1.天津医科大学总医院 天津医科大学 军事科学院卫勤保障技术研究所;2.军事科学院卫勤保障技术研究所;3.天津理工大学;4.天津医科大学总医院;5.天津医科大学;6.天津市口腔医院
1.Tianjin Medical University General Hospital;2.Tianjin Medical University;3.Institute of Military Aid Technology;4.Tianjin University of Technology;5.Tianjin Stomatological Hospital
摘要：目的：体外构建丝素蛋白（SF）、I型胶原(Col-I)和羟基磷灰石(HA)共混体系制备二维复合膜和三维仿生支架，研究其理化性质和生物相容性，探讨其在组织工程支架材料中应用的可行性。 方法：通过在细胞培养小室底部共混SF/ Col-I/HA以及低温3D打印结合真空冷冻干燥法制备二维复合膜及三维支架。通过机械性能测试，电子显微镜和Micro-CT检测材料的理化性质，检测细胞的增殖评估其生物相容性。 结果：通过共混和低温3 D打印获得稳定的二维复合膜及三维多孔结构支架；力学性能具有较好的一致性，孔径、吸水率、孔隙率和弹性模量均符合构建组织工程骨的要求；支架为网格状的白色立方体，内部孔隙连通性较好；羟基磷灰石（HA）均匀分布在复合膜中，细胞黏附在复合膜上，呈扁平状；细胞分布在支架孔壁周围，呈梭形状，生长及增殖良好。 结论：本研究利用SF/ Col-I/HA共混体系成功制备复合膜及三维支架，具有较好的孔连通性与孔结构,有利于细胞和组织的生长以及营养输送其理化性能以及生物相容性符合骨组织工程生物材料的要求。相关研究为骨组织工程支架材料的应用和评估提供了重要的试验数据和理论依据，为临床上实现人体组织器官缺损修复和再生具有重要意义。
Objective: To construct a two-dimensional composite membrane and a three-dimensional biomimetic scaffold by silk fibroin (SF), type I collagen (Col-I) and hydroxyapatite (HA) blends in vitro, and to study its physical and chemical properties and biocompatibility. To explore the feasibility of its application in tissue engineering scaffold materials. Methods: Two-dimensional composite membranes and three-dimensional scaffolds were prepared by blending SF/COL-I/HA at the bottom of the cell culture chamber and low temperature 3D printing combined with vacuum freeze drying. The biocompatibility was evaluated by mechanical properties testing, electron microscopy and Micro-CT to examine the physicochemical properties of the material and to detect cell proliferation. Results: Stable two-dimensional composite membrane and three-dimensional porous structural scaffolds were obtained by blending and low temperature 3D printing. The mechanical properties were consistent. The pore size, water absorption, porosity and elastic modulus were all in accordance with the requirements of constructing tissue engineering bone. The scaffold is a grid-like white cube with good internal pore connectivity; hydroxyapatite (HA) is evenly distributed in the composite membrane, and the cells are attached to the composite membrane in a flat shape; the cells are distributed around the pore walls of the scaffold. The shape of the shuttle is good, and the growth and proliferation are good. Conclusion: In this study, the composite membrane and three-dimensional scaffold prepared by SF/col-I/HA blending system have better pore connectivity and pore structure, which is beneficial to cell and tissue growth and nutrient transport. Its physical and chemical properties and biocompatibility meet the requirements of bone tissue engineering biomaterials. Related research provides important experimental data and theoretical basis for the application and evaluation of bone tissue engineering scaffold materials, which is of great significance for clinically realizing the repair and regeneration of human tissue and organ defects.