Minoxidil decreases collagen I deposition and tissue-like contraction in clubfoot-derived cells: a way to improve conservative treatment of relapsed clubfoot?

Abstract

Minoxidil decreases collagen I deposition and tissue-like contraction in clubfoot-derived cells: a way to improve conservative treatment of relapsed clubfoot?, Connective Tissue Research,

Increased Collagen Crosslinking in Stiff Clubfoot Tissue: Implications for the Improvement of Therapeutic Strategies

“…We isolated, propagated, and characterized cells from the contracted medial side of clubfoot tissue, which we have described in detail [ 21 ]. Briefly, the culture contained 96% of fibroblasts, with an admixture of myofibroblasts and fibrocytes (less than 3%), vascular smooth muscle (less than 0.1%), but no endothelial cells.…”

“…The observed changes in collagen production, deposition, and processing in clubfoot tissue, together with a knowledge of the pathogenesis of more extensively studied fibroproliferative diseases, could form a basis for the development of future targeted treatment strategies. We have recently demonstrated that inhibiting LH2 by Minoxidil in clubfoot-derived cells leads to decreased short-term and long-term levels of deposited collagen, together with reduced tissue-like contraction in a 3D collagen gel model [ 21 ]. To further explore the idea in this study, we tested the therapeutic potential of β-aminopropionitrile (BAPN), an agent that reduces collagen crosslinking by inhibiting members of the LOX family on fibroblast-like cells isolated from clubfoot medial side tissue.…”

“…Tissue samples (M-side and L-side sets) from 5 patients (1 female, 4 males; mean age 43.20 months, SD = 24.40) were mechanically minced and enzymatically digested to isolate primary cell cultures, which were then propagated and characterized, as described previously [ 21 ]. For the experiments, the cells were incubated in DMEM medium (Sigma–Aldrich, Burlington, MA, USA) containing 10% of fetal bovine serum (FBS; Gibco, ThermoFisher Scientific, Waltham, MA, USA) with the addition of 0, 10, 20, and 40 µg/mL of BAPN (beta-aminopropionitrile, CDS007521, Sigma–Aldrich, Burlington, MA, USA; reconstituted in sterile dH2O to 5 mg/mL stock solution).…”

“…To date, only a small number of studies have investigated the possibility of antifibrotic treatment for clubfoot by blocking profibrotic pathways [ 14 , 20 ]; however, no follow-up has been reported. We explored the idea of targeting post-translational modifications of procollagen chains, and we obtained promising results with lysyl hydroxylase inhibition [ 21 ]. Post-translational modifications of collagen affect the way and the quantity in which crosslinks are formed in the newly-created collagen fibers.…”

The possible role of hypoxia in the affected tissue of relapsed clubfoot

“…This could shift clubfoot therapy from symptomatic towards cell-targeted, as has already been described in other fibroproliferative disorders 47 . In addition, the reported stiffness of the fibrotic clubfoot tissue might be alleviated in the future by the application of antifibrotic substances, for example targeting ECM crosslinking enzymes 14 , 48 . Such antifibrotic treatments would facilitate the currently-used conservative therapy by making the collagens of the affected tissue more susceptible to degradation, and thus by making the tissue less stiff.…”