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The proliferative response of equine chondrocytes to bovine lymph node proteins in vitro

Posted on:2008-07-23Degree:M.SType:Thesis
University:Clemson UniversityCandidate:Reed, Alison MarieFull Text:PDF
GTID:2444390005454098Subject:Biology
Abstract/Summary:
Lameness poses a substantial challenge to the horse industry, costing millions of dollars per year in treatments, labor, and loss of use of performance horses. Osteoarthritis is a leading cause of chronic lameness. Treatments that stimulate cartilage development may promote cartilage healing. Many current treatment options for osteoarthritis have limited effectiveness and are expensive. Bovine supramammary lymph nodes are widely available at no cost and contain bioactive growth factors that could promote cartilage healing. The purpose of this study was to determine the proliferative effect of bovine lymph node extract on equine chondrocytes in vitro.; Equine chondrocytes were harvested from the scapular cartilage of adult horses. Lymph node extract (LNE) was prepared by mixing freeze-dried lymph node powder with PBS followed by heat inactivation. Chondrocytes were cultured in vitro, first in DMEM with 10% bovine growth serum (BGS) and antibiotics, then with treatment media containing either BGS or LNE on equal and unequal protein bases. Additionally, LNE fractions from ammonium sulfate precipitation were examined as BGS replacements. The response of the cells to IGF-I and TGF-beta was also examined. DNA accumulation was measured using the CyQUANT DNA assay as a means to quantify cell stimulation, i.e. growth.; When BGS and LNE were compared on equal and unequal protein basis over a range of protein concentrations, there were no statistically significant differences observed in proliferation between BGS- and LNE- treated cells. However, in a separate experiment, cells treated with LNE had significantly higher cell proliferation than cells treated with BGS or different LNE fractions. IGF-I and TGF-beta had no significant effects on cell proliferation.; Overall, the results indicate LNE is capable of stimulating chondrocyte proliferation to an equal or greater degree than BGS, suggesting LNE could promote cartilage healing. Further studies are needed to determine if LNE can be used to develop new therapies for osteoarthritis in horses.
Keywords/Search Tags:LNE, Lymph node, Equine chondrocytes, Bovine, BGS, Promote cartilage healing, Protein
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