Department of Chemical Engineering, Faculty of Engineering, Mersin University, Mersin, Turkey.
J Tissue Eng Regen Med. 2011 Nov;5(10):770-9. doi: 10.1002/term.375. Epub 2010 Dec 29.
A feasibility study was undertaken to examine the potential of biodegradable HEMA-lactate-dextran (HEMA-LLA-D)-based cryogels as scaffolds for cartilage tissue engineering. This was a preliminary in vitro study giving essential information on the biocompatibility of cryogels with cartilage cells. HEMA-lactate (HEMA-LLA) and HEMA-LLA-D were synthesized and characterized by different techniques. Cryogel scaffolds with supermacroporous structures were produced by cryogenic treatment of these macromers. Chondrocytes obtained from bovine articular cartilage were seeded onto cylindrical cryogels and cultured. The samples were examined by several microcopical techniques for cell viability and morphological analyses were performed at two culture points. Histological study of the constructs revealed the cells' growth on the surface and within the scaffolds. Confocal microscopical images demonstrated that the majority of live vs. dead cells had been attached to and integrated with the pores of the scaffold. SEM analysis showed round to oval-shaped chondrocytic cells interconnected with each other by communicating junctions. The chondrocytes rapidly proliferated in the cryogels, manifesting that they fully covered the scaffold surface after 9 days and almost filled the spaces in the pores of the scaffold after 15 days of culture. Chondrocytes secreted significant amount of extracellular matrix in the scaffolds and exhibited highly interconnective morphology. Light and transmission electron microscopy revealed groups of active cartilage cells closely apposed to the cryogel. We concluded that cryogel scaffolds could be excellent candidates for cartilage tissue regeneration with their extraordinary properties, including soft, elastic nature, highly open interconnected pore structure and very rapid, controllable swellability.
进行了一项可行性研究,以考察可生物降解的 HEMA-乳酸-葡聚糖(HEMA-LLA-D)基冷冻凝胶作为软骨组织工程支架的潜力。这是一项初步的体外研究,提供了冷冻凝胶与软骨细胞生物相容性的基本信息。通过不同的技术合成和表征了 HEMA-乳酸(HEMA-LLA)和 HEMA-LLA-D。通过这些大分子的低温处理制备具有超大孔结构的冷冻凝胶支架。从牛关节软骨中获得的软骨细胞接种到圆柱形冷冻凝胶上并进行培养。通过几种显微镜技术检查样品的细胞活力,并在两个培养点进行形态分析。对构建体的组织学研究表明细胞在表面和支架内生长。共聚焦显微镜图像表明,大多数活细胞与死细胞附着在支架的孔上并与支架的孔整合。扫描电子显微镜分析显示,软骨细胞呈圆形到椭圆形,通过通讯连接相互连接。软骨细胞在冷冻凝胶中迅速增殖,表明它们在 9 天后完全覆盖了支架表面,在 15 天的培养后几乎填满了支架孔中的空间。软骨细胞在支架中分泌了大量的细胞外基质,并表现出高度互联的形态。光镜和透射电镜显示,一群活跃的软骨细胞紧密贴合在冷冻凝胶上。我们得出结论,冷冻凝胶支架具有柔软、有弹性的特性、高度开放的互联孔结构以及非常快速、可控的溶胀性,是软骨组织再生的优秀候选材料。