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Chondrocyte-Imprinted Substrates: Promoting MSC Chondrogenesis and Regulating Inflammatory Gene Expression

Authors :

Parisa Madani¹ Sara Derhanbakhsh² Nasrin Salehi³ Farzaneh Safshekan⁴ Javad Mohammadi⁵ Shahin Bonakdar⁶

1- دانشگاه تهران 2- دانشگاه تهران 3- دانشگاه تهران 4- موسسه آموزش عالی احرار 5- دانشگاه تهران 6- انستیتو پاستور ایران

Keywords :

Chondrogenic differentiation،Cell-imprinted substrate،Topography،Cartilage Tissue،Mesenchymal Stem Cells،Tissue engineering

Abstract :

Due to the limitations of using primary chondrocytes, directing stem cells toward chondrogenesis via physical cues has emerged as an efficient strategy for cartilage regeneration. Substrate topography can strongly influence cellular behavior, including differentiation. Here, we investigated the effect of chondrocyte-imprinted substrates on the chondrogenic differentiation of stem cells and dedifferentiated chondrocytes without chemical inducers. Rabbit chondrocytes were isolated, fixed, and used as templates to fabricate patterned polydimethylsiloxane (PDMS) substrates. Rabbit adipose-derived mesenchymal stem cells (ADSCs) and passage 3 dedifferentiated chondrocytes were cultured on these substrates and analyzed using cell staining and real-time PCR for chondrogenic and inflammatory gene expression. Compared to spindle-like control cells, cells on imprinted substrates exhibited round morphologies, enhanced glycosaminoglycan and collagen I/III production, actin fiber rearrangement, and higher expression of chondrogenic genes (collagen II, aggrecan, sox9) with minimal inflammatory gene expression (TNF-α, IL-1, IL-6). These results demonstrate that chondrocyte-imprinted PDMS substrates can effectively induce chondrogenesis without chemical factors, providing a promising tool for cartilage tissue engineering.