脱细胞真皮基质/生物矿化胶原一体化骨软骨支架的制备及其生物相容性的研究  

Study on preparation and biocompatibility of the acellular dermal matrix/biomineralized collagen diphasic osteochondral integrated scaffold

作　　者：郑蕊 杰永生[1] 陈磊[1] 靳少锋 綦惠[1] 孙磊[1] 舒雄[1] ZHENG Rui;JIE Yong-sheng;CHEN Lei;JIN Shao-feng;QI Hui;SUN Lei;SHU Xiong(Beijing Jishuitan Hospital, Beijing Institute of Traumatology and Orthopaedics, Beijing 100035, China)

机构地区：[1]北京积水潭医院/北京市创伤骨科研究所骨库,100035

出　　处：《中国医药生物技术》2019年第2期121-126,共6页Chinese Medicinal Biotechnology

基　　金：北京市属医学科研院所科技发展项目(PXM2017_026275_000004)

摘　　要：目的成功制备脱细胞真皮基质/生物矿化胶原一体化骨软骨支架,并进行理化性质、生物力学与生物相容性评估,为骨软骨缺损修复提供理论依据。方法以物理和化学方法制备牛源脱细胞真皮基质,利用磷酸三钙和胶原,按照不同比例混合,利用自组装和冻干技术,制备以脱细胞真皮基质支架为软骨层,生物矿化胶原为骨层的骨软骨一体化双相支架。通过大体观察、扫描电镜观察、生物力学等检测,对骨软骨一体化支架进行理化性质和力学性能评价。原代培养小鼠软骨细胞和成骨细胞,并分别种植在脱细胞真皮基质和生物矿化胶原双相支架上,利用细胞毒、死/活细胞染色和增殖实验等观察细胞生长情况,测定骨软骨一体化支架的生物相容性。结果大体观察表明支架各层间结合紧密,未出现明显不连续和相互分离。扫描电镜各层内的孔隙结构相互连通且均具有立体多维性,其中脱细胞真皮基质、生物矿化胶原和双相支架的孔径分别为(121.6±8.65)、(98.40±5.56)和(103.2±3.94)μm。同时三组支架的压缩模量分别为(41.05±11.69)、(108.0±12.71)和(84.98±8.51)k Pa,弹性模量分别为(17.24±3.93)、(28.98±3.31)和(21.74±2.92)k Pa。MTT法表明骨软骨一体化支架无细胞毒性。荧光显微镜观察显示,纵切的支架薄片上细胞生长分布均匀,表明支架生物相容性较好,CCK8实验表明支架可以维持良好的细胞活性。结论脱细胞真皮基质/生物矿化胶原骨双相支架中上下层间的理化性能展示了骨软骨组织结构和力学方面的双重仿生,为动物体内实验奠定了基础,有望成为治疗骨软骨缺损修复的一种新手段。Objective To manufacture an acellular dermal matrix/biomineralized collagen diphasic osteochondral scaffold and then evaluate the physical and chemical properties, biomechanics and biocompatibility of the scaffold to provide theoretical basis for the repair of osteochondral defects. Methods Bovine acellular dermal matrix was prepared by physical and chemical methods such as acellular, pepsin and cross-linking. Bone-cartilage integrated biphasic scaffolds with acellular dermal matrix (ADM) as cartilage layer and biomineralized collagen (DCS) as bone layer were prepared by mixing tricalcium phosphate and collagen in different proportions and using self-assembly and freeze-drying technology. We constructed the acellular dermal matrix scaffold (ADM), biomineralized collagen scaffold (DCS), and ADM/DCS scaffold (ADM/DCS), respectively, and then the physical and chemical properties and mechanical properties of osteochondral integrated scaffolds were evaluated by gross observation, scanning electron microscopy and biomechanical testing. The primary cultured chondrocytes and osteoblasts cells of mice were implanted into acellular dermal matrix and biomineralized collagen, respectively. The scaffold was observed by cytotoxicity, dead/living cell staining and proliferation experiments. Results Gross observation showed that the scaffolds were closely interlaminar, without obvious discontinuity and separation. The pore structures in each layer of the scanning electron microscope were interconnected and multidimensional. The pore sizes of ADM, DCS and biphasic scaffolds were (121.6 ± 8.65),(98.40 ± 5.56) and (103.2 ± 3.94)μm, respectively. At the same time, the compression modulus of the three groups of scaffolds were (41.05 ± 11.69),(108.0 ± 12.71) and (84.98 ± 8.51) kPa, and the elastic modulus were (17.24 ± 3.93),(28.98 ± 3.31) and (21.74 ± 2.92) kPa. The result from MTT assay showed that the osteochondral scaffold had no cytotoxicity. Cells grew evenly on the scaffold, indicating that the scaffolds had good bioc

关 键 词：脱细胞真皮基质 生物矿化胶原 双相支架 骨软骨 

分 类 号：R318.08[医药卫生—生物医学工程] R68[医药卫生—基础医学]