Title : 3D in vitro model of Dupuytren’s disease
Abstract:
Dupuytren’s disease (DD) is a common fibrotic disorder of the hand, characterized by progressive thickening and contracture of the palmar and digital fascia. Surgical excision remains the primary treatment; however, there are currently no therapies to prevent disease progression or recurrence. This study aims to develop a 3D in vitro model to test novel antifibrotic therapies. The model is based on decellularized pathological DD tissue seeded with patient-derived fibroblasts, capturing the role of both cellular and extracellular matrix components in disease progression.
Fibrotic DD tissues were obtained from surgical excisions, sectioned, and decellularized. In parallel, primary fibroblasts were isolated from patient samples. The decellularized extracellular matrices (dECMs) were characterized with respect to biochemical composition, collagen structure, and mechanical properties. Fibroblasts were seeded onto the dECMs and cultured stepwise to initially promote proliferation, followed by differentiation into myofibroblasts. We assessed the composition of newly synthesized and deposited ECM by mass spectrometry and compared it with conventional 2D cultivations (standard polystyrene dish and nanofiber membrane). We proved that cells cultured on 3D dECM produce a higher number of fibrosis-associated proteins than those cultured on 2D surfaces. We tested the antifibrotic effect of 0.5mM minoxidil, an inhibitor of collagen crosslinking. The responsiveness to the drug minoxidil was significantly more complex in the 3D model than in 2D cultures. We demonstrated that dECM seeded with DD fibroblasts represents a relevant 3D in vitro model of Dupuytren’s disease. This system provides a reliable platform for antifibrotic drug screening as demonstrated by the testing of minoxidil.
This project was supported by the European Union (project No.CZ.02.01.01/00/22_008/0004562) and co-funded by the Czech Academy of Sciences, Praemium Academiae Grant No. 2202.
