Carbon dioxide-induced decrease in extracellular pH enhances the production of extracellular matrix components by upregulating TGF-β1 expression via CREB activation in human dermal fibroblasts

Mild acidification caused by transcutaneous administration of carbon dioxide (CO2) has been reported to improve some epidermal skin impairments, such as desquamation and inflammation; however, its effects on dermal tissue remain unclear. Here, we examined the effect and mechanism of mild acidity on extracellular matrix (ECM) protein production in normal human dermal fibroblasts (NHDFs). To achieve this, the skin permeability of CO2 and its effect on intradermal pH were evaluated by treating reconstructed human skin equivalents (HSEs) with a CO2-containing formulation. Additionally, NHDFs were cultured in a pH-adjusted medium (pH 6.5). CO2 successfully permeated HSEs and reduced the intradermal pH. Decreased extracellular pH activated CREB, upregulated TGF-beta 1 expression, promoted the production of elastic and collagen fibres, and increased hyaluronan concentration in NHDFs. Additionally, the low pH-induced increase in TGF-beta 1 expression was attenuated via the RNAi-mediated suppression of the expression of CREB1 and proton-sensing G protein-coupled receptors (GPCRs), including GPR4 and GPR65. Moreover, low pH-induced CREB activation was suppressed by the inhibition of the cAMP/PKA and PLC/PKC signalling pathways. Taken together, a CO2-induced decrease in intradermal pH may promote ECM production in NHDFs via the upregulation of TGF-beta 1 expression, which was mediated by the activation of the GPCR signalling pathway and CREB, indicating that CO2 could be used to treat ultraviolet radiation-induced photoaging, intrinsic ageing and ECM deterioration.