Characterization of Regulatory Extracellular Vesicles from Osteoclasts
Open accessAbstract
Extracellular vesicles (EVs), which include exosomes and ectosomes/microvesicles, have emerged as important intercellular regulators. EVs can interact with surface receptors of target cells and can transport luminal components, including messenger RNAs (mRNAs), microRNAs, and enzymes, to the cytosol of the target cell. Here, we show that hematopoietic cells grown in culture shed exosome-like EVs as they differentiate from preosteoclasts into osteoclasts. These EVs were between 25 and 120 nm (mean, 40 nm) in diameter determined by transmission electron microscopy. The exosome-associated markers CD63 and EpCAM were enriched in the isolated EVs while markers of Golgi and endoplasmic reticulum were not detected. Treatment of isolated hematopoietic cells with EVs did not affect their receptor activator of nuclear factor κB–ligand (RANKL)–stimulated differentiation into osteoclasts. However, EVs from osteoclast precursors promoted 1,25-dihydroxyvitamin D 3 –dependent osteoclast formation in whole mouse marrow cultures, and EVs from osteoclast-enriched cultures inhibited osteoclastogenesis in the same cultures. These data suggested that osteoclast-derived EVs are paracrine regulators of osteoclastogenesis. EVs from mature osteoclasts contained receptor activator of nuclear factor κB (RANK). Immunogold labeling showed RANK was enriched in 1 in every 32 EVs isolated from osteoclast-enriched cultures. Depletion of RANK-rich EVs relieved the ability of osteoclast-derived EVs to inhibit osteoclast formation in 1,25-dihydroxyvitamin D 3 –stimulated marrow cultures. In summary, we show for the first time that EVs released by osteoclasts are novel regulators of osteoclastogenesis. Our data suggest that RANK in EVs may be mechanistically linked to the inhibition of osteoclast formation. RANK present in EVs may function by competitively inhibiting the stimulation of RANK on osteoclast surfaces by RANKL similar to osteoprotegerin. RANK-rich EVs may also take advantage of the RANK/RANKL interaction to target RANK-rich EVs to RANKL-bearing cells for the delivery of other regulatory molecules.
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Osteoclasts release extracellular vesicles between 25 and 120 nm in diameter that carry RANK.
“In fact, osteoclasts release EVs (between 25 and 120 nm in diameter) that carry RANK.”
From Bone remodeling: an operational process ensuring survival and bone mechanical competence (2022) · cites this paper as [237]
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- Authors
- Huynh, N., VonMoss, L., Smith, D., Rahman, I., Felemban, M. F., Zuo, J., Rody, W. J., McHugh, K. P., & Holliday, L. S.
- Affiliations
- University of Florida (US)
- University of Florida Health (US)
- Corresponding author
- Holliday, L.S.
- Venue
- Journal of Dental Research
- Publisher
- SAGE Publications
- Published
- 2016-02-23
- Volume
- 95
- Issue
- 6
- Pages
- 673-679
- ISSN
- 0022-0345 (ISSN-L)
- 1544-0591
- PMC
- PMC4924543
- OpenAlex
- W2274581845
- Cited by
- 196
- References
- 32
- Citations by year
- 2026: 12 · 2025: 15 · 2024: 14 · 2023: 16 · 2022: 23 · 2021: 36 · 2020: 23 · 2019: 25 · 2018: 18 · 2017: 11 · 2016: 3
- FWCI
- 9.88
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Journal of Dental Research (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- University of Florida
- National Institutes of Health
- National Institute of Dental and Craniofacial Research (R21DE019862)
- College of Medicine, University of Florida
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bone Metabolism and Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Keywords
- extracellular vesicles, osteoclasts, osteoclastogenesis, RANK, RANKL, paracrine regulation, exosomes, microvesicles, CD63, EpCAM, 1,25-dihydroxyvitamin D3, osteoprotegerin, bone remodeling, intercellular communication
- MeSH terms
- Extracellular Vesicles, Animals, Bone Marrow Cells, Calcitriol, Cell Communication, Cell Differentiation, Cell Survival, Cells, Cultured, Osteoclasts, Biomarkers, Microscopy, Electron, Transmission, Mice, RANK Ligand, Receptor Activator of Nuclear Factor-kappa B
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