纳米晶羟基磷灰石与兔骨髓间充质干细胞复合培养回植体内的成骨性能

背景：骨髓间充质干细胞具有自我更新及多向分化的能力，经诱导后能够向成骨细胞分化，与基质材料结合可以具备形态功能良好的骨缺损修复效应。目的：观察骨髓间充质于细胞与基质材料复合后同体移植修复节段性骨缺损的成骨性能。设计：开放性实验。单位：无锡市第三人民医院细胞实验室。材料：实验于2004—05／2005—03在无锡市第三人民医院细胞实验室完成。选取清洁级6月龄新西兰兔18只，随机数字表法分为细胞支架复合组、单纯支架组、空白对照组，6只／组。支架材料由清华大学材料系生物组提供纳米晶羟基磷灰石胶原材料，具备天然骨微结构特征（组成：羟基磷灰石约57％，胶原约30％，聚乳酸约13％）。方法：①分离培养兔骨髓间充质干细胞，取生长良好的第2代细胞应用EPICS—ALTRA流式细胞仪检测细胞表面抗原表达。②支架材料按实验需求切割成6min×6min×4mm大小，^60Co辐照灭菌，使用前DMEM—LG浸润：收集培养两三代的细胞，调整浓度为10^9L^-1，与纳米晶羟基磷灰石胶原材料于体外联合培养。③各组均建立兔桡骨节段性缺损模型。细胞支架复合组将已备好的复合物采用同体移植的原则嵌入骨缺损中，单纯支架组将空白支架材料嵌入骨缺损中，空白对照组不植入任何材料。各组动物均不作内外固定，严密缝合软组织、皮肤。④扫描电镜下观察各组材料表面结构及细胞附着情况。⑤各组分别于术后4，8，16周行大体观察．组织学分析及X光摄片，了解成骨情况。主要观察指标：①骨髓间充质干细胞分离培养及鉴定情况。②各组扫描电镜观察结果。③术后各组放射学检测结果。④术后各组大体形态观察结果。⑤术后各组组织学检测结果。结果：实验选取清洁级新西兰免18只，全部进入结果分析。①原代培养4h后可?

Bone formation performance by autograft of rabbit mesenchymal stem cells co-cultured with nano basal hydroxyapatite materials

BACKGROUND： Bone bone bone marrow derived mesenchymal stem cells （MSCs） has the capacities of self-renewal and multi-directional differentiation, which can differentiate into osteoblasts after induction. Combined with basal hydroxyapatite materials, MSCs can repair the bone defeet with satisfactory, shape and function.
OBJECTIVE： To observe the bone formation performance of segmental bone defect repaired by autologous transplantation after combination of MSCs and basal hydroxyapatite materials.
DESIGN： Open experiment.
SETTING： Department of Cells, Third People＇s Hospital of Wuxi.
MATERIALS： The experiment was conducted in the Department of Cells, Third People＇s Hospital of Wuxi between May 2004 and March 2005. Eighteen New Zealand rabbits of clean grade, aged 6 months were selected and randomly divided into cytoskcleton group, simple scaffold group and blank control group with 6 rabbits in each group. Nano-collagen basal hydroxyapatite bone were provided by the Department of Material, Tsinghua University, which is characteristic of natural-bone microstrueture. Composition： about 57% were hydroxyapatite,and 30 %. were collagen and 13% were polylactic acid.
METHODS： （1）bone bone marrow derived MSCs of rabbits were isolated and cultured. Well-grown cells of the second generation were inspected of the expression of cell surface antigen （CSA） with EPICS-ALTRA flow cytometry. （2）Scaffold materials were cut into blocks with the size of 6 mm ×6 mm×4 mm, sterilized by ^60Co irradiance and infihrated with DMEM-LG before using. Cells of the 2^ndl and 3^rd generations were collected, cultured, the concentration of which was regulated to 10^9 L^-1. Cells were co-cultured with nano-eollagen basal hydroxyapatite in vitro. （3）Model of segmental defects in radial of rabbits were established in all groups. The prepared compounds in the eytoskeleton compound group were embedded in the bone defect according to autograft principle, and blank scaffold materials were embedded in the bo