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- Author or Editor: John D. Kisiday x
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Abstract
Objective—To compare the mesenchymal stem cell (MSC) yield and chondrogenic and osteogenic differentiation from 5- and 50-mL bone marrow aspirates from horses.
Animals—Six 2- to 5-year-old mixed-breed horses.
Procedures—2 sequential 5-mL aspirates were drawn from 1 ilium or sternebra. A single 50-mL aspirate was drawn from the contralateral ilium, and 2 sequential 50-mL aspirates were drawn from a second sternebra. The MSC yield was determined through the culture expansion process. Chondrogenesis and osteogenesis were evaluated by means of conventional laboratory methods.
Results—The second of the 2 sequential 50-mL sternal aspirates yielded few to no MSCs. Independent of location, the highest density of MSCs was in the first of the 2 sequential 5-mL fractions, although with subsequent culture expansion, the overall yield was not significantly different between the first 5-mL and first 50-mL fractions. Independent of location, chondrogenesis and osteogenesis were not significantly different among fractions. Independent of fraction, the overall cell yield and chondrogenesis from the ilium were significantly higher than that from the sternum.
Conclusions and Clinical Relevance—This study failed to detect an additional benefit of 50-mL aspirates over 5-mL aspirates for culture-expanding MSCs for equine clinical applications. Chondrogenesis was highest for MSCs from ilial aspirates, although it is not known whether chondrogenesis is indicative of activation of other proposed pathways by which MSCs heal tissues.
Abstract
Objective—To develop an in vitro model of cartilage injury in full-thickness equine cartilage specimens that can be used to simulate in vivo disease and evaluate treatment efficacy.
Sample—15 full-thickness cartilage explants from the trochlear ridges of the distal aspect of the femur from each of 6 adult horses that had died from reasons unrelated to the musculoskeletal system.
Procedures—To simulate injury, cartilage explants were subjected to single-impact uniaxial compression to 50%, 60%, 70%, or 80% strain at a rate of 100% strain/s. Other explants were left uninjured (control specimens). All specimens underwent a culture process for 28 days and were subsequently evaluated histologically for characteristics of injury and early stages of osteoarthritis, including articular surface damage, chondrocyte cell death, focal cell loss, chondrocyte cluster formation, and loss of the extracellular matrix molecules aggrecan and types I and II collagen.
Results—Compression to all degrees of strain induced some amount of pathological change typical of clinical osteoarthritis in horses; however, only compression to 60% strain induced significant changes morphologically and biochemically in the extracellular matrix.
Conclusions and Clinical Relevance—The threshold strain necessary to model injury in full-thickness cartilage specimens from the trochlear ridges of the distal femur of adult horses was 60% strain at a rate of 100% strain/s. This in vitro model should facilitate study of pathophysiologic changes and therapeutic interventions for osteoarthritis.
Abstract
Objective—To evaluate the effect of fibrin concentrations on mesenchymal stem cell (MSC) migration out of autologous and commercial fibrin hydrogels.
Sample—Blood and bone marrow from six 2- to 4-year-old horses.
Procedures—Autologous fibrinogen was precipitated from plasma and solubilized into a concentrated solution. Mesenchymal stem cells were resuspended in fibrinogen solutions containing 100%, 75%, 50%, and 25% of the fibrinogen precipitate solution. Fibrin hydrogels were created by mixing the fibrinogen solutions with MSCs and thrombin on tissue culture plates. After incubation for 24 hours in cell culture medium, the MSCs that had migrated onto the tissue culture surface and beyond the boundary of the hydrogels were counted. This procedure was repeated with a commercial fibrin sealant.
Results—Hydrogel-to-surface MSC migration was detected for all fibrin hydrogels. Migration from the 25% autologous hydrogels was 7.3-, 5.2-, and 4.6-fold higher than migration from 100%, 75%, and 50% autologous hydrogels, respectively. The number of migrating cells from 100%, 75%, and 50% autologous hydrogels did not differ significantly. With commercial fibrin sealant, the highest magnitude of migration was from the 25% hydrogels, and it was 26-fold higher than migration from 100% hydrogels. The 75% and 50% hydrogels resulted in migration that was 9.5- and 4.2-fold higher than migration from the 100% hydrogels, respectively.
Conclusions and Clinical Relevance—MSC migration from fibrin hydrogels increased with dilution of the fibrinogen component for both autologous and commercial sources. These data supported the feasibility of using diluted fibrin hydrogels for rapid delivery of MSCs to the surface of damaged tissues.
