Thermogelling chitosan and collagen composite hydrogels initiated with beta-glycerophosphate for bone tissue engineering.
Thermogelling chitosan and collagen composite hydrogels initiated with beta-glycerophosphate for bone tissue engineering.
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DOI：
10.1016/j.biomaterials.2010.01.131
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发表时间：
2010-05
期刊：
影响因子：
14
通讯作者：
Stegemann, Jan P.
中科院分区：
文献类型：
--
作者：
Limin Wang;Stegemann, Jan P.
壳聚糖和I型胶原蛋白是用作细胞载体的天然来源的材料，因为它们能够模拟细胞外环境并指导细胞功能。在这项研究中，β-甘油磷酸盐（β-GP），成骨培养基补充剂和弱碱，用于同时启动纯壳聚糖，纯胶原蛋白，和壳聚糖-胶原蛋白复合材料在生理pH值和温度下的凝胶化。以壳聚糖/胶原蛋白比例为100/0、65/35、25/75和0/100重量%包封在这种水凝胶中的成人骨髓源性干细胞（hBMSC）在包封后第1天表现出高活力，但DNA含量在纯壳聚糖材料中在12天内下降了约一半，而在含有胶原蛋白的材料中增加了两倍。含胶原蛋白的材料压实更强烈，显着比纯壳聚糖凝胶更硬。在单层培养中，hBMSC暴露于β-GP导致细胞代谢活性降低，其随浓度和暴露时间而变化，但洗涤有效地从水凝胶中去除过量的β-GP。材料中壳聚糖的存在导致在有和没有成骨补充剂的培养基中osterix和骨唾液蛋白基因的表达更高。壳聚糖还增加碱性磷酸酶活性和钙沉积在成骨培养基中。壳聚糖-胶原复合材料具有作为细胞包封和递送的基质或作为用于组织修复的原位凝胶形成材料的潜力。
Chitosan and collagen type I are naturally-derived materials used as cell carriers because of their ability to mimic the extracellular environment and direct cell function. In this study beta-glycerophosphate (beta-GP), an osteogenic medium supplement and a weak base, was used to simultaneously initiate gelation of pure chitosan, pure collagen, and chitosan-collagen composite materials at physiological pH and temperature. Adult human bone marrow-derived stem cells (hBMSC) encapsulated in such hydrogels at chitosan/collagen ratios of 100/0, 65/35, 25/75, and 0/100 wt% exhibited high viability at day 1 after encapsulation, but DNA content dropped by about half over 12 days in pure chitosan materials while it increased two-fold in materials containing collagen. Collagen-containing materials compacted more strongly and were significantly stiffer than pure chitosan gels. In monolayer culture, exposure of hBMSC to beta-GP resulted in decreased cell metabolic activity that varied with concentration and exposure time, but washing effectively removed excess beta-GP from hydrogels. The presence of chitosan in materials resulted in higher expression of osterix and bone sialoprotein genes in medium with and without osteogenic supplements. Chitosan also increased alkaline phosphatase activity and calcium deposition in osteogenic medium. Chitosan-collagen composite materials have potential as matrices for cell encapsulation and delivery, or as in situ gel-forming materials for tissue repair.