Enhanced Osteogenesis of Human Mesenchymal Stem Cells by Periodic Heat Shock in Self-Assembling Peptide Hydrogel
Enhanced Osteogenesis of Human Mesenchymal Stem Cells by Periodic Heat Shock in Self-Assembling Peptide Hydrogel
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DOI：
10.1089/ten.tea.2012.0070
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发表时间：
2013-03-01
影响因子：
4.1
通讯作者：
Wang, Sihong
中科院分区：
文献类型：
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作者：
Chen, Jing;Shi, Zhong-Dong;Wang, Sihong
热诱导成骨的机制尚不完全清楚，人间充质干细胞（hMSC）分化的热调节机制也未得到很好的研究。本研究研究了轻度热休克（HS）对自组装肽水凝胶和组织培养板上hMSCs向成骨细胞分化的直接影响。将从人骨髓中分离的hMSCs分别接种于常规培养板（二维培养）和三维（3D） PuraMatrix肽水凝胶（3D培养）表面，在成骨分化过程中，每周1次，41℃高温培养1小时。在分化早期，通过周期性HS培养可增强二维和三维培养的碱性磷酸酶（ALP）活性；在2D和3D培养中，HS均显著增加分化第19天和第27天的钙沉积。在2D培养中，周期性HS也上调了骨特异性基因，第11天上调了骨蛋白（OSX），第19天上调了骨桥蛋白（OP），第25天上调了骨形态发生蛋白2 （BMP2）。在3D PuraMatrix培养中，HS在分化第25天上调了runt相关转录因子2 （Runx2）。热休克蛋白70 （HSP70）在热休克24小时后显著上调。这些结果表明，HS诱导了hMSCs的早期分化，并促进了由hMSCs分化而来的成骨细胞的成熟。因此，温和的HS治疗有可能用于增强骨髓间充质干细胞的骨再生。我们的数据将指导体内加热方案的设计，并进一步研究骨组织工程中MSC成骨的热处理。
The mechanisms for the heat-induced osteogenesis are not completely known and the thermal regulation of human mesenchymal stem cell (hMSC) differentiation is not well studied. In this study, the direct effects of mild heat shock (HS) on the differentiation of hMSCs into osteoblasts in self-assembling peptide hydrogel and on tissue culture plates were investigated. hMSCs isolated from human bone marrow were seeded in conventional culture plates (two-dimensional [2D] culture) and on the surface of three-dimensional (3D) PuraMatrix peptide hydrogel (3D culture), followed by 1 h HS at 41 degrees C once a week during osteogenic differentiation. Alkaline phosphatase (ALP) activity was enhanced in both 2D and 3D cultures via periodic HS at early stage of differentiation; meanwhile, HS significantly increased the calcium deposition at day 19 and 27 of differentiation in both 2D and 3D cultures. The periodic HS also upregulated osteo-specific genes, osterix (OSX) on day 11, osteopontin (OP) on day 19, and bone morphogenetic protein 2 (BMP2) on day 25 in 2D culture. In 3D PuraMatrix culture, the runt-related transcription factor 2 (Runx2) was upregulated by HS on day 25 of differentiation. The heat shock protein 70 (HSP70) was significantly upregulated by HS in differentiated hMSCs analyzed at 24 h after HS. These results demonstrate that HS induced an earlier differentiation of hMSCs and enhanced the maturation of osteoblasts differentiated from hMSCs. Therefore, mild HS treatment may be potentially used to enhance the bone regeneration using hMSCs. Our data will guide the design of in vivo heating protocols and enable further investigations in thermal treatments of MSC osteogenesis for bone tissue engineering.