Mesenchymal Stem Cells Antagonize IFN-Induced Proinflammatory Changes and Growth Inhibition Effects via Wnt/β-Catenin and JAK/STAT Pathway in Human Outer Root Sheath Cells and Hair Follicles

Mesenchymal stem cell therapy (MSCT) has been shown to be a new therapeutic option for treating alopecia areata (AA). Outer root sheath cells (ORSCs) play key roles in maintaining the hair follicle structure and supporting the bulge area. In human ORSCs (hORSCs), the mechanism for this process has not been extensively studied. In this study, we aimed to examine the influence of human hematopoietic mesenchymal stem cells (hHMSCs) in the hORSCs in vitro model of AA and determine the mechanisms controlling efficacy. Interferon-gamma (IFN-γ) pretreatment was used to induce an in vitro model of AA in hORSCs. The effect of MSCT on the viability and migration of hORSCs was examined using co-cultures, the MTT assay, and migration assays. We investigated the expression of molecules related to the Wnt/β-catenin pathway, JAK/STAT pathway, and growth factors in hHMSC-treated hORSCs by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses. hHMSCs increased hORSC viability and migration when they were co-cultured. hHMSCs reverted IFN-γ-induced expression—including NLRP3, ASC, caspase-1, CXCL-9 through 11, IL-1β, and IL-15—and upregulated several growth factors and hair stem cell markers. hHMSCs activated several molecules in the Wnt/β-catenin signaling pathway, such as in the Wnt families, β-catenin, phosphorylated GSK-3β and cyclin D1, and suppressed the expression of DKK1 induced by IFN-γ in hORSCs. hHMSCs suppressed the phosphorylation of JAK1 to 3, STAT1, and STAT3 compared to the controls and IFN-γ-pretreated hORSCs. These results demonstrate that hHMSCs increased hORSC viability and migration in the in vitro AA model. Additionally, MSCT definitely stimulated anagen survival and hair growth in an HF organ culture model. MSCT appeared to be associated with the Wnt/β-catenin and JAK/STAT pathways in hORSCs.

Red Ginseng Water Extract Aggravates Inflammation in Sebocytes and Outer Root Sheath Cells After Treatment With Lipopolysaccharide and Mice With Cutibacterium Acnes-induced Inflammatory Nodules

Ginseng has been known in Korea as a health-supportive herbal medicine from time immemorial. Red ginseng is one of processed ginseng that is produced from white ginseng through steaming and drying. Many protective functions of red ginseng have been reported from various groups in many diseases. In this study, we first investigated whether red ginseng water extract (RG) aggravates inflammation in human sebocytes and outer root sheath (ORS) cells after treatment with lipopolysaccharide (LPS) and mice with Cutibacterium (C.) acnes strain (ATCC 1182)-induced inflammatory nodules. Sebocytes and ORS cells were isolated and cultured from the human scalp. The RG augmented LPS mediated inflammation by increased the mRNA and protein expression of inflammatory cytokines in sebocytes and ORS cells. In addition, RG also showed the increased protein expression of p-NFκB, p-c-jun and p-JNK in the LPS-treated sebocytes and ORS cells. Furthermore, RG upregulated the LPS-induced production of sebum in sebocytes. In addition, RG inhibited improvement of inflammatory nodules and showed the increased expression of inflammatory biomarkers in inflammatory nodules of Cutibacterium acnes injected mice. Collectively, our data strongly suggest that RG is one of the aggravating factors of acne vulgaris. It would be better to stop taking RG in patients with inflammatory acne.

Enhanced Cultivation of Hair Follicle Outer Root Sheath Cells with K + M Culture Medium

Hair follicle stem cells are vital seed cells for hair follicle and tissue engineering skin. However, there is no valid approach to obtain abundant cells within short times in vitro. The purpose of this study was to establish a new medium for culture and expansion of hair follicle outer root sheath (ORS) cells. ORS cells were harvested from human samples and cultured with a medium mixed with keratinocyte serum free medium and mouse embryonic fibroblasts suspension. The proliferation and vitality of ORS cells were evaluated. The cells exhibited typical cobblestone morphology with good adhesion and colony-forming ability. The cells were also characterized through detecting specific markers with immunofluorescent staining and the results showed that partial ORS cells could express hair follicle stem cell markers of CK15, CK19 and FST. At last, ORS cells and fibroblasts were used as seed cells and preliminarily constructed tissue engineering skin successfully. In conclusion, these findings suggested that this new medium could effectively accelerate cell proliferation and maintain the fundamental characteristics of ORS cells.