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Canadian journal of veterinary research = Revue canadienne de recherche veterinaire2006; 70(4); 277-284;

In vitro heterogeneity of osteogenic cell populations at various equine skeletal sites.

Abstract: Bone cell cultures were evaluated to determine if osteogenic cell populations at different skeletal sites in the horse are heterogeneous. Osteogenic cells were isolated from cortical and cancellous bone in vitro by an explant culture method. Subcultured cells were induced to differentiate into bone-forming osteoblasts. The osteoblast phenotype was confirmed by immunohistochemical testing for osteocalcin and substantiated by positive staining of cells for alkaline phosphatase and the matrix materials collagen and glycosaminoglycans. Bone nodules were stained by the von Kossa method and counted. The numbers of nodules produced from osteogenic cells harvested from different skeletal sites were compared with the use of a mixed linear model. On average, cortical bone sites yielded significantly greater numbers of nodules than did cancellous bone sites. Between cortical bone sites, there was no significant difference in nodule numbers. Among cancellous sites, the radial cancellous bone yielded significantly more nodules than did the tibial cancellous bone. Among appendicular skeletal sites, tibial metaphyseal bone yielded significantly fewer nodules than did all other long bone sites. This study detected evidence of heterogeneity of equine osteogenic cell populations at various skeletal sites. Further characterization of the dissimilarities is warranted to determine the potential role heterogeneity plays in differential rates of fracture healing between skeletal sites. Des cultures de cellules osseuses ont été réalisées afin de déterminer si les populations de cellules ostéogéniques provenant de différents sites squelettiques chez le cheval sont hétérogènes. Les cellules ostéogéniques ont été isolées in vitro de l’os cortical et de l’os spongieux par une méthode de culture d’explant. Les cellules cultivées ont été induites à se différencier en ostéoblastes formant de l’os. Le phénotype d’ostéoblaste a été confirmé par épreuve immunohistochimique pour l’ostéocalcine et corroborer par coloration positive des cellules pour la phosphatase alcaline de même que pour le collagène et les glycosaminoglycans. Les nodules osseux ont été colorés par la méthode de von Kossa et dénombrés. Les quantités de nodules produits par les cellules ostéogéniques recueillies des différents sites squelettiques ont été comparées en utilisant un modèle linéaire mixte. En moyenne, les sites d’os cortical ont permis d’obtenir un nombre significativement plus grand de nodules que les sites d’os spongieux. Entre les sites d’os cortical, il n’y avait pas de différence significative dans le nombre de nodules. Parmi les sites d’os spongieux, le site radial a permis de recueillir plus de nodules que le site tibial. Parmi les sites squelettiques appendiculaires, l’os métaphysaire tibial a permis d’obtenir significativement moins de nodules que tous les autres sites d’os longs. La présente étude a permis de démontrer l’hétérogénéité des populations de cellules ostéogéniques à différents sites squelettiques. Une caractérisation supplémentaire des différences observées est de mise afin de déterminer le rôle potentiel joué par l’hétérogénéité dans la variabilité de la guérison des fractures entre les différents sites squelettiques. (Traduit par Docteur Serge Messier)
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Publication Date: 2006-10-18 PubMed ID: 17042380PubMed Central: PMC1562541
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research looks into the diversity of osteogenic cell populations—the cells responsible for bone formation—from different skeletal locations in the horse. The study finds considerable differences when comparing different sites and calls for a more comprehensive examination of these variations.

Research Methodology

  • The study made use of bone cell cultures to look at the heterogeneity of osteogenic cells from varying skeletal locations in a horse.
  • Osteogenic cells were separated from cortical and cancellous (spongy) bone using an explant culture method. The extracted cells were then subcultured—propagated in a culture medium—providing a population large enough to investigate.
  • Methods employed to affirm the osteogenic capability of the cells included immunohistochemical checks for osteocalcin—a protein specifically produced by osteoblasts—and positive staining for alkaline phosphatase, collagen, and glycosaminoglycans. These are all important components of bone cells.
  • Bone nodules were dyed using the von Kossa method—a common staining technique to detect mineralization—to visualize and count.

Results and Conclusions

  • The count revealed that the number of nodules produced by osteogenic cells differed significantly between skeletal sites.
  • Cortical bone locations yielded a higher amount of nodules, on average, than cancellous bone sites. However, no significant difference was seen when comparing cortical bone sites against each other.
  • When comparing cancellous sites, radial cancellous bone saw a higher nodule count compared to tibial cancellous bone.
  • Among appendicular skeletal sites—the limbs—the tibial metaphyseal bone showed a significantly lower nodule count than all other long bone sites.
  • The findings indicate a noticeable heterogeneity of equine osteogenic cell populations across various skeletal sites. The researchers call for further investigation of these disparities to potentially explain differential rates of fracture healing between skeletal sites.

Cite This Article

APA
McD￮ LA, Anderson GI, Wright GM, Ryan DA. (2006). In vitro heterogeneity of osteogenic cell populations at various equine skeletal sites. Can J Vet Res, 70(4), 277-284.

Publication

Canadian journal of veterinary research = Revue canadienne de recherche veterinaire
ISSN: 0830-9000
NlmUniqueID: 8607793
Country: Canada
Language: English
Volume: 70
Issue: 4
Pages: 277-284

Researcher Affiliations

McD￮, Laurie A
  • Departments of Health Management, University of Prince Edward Island, Charlottetown. lmcd￮@upei.ca
Anderson, Gail I
    Wright, Glenda M
      Ryan, Daniel A J

        MeSH Terms

        • Alkaline Phosphatase / metabolism
        • Animals
        • Bone Regeneration
        • Bone Transplantation / veterinary
        • Bone and Bones / cytology
        • Bone and Bones / physiology
        • Cell Differentiation / physiology
        • Cells, Cultured
        • Collagen / metabolism
        • Culture Techniques
        • Fracture Healing / physiology
        • Glycosaminoglycans / metabolism
        • Horses
        • Linear Models
        • Osteoblasts / cytology
        • Osteoblasts / metabolism
        • Osteoblasts / physiology
        • Osteocalcin / metabolism
        • Osteocytes / cytology
        • Osteocytes / metabolism
        • Osteocytes / physiology
        • Osteogenesis / physiology
        • Radius / cytology
        • Tibia / cytology

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        Citations

        This article has been cited 4 times.
        1. Al Naem M, Bourebaba L, Kucharczyk K, Röcken M, Marycz K. Therapeutic mesenchymal stromal stem cells: Isolation, characterization and role in equine regenerative medicine and metabolic disorders. Stem Cell Rev Rep 2020 Apr;16(2):301-322.
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        2. Adams MK, Goodrich LR, Rao S, Olea-Popelka F, Phillips N, Kisiday JD, McIlwraith CW. Equine bone marrow-derived mesenchymal stromal cells (BMDMSCs) from the ilium and sternum: are there differences?. Equine Vet J 2013 May;45(3):372-5.
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        4. Koch TG, Berg LC, Betts DH. Current and future regenerative medicine - principles, concepts, and therapeutic use of stem cell therapy and tissue engineering in equine medicine. Can Vet J 2009 Feb;50(2):155-65.
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