for the regeneration of tissues, and method for the production of porous bodies
CN113041392A (en) * 2021-04-01 2021-06-29 浙江理工大学 Preparation method of silk fibroin-gelatin bioactive composite hydrogel loaded with insulin particles
CN113398338A (en) * 2021-06-30 2021-09-17 华东理工大学 Double-layer repairing film for guiding tissue regeneration and preparation method thereof
CN114028619A (en) * 2021-11-02 2022-02-11 武汉亚洲生物材料有限公司 Double-layer artificial periosteum and preparation method and application thereof
CN114028620A (en) * 2021-11-02 2022-02-11 武汉亚洲生物材料有限公司 Mineralized artificial periosteum and preparation method and application thereof
CN114028611A (en) * 2021-11-02 2022-02-11 武汉亚洲生物材料有限公司 Absorbable double-layer artificial periosteum and preparation method and application thereof

Also Published As

Publication number Publication date
CN101474430B (en) 2013-01-09
CN101474430A (en) 2009-07-08

Similar Documents

Publication Publication Date Title
WO2010081408A1 (en) Bioactive tissue regeneration film and preparation method thereof
JP5289772B2 (en) Cartilage-like structure having multi-layer structure and osteochondral substitute and use thereof
JP5406915B2 (en) Biocompatible implant
WO2011021712A1 (en) Substratum for supplying cells/tissues, cell/tissue supply material and process for production thereof, method for regeneration of tissues, and process for production of porous material
JP2010273847A (en) High-density porous composite
CN104771787A (en) Composite support containing PGA strengthening net, preparation method and applications thereof
KR101429857B1 (en) Method for manufacturing composite bilayer fiber mat for bone hemorrhage application
CN112076350B (en) Biomimetic mineralized hydrogel with nano-micron composite structure and high mineral density as well as preparation method and application thereof
KR102422432B1 (en) silicate-shell hydrogel fiber scaffold and preparation method thereof
CN1210071C (en) Biological active bone tissue inducing regeneration film and preparation method
JP2018536451A (en) Vessel patch using silk matrix and method for producing the same
CN113633825B (en) Preparation method and application of bFGF-loaded heparinized acellular fat material
CN111228578A (en) Drug-loaded silk fibroin bone repair screw and preparation method thereof
JP2005213449A (en) Gelatin sponge
Tan et al. Research on the osteogenesis and biosafety of ECM–Loaded 3D–Printed Gel/SA/58sBG scaffolds
RU2740132C2 (en) Method for application of collagen stimulating coating on endoprosthesis
CN102671243A (en) Quaternized chitosan/siRNA composite particle-loaded skin regeneration material and preparation method thereof
Li et al. 3D printed hydroxyapatite/silk fibroin/polycaprolactone artificial bone scaffold and bone tissue engineering materials constructed with double-transfected bone morphogenetic protein-2 and vascular endothelial growth factor mesenchymal stem cells to repair rabbit radial bone defects
US20190374676A1 (en) A cross-linked structure for tissue regeneration and engineering and the method for synthesising same
Wang et al. Comparison of the Effects of 3D Printing Bioactive Porous Titanium Alloy Scaffolds and Nano-biology for Direct Treatment of Bone Defects
Wang et al. Development of a rapid-shaping and user-friendly membrane with long-lasting space maintenance for guided bone regeneration
Kuevda et al. Application of recellularized non-woven materials from collagen-enriched polylactide for creation of tissue-engineered diaphragm constructs
JP4671596B2 (en) Porous bone repair material and method for producing the same
JP5229937B2 (en) Bone regenerative agent and method for regenerating bone
CN108498861A (en) A kind of preparation method of hydroxyapatite-fibroin-titanium dioxide biomaterial

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 10731042

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 10731042

Country of ref document: EP

Kind code of ref document: A1