Collagen Type II Enhances Chondrogenesis in Adipose Tissue–Derived Stem Cells by Affecting Cell Shape

Ideally, biomaterials have inductive properties, favoring specific lineage differentiation. For chondrogenic induction, these properties have been attributed to collagen type II. However, the underlying mechanisms are largely unknown. This study aimed to investigate whether collagen type II favors chondrogenic induction by affecting cell shape through beta1 integrins and Rho A/Rock signaling. For this purpose, adipose tissue-derived stem cells (ASCs) were encapsulated in collagen type I or II gels and cultured in plain and chondrogenic medium. It was demonstrated that (i) ASCs showed more efficient chondrogenic induction (higher collagen X, aggrecan, sox6, sox9, and collagen II gene expression) in both plain and chondrogenic media in collagen type II versus collagen type I gels; (ii) ASCs showed lower Rock 2 gene expression and a more rounded cell shape in collagen type II versus type I gels when grown in plain medium; (iii) Rock inhibitor (Y27632) more effectively enhanced chondrogenic gene expression of ASCs in collagen type I than in collagen type II gels, and diminished differences in chondrogenic gene expression and cell shape of ASCs between the two gel types; and (iv) beta1 integrins blocking not only reduced the differences of chondrogenic gene expression but also eliminated the differences of Rock 1 and Rock 2 gene expressions and cell shape when comparing ASCs embedded in collagen type I and II gels. We conclude that collagen type II provides the inductive signaling for chondrogenic differentiation in ASCs by evoking a round cell shape through beta1 integrin-mediated Rho A/Rock signaling pathway.