新型骨髓干细胞富集材料富集骨髓细胞的效果

背景：骨髓干细胞富集材料是目前研究热点，制备并筛选一种新型骨髓干细胞富集材料对于组织工程学发展及临床应用至关重要。目的：制备并筛选一种骨组织工程新型骨髓干细胞富集材料，应用选择性细胞滞留技术观察其富集骨髓细胞的效果。方法：用预先制备的人脱钙骨基质与不同体积分数（0.01%，0.05%，0.1%，0.5%，1%）的多聚左旋赖氨酸复合制备多聚左旋赖氨酸-脱钙骨基质材料，应用选择性细胞滞留技术富集骨髓干细胞进行实验。激光显微共聚焦拉曼光谱分析、红外光谱分析鉴定材料成分；三维视频显微镜、扫描电镜观察富集材料孔径、孔隙率、表面及横断面超微结构；对富集前后骨髓有核细胞、纤维母细胞集落形成单位和血小板进行计数分析。结果与结论：多聚左旋赖氨酸-脱钙骨基质富集材料孔隙率高，孔隙内部有大量孔隙相互连通，材料内外表面有乳白色均匀致密的多聚左旋赖氨酸涂层覆盖，并在天然孔隙内形成更小、孔径较均匀的网孔结构。体积分数0.1%多聚左旋赖氨酸制备的富集材料对人骨髓有核细胞、纤维母细胞集落形成单位和血小板的浓度富集倍数分别为（3.18±0.31）、（5.25±1.40）和（3.88±0.68）倍，相应的黏附率分别达（53±12）%，（73±13）%和（34±10）%，对纤维母细胞集落形成单位的选择率为1.41±0.34。结果证实，实验制备并筛选的经多聚左旋赖氨酸修饰的脱钙骨基质具有天然骨海绵状支架网孔结构，对骨髓细胞的选择性滞留率高，是一种较理想的骨髓干细胞富集材料。

Effect of a new-type enriched material on enriching bone marrow stem cells by selective cell retention

BACKGROUND：Enriching material for bone marrow stem cells is a current research hotspot. Preparation and screening of a new type of bone marrow stem cells-enriched material is crucial for the development of tissue engineering and clinical applications. OBJECTIVE：To observe the surface character and effect of a new type of bone marrow stem cells-enriched material on enriching bone marrow stem cells of decalcified bone matrix decorated with poly-L-lysine by selective cellretention technology. METHODS：Decalcified bone matrix was decorated with poly-L-lysine as the scaffold material. Bone marrow mesenchymal stem cells were col ected by using selective cellretention technology. Control group was decalcified bone matrix and experimental groups were Decalcified bone matrix decorated with poly-L-lysine at different concentrations （0.01%, 0.05%, 0.1%, 0.5%, 1%） with freeze-drying method. Material components were analyzed by Raman spectrum and infrared spectrum. Pore size, porosity, surface and cross section were observed under three-dimension video microscope and scanning electron microscope. The number of bone marrow nucleated cells, fibroblast colony-forming units and platelets were counted pre-and post-enrichment. RESULTS AND CONCLUSION：Decalcified bone matrices were covered with wel-distributed and dense poly-L-lysine. The poly-L-lysine/decalcified bone matrix had a high porosity with much more internal y connected pores. After enrichment with enriched material prepared by 0.1%poly-L-lysine, the number of bone marrow nucleated cells, fibroblast colony-forming units and platelets were increased （3.18±0.31）, （5.25±1.40） and （3.88±0.68） times, respectively;the binding efficiency were （53±12）%, （73±13）%and （34±10）%, respectively;the selective rate of fibroblast colony-forming units was 1.41±0.34. The new constructed poly-L-lysine/decalcified bone matrix has good three-dimension space with high selective retention. It is an effective enriching material for bone marrow mesenchymal s