摘要
背景:人工合成的纳米羟基磷灰石具有良好的生物活性和相容性,植入体内后能在短时间内与体内的软硬组织形成紧密结合,因此成为广泛应用的骨组织工程材料。目的:观察豚鼠骨髓间充质干细胞与纳米羟基磷灰石复合多孔陶瓷材料体外复合培养的结合程度,并分析其构建组织工程人工听骨的可行性。方法:采用细胞差速贴壁法分离培养豚鼠骨髓间充质干细胞,使用流式细胞仪检测CD29、CD45、CD44进行间充质干细胞表面标记物鉴定,并检测其骨细胞分化能力。将骨髓间充质干细胞与纳米羟基磷灰石复合多孔陶瓷共培养1,3,5,7,10 d,电镜观察骨髓间充质干细胞与此种支架材料的复合情况。结果与结论:培养3 d后,可见大量骨髓间充质干细胞贴附在材料表层,并且形态稳定,生长旺盛,增殖力强,具有极佳的延伸性;培养5 d后可见材料表面已全部覆盖骨髓间充质干细胞,细胞结合紧密,细胞表面可见大量分泌颗粒,胞体明显增大,边缘完整,呈纤维样伸长;7 d后细胞逐渐从材料表面脱落,仍附在材料表面的细胞出现"星形"改变,"伪足"增多;10 d后细胞呈片状脱落。说明纳米羟基磷灰石材料保持了它良好的生物相容性,利于细胞黏附、增殖,可结合大量的骨髓间充质干细胞,用于构建组织工程人工听骨。
BACKGROUND：Artificially synthetic nano-hydroxyapatite（nHA） has good biocompatibility and bioactivity.It can tightly connect with in vivo tissues closely within a short time after implanted into the body.Therefore nHA has been widely used for bone tissue engineering.OBJECTIVE：To observe the compounded culture degree of guinea pig bone marrow-derived mesenchymal stem cells（MSCs） and nHA,to analyze the feasibility of constructing tissue-engineered artificial otosteon.METHODS：The MSCs were isolated from the bone marrow of guinea pig by differential adhesion method.The surface markers of MSCs were examined CD29,CD44,CD45 by flow cytometric analyses.The differentiated capability of MSCs was also detected.The MSCs were cultured with nHA porous ceramic,and their adhesion on the scaffold was observed by electron microscope at 1,3,5,7,10 days.RESULTS AND CONCLUSION：After three days cultivation,a large amount of MSCs attached on the material surface with stable form,proliferation activity and terrific extensibility;Composite material had all covered MSCs at 5 days,cell surface had a large number of secretary granules,soma amplified obviously,edge of cells was complete and fibroid extension;7 days later cells gradually fell from the material surface,still attached to the material surface and appeared ＂star＂ or ＂pseudopods＂;10 days later,cells flaked off the material surface.It can be explained that nHA maintains a good biocompatibility,which is beneficial for MSCs adhesion,proliferation and combination,nHA can be used to construct tissue-engineered artificial otosteon.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2011年第8期1345-1349,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
新疆维吾尔自治区自然科学基金项目(200821150)~~
