镁基合金支架负载兔源性骨髓间充质干细胞构建组织工程化骨复合支架  

Construction of tissue-engineered bone composite scaffolds by loading rabbit-derived bone marrow mesenchymal stem cells on magnesium-based alloy scaffolds

作　　者：周悦彬 郭洪刚[2] Zhou Yuebin;Guo Honggang(Department of Orthopedics,Tianjin Hospital,Tianjin 300200,China;Department of Orthopedics,General Hospital,Tianjin Medical University,Tianjin 300052,China)

机构地区：[1]天津市天津医院骨科,天津市300200 [2]天津医科大学总医院骨科,天津市300052

出　　处：《中国组织工程研究》2023年第34期5491-5496,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:随着组织工程学的深入研究与快速发展,组织工程支架为骨组织修复提供了新的契机。目的:探讨骨髓间充质干细胞与镁基合金支架共培养后的细胞增殖情况,为组织工程化骨支架构建提供新思路,进而为组织工程支架临床应用提供依据。方法:无菌条件下抽取兔骨髓液,使用全骨髓贴壁法分离培养骨髓间充质干细胞,进行流式细胞学鉴定和成骨分化能力检测。将第3代骨髓间充质干细胞分别接种于镁基合金支架和壳聚糖-聚己内酯-磷酸三钙支架,第3,10天进行激光共聚焦扫描观察细胞生长情况,第1,4,7,10,14天行MTT检测细胞增殖情况。结果与结论:(1)分离培养的细胞高表达CD44和CD90,几乎不表达CD34;(2)经成骨诱导后,细胞形态逐渐变为多角形,Von Kossa染色可见细胞之间钙结节染成黑色,碱性磷酸酶活性较未诱导对照组升高(P<0.05);(3)镁基合金与骨髓间充质干细胞共培养后,共聚焦显微镜扫描显示细胞可在支架上黏附并呈团块状生长,随着时间延长,镁基合金支架较壳聚糖-聚己内酯-磷酸三钙支架上负载的细胞增殖活跃,细胞数量明显增多,MTT检测显示两种材料与骨髓间充质干细胞共培养至10 d时,细胞生长达峰值;(4)结果表明,兔源性骨髓间充质干细胞具有良好的成骨潜能,与镁基合金支架共培养后,细胞在支架表面生长良好,成功构建了组织工程化镁基种植体。BACKGROUND:With the in-depth research and rapid development of tissue engineering,tissue-engineered scaffolds provide a new opportunity for bone tissue repair.OBJECTIVE:To explore the cell proliferation of bone marrow mesenchymal stem cells after co-culture with the magnesium-based alloy scaffolds,to provide a new idea for the construction of tissue-engineered bone scaffolds and provide a basis for the clinical application of tissue-engineered scaffolds.METHODS:The bone marrow fluid of rabbits was extracted under sterile conditions.Bone marrow mesenchymal stem cells were cultured and isolated by the whole bone marrow adherent method.Bone marrow mesenchymal stem cells were identified by flow cytometry and osteogenic differentiation ability was tested.Bone marrow mesenchymal stem cells at passage 3 were seeded on magnesium-based alloy scaffolds and chitosan polycaprolactone tricalcium phosphate scaffolds respectively.The cell growth was observed by laser confocal scanning on days 3 and 10,and cell proliferation was detected using an MTT assay on days 1,4,7,10 and 14,respectively.RESULTS AND CONCLUSION:(1)The cultured cells highly expressed CD44 and CD90,but hardly CD34.(2)The morphology gradually changed to polygonal after osteogenesis induction.Von Kossa staining showed that the calcium nodules between the cells were stained black,and the alkaline phosphatase activity was significantly higher than that of the control group(P<0.05).(3)After co-culture of magnesium-based alloy and bone marrow mesenchymal stem cells,confocal scanning showed that the cells could adhere to the scaffold and grow in clusters.With the extension of time,the magnesium-based alloy scaffold was more active in proliferation than the seed cells loaded on the chitosan polycaprolactone tricalcium phosphate scaffold,and the number of cells was significantly increased.MTT assay showed that when the two materials were co-cultured with bone marrow mesenchymal stem cells for 10 days,the cell growth reached its peak.(4)These results indicate that rabbi

关 键 词：骨髓间充质干细胞 镁基合金 组织工程化骨 细胞增殖 支架 共培养 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R394.2