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Effect of hyaluronan tetrasaccharides on epidermal differentiation in normal human epidermal keratinocytes
Author(s) -
Kage M.,
Tokudome Y.,
Matsunaga Y.,
Hariya T.,
Hashimoto F.
Publication year - 2014
Publication title -
international journal of cosmetic science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.532
H-Index - 62
eISSN - 1468-2494
pISSN - 0142-5463
DOI - 10.1111/ics.12105
Subject(s) - cd44 , keratinocyte , cellular differentiation , microbiology and biotechnology , biology , chemistry , biochemistry , cell , in vitro , gene
Synopsis Objective Hyaluronan ( HA ) plays a role in keratinocyte proliferation and differentiation. In addition, HA has been shown to have different biological activities depending on its molecular weight. It has been reported that HA ‐mediated CD 44 activation regulates keratinocyte differentiation. Therefore, the aim of this study was to investigate the influence of HA tetrasaccharides ( HA 4) on the regulation of keratinocyte differentiation, CD 44 gene expression and CD 44‐phosphorylated protein in human keratinocytes, and compare HA 4 with high molecular weight HA . Methods Normal human epidermal keratinocytes ( NHEK s) were treated at doses of 1 μg mL −1 HA or HA oligosaccharides ( HA 4). After treatment, cell viability was checked using an MTT (3‐(4,5‐di‐methylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide) assay. Each differentiation marker and CD44 mRNA expression was detected by real‐time PCR. Each differentiation marker and CD44‐phosphorylated protein was assessed by Western blotting. Results Hyaluronan and HA4 showed no cytotoxicity up to a dose of 1 μg mL −1 . On day 3 after HA4 treatment, each differentiation marker mRNA and K10 protein level was higher than that of the control. On day 9, late differentiation marker mRNA and protein levels were increased with HA and HA4 treatment. In addition, HA4 treatment increased the expression of CD44 mRNA , CD44‐phosphorylated protein and intracellular calcium concentrations. HA4 enhanced keratinocyte differentiation and increased CD44‐phosphorylated protein levels. Conclusion HA 4 may induce epidermal differentiation through phosphorylation of CD 44.