Lohberger Birgit, Payer Michael, Rinner Beate, Bartmann Christina, Stadelmeyer Elke, Traunwieser Elisabeth, DeVaney Trevor, Jakse Norbert, Leithner Andreas, Windhager Reinhard
Department of Orthopaedic Surgery, Medical University of Graz, Graz, Austria.
J Oral Maxillofac Surg. 2012 Oct;70(10):2309-15. doi: 10.1016/j.joms.2011.06.216. Epub 2011 Sep 21.
The aim of this study was the establishment of a minimally invasive technique of mesenchymal stem cell (MSC) harvesting and a predictable isolation and cultivation method on 2 different bone substitutes used as potential scaffolds.
Human MSCs isolated from the posterior maxilla were characterized by flow cytometric analysis. After in vitro expansion, cells were cultured and differentiated toward osteogenic, adipogenic, and chondrogenic lineages in 2-dimensional cultures and on natural bone mineral of bovine origin and β-tricalcium phosphate scaffolds. Three-dimensional growth was analyzed using live cell staining and confocal laser scanning microscopy.
MSCs from all patients demonstrated the same immunophenotype, with expression of CD73, CD90, and CD105 but no expression of CD45, CD34, CD14, CD11, and HLA-DR. The potential of MSCs for multilineage differentiation along osteogenic, adipogenic, and chondrogenic lines was shown. Based on knowledge of the characteristics of the cells, a method was established to increase MSC expansion efficiency and seeding conditions on each scaffold. Results of the in vitro characterization and laser scanning microscopy visualized the 3-dimensional growth of MSCs on the 2 scaffold types.
The present data showed that intraoral MSCs can be cultured predictably under 2- and 3-dimensional conditions, have proved multiple potencies, and thus seem to be potential candidates for tissue engineering approaches in maxillofacial reconstructions.
本研究的目的是建立一种间充质干细胞（MSC）采集的微创技术，以及一种在两种用作潜在支架的不同骨替代物上可预测的分离和培养方法。
通过流式细胞术分析从后上颌骨分离的人MSC。体外扩增后，细胞在二维培养物中以及在牛源天然骨矿物质和β-磷酸三钙支架上培养并向成骨、成脂和成软骨谱系分化。使用活细胞染色和共聚焦激光扫描显微镜分析三维生长情况。
所有患者的MSC均表现出相同的免疫表型，表达CD73、CD90和CD105，但不表达CD45、CD34、CD14、CD11和HLA-DR。显示了MSC沿成骨、成脂和成软骨谱系进行多谱系分化的潜力。基于对细胞特性的了解，建立了一种提高MSC扩增效率和在每种支架上接种条件的方法。体外表征和激光扫描显微镜的结果显示了MSC在两种支架类型上的三维生长情况。
目前的数据表明，口腔内的MSC可以在二维和三维条件下进行可预测的培养，已证明具有多种潜能，因此似乎是颌面重建组织工程方法的潜在候选者。
