【作者】 张鹏；

【导师】 康斌；

【作者基本信息】 安徽医科大学 ， 外科学（骨外）， 2021， 硕士

【摘要】 目的:通过体外和体内实验研究探索利用模板技术制造的新型的镁基钙磷生物多孔活性支架成骨和成血管能力以及促进骨缺损修复能力。方法:在体外实验中,使用支架浸提液分别与小鼠的前成骨细胞(MC3T3-E1)和人的脐静脉内皮细胞(HUVECs)进行培养。对小鼠的前成骨细胞(MC3T3-E1)进行细胞活性检测,探究本支架对前成骨细胞生物安全性以及增殖效应。然后对小鼠的前成骨细胞(MC3T3-E1)进行碱性磷酸酶表达检测、前成骨细胞体外矿化实验以及相关RNA表达量检测,研究本支架的促成骨效应。对人的脐静脉内皮细胞(HUVECs)进行细胞活性检测,探究本支架对内皮细胞的生物安全性以及增殖效应。然后对内皮细胞(HUVECs)进行内皮细胞成管实验,探索本支架的成血管效应。体内实验采用大鼠作为实验动物,制造大鼠的股骨髁缺损模型并植入本支架,在8周处死小鼠,并进行组织学观察、血管造影以及micro CT,观察新生骨及新生血管生长情况,探究本支架在体内的成骨、成血管效应。结果:体外实验结果表明,镁基钙磷生物多孔活性支架与小鼠前成骨细胞细胞一起培养时可促进细胞向内生长和成骨分化,还有增强矿化,并提高成骨相关基因在细胞内的表达。与人脐静脉内皮细胞一起培养时,有着促进增殖、迁移以及成血管的作用。体内植入后,多孔镁基钙磷生物活性支架随着多孔镁基钙磷生物活性支架的降解而有效刺激血管生成,成骨和重塑,并完美修复大鼠模型中的骨缺损。结论:本研究的表明在本支架体外实验中有优秀的成骨、成血管效应,体内实验中有着优秀的修复骨缺损的能力。更多还原

【Abstract】 Purpose: In vitro and in vivo experimental studies are conducted to explore the osteogenic abilities,angiogenic capabilities and repair ability of bone defect of a new type of calcium-phosphorus coated magnesium based porous bioactive scaffold fabricated through template technology.Methods: In vitro experiments,mouse preosteoblasts(MC3T3-E1)and human umbilical vein endothelial cells(HUVECs)were used and cultured with the scaffold extract liquids.The cell viability of mouse pre-osteoblasts(MC3T3-E1)were tested to explore the biological safety and proliferation effects of the scaffold on pre-osteoblasts.Then,alkaline phosphatase expression detection,pre-osteoblast in vitro mineralization experiment and related RNA expression detection were performed on mouse pre-osteoblasts(MC3T3-E1)to explore the osteogenic promotion effects.The cell viability test of human umbilical vein endothelial cells(HUVECs)was carried out toexplore the biological safety and proliferation effects of the scaffold on endothelial cells.Then,the endothelial cell tube formation experiment was performed on the endothelial cells(HUVECs)to explore the angiogenic promotion effects.In vivo experiments,rats were used as experimental animals.The scaffold was implanted in the femoral condyle defect model of rats.The rats were sacrificed at 8 weeks,and histological observation,angiography and micro CT were performed to observe the growth of new bone and new blood vessels.The comparison between the two groups explore the osteogenesis and angiogenesis effects of the scaffold in the body.Results:The results of in vitro experiments showed that the extract liquids of the the porous magnesium-based calcium-phosphorus bioactive coated scaffold can promote cell growth and osteogenic differentiation and can also enhance mineralization and improve osteogenic related genes expression when cultured with mouse pre-osteoblast.While cultured with human umbilical vein endothelial cells,the results shows that the porous magnesium-based calcium-phosphorus bioactive coated scaffold can promote migration,proliferation and angiogenesis of HUVECs.In vivo,the porous magnesium-based calcium-phosphorus bioactive coated scaffold can effectively stimulates angiogenesis,osteogenesis and perfectly repairs the bone defect in the rat models as the porous magnesium-based calcium-phosphorus bioactive scaffold degraded.Conclusion: This study shows that the scaffold has excellent osteogenesis and angiogenesis promotion effects in vitro experiments,and has excellent ability to repair bone defects in vivo experiments.更多还原