Effects of oxidized LDL versus IL‐1ß/TNF‐ɑ/INFɣ on human gingival mesenchymal stem cells properties

Abstract

AbstractAimsOxidized low‐density lipoprotein (oxLDL) is an important player in the course of metabolic inflammatory diseases. oxLDL was identified in the gingival crevicular fluid, denoting possible associations between oxLDL‐induced inflammation and periodontal disease. The current investigation compared for the first‐time direct effects of oxLDL to a cytokine cocktail of IL‐1ß/TNF‐ɑ/INF‐γ on gingival mesenchymal stem cells' (G‐MSCs) attributes.MethodsHuman third passage G‐MSCs, isolated from connective tissue biopsies (n = 5) and characterized, were stimulated in three groups over 7 days: control group, cytokine group (IL‐1β[1 ng/mL], TNF‐α[10 ng/mL], IFN‐γ[100 ng/mL]), or oxLDL group (oxLDL [50 μg/mL]). Next Generation Sequencing and KEGG pathway enrichment analysis, stemness gene expression (NANOG/SOX2/OCT4A), cellular proliferation, colony‐formation, multilinear potential, and altered intracellular pathways were investigated via histochemistry, next‐generation sequencing, and RT‐qPCR.ResultsG‐MSCs exhibited all mesenchymal stem cells' characteristics. oxLDL group and cytokine group displayed no disparities in their stemness markers (p > .05). Next‐generation‐sequencing revealed altered expression of the TXNIP gene in response to oxLDL treatment compared with controls (p = .04). Following an initial boosting for up to 5 days by inflammatory stimuli, over 14 day, cellular counts [median count ×10−5 (Q25/Q75)] were utmost in control – [2.6607 (2.0804/4.5357)], followed by cytokine – [0.0433 (0.0026/1.4215)] and significantly lowered in the oxLDL group [0.0274 (0.0023/0.7290); p = .0047]. Osteogenic differentiation [median relative Ca2+ content(Q25/Q75)] was significantly lower in cytokine – [0.0066 (0.0052/0.0105)] compared to oxLDL – [0.0144 (0.0108/0.0216)] (p = .0133), with no differences notable for chondrogenic and adipogenic differentiation (p > .05).ConclusionsWithin the current investigation's limitations, in contrast to cytokine‐mediated inflammation, G‐MSCs appear to be minimally responsive to oxLDL‐mediated metabolic inflammation, with little negative effect on their differentiation attributes and significantly reduced cellular proliferation.