基于3D打印的双相磷酸钙胶原改性支架  

Biphasic calcium phosphate scaffolds modified with collagen based on three-dimensional printing technology

作　　者：张海鸽 索海瑞[1,2] 王玲 徐铭恩[1,2] Zhang Haige;Suo Hairui;Wang Ling;Xu Mingen(Zhejiang Provincial Key Lab of Medical Information and Three-Dimensional Bio-Printing,Hangzhou Dianzi University,Hangzhou 310018,Zhejiang Province,China;College of Automation,Hangzhou Dianzi University,Hangzhou 310018,Zhejiang Province,China)

机构地区：[1]杭州电子科技大学浙江省医学信息与生物三维打印重点实验室,浙江省杭州市310018 [2]杭州电子科技大学自动化学院,浙江省杭州市310018

出　　处：《中国组织工程研究》2019年第30期4780-4786,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(61675059),项目负责人:王玲;国家重点研发计划(2017YFC1103400),项目负责人:徐铭恩~~

摘　　要：背景:制作出类似天然骨成分和机械强度、表面活性良好且具有多孔结构的三维胶原/双相磷酸钙支架是骨组织工程研究的一个难点。目的:制备兼具高孔隙率、高机械强度和高表面生物活性的三维多孔骨组织支架。方法:以质量比为60∶40的羟基磷灰石与β-磷酸三钙为浆料,利用3D打印技术制备双相磷酸钙支架,高温烧结后,分别以0.5,1.0,1.5g/L的Ⅰ型胶原溶液对支架进行涂覆处理,制备胶原/双相磷酸钙支架,通过表观形貌、孔隙率、吸水率、机械性能测试筛选最佳Ⅰ型胶原溶液涂覆质量浓度,进行细胞实验。将大鼠骨髓间充质干细胞分别接种于双相磷酸钙支架与胶原/双相磷酸钙支架上,培养1,7d,Calcein-AM染色观察细胞活性;培养1,3,7d,Alamar Blue法检测细胞增殖。结果与结论:①当Ⅰ型胶原溶液涂覆质量浓度为0.5g/L时,胶原/双相磷酸钙支架具有良好的孔隙率与吸水率,抗压强度为(4.99±0.15)MPa,压缩模量为(95.24±0.57)MPa,符合天然松质骨的机械强度要求,细胞实验选择此涂覆质量浓度;②两种支架都支持骨髓间充质干细胞的生长,培养至7d时,细胞在胶原/双相磷酸钙支架表面已基本长满,大多呈梭形或星形,细胞伸展良好,排列紧密,在支架上黏附形成网状结构,但双相磷酸钙支架上还有部分区域无细胞黏附;③两组支架中培养1,3d的细胞增殖比较差异无显著性意义(P>0.05),胶原/双相磷酸钙支架上培养7d的细胞增殖快于双相磷酸钙支架(P<0.05);④结果表明在保证孔隙率、高机械强度的情况下,Ⅰ型胶原溶液涂覆可提高双相磷酸钙支架的生物活性。BACKGROUND: Three-dimensional collagen/biphasic calcium phosphate scaffolds with natural bone composition and mechanical strength, good surface activity and porous structure are a difficult point in bone tissue engineering research. OBJECTIVE: To manufacture a three-dimensional porous bone tissue scaffold with high porosity, high mechanical strength and high surface biological activity. METHODS: Biphasic calcium phosphate scaffold was manufactured by three-dimensional printing technology using hydroxyapatite and β-tricalcium phosphate with a mass ratio of 60∶40 as slurry. After high temperature sintering, the biphasic calcium phosphate scaffolds were coated with type I collagen at the concentrations of 0.5, 1.0 and 1.5 g/L. The optimal concentration of type I collagen solution was screened by apparent morphology, porosity, water absorption and mechanical properties to conduct cell experiments. Rat bone marrow mesenchymal stem cells were seeded onto biphasic calcium phosphate scaffolds and collagen/biphasic calcium phosphate scaffolds, separately. Cell viability was observed by Calcein-AM staining at 1 and 7 days;cell proliferation was detected by Alamar Blue method at 1, 3 and 7 days. RESULTS AND CONCLUSION: When the mass concentration of type I collagen solution was 0.5 g/L, the collagen/biphasic calcium phosphate scaffold had good porosity and water absorption, the compressive strength was (4.99±0.15) MPa and the compressive modulus was (95.24±0.57) MPa, which met the mechanical strength requirements of natural cancellous bone. Therefore, the coating mass concentration 0.5 g/L was selected for cell experiments. Both kinds of scaffolds supported the growth of mesenchymal stem cells. After 7 days of culture, cells almost covered the surface of the collagen/biphasic calcium phosphate scaffold. Most of them were fusiform or star-shaped, and the cells were well stretched, arranged closely and adhered to form network structure. However, there was still no cell adhesion in some areas of the biphasic calcium phosp

关 键 词：3D打印 双相磷酸钙 胶原 涂覆 细胞活性 生物材料 骨组织 胶原/双相磷酸钙支架 细胞增殖 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]