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Topic:Bone Marrow
Bone marrow in horses is a spongy tissue found within the cavities of certain bones, such as the pelvis, ribs, and femur. It is a vital component of the hematopoietic system, responsible for the production of blood cells, including red blood cells, white blood cells, and platelets. These cells are crucial for oxygen transport, immune function, and blood clotting, respectively. Bone marrow consists of two types: red marrow, which is actively involved in blood cell production, and yellow marrow, which primarily stores fat. The composition and activity of bone marrow can change with age, health status, and physiological demands. This page compiles peer-reviewed research studies and scholarly articles that explore the structure, function, and clinical implications of bone marrow in equine health and disease.
Evaluation of intra-articular mesenchymal stem cells to augment healing of microfractured chondral defects. This study evaluated intra-articular injection of bone marrow-derived mesenchymal stem cells (BMSCs) to augment healing with microfracture compared with microfracture alone. Methods: Ten horses (aged 2.5 to 5 years) had 1-cm2 defects arthroscopically created on both medial femoral condyles of the stifle joint (analogous to the human knee). Defects were debrided to subchondral bone followed by microfracture. One month later, 1 randomly selected medial femorotibial joint in each horse received an intra-articular injection of either 20 × 10(6) BMSCs with 22 mg of hyaluronan or 22 mg of hyalurona...
Lateral ridge augmentation using an equine flex bone block infused with recombinant human platelet-derived growth factor BB: a clinical and histologic study. This paper reports on a patient who underwent horizontal ridge augmentation using recombinant human platelet-derived growth factor BB (rhPDGF-BB) in combination with a collagenate equine block. Ninety days after the first surgery, 8 mm of new bone was noted on a computed tomography scan, and three 5-mm implants were placed. Histology, performed using confocal laser scanning microscopy, showed a large amount of newly formed bone well characterized with osteon and resorption lacunae, which demonstrated the intense bone remodeling. This study supports the use of rhPDGF-BB in combination with allo...
Osteogenic differentiation of equine cord blood multipotent mesenchymal stromal cells within coralline hydroxyapatite scaffolds in vitro. To investigate the osteogenic differentiation potential of equine umbilical cord blood-derived multipotent mesenchymal stromal cells (CB-MSC) within coralline hydroxyapatite scaffolds cultured in osteogenic induction culture medium. Methods: Scaffolds seeded with equine CB-MSC were cultured in cell expansion culture medium (control) or osteogenic induction medium (treatment). Cell viability and distribution were confirmed by the MTT cell viability assay and DAPI nuclear fluorescence staining, respectively. Osteogenic differentiation was evaluated after 10 days using reverse transcription polym...
Chondrogenic effects of exogenous retinoic acid or a retinoic acid receptor antagonist (LE135) on equine chondrocytes and bone marrow-derived mesenchymal stem cells in monolayer culture. To determine effects of various concentrations of retinoic acid (RA) or a synthetic RA receptor antagonist (LE135) on equine chondrocytes or bone marrow-derived equine mesenchymal stem cells (BMDMSCs) in monolayer cultures. Methods: Articular cartilage and BMDMSCs from 5 clinically normal horses. Methods: Monolayers of chondrocytes cultured in standard media and of BMDMSCs cultured in chondrogenic media were treated with RA at concentrations of 0, 0.1, 1, or 10 μM or LE135 at concentrations of 0, 0.1, 1, or 10 μM on day 0. On days 7 and 14, samples were analyzed for DNA concentration, chondr...
Immunophenotype and gene expression profiles of cell surface markers of mesenchymal stem cells derived from equine bone marrow and adipose tissue. Bone marrow and adipose tissue are the two main sources of mesenchymal stem cell (MSC). The aim of this work was to analyse the immunophenotype of 7 surface markers and the expression of a panel of 13 genes coding for cell surface markers in equine bone marrow and adipose tissue-derived MSCs obtained from 9 horses at third passage. The tri-lineage differentiation was confirmed by specific staining. Equine MSCs from both sources were positive for the MSC markers CD29 and CD90, while were negative for CD44, CD73, CD105, CD45 and CD34. The gene expression of these molecules was also evaluated by ...
Acute myeloid leukaemia (M6B: pure acute erythroid leukaemia) in a Thoroughbred foal. A 10-week-old Thoroughbred filly was referred for anaemia of 4 weeks' duration. Haematology revealed severe anaemia and panleucopenia. Cytological examination of bone marrow smears revealed a myeloid to erythroid ratio <0.02:1 (reference range 0.5-2.4:1.0) and an abundance of erythroid precursor cells. The erythroid cell population included rubriblasts, prorubricytes and rubricytes, with only scant numbers of metarubricytes present. There were numerous mitotic erythroid cells, some of which were atypical and megaloblastic. These cytomorphological changes are consistent with pure acute eryth...
Comparative study of equine bone marrow and adipose tissue-derived mesenchymal stromal cells. Mesenchymal stromal cells (MSCs) represent an attractive source for regenerative medicine. However, prior to their application, fundamental questions regarding molecular characterisation, growth and differentiation of MSCs must be resolved. Objective: To compare and better understand the behaviour of equine MSCs obtained from bone marrow (BM) and adipose tissue (AT) in culture. Methods: Five horses were included in this study. Proliferation rate was measured using MTT assay and cell viability; apoptosis, necrosis and late apoptosis and necrosis were evaluated by flow cytometry. The mRNA expres...
Impact of successive freezing-thawing cycles on 3-T magnetic resonance images of the digits of isolated equine limbs. To assess the impact of cycles of freezing and thawing on magnetic resonance (MR) images (obtained by use of a 3-T magnet) of equine feet examined ex vivo. Methods: 9 forelimbs from 9 horse cadavers. Methods: 9 forefeet underwent MR imaging first at ambient temperature within 12 hours after the horses' death and then after each freezing-thawing cycle. Three digits underwent freezing and thawing (at 4°C for 36 hours) 2 times, 3 digits underwent freezing and thawing (at 4°C for 36 hours) once and rescanning after 24 hours at ambient temperature, and 3 digits underwent freezing and thawing at a...
Direct delayed human adenoviral BMP-2 or BMP-6 gene therapy for bone and cartilage regeneration in a pony osteochondral model. To evaluate healing of surgically created large osteochondral defects in a weight-bearing femoral condyle in response to delayed percutaneous direct injection of adenoviral (Ad) vectors containing coding regions for either human bone morphogenetic proteins 2 (BMP-2) or -6. Methods: Four 13mm diameter and 7mm depth circular osteochondral defects were drilled, 1/femoral condyle (n=20 defects in five ponies). At 2 weeks, Ad-BMP-2, Ad-BMP-6, Ad-green fluorescent protein (GFP), or saline was percutaneously injected into the central drill hole of the defect. Quantitative magnetic resonance imaging (...
Implantation of bone marrow-derived mesenchymal stem cells demonstrates improved outcome in horses with overstrain injury of the superficial digital flexor tendon. Mesenchymal stem (progenitor; stromal) cell (MSC) therapy has gained popularity for the treatment of equine tendon injuries but without reports of long-term follow-up. Objective: To evaluate the safety and reinjury rate of racehorses after intralesional MSC injection in a large study of naturally occurring superficial digital flexor tendinopathy and to compare these data with those published for other treatments. Methods: Safety was assessed clinically, ultrasonographically, scintigraphically and histologically in a cohort of treated cases: 141 client-owned treated racehorses followed-up for a...
Evaluation of senescence in mesenchymal stem cells isolated from equine bone marrow, adipose tissue, and umbilical cord tissue. Mesenchymal stem cells (MSCs) from adult and neonatal tissues are intensively investigated for their use in regenerative medicine. The purpose of this study was to compare the onset of replicative senescence in MSCs isolated from equine bone marrow (BMSC), adipose tissue (ASC), and umbilical cord tissue (UCMSC). MSC proliferation (cell doubling), senescence-associated β-galactosidase staining, telomere length, Sox-2, and lineage-specific marker expression were assessed for MSCs harvested from tissues of 4 different donors. The results show that before senescence ensued, all cell types prolife...
The effect of environmental storage conditions on bone marrow fat determination in three species. Diagnostic laboratories are frequently required to assess the antemortem nutritional condition of deceased animals. The percentage of fat in the bone marrow is used to diagnose starvation because this fat depot is typically the last in the body to be depleted. Diagnosticians rely on measurement of bone marrow adipose content using fat solvent-extraction methods; however, the effects of tissue storage conditions before processing have not been fully assessed. The current study focuses on evaluating the effects of 3 storage conditions (refrigeration [4 °C], freezing [-20 °C], and ambient tempe...
Regenerative medicine for the treatment of musculoskeletal overuse injuries in competition horses. Tissue repair in musculoskeletal injuries is often a slow and sometimes incomplete process. Regenerative medicine based on the use of growth factors (GFs) and cell therapy is aimed at improving the quality and speed of tendon and ligament healing. The aim of this study was to evaluate the potential for the administration of a combination of autologous platelet-rich plasma (PRP) and freshly isolated bone marrow mononucleated cells (BMMNCs) in 13 competition horses affected by overuse musculoskeletal injuries (suspensory ligament desmopathy and superficial flexor tendinopathy) and refractory to ...
The regenerative medicine laboratory: facilitating stem cell therapy for equine disease. This article focuses on the emerging field of equine regenerative medicine with an emphasis on the use of mesenchymal stem cells (MSCs) for orthopedic diseases. We detail laboratory procedures and protocols for tissue handling and MSC isolation, characterization, expansion, and cryopreservation from bone marrow, fat, and placental tissues. We provide an overview of current clinical uses for equine MSCs and how MSCs function to heal tissues. Current laboratory practices in equine regenerative medicine mirror those in the human field. However, the translational use of autologous and allogeneic M...
Evaluation of equine peripheral blood apheresis product, bone marrow, and adipose tissue as sources of mesenchymal stem cells and their differentation potential. To evaluate effects of apheresis on mesenchymal stem cells (MSCs) and compare those MSCs with MSCs obtained from adipose tissue or bone marrow (BM). Methods: Samples obtained from 6 adult horses. Methods: Samples of blood from a peripheral vein, adipose tissue, and BM aspirate were obtained from each horse. Samples were processed via apheresis of blood and techniques reported elsewhere for adipose tissue and BM. Cultures were maintained until adherence and subsequently were subjected to differentiation protocols to evaluate adipogenic, osteoblastogenic, and chondrogenic potential. Results: Aph...
Comparison of equine bone marrow-, umbilical cord matrix and amniotic fluid-derived progenitor cells. The aim of the study was to compare in vitro the stemness features of horse progenitor cells derived from bone marrow (BM-MSCs), amniotic fluid (AF-MSCs) and umbilical cord matrix (EUC-MSCs). It has been suggested that there may be a stem cell population within both umbilical cord matrix and amniotic fluid. However, little knowledge exists about the characteristics of these progenitor cells within these sources in the equine species. This study wanted to investigate an alternative and non-invasive stem cell source for the equine tissue engineering and to learn more about the properties of thes...
Magnetic resonance imaging features of serous atrophy of bone marrow fat in the distal limb of three horses. Emaciated human patients have changes in the fat content in medullary bone that are consistent with serous atrophy of the bone marrow histologically. Serous atrophy has been identified at postmortem examination in horses; however, the magnetic resonance (MR) characteristics have not been documented. Herein we describe the abnormalities of the bone marrow and medullary bone detected by low-field and high-field MR imaging of the distal limbs of three emaciated horses. These low- and high-field MR imaging abnormalities are characterized by a decrease in signal intensity on T1-weighted images in c...
Expansion of mesenchymal stem cells on fibrinogen-rich protein surfaces derived from blood plasma. Mesenchymal stem cells (MSCs) are present in low density in bone marrow and culture expansion is necessary to obtain sufficient numbers for many proposed therapies. Researchers have characterized MSC growth on tissue culture plastic (TCP), although few studies have explored proliferation on other growth substrates. Using adult equine MSCs, we evaluated proliferation on fibrinogen-rich precipitate (FRP) surfaces created from blood plasma. When seeded at 1 × 10(4) cells/cm(2) and passaged five times over 10 days, MSCs on FRP in medium containing fibroblast growth factor 2 (FGF2) resulted in a â...
Cloning and tissue expression of the equine transferrin receptor. Characterization of anemia in horses presents a challenge, as they do not release reticulocytes into peripheral blood. Transferrin receptor (TfR) expression is highest on erythroid cells in people and rats, and measurement of a soluble serum form (sTfR) is used to quantify erythropoiesis in these species. We hypothesized that equine TfR (eTfR) expression is similar in quantity and distribution to that in these other species and thus has potential for characterization of the regenerative response in anemic horses. Objective: This study was conducted to clone and sequence the eTfR gene and measu...
Periodontal regeneration capacity of equine particulate bone in canine alveolar bone defects. This study was performed to evaluate the periodontal wound healing effect of particulate equine bone mineral on canine alveolar bone defects. Methods: Twelve adult male beagle dogs were used as study subjects. The mandibular second and fourth premolars were extracted prior to the experimental surgery, and the extraction sites were allowed to heal for 8 weeks. After periodontal probing, two-walled defects were created at the mesial and distal sides of the mandibular third premolars bilaterally, and the defects were filled with equine particulate bone with collagen membrane or bovine particulate...
Resolution of lesions on STIR images is associated with improved lameness status in horses. Magnetic resonance (MR) imaging is important in diagnosing musculoskeletal injuries in horses. However, there is still much to learn regarding the significance of lesions identified in equine MR images. Of particular importance is the clinical significance of signal change as a function of pulse sequence. We hypothesized that a resolution of tendon, ligament, and bone marrow lesions on short-tau inversion recovery (STIR) images would be associated with a return to soundness, and that a persistence of tendon and ligament lesions on only T1-weighted (T1w) gradient recalled echo (GRE) images woul...
Analysis of CD14 expression levels in putative mesenchymal progenitor cells isolated from equine bone marrow. A long-term goal of mesenchymal progenitor cell (MPC) research is to identify cell-surface markers to facilitate MPC isolation. One reported MPC feature in humans and other species is lack of CD14 (lipopolysaccharide receptor) expression. The aim of this study was to evaluate CD14 as an MPC sorting marker. Our hypothesis was that cells negatively selected by CD14 expression would enrich MPC colony formation compared with unsorted and CD14-positive fractions. After validation of reagents, bone marrow aspirate was obtained from 12 horses. Fresh and cultured cells were analyzed by flow cytometry ...
Comparison of the osteogenic potential of equine mesenchymal stem cells from bone marrow, adipose tissue, umbilical cord blood, and umbilical cord tissue. To determine the optimal osteogenic source of equine mesenchymal stem cells (eMSCs) and optimize collection of and expansion conditions for those cells. Methods: 10 adult Quarter Horses and 8 newborn Thoroughbred foals. Methods: eMSCs were isolated from bone marrow (BM), adipose tissue, and umbilical cord blood and tissue, and the osteogenic potential of each type was assessed. Effects of anatomic site, aspiration volume, and serum type on eMSC yield from BM were investigated. Results: BM-eMSCs had the highest overall expression of the osteogenic genes Cbfa1, Osx, and Omd and staining for ALP ...
Optimisation of bone marrow aspiration from the equine sternum for the safe recovery of mesenchymal stem cells. Mesenchymal stem cell (MSC) therapy for orthopaedic disease is being used with increasing frequency; there is a need to define a safe, reliable and effective technique for the recovery of MSCs from the sternum of the horse. Objective: To describe an optimised safe technique for obtaining bone marrow-derived MSCs from the sternum of the Thoroughbred horse. Methods: The anatomical relationship of the sternum with the heart and internal anatomy was demonstrated in cadavers. Sternal anatomy was evaluated ultrasonographically and after midline sectioning. Sternebrae were examined histologically aft...
Concentrated bone marrow aspirate improves full-thickness cartilage repair compared with microfracture in the equine model. The purpose of this study was to compare the outcomes of treatment with bone marrow aspirate concentrate, a simple, one-step, autogenous, and arthroscopically applicable method, with the outcomes of microfracture with regard to the repair of full-thickness cartilage defects in an equine model. Methods: Extensive (15-mm-diameter) full-thickness cartilage defects were created on the lateral trochlear ridge of the femur in twelve horses. Bone marrow was aspirated from the sternum and centrifuged to generate the bone marrow concentrate. The defects were treated with bone marrow concentrate and mic...
Isolation of equine bone marrow-derived mesenchymal stem cells: a comparison between three protocols. There is a need to assess and standardise equine bone marrow (BM) mesenchymal stem cell (MSC) isolation protocols in order to permit valid comparisons between therapeutic trials at different sites. Objective: To compare 3 protocols of equine BM MSC isolation: adherence to a plastic culture dish (Classic) and 2 gradient density separation protocols (Percoll and Ficoll). Methods: BM aspirates were harvested from the sternum of 6 mares and MSCs isolated by all 3 protocols. The cell viability after isolation, MSC yield, number of MSCs attained after 14 days of culture and the functional characteri...
Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage. To quantify the structural characteristics and nanomechanical properties of aggrecan produced by adult bone marrow stromal cells (BMSCs) in peptide hydrogel scaffolds and compare to aggrecan from adult articular cartilage. Methods: Adult equine BMSCs were encapsulated in 3D-peptide hydrogels and cultured for 21 days with TGF-β1 to induce chondrogenic differentiation. BMSC-aggrecan was extracted and compared with aggrecan from age-matched adult equine articular cartilage. Single molecules of aggrecan were visualized by atomic force microscopy-based imaging and aggrecan nanomechanical stiffness...
Third metacarpal condylar fatigue fractures in equine athletes occur within previously modelled subchondral bone. Bone modelling and remodelling reduce the risk of fatigue fractures; the former by adapting bone to its loading circumstances, the latter by replacing fatigued bone. Remodelling transiently increases porosity because of the normal delay in onset of the formation phase of the remodelling sequence. Protracted intense loading suppresses remodelling leaving modelling as the only means of maintaining bone strength. We therefore hypothesized that race horses with fatigue fractures of the distal third metacarpal bone (MC3) will have reduced porosity associated with suppressed remodelling while contin...
Coculture of equine mesenchymal stem cells and mature equine articular chondrocytes results in improved chondrogenic differentiation of the stem cells. Bone marrow derived mesenchymal stem cells (MSCs) can be used to repair articular cartilage defects, these cells should be properly stimulated so that they could differentiate morphologically and hold cellular synthetic features closer to maturely differentiated chondrocytes. It is well known that tissue specific environment plays an important role in cell fate determination. Once improved isolation, proliferation and differentiation protocols have been developed, the likelihood of spontaneous differentiation of MSCs into divergent lineages will be reduced, thus increasing their value for cart...
Gelatinous marrow transformation and hematopoietic atrophy in a miniature horse stallion. Gelatinous marrow transformation, or serous atrophy of bone marrow fat, has been noted in livestock, laboratory animals, and wildlife in association with an inadequate plane of nutrition, inanition, or intoxication. This is a report of gelatinous marrow transformation and hematopoietic marrow atrophy in a 5-year-old miniature horse stallion. The horse had oral malformations leading to poor food assimilation and emaciation. A bone marrow biopsy obtained to investigate persistent anemia and leukopenia showed hematopoietic atrophy and replacement of fat with a granular extracellular substance, wh...
