CN103638559A - Water-insoluble ultrafine fibroin powder/polylactic acid composite porous scaffold material and preparation method thereof - Google Patents
Water-insoluble ultrafine fibroin powder/polylactic acid composite porous scaffold material and preparation method thereof Download PDFInfo
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Abstract
本发明属于生物医用高分子材料领域，涉及一种组织工程用非水溶性超细丝素粉体增强的高分子复合多孔支架材料及其制备方法，其制备方法包括以下步骤：将聚乳酸溶解于二氧六环或二氧六环/二甲基亚砜混合溶剂中，得到聚乳酸溶液，再将一定质量的非水溶性超细丝素粉体加入聚乳酸溶液中，混合均匀后超声脱泡，然后将上述聚乳酸混合溶液注入聚四氟乙烯模具中，在-10~-50℃下冷冻1~5h，最后于冻干机中在-45~-55℃下冻干，得到非水溶性超细丝素粉体/聚乳酸复合多孔支架材料。本发明制备工艺简单，条件易于控制，所制备的多孔支架材料具有更好的生物相容性，孔隙率高，亲水性强，有利于细胞的粘附、生长与繁殖，而且具有较好的力学强度，可广泛应用于组织工程领域。
The invention belongs to the field of biomedical polymer materials, and relates to a polymer composite porous scaffold material reinforced by non-water-soluble ultrafine silk fibroin powder for tissue engineering and a preparation method thereof. The preparation method comprises the following steps: dissolving polylactic acid in Dioxane or dioxane/dimethyl sulfoxide mixed solvent to obtain a polylactic acid solution, then add a certain quality of non-water-soluble superfine silk fibroin powder into the polylactic acid solution, mix well and then ultrasonically defoam , and then inject the above polylactic acid mixed solution into a polytetrafluoroethylene mold, freeze at -10~-50°C for 1~5h, and finally freeze dry at -45~-55°C in a lyophilizer to obtain a water-insoluble Superfine silk fibroin powder/polylactic acid composite porous scaffold material. The preparation process of the present invention is simple, the conditions are easy to control, the prepared porous scaffold material has better biocompatibility, high porosity, strong hydrophilicity, is beneficial to the adhesion, growth and reproduction of cells, and has better The mechanical strength can be widely used in the field of tissue engineering.
Description
技术领域 technical field
本发明属生物医用高分子材料领域，具体涉及一种组织工程用非水溶性丝素粉体/聚乳酸复合多孔支架材料及其制备方法。 The invention belongs to the field of biomedical polymer materials, in particular to a non-water-soluble silk fibroin powder/polylactic acid composite porous scaffold material for tissue engineering and a preparation method thereof.
背景技术 Background technique
组织工程支架材料是指能与组织活体细胞结合并且能植入生物体的材料，它是组织工程化组织的最基本构架。生物可降解高分子材料是组织工程支架材料中目前研究较多的一类材料，它具有良好生物相容性，而且植入生物体后，细胞在其表面粘附、增殖形成组织的过程中的同时，在体液、酶、细胞等的作用下发生降解，变成小分子物质被吸收或通过新陈代谢排出体外。聚乳酸是一种生物可降解的脂肪族聚酯，其降解产物为乳酸，无毒且可随人体新陈代谢排出体外，在组织工程支架材料中备受关注，但因其亲水性差、降解速率慢、细胞亲和性差等缺点限制了其在生物组织工程方面的应用。 Tissue engineering scaffold materials refer to materials that can be combined with tissue living cells and can be implanted into organisms. It is the most basic structure of tissue engineering tissues. Biodegradable polymer materials are currently a class of materials that have been studied more in tissue engineering scaffold materials. At the same time, it degrades under the action of body fluids, enzymes, cells, etc., and becomes small molecular substances to be absorbed or excreted through metabolism. Polylactic acid is a biodegradable aliphatic polyester, and its degradation product is lactic acid, which is non-toxic and can be excreted with human metabolism. It has attracted much attention in tissue engineering scaffold materials, but because of its poor hydrophilicity and slow degradation rate , poor cell affinity and other shortcomings limit its application in biological tissue engineering.
天然高分子，如蛋白质，多糖等由于具有良好的生物相容性，生物可降解性，在临床修复和组织工程支架等方面有广泛的应用前景。近年来，人们利用蛋白质及多糖对聚乳酸进行改性，制备各种聚乳酸复合多孔支架材料。中国专利公告号CN 101703808A，公告日是2010年5月12日，名称为‘一种多孔胶原/聚乳酸支架’中公开了一种多孔胶原/聚乳酸支架。该方法将胶原蛋白、聚乳酸和表皮细胞生长因子EGF的丙酮混合溶液在低温下冻干，制备多孔复合支架。由于采用在低温下不能结冰的丙酮，利用冻干并不能很好控制多孔材料的孔径。中国专利公告号CN 1958081A，公告日是2008年11月12日，名称为‘壳聚糖-聚乳酸共聚物三维多孔支架制备方法’中公开了一种壳聚糖-聚乳酸共聚物三维多孔支架制备方法，将壳聚糖溶解于乳酸中，加入氯化钠或氯化钾，然后真空升温共聚，并将固态物用去离子水进行浸泡洗涤后冷冻干燥，得到壳聚糖-聚乳酸共聚物多孔支架。但这种化学反应的程度和聚乳酸的接枝率不易控制。中国专利公告号CN 101053670，公告日是2007年10月17日，名称为‘一种丝素和聚乳酸复合膜及其制备方法’公开了一种利用丝素溶液和聚乳酸溶液混合干燥制备丝素/聚乳酸复合膜的方法，这种复合膜可用于创面保护膜、人工皮肤等方面。但需要对丝素蛋白进行精炼及溶解等处理得到水溶性丝素，丝素分子量小，易从所制备的复合膜中溶出。 Natural polymers, such as proteins and polysaccharides, have broad application prospects in clinical repair and tissue engineering scaffolds due to their good biocompatibility and biodegradability. In recent years, people have used proteins and polysaccharides to modify polylactic acid to prepare various polylactic acid composite porous scaffold materials. Chinese patent announcement number CN 101703808A, date of announcement is May 12, 2010, and a kind of porous collagen/polylactic acid scaffold is disclosed in the name "a kind of porous collagen/polylactic acid scaffold". In the method, the acetone mixed solution of collagen, polylactic acid and epidermal growth factor EGF is freeze-dried at low temperature to prepare a porous composite scaffold. Due to the use of acetone, which cannot freeze at low temperatures, the pore size of the porous material cannot be well controlled by freeze-drying. Chinese Patent Announcement No. CN 1958081A, dated November 12, 2008, titled 'Preparation Method of Chitosan-Polylactic Acid Copolymer Three-Dimensional Porous Scaffold' discloses a chitosan-polylactic acid copolymer three-dimensional porous scaffold The preparation method is to dissolve chitosan in lactic acid, add sodium chloride or potassium chloride, then raise the temperature in vacuum to copolymerize, soak and wash the solid matter with deionized water, and then freeze-dry to obtain chitosan-polylactic acid copolymer Porous scaffold. However, the degree of this chemical reaction and the grafting rate of PLA are not easy to control. Chinese Patent Announcement No. CN 101053670, dated October 17, 2007, titled 'A Composite Film of Silk Fibroin and Polylactic Acid and Its Preparation Method' discloses a method for preparing silk by mixing and drying silk fibroin solution and polylactic acid solution. The method of plain/polylactic acid composite film, this composite film can be used in aspects such as wound surface protective film, artificial skin. However, it is necessary to refine and dissolve silk fibroin to obtain water-soluble silk fibroin, which has a small molecular weight and is easy to dissolve from the prepared composite film.
发明内容 Contents of the invention
本发明的目的在于提供一种非水溶性丝素粉体增强聚乳酸复合多孔支架材料及其制备方法，该方法是将通过物理研磨得到的非水溶性超细丝素粉体与聚乳酸溶液混合，经过冷冻干燥，制备非水溶性超细丝素粉体/聚乳酸复合多孔支架材料。 The object of the present invention is to provide a kind of water-insoluble silk fibroin powder reinforced polylactic acid composite porous scaffold material and its preparation method, the method is to mix the water-insoluble superfine silk fibroin powder obtained by physical grinding with polylactic acid solution , after freeze-drying, the non-water-soluble superfine silk fibroin powder/polylactic acid composite porous scaffold material was prepared.
本发明是采用以下技术方案加以实现的，非水溶性超细丝素粉体/聚乳酸复合多孔支架材料及其制备方法，其制备方法包括以下步骤： The present invention is realized by adopting the following technical scheme, non-water-soluble superfine silk fibroin powder/polylactic acid composite porous scaffold material and its preparation method, and its preparation method comprises the following steps:
1） 在室温下，将聚乳酸溶解于二氧六环或二氧六环/二甲基亚砜的混合溶剂中，得到重量百分含量为3 ~ 10%的聚乳酸溶液； 1) Dissolve polylactic acid in dioxane or a mixed solvent of dioxane/dimethyl sulfoxide at room temperature to obtain a polylactic acid solution with a weight percentage of 3 to 10%;
2） 将非水溶性超细丝素粉体加入聚乳酸溶液中，混合均匀，超声脱泡后注入聚四氟乙烯模具中，于-10 ~ -50℃下冷冻1 ~ 5h，最后于冻干机中在-45 ~ -55℃下冻干，得到本发明的非水溶性超细丝素粉体/聚乳酸复合多孔支架材料。 2) Add the non-water-soluble superfine silk fibroin powder into the polylactic acid solution, mix evenly, pour it into a polytetrafluoroethylene mold after ultrasonic defoaming, freeze at -10 ~ -50°C for 1 ~ 5h, and finally freeze-dry Freeze-dry in the machine at -45~-55°C to obtain the non-water-soluble superfine silk fibroin powder/polylactic acid composite porous scaffold material of the present invention.
上述的二氧六环/二甲基亚砜混合溶剂是指二氧六环与二甲基亚砜按体积比为99:1 ~ 50:50混合得到。 The above-mentioned dioxane/dimethyl sulfoxide mixed solvent refers to the mixture of dioxane and dimethyl sulfoxide in a volume ratio of 99:1 to 50:50.
上述非水溶性丝素粉体的加入量是按非水溶性丝素粉体与聚乳酸的质量比为5:95 ~ 50:50计量。 The addition amount of the above-mentioned non-water-soluble silk fibroin powder is measured according to the mass ratio of the non-water-soluble silk fibroin powder and polylactic acid of 5:95 to 50:50.
上述的聚乳酸是指聚L-乳酸、聚D-乳酸或聚D,L-乳酸，聚乳酸的分子量为1×105 ~ 5×105。 The polylactic acid mentioned above refers to poly L-lactic acid, poly D-lactic acid or poly D,L-lactic acid, and the molecular weight of polylactic acid is 1×10 5 ~5×10 5 .
上述的非水溶性超细丝素粉体的平均粒径为2.4μm。 The average particle diameter of the above-mentioned water-insoluble superfine silk fibroin powder is 2.4 μm.
由于采用上述技术方案，本发明的制备方法与现有技术相比，具有如下优点和有益效果：丝素是天然蛋白质，具有良好的生物相容性及生物可降解性，将丝素纤维通过物理研磨的方法得到的超细丝素粉体保留了丝素蛋白原有的生化结构及活性的完整性，以及良好的力学性能，克服了水溶性丝素分子量小，强度不足及易溶出的缺点。该制备方法工艺简单，条件温和，得到的复合多孔支架材料具有良好的力学性能，亲水性好，孔隙率高，改善了聚乳酸支架的亲水性，增加了细胞粘附力，为细胞在其表面生长、增殖和分化提供了良好的微环境，可广泛应用于组织工程支架材料。 Due to the above-mentioned technical scheme, compared with the prior art, the preparation method of the present invention has the following advantages and beneficial effects: silk fibroin is a natural protein with good biocompatibility and biodegradability, and the silk fibroin fiber is physically The ultra-fine silk fibroin powder obtained by the grinding method retains the original biochemical structure and active integrity of silk fibroin, as well as good mechanical properties, and overcomes the shortcomings of water-soluble silk fibroin such as small molecular weight, insufficient strength and easy dissolution. The preparation method has a simple process and mild conditions, and the obtained composite porous scaffold material has good mechanical properties, good hydrophilicity, and high porosity, which improves the hydrophilicity of the polylactic acid scaffold, increases cell adhesion, and provides support for cells in Its surface growth, proliferation and differentiation provide a good microenvironment, which can be widely used in tissue engineering scaffold materials.
附图说明 Description of drawings
图1为本发明复合多孔支架材料所用的非水溶性丝素粉体的粒径分布图。 Fig. 1 is a particle size distribution diagram of the water-insoluble silk fibroin powder used in the composite porous scaffold material of the present invention.
图2为本发明复合多孔支架材料的吸水率随时间的变化图。 Fig. 2 is a graph showing the water absorption of the composite porous scaffold material of the present invention as a function of time.
图3为本发明复合多孔支架材料的压缩强度图。 Fig. 3 is a graph of the compressive strength of the composite porous scaffold material of the present invention.
图4 为本发明复合多孔支架材料的扫描电镜图。 Figure 4 is a scanning electron micrograph of the composite porous scaffold material of the present invention.
具体实施方式 Detailed ways
下面结合实施例和附图对本发明作进一步详细的描述。 The present invention will be further described in detail below in conjunction with the embodiments and the accompanying drawings.
实施例1Example 1
分别称取0.3g聚D,L-乳酸(分子量为1×105)和9.7g二氧六环，加到100ml烧杯中，磁力搅拌1h。待聚乳酸完全溶解后，称取0.045g丝素粉体加入聚乳酸溶液中，继续搅拌3h，混合均匀后，超声30min脱泡，倒入聚四氟乙烯模具中，放入-20℃的冰箱中冷冻1小时，然后在-45 ~ -55℃温度下冻干，脱模得到超细丝素粉体/聚乳酸复合多孔支架材料。所制备的复合多孔支架材料的孔隙率为90.5%。图2a为所制备的复合多孔材料的吸水率随时间的变化曲线，图3a为所制备的复合多孔支架材料压缩强度，图4a为所制备的复合多孔支架材料的扫描电镜图。 Weigh 0.3g of poly-D,L-lactic acid (molecular weight: 1×10 5 ) and 9.7g of dioxane, respectively, add them into a 100ml beaker, and stir for 1 hour under magnetic force. After the polylactic acid is completely dissolved, weigh 0.045g silk fibroin powder and add it to the polylactic acid solution, continue to stir for 3 hours, after mixing evenly, ultrasonically defoam for 30 minutes, pour it into a polytetrafluoroethylene mold, and put it in a refrigerator at -20°C Freeze in medium for 1 hour, then freeze-dry at -45~-55°C, and demould to obtain superfine silk fibroin powder/polylactic acid composite porous scaffold material. The porosity of the prepared composite porous scaffold material was 90.5%. Figure 2a is the water absorption curve of the prepared composite porous material over time, Figure 3a is the compressive strength of the prepared composite porous scaffold material, and Figure 4a is the scanning electron microscope image of the prepared composite porous scaffold material.
实施例2Example 2
分别称取0.5g聚L-乳酸(分子量为1.8×105)和9.5g二氧六环与二甲基亚砜混合溶剂（80/20，体积比），加到100ml烧杯中，磁力搅拌1h。待聚乳酸完全溶解后，称取0.1g丝素粉体加入聚乳酸溶液中，继续搅拌3h，混合均匀后，超声30min脱泡，倒入聚四氟乙烯模具中，放入-30℃的冰箱中冷冻2小时，然后在-45 ~ -55℃温度下冻干，脱模得到超细丝素粉体/聚乳酸复合多孔支架材料。所制备的复合多孔支架材料的孔隙率为88.6%。 Weigh 0.5g of poly-L-lactic acid (molecular weight: 1.8×10 5 ) and 9.5g of mixed solvent of dioxane and dimethyl sulfoxide (80/20, volume ratio), add them into a 100ml beaker, and stir for 1h . After the polylactic acid is completely dissolved, weigh 0.1g silk fibroin powder and add it to the polylactic acid solution, continue to stir for 3 hours, after mixing evenly, ultrasonically defoam for 30 minutes, pour it into a polytetrafluoroethylene mold, and put it in a refrigerator at -30°C Freeze in medium for 2 hours, then freeze-dry at -45~-55°C, and demould to obtain superfine silk fibroin powder/polylactic acid composite porous scaffold material. The porosity of the prepared composite porous scaffold material was 88.6%.
实施例3Example 3
分别称取0.6g聚D-乳酸(分子量为1×105)和5.4g二氧六环与二甲基亚砜混合溶剂（90/10，体积比），加到100ml烧杯中，磁力搅拌2h。待聚乳酸完全溶解后，称取0.18g丝素粉体加入聚乳酸溶液中，继续搅拌3h，混合均匀后，超声40min脱泡，倒入聚四氟乙烯模具中，放入-40℃的冰箱中冷冻4小时，然后在-45 ~ -55℃温度下冻干，脱模得到超细丝素粉体/聚乳酸复合多孔支架材料。所制备的复合多孔支架材料的孔隙率为89.7%。图2b为所制备的复合多孔材料的吸水率随时间的变化曲线，图3b为所制备的复合多孔支架材料压缩强度，图4b为所制备的复合多孔支架材料的扫描电镜图。 Weigh 0.6g of poly-D-lactic acid (molecular weight: 1×10 5 ) and 5.4g of mixed solvent of dioxane and dimethyl sulfoxide (90/10, volume ratio), add them into a 100ml beaker, and stir for 2h . After the polylactic acid is completely dissolved, weigh 0.18g silk fibroin powder and add it to the polylactic acid solution, continue to stir for 3 hours, after mixing evenly, ultrasonically defoam for 40 minutes, pour it into a polytetrafluoroethylene mold, and put it in a refrigerator at -40°C Freeze in medium for 4 hours, then freeze-dry at -45~-55°C, and demould to obtain superfine silk fibroin powder/polylactic acid composite porous scaffold material. The porosity of the prepared composite porous scaffold material was 89.7%. Figure 2b is the water absorption curve of the prepared composite porous material over time, Figure 3b is the compressive strength of the prepared composite porous scaffold material, and Figure 4b is the scanning electron microscope image of the prepared composite porous scaffold material.
实施例4Example 4
分别称取0.6g聚D-乳酸(分子量为3.2×105)和6.9g二氧六环与二甲基亚砜混合溶剂（60/40，体积比），加到100ml烧杯中，磁力搅拌2h。待聚乳酸完全溶解后，称取0.3g丝素粉体加入聚乳酸溶液中，继续搅拌3h，混合均匀后，超声30min脱泡，然后倒入聚四氟乙烯模具中，放入-50℃的冰箱中冷冻2小时，最后在-45 ~ -55℃温度下冻干，脱模得到超细丝素粉体/聚乳酸复合多孔支架材料。所制备的复合多孔支架材料的孔隙率为87.3%。图4d为所制备的复合多孔支架材料的扫描电镜图。 Weigh 0.6g of poly D-lactic acid (molecular weight: 3.2×10 5 ) and 6.9g of dioxane and dimethyl sulfoxide mixed solvent (60/40, volume ratio), add them into a 100ml beaker, and magnetically stir for 2h . After the polylactic acid is completely dissolved, weigh 0.3g silk fibroin powder and add it to the polylactic acid solution, continue to stir for 3 hours, and after mixing evenly, ultrasonically defoam for 30 minutes, then pour it into a polytetrafluoroethylene mold, and put it in a -50°C Freeze in the refrigerator for 2 hours, and finally freeze-dry at a temperature of -45 to -55°C, and demould to obtain a superfine silk fibroin powder/polylactic acid composite porous scaffold material. The porosity of the prepared composite porous scaffold material was 87.3%. Figure 4d is a scanning electron micrograph of the prepared composite porous scaffold material.
实施例5Example 5
分别称取0.4g聚D,L-乳酸(分子量为5×105)和9.6g二氧六环，加到100ml烧杯中，磁力搅拌3h。待聚乳酸完全溶解后，称取0.16g丝素粉体加入聚乳酸溶液中，继续搅拌3h，混合均匀后，超声20min脱泡，然后倒入聚四氟乙烯模具中，放入-30℃的冰箱中冷冻4小时，最后在-45 ~ -55℃温度下冻干，脱模得到超细丝素粉体/聚乳酸复合多孔支架材料。所制备的复合多孔支架材料的孔隙率为91.6%。 Weigh 0.4g of poly-D,L-lactic acid (molecular weight: 5×10 5 ) and 9.6g of dioxane, respectively, add them into a 100ml beaker, and stir magnetically for 3h. After the polylactic acid is completely dissolved, weigh 0.16g of silk fibroin powder and add it to the polylactic acid solution, and continue to stir for 3 hours. Freeze in the refrigerator for 4 hours, and finally freeze-dry at a temperature of -45 to -55°C, and demould to obtain a superfine silk fibroin powder/polylactic acid composite porous scaffold material. The porosity of the prepared composite porous scaffold material was 91.6%.
实施例6Example 6
分别称取0.6g聚D,L-乳酸(分子量为1.2×105)和5.4g二氧六环与二甲基亚砜混合溶剂（50/50，体积比），加到100ml烧杯中，磁力搅拌2h。待聚乳酸完全溶解后，称取0.3g丝素粉体加入聚乳酸溶液中，继续搅拌3h，混合均匀后，超声40min脱泡，倒入聚四氟乙烯模具中，放入-40℃的冰箱中冷冻4小时，然后在-45 ~ -55℃温度下冻干，脱模得到超细丝素粉体/聚乳酸复合多孔支架材料。所制备的复合多孔支架材料的孔隙率为88.5%。图2c为所制备的复合多孔材料的吸水率随时间的变化曲线，图3c为所制备的复合多孔支架材料压缩强度，图4c为所制备的复合多孔支架材料的扫描电镜图。 Weigh 0.6g of poly D,L-lactic acid (molecular weight: 1.2×10 5 ) and 5.4g of mixed solvent of dioxane and dimethyl sulfoxide (50/50, volume ratio), add them into a 100ml beaker, and magnetically Stir for 2h. After the polylactic acid is completely dissolved, weigh 0.3g silk fibroin powder and add it to the polylactic acid solution, continue to stir for 3 hours, after mixing evenly, ultrasonically defoam for 40 minutes, pour it into a polytetrafluoroethylene mold, and put it in a refrigerator at -40°C Freeze in medium for 4 hours, then freeze-dry at -45~-55°C, and demould to obtain superfine silk fibroin powder/polylactic acid composite porous scaffold material. The porosity of the prepared composite porous scaffold material was 88.5%. Figure 2c is the water absorption curve of the prepared composite porous material with time, Figure 3c is the compressive strength of the prepared composite porous scaffold material, and Figure 4c is the scanning electron microscope image of the prepared composite porous scaffold material.
实施例7Example 7
分别称取0.4g聚L-乳酸(分子量为4×105)和7.6g二氧六环与二甲基亚砜混合溶剂（99/1，体积比），加到100ml烧杯中，磁力搅拌2h。待聚乳酸完全溶解后，称取0.12g丝素粉体加入聚乳酸溶液中，继续搅拌3h，混合均匀后，超声30min脱泡，然后倒入聚四氟乙烯模具中，放入-30℃的冰箱中冷冻2小时，最后在-45 ~ -55℃温度下冻干，脱模得到超细丝素粉体/聚乳酸复合多孔支架材料。所制备的复合多孔支架材料的孔隙率为89.4%。 Weigh 0.4g of poly-L-lactic acid (molecular weight: 4×10 5 ) and 7.6g of mixed solvent of dioxane and dimethyl sulfoxide (99/1, volume ratio), add them into a 100ml beaker, and stir for 2h . After the polylactic acid is completely dissolved, weigh 0.12g silk fibroin powder and add it to the polylactic acid solution, continue to stir for 3 hours, after mixing evenly, ultrasonically defoam for 30 minutes, then pour it into a polytetrafluoroethylene mold, and put it in a -30°C Freeze in the refrigerator for 2 hours, and finally freeze-dry at a temperature of -45 to -55°C, and demould to obtain a superfine silk fibroin powder/polylactic acid composite porous scaffold material. The porosity of the prepared composite porous scaffold material was 89.4%.
从图1中可以看出，通过物理研磨的方法得到的超细丝素粉体的平均粒径在2.4μm左右，从图2及图3中可以看出，非水溶性丝素粉体的加入及随着加入量的增加，所制备的复合多孔支架材料的亲水性及力学强度明显增强。因此，超细丝素粉体的加入改善了复合多孔材料的细胞粘附力，为细胞在其表面生长、增殖和分化提供了良好的微环境。 It can be seen from Figure 1 that the average particle size of ultrafine silk fibroin powder obtained by physical grinding is about 2.4 μm. It can be seen from Figure 2 and Figure 3 that the addition of water-insoluble silk fibroin powder And with the increase of the added amount, the hydrophilicity and mechanical strength of the prepared composite porous scaffold material are obviously enhanced. Therefore, the addition of ultrafine silk fibroin powder improves the cell adhesion of the composite porous material and provides a good microenvironment for the growth, proliferation and differentiation of cells on its surface.
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