Munirah S, Kim S H, Ruszymah B Hi, Khang G
Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia.
Eur Cell Mater. 2008 Feb 21;15:41-52. doi: 10.22203/ecm.v015a04.
Our preliminary results indicated that fibrin and poly(lactic-co-glycolic acid) (PLGA) hybrid scaffold promoted early chondrogenesis of articular cartilage constructs in vitro. The aim of this study was to evaluate in vivo cartilaginous tissue formation by chondrocyte-seeded fibrin/PLGA hybrid scaffolds. PLGA scaffolds were soaked carefully, in chondrocyte-fibrin suspension, and polymerized by dropping thrombin-calcium chloride (CaCl2) solution. PLGA-seeded chondrocytes were used as a control. Resulting constructs were implanted subcutaneously, at the dorsum of nude mice, for 4 weeks. Macroscopic observation, histological evaluation, gene expression and sulphated-glycosaminoglycan (sGAG) analyses were performed at each time point of 1, 2 and 4 weeks post-implantation. Cartilaginous tissue formation in fibrin/PLGA hybrid construct was confirmed by the presence of lacunae and cartilage-isolated cells embedded within basophilic ground substance. Presence of proteoglycan and glycosaminoglycan (GAG) in fibrin/PLGA hybrid constructs was confirmed by positive Safranin O and Alcian Blue staining. Collagen type II exhibited intense immunopositivity at the pericellular matrices. Chondrogenic properties were further demonstrated by the expression of gene encoded cartilage-specific markers, collagen type II and aggrecan core protein. The sGAG production in fibrin/PLGA hybrid constructs was higher than in the PLGA group. In conclusion, fibrin/PLGA hybrid scaffold promotes cartilaginous tissue formation in vivo and may serve as a potential cell delivery vehicle and a structural basis for articular cartilage tissue-engineering.
我们的初步结果表明,纤维蛋白与聚乳酸-羟基乙酸共聚物(PLGA)混合支架可促进关节软骨构建体在体外的早期软骨形成。本研究的目的是评估接种软骨细胞的纤维蛋白/PLGA混合支架在体内软骨组织的形成情况。将PLGA支架小心地浸泡在软骨细胞-纤维蛋白悬液中,通过滴加凝血酶-氯化钙(CaCl₂)溶液使其聚合。接种PLGA的软骨细胞用作对照。将所得构建体皮下植入裸鼠背部4周。在植入后1、2和4周的每个时间点进行宏观观察、组织学评估、基因表达和硫酸化糖胺聚糖(sGAG)分析。纤维蛋白/PLGA混合构建体中存在陷窝以及嵌入嗜碱性基质中的软骨分离细胞,证实了软骨组织的形成。纤维蛋白/PLGA混合构建体中蛋白聚糖和糖胺聚糖(GAG)的存在通过番红O和阿尔新蓝染色阳性得以证实。II型胶原在细胞周围基质处呈现强烈的免疫阳性。软骨特异性标志物II型胶原和聚集蛋白聚糖核心蛋白编码基因的表达进一步证明了软骨生成特性。纤维蛋白/PLGA混合构建体中的sGAG产量高于PLGA组。总之,纤维蛋白/PLGA混合支架可促进体内软骨组织的形成,并可能作为一种潜在的细胞递送载体和关节软骨组织工程的结构基础。