Jin Guang-Zhen, Kim Hae-Won
Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Cheonan, South Korea.
Department of Biomaterials Science, College of Dentistry, Dankook University, Cheonan, South Korea.
J Tissue Eng. 2018 Oct 8;9:2041731418802438. doi: 10.1177/2041731418802438. eCollection 2018 Jan-Dec.
Dedifferentiation of chondrocytes remains a major problem in cartilage tissue engineering. The development of hydrogels that can preserve chondrogenic phenotype and prevent chondrocyte dedifferentiation is a meaningful strategy to solve dedifferentiation problem of chondrocytes. In the present study, three gels were prepared (alginate gel (Alg gel), type I collagen gel (Col gel), and their combination gel (Alg/Col gel)), and the in vitro efficacy of chondrocytes culture while preserving their phenotypes was investigated. While Col gel became substantially contracted with time, the cells encapsulated in Alg gel preserved the shape over the culture period of 14 days. The mechanical and cell-associated contraction behaviors of Alg/Col gel were similar to those of Alg. The cells in Alg and Alg/Col gels exhibited round morphology, whereas those in Col gel became elongated (i.e. fibroblast-like) during cultures. The cells proliferated with time in all gels with the highest proliferation being attained in Col gel. The expression of chondrogenic genes, including , type II collagen, and aggrecan, was significantly up-regulated in Alg/Col gel and Col gel, particularly in Col gel. However, the chondrocyte dedifferentiation markers, type I collagen and alkaline phosphatase (), were also expressed at significant levels in Col gel, which being contrasted with the events in Alg and Alg/Col gels. The current results suggest the cells cultured in hydrogels can express chondrocyte dedifferentiation markers as well as chondrocyte markers, which draws attention to choose proper hydrogels for chondrocyte-based cartilage tissue engineering.
软骨细胞去分化仍然是软骨组织工程中的一个主要问题。开发能够保留软骨生成表型并防止软骨细胞去分化的水凝胶是解决软骨细胞去分化问题的一种有意义的策略。在本研究中,制备了三种凝胶(海藻酸钠凝胶(Alg凝胶)、I型胶原凝胶(Col凝胶)及其复合凝胶(Alg/Col凝胶)),并研究了在保留软骨细胞表型的同时进行体外培养的效果。虽然Col凝胶随时间显著收缩,但包裹在Alg凝胶中的细胞在14天的培养期内保持了形状。Alg/Col凝胶的力学和细胞相关收缩行为与Alg凝胶相似。在培养过程中,Alg凝胶和Alg/Col凝胶中的细胞呈现圆形形态,而Col凝胶中的细胞则变得细长(即成纤维细胞样)。所有凝胶中的细胞均随时间增殖,其中Col凝胶中的增殖率最高。包括II型胶原和聚集蛋白聚糖在内的软骨生成基因的表达在Alg/Col凝胶和Col凝胶中显著上调,尤其是在Col凝胶中。然而,软骨细胞去分化标志物I型胶原和碱性磷酸酶()在Col凝胶中也有显著表达,这与Alg凝胶和Alg/Col凝胶中的情况形成对比。目前的结果表明,在水凝胶中培养的细胞既能表达软骨细胞去分化标志物,也能表达软骨细胞标志物,这提醒人们在基于软骨细胞的软骨组织工程中要选择合适的水凝胶。