Ye Qing, Chen Kaining, Huang Wu, He Yunsong, Nong Mingshan, Li Chunxiang, Liang Tiansen
Department of Orthopedics, The General Hospital of the Armed Police Force of Guangxi, Nanning, Guangxi 530003, P.R. China ; Center of Tissue Engineering Research and Application, The General Hospital of the Armed Police Force of Guangxi, Nanning, Guangxi 530003, P.R. China.
Department of Neurology, The General Hospital of the Armed Police Force of Guangxi, Nanning, Guangxi 530003, P.R. China.
Exp Ther Med. 2015 Jan;9(1):25-32. doi: 10.3892/etm.2014.2067. Epub 2014 Nov 12.
Poly-L-lysine (PLL) is commonly used as an adhibiting agent due to its good viscosity, and demineralized bone matrix (DBM) is a common enriched matrix for selective cell retention technology. Therefore, the aim of this study was to use PLL to coat the surface and interspaces of DBM to form a novel type of enriched matrix [DBM coated with PLL (PLL-DBM)], in order to effectively improve the enrichment effects of bone marrow stem cells and enhance their osteogenic ability. Electron microscope scanning and the infrared spectrum were used to observe the structure of PLL-DBM and the optimal conditions for the combination of PLL and DBM. Enriching effects on bone marrow nucleated cells (NCs) and platelets (PLTs) were detected with an automated hematology analyzer. The osteogenesis of the following four groups was assessed with a grafting bone model in a goat spinal transverse process: IA, tissue engineered bone (TEB) fabricated following enrichment of bone marrow with PLL-DBM; IB, autogenous iliac bone; IIC, TEB fabricated following enrichment of bone marrow with DBM; IID, blank DBM. The goats were sacrificed in one batch at week 16 after the surgery and the fusion specimens were examined using X-ray and three-dimensional computed tomography (CT). In addition, the CT value was determined and the histology and biomechanics were analyzed in order to evaluate the osteogenic ability. The results showed that PLL and DBM combined well and that PLL-DBM exhibited a natural mesh pore structure. The fold enrichment of NCs and PLTs with PLL-DBM was significantly higher than that with DBM. The fusion effects of the IA and IB groups were similar and significantly enhanced compared with those of the IIC and IID groups. The results confirmed that PLL-DBM is an ideal enriched matrix for bone marrow stem cells, and TEB rapidly fabricated by PLL-DBM intraoperatively enriched bone marrow stem cells exhibits an improved osteogenic ability.
聚-L-赖氨酸（PLL）因其良好的粘性常被用作黏附剂，而脱矿骨基质（DBM）是选择性细胞保留技术中常用的富集基质。因此，本研究旨在用PLL包被DBM的表面和间隙，形成一种新型的富集基质[PLL包被的DBM（PLL-DBM）]，以有效提高骨髓干细胞的富集效果并增强其成骨能力。采用电子显微镜扫描和红外光谱观察PLL-DBM的结构以及PLL与DBM结合的最佳条件。用自动血液分析仪检测对骨髓有核细胞（NCs）和血小板（PLTs）的富集效果。在山羊脊柱横突的植骨模型中评估以下四组的成骨情况：IA组，用PLL-DBM富集骨髓后制备的组织工程骨（TEB）；IB组，自体髂骨；IIC组，用DBM富集骨髓后制备的TEB；IID组，空白DBM。术后16周将山羊一次性处死，用X射线和三维计算机断层扫描（CT）检查融合标本。此外，测定CT值并分析组织学和生物力学以评估成骨能力。结果表明，PLL与DBM结合良好，PLL-DBM呈现出天然的网状孔隙结构。PLL-DBM对NCs和PLTs的富集倍数显著高于DBM。IA组和IB组的融合效果相似，与IIC组和IID组相比显著增强。结果证实，PLL-DBM是骨髓干细胞理想的富集基质，术中用PLL-DBM快速制备的TEB富集骨髓干细胞后成骨能力得到提高。
