丝素蛋白复合胶原蛋白支架的制备及性能研究

发布时间：2018-03-30 02:01

本文选题：软骨组织工程　切入点：丝素蛋白　出处：《中国修复重建外科杂志》2014年07期

【摘要】：目的制备不同质量比的丝素蛋白复合胶原蛋白支架,分析其综合性能,优选软骨组织工程支架材料。方法按丝素蛋白∶胶原蛋白不同质量比,采用真空冷冻干燥法制备3种复合支架,分别为4∶2(A组)、4∶4(B组)和4∶8(C组);检测每组复合支架的孔隙率、吸水膨胀率、力学性能及孔径。采用密度梯度离心法分离培养4周龄雄性Wistar大鼠BMSCs,取第3代BMSCs接种于各组复合支架,培养1、3、5、7、9、11、13 d,MTT检测支架内细胞增殖情况;培养14 d采用HE染色和扫描电镜观察细胞在支架内部增殖、分布情况。结果 A、B、C组支架孔隙率分别为94.6%±1.6%、80.6%±1.1%、60.6%±1.0%,A组显著高于B、C组,B组高于C组(P0.05)。吸水膨胀率分别为1 523.7%±186.6%、1 091.0%±151.6%、659.6%±161.4%,A组显著高于B、C组,但组间比较差异无统计学意义(F=6.67,P=0.08);各组支架均具有较好的黏弹性,A、B、C组弹性模量分别为(23.1±2.5)、(25.1±2.3)、(29.8±2.6)kPa,组间比较差异无统计学意义(F=2.00,P=0.28),符合关节软骨弹性模量值(4~35 kPa);A、B、C组支架孔径分别为(103±12)、(80±15)、(60±16)μm,3组间比较差异无统计学意义(F=2.22,P=0.26)。MTT检测示,7、9、11、13 d时A组细胞增殖高于B、C组(P0.05)。培养14 d,扫描电镜观察示A组支架孔径大小较一致,相通性好,细胞在支架内部生长良好,伸展充分,基质分泌多;B、C组支架孔径较小,孔隙间相通性差,细胞在内部生长较差;HE染色和扫描电镜示支架内部A组细胞数量明显多于B、C组。结论以丝素蛋白∶胶原蛋白质量比为4∶2制备复合支架具有合适的孔隙率、吸水膨胀率、弹性模量和孔径,细胞在支架内部增殖更好,是软骨组织工程的理想支架材料。
[Abstract]:Objective to prepare fibroin composite collagen scaffolds with different mass ratios, analyze their comprehensive properties and select cartilage tissue engineering scaffolds. Three kinds of composite scaffolds were prepared by vacuum freeze-drying (group 4: 2A, group 4: 4: 4B) and group C (group C, 4: 8). The porosity and water swelling rate of each group of composite scaffolds were measured. Mechanical properties and pore diameter. BMSCs of 4-week-old male Wistar rats were isolated and cultured by density gradient centrifugation. The third generation of BMSCs was inoculated into each group of composite scaffolds. The cell proliferation in the scaffold was observed by HE staining and scanning electron microscope on the 14th day of culture. Results the porosity of stent in group A was 94.6% 卤1.60.The porosity of group A was significantly higher than that of group B (P 0.05). The swelling rate of group A was 1 523.7% 卤186.6% 卤151.6% 卤659.6% 卤161.4%, respectively, and that of group A was significantly higher than that of group B (P < 0.05), and that of group A was significantly higher than that of group B (P < 0.05), and that of group A was significantly higher than that of group B (P < 0.05), and that of group A was significantly higher than that of group B (P < 0.05), and that of group A was significantly higher than that of group B (P < 0.05). However, there was no significant difference between the two groups, and the elastic modulus of each group was 23.1 卤2.5kPa29.8 卤2.6kPa.There was no significant difference between the two groups, and there was no significant difference between the two groups, which was consistent with the elastic modulus of articular cartilage and the diameter of the scaffold in ABC group. There was no significant difference among the three groups (103 卤12), 80 卤15 (60 卤16) 渭 m ~ (-1). MTT assay showed that the proliferation of cells in group A was higher than that in group A at day 1113. The diameter of stents in group A was the same as that in group A after 14 days of culture, and the diameter of stents in group A was the same as that in group A by scanning electron microscopy (SEM), the results showed that the cell proliferation of group A was higher than that of group B at 1113 d. The cells grew well and stretched well in the scaffold. The pore diameter of the scaffold was smaller and the interpore permeability was poor in the group of multi-secretory matrix. The results of HE staining and SEM showed that the number of cells in group A was significantly higher than that in group B (C). Conclusion the composite scaffold prepared by 4:2 ratio of silk fibroin to collagen has a suitable porosity and water swelling rate. With elastic modulus and pore diameter, cells proliferate better in the scaffold, which is the ideal scaffold material for cartilage tissue engineering.
【作者单位】：天津医科大学研究生院;军事医学科学院卫生装备研究所;天津医科大学总医院骨科;
【基金】：国家自然科学基金资助项目(11072266)~~
【分类号】：R318.08

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