矿化胶原与脂肪源间充质干细胞的生物相容性及对其成骨分化的影响

发布时间：2018-02-09 20:22

本文关键词： 矿化胶原 Ⅰ型胶原羟基磷灰石间充质干细胞成骨分化　出处：《北京协和医学院》2016年博士论文　论文类型：学位论文

【摘要】：研究背景：虽然骨组织有一定的再生和修复能力,但仍不足以代偿病理性骨折、创伤等因素造成的较大骨量缺损。周围血运不足、感染或全身性疾病等因素也会增加骨缺损治疗的难度。骨移植材料可分为自体骨、异体骨和人工骨材料,自体骨和异体骨均因各自的缺陷而导致在临床上应用受限,因此人工骨材料逐渐成为再生医学领域新的研究热点。矿化胶原(Mineralized collagen, MC)是模拟骨基质化学组成和微观结构的仿生复合材料,其与人脂肪源间充质干细胞(human adipose-derived stem cells, hADSCs)的研究目前鲜有报道。本研究将矿化胶原与I型胶原和羟基磷灰石进行对比,探究三者对hADSCs的生物相容性以及成骨分化的影响。研究方法：制备矿化胶原、胶原、羟基磷灰石三种材料的圆片,通过X射线衍射(X-ray diffraction, XRD)、FTIR (Fourier transform infrared spectroscopy, FTIR)检测材料物相组成和分子结构,通过接触角评估材料可湿润性,通过环境扫描电子显微镜(ESEM, environment scanning electron microscope)评估材料的表面形貌和细胞在材料上的贴附和生长状态；利用WST-8法观察细胞增殖；利用碱性磷酸酶(alkaline phosphatase, ALP)活性检测hADSCs成骨早期分化；通过real time PCR比较成晋标志性基因的表达。研究结果：矿化胶原具有与天然骨基质类似的化学组成和微观结构,hADSCs在其上表现出良好的贴附和生长行为。相比I型胶原和羟基磷灰石,矿化胶原更有利于hADSCs的增殖,并且成骨分化早期(RUNX2、ALP、COL1A1)和晚期(IBSP、OCN、OPN)的标志性基因均有不同程度的表达升高。结论：本研究表明相较于I型胶原和羟基磷灰石,矿化胶原对hADSCs表现出更好的生物相容性和成骨诱导能力,其与种子细胞hADSCs在骨组织工程领域有潜在的应用价值。
[Abstract]:Background: although bone tissue has a certain ability to regenerate and repair, it is still not sufficient to compensate for the large bone defect caused by pathological fracture, trauma and other factors. Infection or systemic diseases can also increase the difficulty in the treatment of bone defects. Bone graft materials can be divided into autologous bone, allograft and artificial bone materials, and both autogenous bone and allogeneic bone are limited in clinical application due to their respective defects. Therefore, artificial bone materials have gradually become a new research hotspot in regenerative medicine. Mineralized collagen Mineralized collagenesis (MCMC) is a biomimetic composite which simulates the chemical composition and microstructure of bone matrix. There are few reports on the comparison of mineralized collagen with type I collagen and hydroxyapatite in comparison with human adipose-derived stem cells (hADSCss) of human adipose derived mesenchymal stem cells (hADSCs). To explore the effects of the three factors on the biocompatibility and osteogenic differentiation of hADSCs. Methods: to prepare mineralized collagen, collagen and hydroxyapatite wafers. The phase composition and molecular structure of the material were examined by X-ray diffraction and FTIR transform infrared spectroscopy. The wettability of the material was evaluated by contact angle. The surface morphology and cell adhesion and growth state of the material were evaluated by environment scanning electron microscope. the cell proliferation was observed by WST-8 method, and the early differentiation of hADSCs osteogenesis was detected by alkaline phosphatase (ALP) activity. The expression of signature gene in Chengjin was compared by real time PCR. The results showed that mineralized collagen had similar chemical composition and microstructure to natural bone matrix and showed good adhesion and growth behavior. Compared with type I adhesive, mineralized collagen exhibited good adhesion and growth behavior. Primary and hydroxyapatite, Mineralized collagen is more conducive to the proliferation of hADSCs, and the expression of the iconic genes of RUNX2ALPPAL1A1) and late stage of hADSCs OCN-OPN are increased in different degrees. Conclusion: compared with type I collagen and hydroxyapatite, this study shows that, Mineralized collagen has better biocompatibility and osteogenic induction ability to hADSCs, and it has potential application value in bone tissue engineering field with seed cell hADSCs.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R318.08;R687

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