Froelich Katrin, Setiawan Lydia E, Technau Antje, Tirado Mario Ramos, Hackenberg Stephan, Hagen Rudolf, Staudenmaier Rainer, Kleinsasser Norbert H
Department of Otorhinolaryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University of Wuerzburg, 97080 Wuerzburg, Germany.
Int J Artif Organs. 2012 Dec;35(12):1047-60. doi: 10.5301/ijao.5000132.
Chondrogenic differentiation of adipose-derived stem cells (ASCs) has proven to be feasible. To compensate for laryngeal palsy or cartilage defects after surgery or trauma using tissue engineering, a formable and stable scaffold material is mandatory.
ASCs were seeded in fibrin-polyurethane scaffolds and cultured in chondrogenic differentiation medium adding the growth factors TGF-b1, TGF-b3, and BMP-2 for up to 35 days.
Histological examination showed acid glycosaminoglycans in the extracellular matrix in all groups. Immunofluorescence presented positive staining for collagen II, aggrecan, and SOX-9 in the TGF-b1-, TGF-b3-, and BMP-2-group. With Real-time PCR analyses, chondrogenic differentiation became apparent by the expression of the specific genes COL2A1 (collagen II), AGC 1 (aggrecan), and SOX-9, whereas collagen II expression was low in all groups compared to bone marrow-derived stem cells (BMSC) due to reduced chondrogenic ability.
These findings demonstrate the general ability of ASCs to differentiate into matrix-producing chondrocytes in fibrin-polyurethane scaffolds. However, further experiments are necessary to enhance this chondrogenic potential of ASCs seeded in fibrin-polyurethane scaffolds in order to produce a suitable regeneration method for treating cartilage defects or an implantable medialization material for vocal cord palsy.
脂肪来源干细胞(ASCs)向软骨细胞分化已被证明是可行的。为了利用组织工程技术弥补手术或创伤后喉麻痹或软骨缺损,一种可成型且稳定的支架材料是必不可少的。
将ASCs接种于纤维蛋白-聚氨酯支架中,并在添加生长因子转化生长因子-β1(TGF-β1)、转化生长因子-β3(TGF-β3)和骨形态发生蛋白-2(BMP-2)的软骨分化培养基中培养长达35天。
组织学检查显示所有组的细胞外基质中均有酸性糖胺聚糖。免疫荧光显示TGF-β1组、TGF-β3组和BMP-2组中Ⅱ型胶原蛋白、聚集蛋白聚糖和SRY-box转录因子9(SOX-9)呈阳性染色。通过实时聚合酶链反应(PCR)分析,特定基因Ⅱ型胶原蛋白α1链(COL2A1)、聚集蛋白聚糖1(AGC 1)和SOX-9的表达表明软骨分化明显,但由于软骨生成能力降低,与骨髓来源干细胞(BMSC)相比,所有组中Ⅱ型胶原蛋白的表达均较低。
这些发现证明了ASCs在纤维蛋白-聚氨酯支架中分化为产生基质的软骨细胞的一般能力。然而,有必要进一步开展实验以增强接种于纤维蛋白-聚氨酯支架中的ASCs的软骨生成潜能,从而开发出一种合适的治疗软骨缺损的再生方法或一种用于声带麻痹的可植入的喉内移植物材料。