Background: Impaired wound healing is one of the complications of diabetes. Carvacrol, a natural substance, shows promising results in diabetic wound healing despite the fact that its therapeutic mechanisms are not fully understood. Objectives: The aim of this study was to investigate the molecular alterations caused by carvacrol intervention in human dermal fibroblasts (HDFs) cultured at a high-glucose condition. Methods: The HDFs were incubated in different glucose concentrations, and cell viability was assessed. In addition, carvacrol cytotoxicity was determined. Then, the HDFs were incubated in 50 mM glucose prior to treatment. After that, the cells were divided into 4 groups: Controls, high-glucose (50 mM), carvacrol-treated (9 µM), and high-glucose carvacrol-treated for 24 h. Cell proliferation and migration were examined. Furthermore, superoxide dismutase (SOD) production, collagen deposition, and RNA levels of TGFβ1, ACTA2, and miR-155 were investigated. Results: The in vitro scratch assay revealed that the fibroblast migration, which was reduced at high-glucose concentration, was reversed due to the carvacrol intervention during 12 h and 24 h (P < 0. 01 and P < 0. 001, respectively). Collagen deposition and SOD synthesis showed an increase in treated cells (P < 0. 001). Both TGFβ1 and ACTA2 mRNA expressions were elevated due to the carvacrol treatment, while the miR-155 level decreased (P < 0. 001). Conclusions: A high level of glucose impaired the cellular function of human dermal fibroblast. Carvacrol reversed the adverse effects of high-glucose stress and promoted wound healing through greater cell migration, collagen deposition, and levels of TGFb1 and ACTA2 gene expression. It showed inhibitory effects against miR-155, which is known for its negative role in diabetic wound healing.

Background: Rheumatic heart disease (RHD) is exacerbated by chronic inflammation that stimulates the release of proinflammatory cytokines, most notably transforming growth factor-beta 1 (TGF-β1), which promotes myofibroblast differentiation. This study aims to determine the optimal dosage of Lisinopril, an angiotensin-converting enzyme inhibitor, for mitigating the fibrotic changes associated with RHD. Methods: This in vitro, posttest-only control group study involved obtaining valvular interstitial cells from the heart valves of 25 male New Zealand rabbits (Oryctolagus cuniculus). Valvular interstitial cells were divided into 5 groups: a control group exposed to TGF-β1, and 4 experimental groups exposed to various Lisinopril doses (1 µM, 10 µM, and 100 µM) in addition to TGF-β1. The effect of Lisinopril on myofibroblast differentiation was assessed by measuring alpha-smooth muscle actin (αSMA) expression through immunocytochemical methods. Statistical significance was determined using an independent T-test with a P value of less than 0. 050. Results: Independent T-tests conducted on 25 male Oryctolagus cuniculus demonstrated significantly lower αSMA expression in the groups treated with various Lisinopril doses (1 µM, 10 µM, and 100 µM) compared with the TGF-β1-induced control group (P<0. 050). The most significant reduction in αSMA expression was observed in the group treated with the highest Lisinopril dose of 100 µM. Conclusion: Lisinopril demonstrates a significant ability to inhibit TGF-β1-induced myofibroblast differentiation in rabbit valve interstitial cells, with the 100 µM dose proving most effective. These results suggest that Lisinopril may have the potential to curb RHD progression, warranting further investigations in vivo.

Objective: Stromal elements play a key role in growth and development of different neoplasms. Myofibroblasts are the major components and occur in stromal tissue during carcinogenesis processes. The purpose of this study was to review the frequency and the distribution pattern of myofibroblasts (aSMA-positive) in the stroma of squamous epithelial carcinoma and to compare values with those for with oral dysplasia and hyperkeratosis.Materials and Methods: we evaluated aSMA protein frequency in hyperkeratosis (n=18), oral epithelial dysplasia (n=18) and oral squamous cell carcinoma (n=18) using immunohistochemistry. Results: αSMA-positive expression was observed in 67% of OSCC tissue samples with network and spindle patterns, whereas it was seen in 22% with a focal pattern in dysplasia and in 6% with a scanty pattern in hyperkeratosis cases.Conclusion: These findings suggest that an increase in number of myofibroblasts and change in their distribution pattern occurs during carcinogenesis which can be an expression of their role in tumor invasive characteristics.

Objective: To study the therapeutic effect of mesenchymal stem cells (MSC) on induced liver fibrosis in mice model and to uncover its mechanism.Materials and Methods: we infused GFP+ MSCs of male into the tail vein of female mice that received CCl4 injection biweekly to induce liver fibrosis. MSC which were derived from bone marrow obtained from femoral and tibial bones, isolated and proliferated in culture. They were characterized morphologically and by their potential of differentiation to osteo and adipo lineage. Animals were divided into 6 groups: control, CCl4 4 week, CCL4 8 week, CCl4 4week plus MSC, CCl4 8 week plus MSC, CCL4 plus opportunity to regeneration. Liver tissue was examined histopathologically and by software to quantify collagene deposition as the stage of liver fibrosis. Lipid peroxidation was quantified as a marker of liver injury level. Gene expression ratio of the collagen (type I), TIMP1, MMP-9, MMP-13, alph SMA was detected. Immunostaining were done to show homing and differentiation to hepatocyte of the cells and presentation of Hepatic stellate cells (HSC) liver functions (serum ALT and AST) were estimated for all groups.Results: Quantitative RT-PCR analysis showed administration of MSCs has a significant antifibrotic effect as evidenced by the significant decrease in liver collagen and increase MMP13 gene expression in the CCl4/MSC group compared to the CCl4 group, 4 weeks after transplantation. The expression of αSMA (smooth muscle actin) and TIMP also reduced in CCl4/MSC group. However, this was statistically nonsignificant. Additionally, the expression of MMP9 significant increase in CCl4 treated groups; however, there was no significant change after MSC injection. The reduction in liver collagen confirmed histopathologically by quantitative analysis. Moreover, lipid peroxidation content in transplanted group decreases significantly. Immunostaining of transplanted cells showed GFP-positive cells in the liver and some of them expressed albumin or αSMA.Conclusion: MSCs can enhance recovery of CCl4-injured mouse liver through their influence in reduction collagen deposition by possible effect on matrix metalloptoteases and their capacity to differentiate into hepatocyte-like cells.

Introduction: Understanding the key role of the tumor microenvironment in specifying molecular markers of breast cancer subtypes is of a high importance in diagnosis and treatment. Therefore, the possibility of interconversion of luminal states and their specific markers alteration under the control of tumor microenvironment (TME), particularly cancer-associated fibroblasts (CAFs) deserves to be further investigated. Methods: To activate normal human fibroblasts, liquid overlay technique or nemosis was used and α-SMA protein expression, CAFs marker, in fibroblastic spheroids was measured by blotting. The luminal A, MCF-7, and luminal B, MDA-MB 361, cell lines were treated with normal and spheroidal/activated fibroblast conditioned medium for 48 hours. The morphological changes of both luminal A and B cells were evaluated by invert light microscopy and analyzed through the shape factor formula. Moreover, chemo-sensitivity, proliferation, and changes in ER-related and proliferative genes expression levels were assessed respectively via MTT assay, Ki67 expression Immunofluorescence assay, real time PCR and Annexin V-FITC techniques. Results: Activated (spheroidal) fibroblasts, expressed αSMA marker two folds more than monolayer cultured fibroblasts. Our study indicated a significant increase in IC50 of both luminal A and B cell lines after being treated with conditioned medium particularly in treated group with spheroidal conditioned medium. Studying Morphological changes using shape factor formula demonstrated more aggressiveness with gaining mesenchymal features in both luminal A and B subtypes by increasing exposure time. Changes in the expression of Ki67 were observed following treatment with fibroblastic and spheroidal paracrine secretome. Driven Data from Ki67 assay supports the luminal A and B interconversion by elevated Ki67 expression in luminal A and lowered Ki67 expression in luminal B. Gene expression analysis revealed that anti-apoptotic Bcl2 gene expression in both luminal types treated with condition medium has been increased though there has seen no interchange in expression of ER-related and proliferative genes between luminal A (MCF7) and luminal B (MDA-MB361) subtypes, the results of Annexin V-FITC flow cytometry test indicated a decrease in the population of both early and late apoptotic cells in groups treated with both fibroblastic and spheroidal condition medium compared to of control group. Conclusion: Under the paracrine influence of fibroblast cells, both luminal A (MCF7) and luminal B (MDA-MB) subtypes of breast cancer gained invasive, anti-apoptotic, and chemoresistance features which are mostly increased by activated(spheroidal) fibroblasts conditioned medium mimicking CAFs. There was no strong proof for interconversion of luminal A and luminal B which share more similarities among breast cancer molecular subtypes.