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April 24, 2026Cureus0 citationsOpen Access

Multi-species Microbial Modulation of the RANK/RANKL (Receptor Activator of Nuclear Factor κB/Receptor Activator of Nuclear Factor κB Ligand) Pathway Induces Osteoclastogenic Priming in Apical Periodontitis

VGVictoria Alejandra Gutiérrez-LópezRMRita Elizabeth Martínez-MartínezJAJosé Manuel Alderete-Hernández

Key Points

  • The study aims to assess how microbial species related to apical periodontitis impact osteoclastogenic priming in human macrophages via the RANK/RANKL pathway.
  • Stimulated THP-1 monocytes/macrophages with live bacteria and RANKL.
  • Analyzed RANK/RANKL/OPG expression using flow cytometry.
  • Measured cytokine levels via cytometric bead arrays.
  • Bacteria like Enterococcus and Candida increased RANK⁺ and RANKL⁺ cell populations.
  • Sustained RANK/RANKL expression was promoted by Enterococcus and Candida.
  • Significant early increases in TNF-α, IL-1β, and IL-6 were observed following microbial stimulation.

Abstract

Background: Apical periodontitis (AP) is a prevalent inflammatory condition affecting the periapical tissues surrounding the tooth root, often initiated by microbial invasion. Objective: The objective of this study is to evaluate the osteoclastogenic priming effects of microbial species associated with primary and secondary AP on human macrophages and to determine their capacity to modulate the RANK/RANKL-OPG (receptor activator of nuclear factor kappa-B, receptor activator of nuclear factor kappa-B ligand, osteoprotegerin) pathway​. Materials and methods: THP-1 monocytes/macrophages were stimulated with live bacteria, with or without RANKL, and analyzed for RANK/RANKL/OPG expression by flow cytometry. Cytokine levels (interleukin-1β (IL-1β), tumoral necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), interleukin-12p70 (IL-12p70)) were measured using cytometric bead arrays. Results: Macrophages challenged with Enterococcus, Streptococcus, Actinomyces, and Candida exhibited an increase in RANK⁺, RANKL⁺, and RANK⁺RANKL⁺ cellular populations. Enterococcus and Candida promoted sustained RANK/RANKL expression across conditions, while Streptococcus and Actinomyces elicited significant double-positive cell expansion. Early microbial stimulation triggered robust TNF-α, IL-1β, and IL-6 production, peaking with Streptococcus intermedius, Enterococcus faecalis, and Actinomyces, followed by a decrease at 96 hours. In contrast, IL-8 secretion remained high among species and time points, with Actinomyces inducing the strongest response. Conclusion: Our data suggest that AP-associated bacteria can induce osteoclastogenic priming and promote the release of pro-inflammatory cytokines via RANK/RANKL activation, indicating a potential role in the early processes leading to periapical bone resorption.

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Cite This Study

Gutiérrez-López et al. (2026) studied this question.

synapsesocial.com/papers/69eb084f553a5433e34b35d8https://doi.org/10.7759/cureus.107487
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Also Consider

Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context:

  1. 1The global prevalence of apical periodontitis: a systematic review and meta‐analysis2020 · 575 citations
  2. 2Cell Wall Mannan of Candida Attenuates Osteogenic Differentiation by Human Dental Pulp Cells2022 · 4 citations
  3. 3Molecular analysis of the root canal microbiota associated with endodontic treatment failures2008 · 167 citations
  4. 4Association of Enterococcus faecalis With Different Forms of Periradicular Diseases2004 · 731 citations
  5. 5TNF-α induces osteoclastogenesis by direct stimulation of macrophages exposed to permissive levels of RANK ligand2000 · 1,436 citations