10.6084/m9.figshare.5182630.v1
van Kilsdonk J.W.J.
van Kilsdonk J.W.J.
Jansen P.A.M.
Jansen
P.A.M.
van den Bogaard E.H.
van den Bogaard E.H.
Bos C.
Bos
C.
Bergers M.
Bergers
M.
Zeeuwen P.L.J.M.
Zeeuwen
P.L.J.M.
Schalkwijk J.
Schalkwijk
J.
Supplementary Material for: The Effects of Human Beta-Defensins on Skin Cells in vitro
Karger Publishers
2017
Keratinocytes
Fibroblasts
β-Defensins
Wound healing
2017-07-07 07:58:16
Journal contribution
https://karger.figshare.com/articles/journal_contribution/Supplementary_Material_for_The_Effects_of_Human_Beta-Defensins_on_Skin_Cells_in_vitro/5182630
<p><b><i>Background:</i></b> Defensins are antimicrobial peptides that
exert immunomodulatory and chemotactic functions. Based on these
properties and their high expression levels in the skin, they are likely
to affect skin inflammation, infection, and wound healing. This may
lead to therapeutic applications in (burn) wound healing. <b><i>Objective:</i></b>
We aimed to investigate the effects of human β-defensins (hBDs) on
keratinocytes and fibroblasts, 2 major skin cell types involved in skin
regeneration. <b><i>Methods:</i></b> Monolayer keratinocyte and
fibroblast cultures were exposed to recombinant hBDs, and we
overexpressed hBD2 and hBD3 in keratinocytes of reconstructed epidermal
equivalents by lentiviral transduction. The effects were measured by
immunohistochemistry, quantitative real-time PCR, and migration assays.
Kinome analyses were performed on cultured keratinocytes to investigate
the signal transduction events elicited by hBD stimulation. <b><i>Results:</i></b>
We found that hBD3 induced the expression of cytokines and chemokines
in keratinocytes, which was not observed in fibroblasts. hBD2, however,
stimulated cell migration only in fibroblasts, which was not found for
hBD3. Both defensins are likely to exert receptor-mediated effects in
keratinocytes, as witnessed by changes in protein kinase activation
following stimulation by hBD2 and hBD3. Kinome analysis suggested that
protein kinase C activation was a common event for both defensins. We
observed, however, considerable differences in keratinocyte responses
between stimulation by exogenous recombinant defensins and endogenous
defensins expressed following lentiviral transduction. <b><i>Conclusion:</i></b>
Defensins exert modest biological effects on skin cells that are
potentially beneficial in wound healing, but many questions regarding
the biological mechanisms of action and relevance for the in vivo
situation are still remaining.</p>