Hanieh Sadat Hashemi Motahar
1 
, Mojtaba Tajbakhsh
2, Mahboobeh Tavassoli
3, Shahin Bonakdar
4* , Mohammad Reza Sadeghi
1,5* 1 Department of Molecular Medicine, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran
2 Orthopedic Clinic, Mardom Hospital, Tehran, Iran
3 Obstetrics and Gynecology Clinic, Mardom Hospital, Tehran, Iran
4 National Cell Bank Department, Pasteur Institute of Iran, Tehran, Iran
5 Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz University of Medical Sciences, Tabriz, Iran
Abstract
Introduction: The objective of this study is twofold: first, to investigate the relationship between chondrocyte morphology and their own gene and protein expression profiles in healthy and osteoarthritic (OA) cartilage; and second, to assess whether replicating the morphology of OA chondrocytes (OACs) can induce a hypertrophic expression pattern in MSCs.
Methods: Polydimethylsiloxane (PDMS) substrates were fabricated to replicate the morphologies of human OACs and healthy chondrocytes (HCs). MSCs were cultured on these imprinted substrates, and differentiation was assessed using real-time PCR, immunocytochemistry, Alcian blue/Safranin O staining, and scanning electron microscopy (SEM).
Results: SEM and optical microscopy revealed that OACs had a larger surface area than HCs. Real-time PCR analysis showed morphology-dependent variations in the expression of cartilage- and OA-related markers, with statistically significant differences observed only for SOX9. Immunofluorescence analysis of collagen types I and II supported these findings, though visual inspection of the staining did not indicate any significant changes.
Conclusion: The results show that OACs-imprinted substrates can be effectively combined with other methods to improve in vitro models of OA. This offers a useful tool for exploring disease mechanisms and potential therapies.