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. 2020 Aug;47(8):991-1005.
doi: 10.1111/jcpe.13323. Epub 2020 Jun 11.

TLR3 ligation affects differentiation and stemness properties of gingival mesenchymal stem/progenitor cells

Mohamed Mekhemar  1 , Johannes Tölle  1 , Christof Dörfer  1 , Karim Fawzy El-Sayed  1   2
Affiliations

TLR3 ligation affects differentiation and stemness properties of gingival mesenchymal stem/progenitor cells

Mohamed Mekhemar et al. J Clin Periodontol. 2020 Aug.

Abstract

Aim: Toll-like receptors are key players in mesenchymal stem/progenitor cells' micro-environmental crosstalk, endorsing various biological reactions. For the first time, this study investigates the effects of TLR3-ligation on gingival mesenchymal stem/progenitor cells (G-MSCs) stemness and differentiation properties.

Material and methods: G-MSCs (n = 5) were isolated, sorted using anti-STRO-1 antibodies,and sowed on culture dishes to generate colony-forming units (CFUs), and their stem/progenitor cells' features and TLR3 expression were characterized. Subsequently, TLR3 activation of G-MSCs via Poly (I:C) was done, followed by an analysis of the expression of pluripotency-related factors, mesenchymal stemness-associated surface markers, and the ability to form CFUs and multilineage differentiation, using qualitative and quantitative histochemistry and RT-PCR.

Results: G-MSCs demonstrated all predefined stem/progenitor cells' characteristics and TLR3 expression. TLR3-activated G-MSCs showed a significantly reduced ability to form CFUs and pluripotency transcriptional factors expression. Mesenchymal stem/progenitor cell-associated surface markers and multilinear differentiation potential were significantly higher following TLR3 ligation (p < .05, Wilcoxon signed rank test).

Conclusions: TLR3-mediated activation maintains the mesenchymal stem/progenitor cells phenotype and drives G-MSCs' differentiation and commitment, with a shift away from an undifferentiated pluripotent cellular phenotype. This distinctive modulation could influence potential therapeutic applications of G-MSCs.

Keywords: gingiva; inflammation; polymerase chain reaction; stem cells; stemness; toll-like receptor.

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