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首页> 外文期刊>Acta biomaterialia >Initial cell pre-cultivation can maximize ECM mineralization by human mesenchymal stem cells on silk fibroin scaffolds.
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Initial cell pre-cultivation can maximize ECM mineralization by human mesenchymal stem cells on silk fibroin scaffolds.

机译：初始细胞的预培养可以最大程度地通过人丝蛋白支架上的人间充质干细胞使ECM矿化。

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Fast remineralization of bone defects by means of tissue engineering is one of many targets in orthopedic regeneration. This study investigated the influence of a range of pre-culture durations for human bone marrow derived mesenchymal stem cells (hMSC) before inducing differentiation into osteoblast-like cells. The aim was to find the conditions that lead to maximal extracellular matrix (ECM) mineralization, in terms of both amount and best distribution. Additionally, the influence of silk fibroin scaffold pore size on mineralization was assessed. The formation of mineralized ECM by hMSCs cultured in osteogenic medium on silk fibroin scaffolds was monitored and quantified for up to 72 days in culture using non-invasive time-lapse micro-computed tomography (micro-CT). ECM mineralization increased linearly 3 weeks after the beginning of the experiment with addition of differentiation medium. Biochemical end-point assays measured the amount of DNA, calcium deposits, alkaline phosphatase activity and cell metabolic activity to corroborate the hypothesis that an initial pre-culture period of hMSCs on silk fibroin scaffolds can accelerate mineralized ECM formation. According to the micro-CT analysis mineralization on silk fibroin scaffolds with pores of 112-224 mum diameter was most efficient with an initial cell pre-culture period of 9 days, showing 6.87+/-0.81x higher mineralization values during the whole cultivation period than without an initial cell pre-culture period.
机译：通过组织工程技术快速再矿化骨缺损是骨科再生的众多目标之一。这项研究在诱导分化为成骨细胞样细胞之前，研究了一系列预培养时间对人骨髓源间充质干细胞（hMSC）的影响。目的是找到在数量和最佳分布方面导致最大的细胞外基质（ECM）矿化的条件。此外，评估了丝素蛋白支架的孔径对矿化的影响。使用无创延时微电脑断层扫描（micro-CT）监测并量化了在丝素蛋白支架上的成骨培养基中培养的hMSC的矿化ECM的形成情况，并长达72天进行了定量。实验开始后3周，加入分化培养基后，ECM矿化呈线性增加。生化终点测定法测量了DNA的量，钙沉积，碱性磷酸酶活性和细胞代谢活性，从而证实了hMSC在丝素蛋白支架上的初始预培养阶段可以加速矿化ECM形成的假说。根据Micro-CT分析，在孔径为112-224微米的丝素蛋白支架上进行矿化是最有效的，初始细胞预培养期为9天，在整个培养期间显示出更高的矿化值6.87 +/- 0.81x比没有最初的细胞预培养期。

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