A brushite-forming calcium phosphate cement (CPC) was mechanically stabilized by addition of poly (l-lactid-co-glycolide; PLGA) fibers (≤10% w/w). It proved highly biocompatible and its fiber component enhanced bone formation in a sheep lumbar vertebroplasty model. However, possible effects on the osteogenic differentiation of resident mesenchymal stem cells (MSCs) remained unexplored. The present study used a novel approach, simultaneously analyzing the influence of a solid CPC scaffold and its relatively low PLGA proportion (a mimicry of natural bone) on osteogenic, chondrogenic, and adipogenic differentiation, as well as the pluripotency of human adipose tissue-derived mesenchymal stem cells (hASCs). hASCs were cultured on CPC discs with/without PLGA fibers (5% and 10%) in the absence of osteogenic medium for 3, 7, and 14 d. Gene expression of osteogenic markers (Runx2, osterix, alkaline phosphatase, collagen I, osteonectin, osteopontin, osteocalcin), chondrogenic markers (collagen II, Sox9, aggrecan), adipogenic markers (PPARG, Leptin, and FABP4), and pluripotency markers (Nanog, Tert, Rex) was analyzed by RT-PCR. The ability of hASCs to synthesize alkaline phosphatase was also evaluated. Cell number and viability were determined by fluorescein diacetate/propidium iodide staining. Compared to pure CPC, cultivation of hASCs on fiber-reinforced CPC transiently induced the gene expression of Runx2 and osterix (day 3), and long-lastingly augmented the expression of alkaline phosphatase (and its enzyme activity), collagen I, and osteonectin (until day 14). In contrast, augmented expression of all chondrogenic, adipogenic, and pluripotency markers was limited to day 3, followed by significant downregulation. Cultivation of hASCs on fiber-reinforced CPC reduced the cell number, but not the proportion of viable cells (viability > 95%). The PLGA component of fiber-reinforced, brushite-forming CPC supports long-lasting osteogenic differentiation of hASCs, whereas chondrogenesis, adipogenesis, and pluripotency are initially augmented, but subsequently suppressed. In view of parallel animal results, PLGA fibers may represent an interesting clinical target for future improvement of CPC- based bone regeneration.

中文翻译：

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形成透钙磷石的磷酸钙水泥的聚（1-丙交酯-乙交酯； PLGA）纤维成分可诱导人脂肪组织来源的干细胞的成骨分化。

通过添加聚（1-丙交酯-乙交酯； PLGA）纤维（≤10％w / w），使透钙磷石形成的磷酸钙水泥（CPC）机械稳定。它被证明具有高度的生物相容性，并且其纤维成分增强了绵羊腰椎椎体成形术模型中的骨形成。但是，尚未探讨可能对居民间充质干细胞（MSCs）成骨分化的影响。本研究采用一种新颖的方法，同时分析了固体CPC支架及其相对较低的PLGA比例（天然骨的模拟物）对成骨，软骨形成和成脂分化以及人类脂肪组织来源的多能性的影响。间充质干细胞（hASCs）。在没有成骨培养基的情况下，在有/无PLGA纤维（5％和10％）的CPC圆盘上培养hASC 3、7和14 d。成骨标记物（Runx2，osterix，碱性磷酸酶，胶原I，骨连接蛋白，骨桥蛋白，骨钙素），软骨形成标记物（胶原蛋白II，Sox9，聚集蛋白聚糖），成脂标记物（PPARG，Leptin和FABP4）和多能性标记物（Nanog）的基因表达，Tert，Rex）通过RT-PCR分析。还评估了hASCs合成碱性磷酸酶的能力。通过荧光素二乙酸盐/碘化丙啶染色确定细胞数目和活力。与纯CPC相比，在纤维增强的CPC上培养hASC会短暂诱导Runx2和osterix的基因表达（第3天），并持久地增强碱性磷酸酶（及其酶活性），胶原I和骨连接蛋白的表达（直到第14天）。相反，所有软骨形成，脂肪形成和多能性标志物的增强表达仅限于第3天，其次是大幅下调。在纤维增强的CPC上培养hASC可减少细胞数量，但不能减少存活细胞的比例（存活率> 95％）。纤维增强的透钙磷石形成的CPC的PLGA成分支持hASC的持久成骨分化，而软骨形成，脂肪形成和多能性最初得到增强，但随后被抑制。鉴于平行的动物结果，PLGA纤维可能代表了基于CPC的骨再生未来改进的有趣的临床目标。脂肪形成和多能性最初增强，但随后被抑制。鉴于平行的动物结果，PLGA纤维可能代表了基于CPC的骨再生未来改进的有趣的临床目标。脂肪形成和多能性最初增强，但随后被抑制。鉴于平行的动物结果，PLGA纤维可能代表了基于CPC的骨再生未来改进的有趣的临床目标。