Burnsed Olivia A, Schwartz Zvi, Marchand Katherine O, Hyzy Sharon L, Olivares-Navarrete René, Boyan Barbara D
Wallace H. Coulter Department of Biomedical Engineering and Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA, USA.
Department of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA, USA; Department of Periodontics, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA.
Acta Biomater. 2016 Oct 1;43:139-149. doi: 10.1016/j.actbio.2016.07.034. Epub 2016 Jul 20.
Limited supplies of healthy autologous or allogeneic cartilage sources have inspired a growing interest in xenogeneic cartilage matrices as biological scaffolds for cartilage tissue engineering. The objectives of this study were to determine if shark and pig cartilage extracellular matrix (ECM) hydrogels can stimulate chondrocytic differentiation of mesenchymal stem cells (MSCs) without exogenous growth factors and to determine if the soluble factors retained by these ECM hydrogels are responsible. Human MSCs cultured on hydrogels from shark skull cartilage, pig articular cartilage, and pig auricular cartilage ECM had increased expression of chondrocyte markers and decreased secretion of angiogenic factors VEGF-A and FGF2 in comparison to MSCs cultured on tissue culture polystyrene (TCPS) at one week. MSCs grown on shark ECM gels had decreased type-1 collagen mRNA as compared to all other groups. Degradation products of the cartilage ECM gels and soluble factors released by the matrices increased chondrogenic and decreased angiogenic mRNA levels, indicating that the processed ECM retains biochemically active proteins that can stimulate chondrogenic differentiation. In conclusion, this work supports the use of cartilage matrix-derived hydrogels for chondrogenic differentiation of MSCs and cartilage tissue engineering. Longer-term studies and positive controls will be needed to support these results to definitively demonstrate stimulation of chondrocyte differentiation, and particularly to verify that calcification without endochondral ossification does not occur as it does in shark cartilage.
The objectives of this study were to determine if shark and pig cartilage extracellular matrix (ECM) hydrogels can stimulate chondrocytic differentiation of mesenchymal stem cells (MSCs) without exogenous growth factors and to determine if the soluble factors retained by these ECM hydrogels are responsible for this induction. Sharks are an especially interesting model for cartilage regeneration because their entire skeleton is composed of cartilage and they do not undergo endochondral ossification. Culturing human MSCs on porcine and shark cartilage ECM gels directly, with ECM gel conditioned media, or degradation products increased mRNA levels of chondrogenic factors while decreasing angiogenic factors. These studies indicate that xenogeneic cartilage ECMs have potential as biodegradable scaffolds capable of stimulating chondrogenesis while preventing angiogenesis for regenerative medicine applications and that ECM species selection can yield differential effects.
健康的自体或异体软骨来源供应有限，这激发了人们对异种软骨基质作为软骨组织工程生物支架的兴趣日益浓厚。本研究的目的是确定鲨鱼和猪软骨细胞外基质（ECM）水凝胶是否能在无外源性生长因子的情况下刺激间充质干细胞（MSC）向软骨细胞分化，并确定这些ECM水凝胶保留的可溶性因子是否起作用。与在组织培养聚苯乙烯（TCPS）上培养的MSC相比，在来自鲨鱼头骨软骨、猪关节软骨和猪耳软骨ECM的水凝胶上培养的人MSC在一周时软骨细胞标志物的表达增加，血管生成因子VEGF-A和FGF2的分泌减少。与所有其他组相比，在鲨鱼ECM凝胶上生长的MSC的I型胶原mRNA减少。软骨ECM凝胶的降解产物和基质释放的可溶性因子增加了软骨生成相关mRNA水平并降低了血管生成相关mRNA水平，表明经过处理的ECM保留了能够刺激软骨生成分化的生物化学活性蛋白。总之，这项工作支持使用软骨基质衍生的水凝胶进行MSC的软骨生成分化和软骨组织工程。需要进行长期研究和设置阳性对照来支持这些结果，以明确证明对软骨细胞分化的刺激作用，特别是要验证是否不会像在鲨鱼软骨中那样发生无软骨内成骨的钙化现象。
本研究的目的是确定鲨鱼和猪软骨细胞外基质（ECM）水凝胶是否能在无外源性生长因子的情况下刺激间充质干细胞（MSC）向软骨细胞分化，并确定这些ECM水凝胶保留的可溶性因子是否负责这种诱导作用。鲨鱼是软骨再生的一个特别有趣的模型，因为它们的整个骨骼由软骨组成，并且它们不会经历软骨内成骨。将人MSC直接培养在猪和鲨鱼软骨ECM凝胶上、用ECM凝胶条件培养基或降解产物培养，可增加软骨生成因子的mRNA水平，同时降低血管生成因子的水平。这些研究表明，异种软骨ECM有潜力作为可生物降解支架，能够在再生医学应用中刺激软骨生成同时防止血管生成，并且ECM物种的选择可能会产生不同的效果。
