ROCK inhibitor(Devgen)

Male Sprague Dawley rats were used for mature hepatocytes (HEP) isolation to induce CLiP using 3 small molecules cocktail (ROCK-inhibitor, TGF beta-1 inhibitor, GSK3 inhibitor). Harvested rat CLiP (1 × 10⁶) were transplanted to the posterior lobe of the liver (30 % of whole liver) of Nagase analbuminemic rats (NAR) through portal vein injection. One week later, portal venous branch to non-transplanted lobe was ligated to induce liver regeneration in CLiP bearing liver lobe (portal branch ligation: PBL). Only CLiP Tx or HEP Tx of same dose were used as control groups. Serial measurement of serum albumin level was measured by ELISA method. All NARs were subject to immunosuppression therapy using tacrolimus s.q. 3 days per week.

After HEP Tx especially with PBL, serum albumin level was significantly elevated. However, even with PBL, CLiP Tx group did not show significant increase in serum albumin. CLiP were surrounded by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) positive cells in portal veins suggesting that CLiP were exposed to the risk of innate immunity.

Although CLiP respond to growth factors and proliferate better than HEP in vitro, their behavior in vivo in response to liver regenerative stimulus were not similar to in vitro probably because of less immaturity of receptors and/or high immunogenicity as compared to HEP.

Keloid disease is a common fibroproliferative skin disorder characterized by excessive scar tissue formation and frequent recurrence. Limited therapies and study models hinder progress in addressing this unmet clinical need. AMA0825, a ROCK (Rho-associated protein kinase) inhibitor, has shown promising antifibrotic and antiproliferative effects in other fibrotic conditions. This study investigated the therapeutic potential of AMA0825 using in vitro, ex vivo, and a 3-dimensional spheroid model of keloid disease, which partially reflects features of the keloid microenvironment. AMA0825 demonstrated potent antiproliferative activity against keloid fibroblasts, with a half-maximal growth inhibitory concentration of 28.19 ± 1.6 nM, significantly outperforming dexamethasone (half-maximal growth inhibitory concentration = 35.35 ± 2.6 μM) and triamcinolone (half-maximal growth inhibitory concentration = 37.84 ± 3 μM). This effect was confirmed by decreased Ki-67 expression and cell cycle arrest at the G1 phase. In the 3-dimensional spheroid model, AMA0825 effectively inhibited cell proliferation at nanomolar concentrations, exceeding the efficacy of dexamethasone. Although AMA0825 did not demonstrate significant antifibrotic activity at lower concentrations, it exhibited antifibrotic effects at higher concentrations. In addition, synergistic effects were observed when combined with dexamethasone. This study highlights the potential of ROCK inhibitors, particularly AMA0825, as an antiproliferative agent for keloid disease and underscores the value of 3-dimensional spheroid models for evaluating alternative therapeutic strategies.