Plastic Surgery Research Laboratory, Division of Plastic Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Tissue Eng Part A. 2011 Feb;17(3-4):301-9. doi: 10.1089/ten.TEA.2010.0322. Epub 2010 Oct 12.
The objective of this study was to fabricate hydrogel matrix-engineered biosynthetic cartilage using a porous poly(vinyl alcohol) hydrogel (PVA-H) and articular chondrocytes. Chondrocytes were suspended in fibrin gel (FG) or saline carriers and injected into porous PVA-H discs and three-layered constructs (PVA-H between devitalized cartilage). After implantation in nude mice, PVA discs were explanted at 6 weeks and subjected to creep testing for a 20 h period. The three-layered constructs were explanted at 12 weeks and subjected to tensile testing to determine the strength of the interface between the engineered hydrogel and devitalized cartilage. Histological analysis revealed PVA-H porous channels occupied by chondrocytes. Extracellular matrix was identified by Safranin-O and toluidine blue stains. Immunohistochemical analysis revealed a positive stain for COL II and scant staining for COL I. Creep and relaxation response of PVA-FG-chondrocyte constructs was similar to that of native cartilage. The presence of cells and FG significantly enhanced the integration strength of layered constructs (p < 0.05). These results demonstrate that porous PVA-H in combination with FG and chondrocytes provides a favorable microenvironment for tissue engineering of articular cartilage, creating a biosynthetic construct that can adhere to native devitalized articular cartilage utilizing hydrogel matrix-engineered technology.
本研究的目的是利用多孔聚(醇)水凝胶(PVA-H)和关节软骨细胞制造水凝胶基质工程化生物合成软骨。软骨细胞悬浮于纤维蛋白凝胶(FG)或盐载体中,并注射到多孔 PVA-H 圆盘和三层结构(PVA-H 在去活软骨之间)中。在裸鼠体内植入后,6 周时取出 PVA 圆盘,并进行 20 小时的蠕变测试。12 周时取出三层结构,并进行拉伸测试,以确定工程化水凝胶与去活软骨之间的界面强度。组织学分析显示 PVA-H 多孔通道被软骨细胞占据。通过番红 O 和甲苯胺蓝染色鉴定细胞外基质。免疫组织化学分析显示 COL II 呈阳性染色,COL I 呈少量染色。PVA-FG-软骨细胞构建体的蠕变和松弛响应类似于天然软骨。细胞和 FG 的存在显著增强了分层结构的整合强度(p<0.05)。这些结果表明,多孔 PVA-H 与 FG 和软骨细胞结合为关节软骨的组织工程提供了有利的微环境,利用水凝胶基质工程技术制造了能够附着于天然去活关节软骨的生物合成构建体。