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

Karim Fawzy El-Sayed; Elena Mahlandt; Kristina Schlicht; Kim Enthammer; Johannes Tölle; Juliane Wagner; Katharina Hartmann; Peter R Ebeling; Christian Graetz; Mathias Laudes; Christof E Dörfer; Dominik M Schulte

doi:10.1111/jre.13319

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

J Periodontal Res. 2024 Jul 2. doi: 10.1111/jre.13319. Online ahead of print.

Authors

Karim Fawzy El-Sayed^{1

2

3}, Elena Mahlandt⁴, Kristina Schlicht⁴, Kim Enthammer⁴, Johannes Tölle^{1

5

6}, Juliane Wagner⁷, Katharina Hartmann⁴, Peter R Ebeling⁸, Christian Graetz¹, Mathias Laudes^{4

9}, Christof E Dörfer¹, Dominik M Schulte^{4

8

9}

Affiliations

¹ Clinic for Conservative Dentistry and Periodontology, University Hospital of Schleswig-Holstein, Kiel, Germany.
² Oral Medicine and Periodontology Department, Faculty of Dentistry, Cairo University, Cairo, Egypt.
³ Stem Cells and Tissue Engineering Unit, Faculty of Dentistry, Cairo University, Cairo, Egypt.
⁴ Institute of Diabetes and Clinical Metabolic Research, University Hospital of Schleswig-Holstein, Kiel, Germany.
⁵ Department of Dermatology, University Hospital Schleswig-Holstein, Kiel, Germany.
⁶ Institute of Immunology, University Hospital Schleswig-Holstein, Kiel, Germany.
⁷ Department of Oral and Maxillofacial Surgery, University Hospital of Schleswig-Holstein, Kiel, Germany.
⁸ Department of Medicine, School of Clinical Sciences at Monash Health, Monash University, Melbourne, Victoria, Australia.
⁹ Division of Endocrinology, Diabetes and Clinical Nutrition, Department of Internal Medicine I, University Hospital Schleswig-Holstein, Kiel, Germany.

PMID: 38952262
DOI: 10.1111/jre.13319

Abstract

Aims: Oxidized 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.

Methods: Human 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.

Results: G-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).

Conclusions: Within 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.

Keywords: OxLDL; cytokines; inflammation; periodontitis; stem cells.