Equine osteoclast-like cells generated in vitro demonstrate similar characteristics to directly isolated mature osteoclasts

Gray, A.W.; Davies, Malcolm; Jeffcott, L. B.

doi:10.1053/rvsc.1999.0367

Cited by 5 publications

(6 citation statements)

References 32 publications

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“…PBMCs and bone marrow derived monocytes/macrophages (BMM) are ubiquitously used for in-vitro osteoclastogenesis. Previous investigations have presented in-vitro OC formation from different mammalian species such as humans [29], primates [30], rodents [31], dogs [32], cats [33,34], rabbits [35], horses [36], and pigs [37]. To the best of our knowledge, the current work is the first to report bears' OC formation in-vitro.…”

Section: Discussionmentioning

confidence: 81%

Hibernating bear serum hinders osteoclastogenesis in-vitro

et al. 2020

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Bears do not suffer from osteoporosis during hibernation, which is associated with long-term inactivity, lack of food intake, and cold exposure. However, the mechanisms involved in bone loss prevention have scarcely been elucidated in bears. We investigated the effect of serum from hibernating Japanese black bears (Ursus thibetanus japonicus) on differentiation of peripheral blood mononuclear cells (PBMCs) to osteoclasts (OCs). PBMCs collected from 3 bears were separately cultured with 10% serum of 4 active and 4 hibernating bears (each individual serum type was assessed separately by a bear PBMCs), and differentiation were induced by treatment with macrophage colony stimulating factor (M-CSF) and receptor activator of NF-kB ligand (RANKL). PBMCs that were cultured with the active bear serum containing medium (ABSM) differentiated to multi-nucleated OCs, and were positive for TRAP stain. However, cells supplemented with hibernating bear serum containing medium (HBSM) failed to form OCs, and showed significantly lower TRAP stain (p < 0.001). On the other hand, HBSM induced proliferation of adipose derived mesenchymal stem cells (ADSCs) similarly to ABSM (p > 0.05), indicating no difference on cell growth. It was revealed that osteoclastogenesis of PBMCs is hindered by HBSM, implying an underlying mechanism for the suppressed bone resorption during hibernation in bears. In addition, this study for the first time showed the formation of bears' OCs in-vitro.

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Section: Discussionmentioning

confidence: 81%

Hibernating bear serum hinders osteoclastogenesis in-vitro

et al. 2020

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“…Equine osteoclasts have been studied in vitro in only a limited number of studies to date [62][63][64][65][66]83 despite their important role in subchondral bone remodelling events in growing horses 67,68 and focal subchondral resorption with repetitive exercise in adults that may precipitate fractures. 12,13,92 Equine osteoclasts differentiated from sternal bone marrow aspirates have been investigated but the osteoclast counts were not reported.…”

Section: Discussionmentioning

confidence: 99%

“…In the past, equine osteoclast resorption has been studied on dentine slices, a commercial substrate, and, although small circular pits were observed; variation in their number and difficulties with the toluidine blue staining technique was mentioned and it was not quantified. 62,63,65 It is interesting to note that equine bone slices have been employed as a substrate in two previous studies of osteoclasts from other species: chick embryo osteoclasts had barely detectable resorption (mean of 0.003%) 95 and human osteoclasts were reported to resorb equine bone, but it was not quantified. 96 Combined, these results reveal that equine osteoclasts can now be differentiated from sternal bone marrow aspirates in standing horses and cultured on equine bone slices to quantify their function (resorptive response).…”

Section: Discussionmentioning

confidence: 99%

“…The duration of the cultures, 14 days for the pure osteoclast cultures and 22 days for the osteoclasts cultured on bone, was informed by prior investigations in horses and other species 62,63,65,75 and established in our laboratory by many preliminary experiments (data not published). We first observed the appearance of early osteoclast phenotypes on d7 of differentiation in the pure cultures from haematopoietic stem cells.…”

Section: Discussionmentioning

confidence: 99%

“…Only a few laboratories have cultured equine osteoclasts to date. In these studies, haematopoietic stem cells were differentiated into mature osteoclasts from post mortem femoral bone marrow [62][63][64][65] and, more recently and conveniently, from standing equine sternal bone marrow aspirates. 66 However, no study to date has investigated equine osteoclast growth and activity on equine bone slices, their natural substrate, to our knowledge.…”

Section: Introductionmentioning

confidence: 99%

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Anin vitromodel for discovery of osteoclast specific biomarkers towards identification of racehorses at risk for catastrophic fractures

Malek

Richard

Beauchamp

2022

Equine Veterinary Journal

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Background Focal bone microcracks with osteoclast recruitment and bone lysis, may reduce fracture resistance in racehorses. As current imaging does not detect all horses at risk for fracture, the discovery of novel serum biomarkers of bone resorption or osteoclast activity could potentially address this unmet clinical need. The biology of equine osteoclasts on their natural substrate, equine bone, has never been studied in vitro and may permit identification of specific biomarkers of their activity. Objectives (1) Establish osteoclast cultures on equine bone, (2) Measure biomarkers (tartrate resistant acid phosphatase isoform 5b [TRACP‐5b] and C‐terminal telopeptide of type I collagen [CTX‐I]) in vitro and (3) Study the effects of inflammation. Study design In vitro experiments. Methods Haematopoietic stem cells, from five equine sternal bone marrow aspirates, were differentiated into osteoclasts and cultured either alone or on equine bone slices, with or without a pro‐inflammatory stimulus (IL‐1β or LPS). CTX‐I and TRACP‐5b were immunoassayed in the media. Osteoclast numbers and bone resorption area were assessed. Results TRACP‐5b increased over time in osteoclast cultures without bone (p < 0.0001) and correlated with osteoclast number (r = 0.63, p < 0.001). CTX‐I and TRACP‐5b increased with time for cultures with bone (p = 0.002; p = 0.02 respectively), correlated with each other (r = 0.64, p < 0.002) and correlated with bone resorption (r = 0.85, p < 0.001; r = 0.82, p < 0.001 respectively). Inflammation had no measurable effects. Main limitations Specimen numbers limited. Conclusions Equine osteoclasts were successfully cultured on equine bone slices and their bone resorption quantified. TRACP‐5b was shown to be a biomarker of equine osteoclast number and bone resorption for the first time; CTX‐I was also confirmed to be a biomarker of equine bone resorption in vitro. This robust equine specific in vitro assay will help the study of osteoclast biology.

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