Vo T N, Shah S R, Lu S, Tatara A M, Lee E J, Roh T T, Tabata Y, Mikos A G
Department of Bioengineering, Rice University, P.O. Box 1892, MS 142, Houston, TX, 77251-1892, USA.
Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.
Biomaterials. 2016 Mar;83:1-11. doi: 10.1016/j.biomaterials.2015.12.026. Epub 2015 Dec 31.
The present work investigated the osteogenic potential of injectable, dual thermally and chemically gelable composite hydrogels for mesenchymal stem cell (MSC) delivery in vitro and in vivo. Composite hydrogels comprising copolymer macromers of N-isopropylacrylamide were fabricated through the incorporation of gelatin microparticles (GMPs) as enzymatically digestible porogens and sites for cellular attachment. High and low polymer content hydrogels with and without GMP loading were shown to successfully encapsulate viable MSCs and maintain their survival over 28 days in vitro. GMP incorporation was also shown to modulate alkaline phosphatase production, but enhanced hydrogel mineralization along with higher polymer content even in the absence of cells. Moreover, the regenerative capacity of 2 mm thick hydrogels with GMPs only, MSCs only, or GMPs and MSCs was evaluated in vivo in an 8 mm rat critical size cranial defect for 4 and 12 weeks. GMP incorporation led to enhanced bony bridging and mineralization within the defect at each timepoint, and direct bone-implant contact as determined by microcomputed tomography and histological scoring, respectively. Encapsulation of both GMPs and MSCs enabled hydrogel degradation leading to significant tissue infiltration and osteoid formation. The results suggest that these injectable, dual-gelling cell-laden composite hydrogels can facilitate bone ingrowth and integration, warranting further investigation for bone tissue engineering.
本研究探讨了可注射的、具有热和化学双重凝胶化特性的复合水凝胶在体外和体内用于间充质干细胞(MSC)递送时的成骨潜力。通过掺入明胶微粒(GMPs)作为可酶解的致孔剂和细胞附着位点,制备了包含N-异丙基丙烯酰胺共聚物大分子单体的复合水凝胶。结果表明,含和不含GMP负载的高、低聚合物含量水凝胶能够成功包封活的MSC,并在体外维持其存活28天。还发现,即使在没有细胞的情况下,掺入GMP也能调节碱性磷酸酶的产生,但随着聚合物含量的增加,水凝胶矿化增强。此外,在8毫米大鼠临界尺寸颅骨缺损模型中,对仅含GMP、仅含MSC或同时含GMP和MSC的2毫米厚水凝胶进行了4周和12周的体内再生能力评估。在每个时间点,掺入GMP均导致缺损内骨桥接和矿化增强,分别通过微型计算机断层扫描和组织学评分确定了直接的骨-植入物接触。同时包封GMP和MSC可使水凝胶降解,导致显著的组织浸润和类骨质形成。结果表明,这些可注射的、双凝胶化的载细胞复合水凝胶可促进骨长入和整合,值得进一步开展骨组织工程研究。