Pellet Culture Elicits Superior Chondrogenic Redifferentiation than Alginate-Based Systems
Pellet Culture Elicits Superior Chondrogenic Redifferentiation than Alginate-Based Systems
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DOI：
10.1002/btpr.186
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发表时间：
2009-07-01
影响因子：
2.9
通讯作者：
Fickert, Stefan
中科院分区：
文献类型：
--
作者：
Bernstein, Peter;Dong, Meng;Fickert, Stefan
尽管颗粒培养和软骨细胞包封成凝胶状生物材料已经导致软骨组织工程的重大进展，但在这些培养过程中细胞分化行为的定量比较表征尚未进行。因此，我们的研究旨在回答以下问题：软骨细胞的再分化途径是否会因藻酸盐生物材料类型（纯藻酸盐、藻酸盐-纤维蛋白、藻酸盐-壳聚糖）的轻微变化而改变？与无生物材料（颗粒）三维培养相比，这些细胞的行为如何？用海藻酸盐、海藻酸盐-壳聚糖、海藻酸盐-纤维蛋白珠和微球培养健康成年猪膝关节的单层扩张软骨细胞，培养时间长达4周。采用定量PCR和免疫组织学方法评估软骨形成标志物。海藻酸纤维蛋白包被的软骨细胞表现为单层软骨细胞。海藻酸盐和海藻酸-壳聚糖包封导致低软骨标记基因表达。虽然所有3d培养的软骨细胞都显示出相当数量的Sox9表达，但只有颗粒培养才能产生足够的Collagen II表达。这使得海藻酸盐培养的软骨组织工程结构的使用受到质疑。纤维蛋白的加入不利于软骨分化。Sox9和胶原蛋白H的表现不同，取决于周围的3d环境。(C) 2009美国化学工程师学会。掠夺。科学通报，25 (5):1146-1152,2009
Although pellet culture and encapsulation of chondrocytes into gel-like biomaterials have lead to major advances in cartilage tissue engineering, a quantitative comparative characterization of cellular differentiation behavior during those cultivation procedures has not yet been performed. Our study therefore aimed at answering the following question: is the redifferentiation pathway of chondrocytes altered by slight changes in the type of alginate biomaterial (pure alginate, alginate-fibrin, alginate-chitosan) and how do the cells behave in comparison to biomaterial-free (pellet) three-dimensional culturing? Monolayer-expanded chondrocytes from healthy adult porcine knee joints were cultivated in alginate, alginate-chitosan, alginate-fibrin beads and as pellets up to 4 weeks. Quantitative PCR and Immunohistology were used to assess chondrogenic markers. Alginate-fibrin-encapsulated chondrocytes behaved almost like monolayer chondrocytes. Alginate- and alginate-chitosan encapsulation lead to a low chondrogenic marker gene expression. Although all 3D-cultured chondrocytes showed a considerable amount of Sox9 expression, only pellet cultivation lead to a sufficient Collagen II expression. This puts the usage of alginate-cultivated cartilage tissue engineering constructs under question. Fibrin addition is not beneficial for chondrogenic differentiation. Sox9 and Collagen H behave differently, depending upon the surrounding 3D-environment. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 25: 1146-1152, 2009