Establishment and validation of an efficient method for the 3D culture of osteoclasts in vitro.

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  • Additional Information
    • Source:
      Publisher: Elsevier Country of Publication: England NLM ID: 0354422 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-176X (Electronic) Linking ISSN: 03005712 NLM ISO Abbreviation: J Dent Subsets: MEDLINE
    • Publication Information:
      Publication: Kidlington : Elsevier
      Original Publication: Bristol, Eng., Wright.
    • Subject Terms:
      Osteoclasts*/cytology ; Hydrogels* ; Cell Culture Techniques*; Animals ; Cell Differentiation ; Collagen ; Mice ; Cell Culture Techniques, Three Dimensional/methods ; Macrophages/cytology ; Cathepsin K ; Cytokines/metabolism ; Cells, Cultured
    • Abstract:
      Introduction: Osteoclasts (OCs) play a crucial role in maintaining bone health. Changes in OC activity are linked to different bone diseases, making them an intriguing focus for research. However, most studies on OCs have relied on 2D cultures, limiting our understanding of their behavior. Yet, there's a lack of knowledge regarding platforms that effectively support osteoclast formation in 3D cultures.
      Methods: In our investigation, we explored the capacity of collagen and GelMA hydrogels to facilitate osteoclast development in 3D culture settings. We assessed the osteoclast development by using different hydrogels and cell seeding strategies and optimizing cell seeding density and cytokine concentration. The osteoclast development in 3D cultures was further validated by biochemical assays and immunochemical staining.
      Results: Our findings revealed that 0.3 % (w/v) collagen was conducive to osteoclast formation in both 2D and 3D cultures, demonstrated by increased multinucleation and higher TRAP activity compared to 0.6 % collagen and 5 % to 10 % (w/v) GelMA hydrogels. Additionally, we devised a "sandwich" technique using collagen substrates and augmented the initial macrophage seeding density and doubling cytokine concentrations, significantly enhancing the efficiency of OC culture in 3D conditions. Notably, we validated osteoclasts derived from macrophages in our 3D cultures express key osteoclast markers like cathepsin K and TRAP.
      Conclusions: To conclude, our study contributes to establishing an effective method for cultivating osteoclasts in 3D environments in vitro. This innovative approach not only promises a more physiologically relevant platform to study osteoclast behavior during bone remodeling but also holds potential for applications in bone tissue engineering.
      Clinical Significance: This study introduces an efficient method for cultivating osteoclasts in 3D environments in vitro. It offers a more physiologically relevant platform to investigate osteoclast behavior and holds promise to advance research in bone biology and regenerative dentistry.
      Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
      (Copyright © 2024 Elsevier Ltd. All rights reserved.)
    • Contributed Indexing:
      Keywords: 3D culture; Hydrogels; In vitro model; Osteoclasts
    • Accession Number:
      0 (Hydrogels)
      9007-34-5 (Collagen)
      EC 3.4.22.38 (Cathepsin K)
      0 (Cytokines)
    • Publication Date:
      Date Created: 20240325 Date Completed: 20240424 Latest Revision: 20240614
    • Publication Date:
      20240614
    • Accession Number:
      10.1016/j.jdent.2024.104957
    • Accession Number:
      38527517