Efficacy of probiotic Streptococcus thermophilus in counteracting TGF-β1-induced fibrotic response in normal human dermal fibroblasts

Abstract

Background Abnormal and deregulated skin wound healing associated with prolonged inflammation may result in dermal fibrosis. Since the current therapeutic strategies revealed unsatisfactory, the investigation of alternative approaches such as those based on the use of specific probiotic strains could provide promising therapeutic options. In this study, we aimed to evaluate whether the lysate from S. thermophilus could antagonize the fibrogenic effects of TGF-β1 in normal human dermal fibroblasts (NHDF). Methods NHDF were exposed to TGF-β1 to establish a fibrotic phenotype. Proliferation rate and cell number were measured using the IncuCyte® Live Cell Imager system and the trypan blue dye exclusion test. Phenoconversion markers (α-SMA and fibronectin) and collagen I levels were assessed by western blot and immunofluorescence. The mRNA levels of TGF-β1 were evaluated by RT-PCR. The Smad2/3 phosphorylation level as well as β-catenin and PPARγ expression, were assessed by western blot. The cell contractility function and migration of NHDF were studied using collagen gel retraction assay, and scratch wound healing assay, respectively. The effects of S. thermophilus lysate, alone or combined with TGF-β1, were evaluated on all of the above-listed parameters and markers associated with TGF-β1-induced fibrotic phenotype. Results Exposure to the S. thermophilus lysate significantly reduced the key mediators and events involved in the abnormal activation of myofibroblasts by TGF-β1 within the fibrotic profile. The S. thermophilus treatment significantly reduced cell proliferation, migration, and myo-differentiation. In addition, the treatment with probiotic lysate reduced the α-SMA, fibronectin, collagen-I expression levels, and affected the collagen contraction ability of activated dermal fibroblasts. Moreover, the probiotic targeted the TGF-β1 signaling, reducing Smad2/3 activation, TGF-β1 mRNA level, and β-catenin expression through the upregulation of PPARγ. Conclusion This is the first report showing that S. thermophilus lysate had a remarkable anti-fibrotic effect in TGF-β1-activated NHDF by inhibiting Smad signaling. Notably, the probiotic was able to reduce β-catenin and increase PPARγ levels. The findings support our point that S. thermophilus may help prevent or treat hypertrophic scarring and keloids.

Looking for the Ideal Probiotic Healing Regime

“…Mature fibroblasts, after being differentiated into myofibroblasts, migrate towards the granulation tissue and, driven by TGF-β, are ready to express α-smooth muscle actin (α-SMA), which contracts the wound. L. plantarum GMNL-6, L. paracasei GMNL-653, and S. thermophilus DSM 24731 were found to exert an earlier, dose-dependent inhibitory effect on α-SMA expression, in parallel with the presence of structured collagen fiber, probably contributing to the prevention of excessive skin fibrosis [40,56].…”

“…In addition, the treatment with parabiotic lysate reduced the α-SMA, fibronectin, and collagen-I expression levels and affected the collagen contraction ability of activated dermal fibroblasts. Moreover, parabiotic S. thermophilus targeted the TGF-β1 signaling, reducing Smad2/3 activation, TGF-β1 mRNA level, and β-catenin expression through the up-regulation of PPARγ [56].…”

Evaluation of the Effectiveness of an Innovative Polycomponent Formulation on Adult and Aged Human Dermal Fibroblasts

“…To evaluate the contraction ability of adult or aged HDFs, the fibroblasts were put into three-dimensional collagen lattices as previously described [ 22 ]. Briefly, acid-extracted type I collagen (5 mg/mL) mixture was made on ice with rat tail collagen I (Enzo Life Sciences, Lausen, Switzerland) in acetic acid 0.2%.…”

Probiotics in Wound Healing

“…The L. plantarum and Streptococcus thermophilus probiotic strains were found to initiate earlier collagen III synthesis and deposition through TGF-β and then replace it with type I collagen. In addition, lactobacilli proved to accelerate keratinocyte migration and proliferation [ 63 ].…”