Background:Keloids are benign fibroproliferative skin lesions that are difficult to treat and become a lifetime predicament for patients.Several treatment modalities have been put forth,but as yet no satisfactory appr...Background:Keloids are benign fibroproliferative skin lesions that are difficult to treat and become a lifetime predicament for patients.Several treatment modalities have been put forth,but as yet no satisfactory approach to the prevention or treatment of keloids has been identified.The process of epithelial-to-mesenchymal transition(EMT)has been implicated in keloid scarring,as keloid keratinocytes display an EMT-like phenotype.This study investigated the potential of pirfenidone,an antifibrotic agent,to counteract EMT-like alterations in keloid keratinocytes,including gene expression,cell migratory and proliferative functions.Methods:Normal and keloid keratinocytes were isolated from discarded normal skin tissues and from resected keloid tissues,respectively.Cells were quiesced for 24 h without epidermal growth factor DS-Qi1MCDigital and were exposed to transforming growth factor-beta1(TGF-β1;10 ng/mL),with or without pirfenidone(400μg/mL),for an additional 24 h.The effects of pirfenidone on cytotoxicity,cell migration,cell proliferation,and on expression of genes and proteins involved in EMT were assayed.Statistical significance was determined by two-way ANOVA using Sigma Plot.Results:We found that pirfenidone did not elicit any cytotoxic effect at concentrations up to 1000μg/mL.A statistically significant dose-dependent decrease in basal cell proliferation rate was noted in both normal and keloid keratinocytes when exposed to pirfenidone at concentrations ranging from 200 to 1000μg/mL.Pirfenidone significantly decreased basal cell migration in both normal and keloid keratinocytes,but a significant decrease in TGF-β1-induced cell migration was seen only in keloid keratinocytes.Significant inhibition of the expression of TGF-β1-induced core EMT genes,namely hyaluronan synthase 2,vimentin,cadherin-11,and wingless-type MMTV integration site family,member 5A along with fibronectin-1,was observed in both normal and keloid keratinocytes treated with pirfenidone.In addition,the protein levels of vimentin and fibronectin were significantly reduced by pirfenidone(400μg/mL)in both normal and keloid keratinocytes.Conclusions:For the first time,this study shows the efficacy of pirfenidone in inhibiting the EMTlike phenotype in keratinocytes derived from keloids,suggesting that pirfenidone may counteract a critical contributor of keloid progression and recurrence.展开更多
文摘Background:Keloids are benign fibroproliferative skin lesions that are difficult to treat and become a lifetime predicament for patients.Several treatment modalities have been put forth,but as yet no satisfactory approach to the prevention or treatment of keloids has been identified.The process of epithelial-to-mesenchymal transition(EMT)has been implicated in keloid scarring,as keloid keratinocytes display an EMT-like phenotype.This study investigated the potential of pirfenidone,an antifibrotic agent,to counteract EMT-like alterations in keloid keratinocytes,including gene expression,cell migratory and proliferative functions.Methods:Normal and keloid keratinocytes were isolated from discarded normal skin tissues and from resected keloid tissues,respectively.Cells were quiesced for 24 h without epidermal growth factor DS-Qi1MCDigital and were exposed to transforming growth factor-beta1(TGF-β1;10 ng/mL),with or without pirfenidone(400μg/mL),for an additional 24 h.The effects of pirfenidone on cytotoxicity,cell migration,cell proliferation,and on expression of genes and proteins involved in EMT were assayed.Statistical significance was determined by two-way ANOVA using Sigma Plot.Results:We found that pirfenidone did not elicit any cytotoxic effect at concentrations up to 1000μg/mL.A statistically significant dose-dependent decrease in basal cell proliferation rate was noted in both normal and keloid keratinocytes when exposed to pirfenidone at concentrations ranging from 200 to 1000μg/mL.Pirfenidone significantly decreased basal cell migration in both normal and keloid keratinocytes,but a significant decrease in TGF-β1-induced cell migration was seen only in keloid keratinocytes.Significant inhibition of the expression of TGF-β1-induced core EMT genes,namely hyaluronan synthase 2,vimentin,cadherin-11,and wingless-type MMTV integration site family,member 5A along with fibronectin-1,was observed in both normal and keloid keratinocytes treated with pirfenidone.In addition,the protein levels of vimentin and fibronectin were significantly reduced by pirfenidone(400μg/mL)in both normal and keloid keratinocytes.Conclusions:For the first time,this study shows the efficacy of pirfenidone in inhibiting the EMTlike phenotype in keratinocytes derived from keloids,suggesting that pirfenidone may counteract a critical contributor of keloid progression and recurrence.
