Fenotipske i funkcionalne karakteristike menzenhimalnih stromalnih matičnih ćelija izolovanih iz zdravog i inflamiranog tkiva humane gingive/

Abstract

Dokazano je da mezenhimalne stromalne matične ćelije (GMSC) izolovane iz gingivalnog tkiva imaju važnu ulogu u procesu parodontopatije. Međutim, kako je P. Gingivalis jedan od ključnih etioloških faktora u nastanku bolesti parodoncijuma, njegov uticaj na zdravu (H) i parodontopatijom zahvaćenu gingivu (P) nije dovoljno poznat. U cilju rješavanja ovog problema, uspostavili smo 10 H-GMSC i 12 P-GMSC linija. Nije bilo značajnih razlika u morfologiji, diferencijaciji u hondrondroblaste i adipocite, ekspresiji karakterističnih MSC markera, uključujući i pericitne markere NG2 i PDGFR između H- i P- GMSC linija. Međutim, proliferacija, veličina ćelija i osteogeni potencijal su bili veći kod P- GMSC linija, za razliku od njihove manje sposobnosti inhibicije proliferacije mononuklearnih ćelija periferne krvi. P. Gingivalis je povećao mRNA ekspresiju IL-6, IL-8, MCP-1, Gro-α, Rantes, TLR-2, HIF-1α, OPG, MMP-3, SDF-1, HGF i IP-10 u P-GMSC, dok je ekspresija samo IL-6, MCP-1 i GRO-Α bila povećana kod H-GMSC. Ekspresija MCP-1, Rantes, IP-10 i HGF je bila značajno veća u P-GMSC-u u odnosu na H-GMSC, ali je ekspresija IDO-1 bila manja. Nije bilo značajnih promjena u ekspresiji TLR-3, TLR- 4, TGF-β, LAP, IGFBP4 i TIMP-1 u oba tipa GMSC. Zaključno, naši rezultati ukazuju da P-GMSC zadržavaju svoja pro-inflamatorna svojstva u kulturi, imaju manji imunosupresivni potencijal nego H-GMSC a ekspresija gena povezanih sa regeneracijom tkiva je poremećena. Sve ove funkcije i genska ekspresija su bile značajno potencirane ili modifikovane nakon tretmana ćelijskih kultura sa P. Gingivalis-om.

Gingiva-Derived Mesenchymal Stromal Cells (GMSCs) play significant role in the pathogenesis of chronic periodontitis. However, little is known how P.gingivalis, one of the key etiological factor of periodontal disease, influences GMSC established from healthy (H) and periodontitis (P) gingiva. To better understand this problem, 10 H-GMSC and 12 P-GMSC lines were established. The obtained results showed that morphology, the potential of the lines to differentiate into chondroblasts and adipocytes, and phenotypic profile of MSCS, including the expression of pericyte markers (NG2 and PDGFR), did not different between H- and P-GMSC lines. Contrary, P-GMSC showed a lower capability to inhibit the proliferation of peripheral blood mononuclear cells. P. gingivalis augmented the expression of IL-6, IL-8, GRO-α, MCP-1, TLR-2, RANTES, OPG, HIF-1α, SDF-1, HGF, MMP-3, and IP-10 in P-GMSCs at the level of mRNA, IL-6, MCP-1 and GRO-α were up-regulated in H-GMSCs. The expression of MCP- 1, RANTES, IP-10, and HGF in P-GMSCs was statistically significantly increased in comparison with H-GMSCs. In contrast, the expression of IDO-1 was down-regulated. The expression of TLR-3, TLR-4, TGF-β, LAP, IGFBP4, and TIMP-1 in both H- and P- GMSCs was not significantly modified. In conclusion, our results imply that P-GMSCs in culture keep characteristics in culture, and have a lower capacity to suppress lymphocyte proliferation than H-GMSC. In addition, the expression of genes associated with tissue regeneration was impaired in P-GMSC. P. gingivalis was able to potentiate all these functions and modulate the expression of many investigated genes.