Phenotypic expression of equine articular chondrocytes grown in three-dimensional cultures supplemented with supraphysiologic concentrations of insulin-like growth factor-1.
Abstract: To assess the effects of supraphysiologic concentrations of insulin-like growth factor-1 (IGF-1) on morphologic and phenotypic responses of chondrocytes. Methods: Articular cartilage obtained from 2 young horses. Methods: Chondrocytes were suspended in fibrin cultures and supplemented with 25, 12.5, or 0 mg of IGF-1/ml of fibrin. Chondrocyte morphology and phenotypic expression were assessed histologically, using H&E and Alcian blue stains, immunoreaction to collagen type I and II, and in situ hybridization. Proteoglycan content, synthesis, and monomer size were analyzed. The DNA content was determined by bisbenzimide-fluorometric assay, and elution of IGF-1 into medium was determined by IGF-1 radioimmunoassay. Results: Both 12.5 and 25 kg of IGF-1/ml enhanced phenotypic expression of chondrocytes without inducing detrimental cellular or metabolic effects. Highest concentration of IGF-1 (25 microg/ml) significantly increased total DNA content, glycosaminoglycan (GAG) content, GAG synthesis, and size of proteoglycan monomers produced, compared with cultures supplemented with 12.5 microg of IGF-1/ml or untreated cultures. Histologic examination confirmed these biochemical effects. Matrix metachromasia, type-II collagen in situ hybridization and immunoreaction were increased in cultures treated with 25 microg of IGF-1/ml, compared with cultures supplemented with 12.5 microg of IGF-1/ml or untreated cultures. Conclusions: Chondrocytes exposed to high concentrations of IGF-1 maintained differentiated chondrocyte morphology and had enhanced synthesis of matrix molecules without inducing apparent detrimental effects on chondrocyte metabolism. These results suggest that application of such composites for in vivo use during cartilage grafting procedures should provide an anabolic effect on the grafted cells.
Publication Date: 2002-02-15 PubMed ID: 11843134DOI: 10.2460/ajvr.2002.63.301Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The study explores the impact of high concentrations of insulin-like growth factor-1 (IGF-1) on horse chondrocytes, which are cells in cartilage. Results found increased positive effects on cartilage cell appearance and characteristics without introducing harmful effects, suggesting potential use in cartilage graft procedures.
Methodology
- Chondrocytes, or cartilage cells, were extracted from the articular cartilage of two young horses.
- The cells were suspended in fibrin cultures and treated with IGF-1 in varying concentrations (25, 12.5, or 0 mg per ml of fibrin).
- Assessments were made using multiple techniques, among them histology (H&E and Alcian blue stains), immunoreaction to collagen type I and II, and in situ hybridization.
- Detailed analyses on proteoglycan content, monomer size, and synthesis were undertaken.
- DNA content within cells was determined by a bisbenzimide-fluorometric assay.
- The release of IGF-1 into the surrounding medium was quantified using a radioimmunoassay.
Results
- Phenotypic expression of chondrocytes was enhanced with IGF-1 treatment without inducing detrimental cellular or metabolic effects.
- The highest tested concentration of IGF-1 (25 micrograms/ml) spurred significant increases in DNA content, glycosaminoglycan (GAG) content, GAG synthesis, and the size of proteoglycan monomers produced. These increases were compared with cultures supplemented with lower concentrations of IGF-1 and untreated cultures.
- Increased matrix metachromasia, type-II collagen in situ hybridization, and immunoreaction were observed in cultures that received 25 micrograms of IGF-1 per millilitre. This was in comparison to cultures treated with lower concentrations of IGF-1 as well as untreated cultures.
Conclusion
- High concentrations of IGF-1 maintained chondrocyte morphology and increased the synthesis of matrix molecules without introducing harmful metabolic effects.
- The study findings suggest the potential application of such composites for in vivo use during cartilage grafting operations, expected to provide anabolic or constructive effects on the grafted cells.
Cite This Article
APA
Fortier LA, Nixon AJ, Lust G.
(2002).
Phenotypic expression of equine articular chondrocytes grown in three-dimensional cultures supplemented with supraphysiologic concentrations of insulin-like growth factor-1.
Am J Vet Res, 63(2), 301-305.
https://doi.org/10.2460/ajvr.2002.63.301 Publication
Researcher Affiliations
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
MeSH Terms
- Animals
- Cartilage, Articular / cytology
- Cell Culture Techniques / methods
- Cell Culture Techniques / veterinary
- Cells, Cultured
- Chondrocytes / cytology
- Culture Media
- DNA / analysis
- Glycosaminoglycans / analysis
- Horses
- Insulin-Like Growth Factor I
- Phenotype
Grant Funding
- AR08360 / NIAMS NIH HHS
Citations
This article has been cited 13 times.- Thampi P, Samulski RJ, Grieger JC, Phillips JN, McIlwraith CW, Goodrich LR. Gene therapy approaches for equine osteoarthritis. Front Vet Sci 2022;9:962898.
- Ekram S, Khalid S, Salim A, Khan I. Regulating the fate of stem cells for regenerating the intervertebral disc degeneration. World J Stem Cells 2021 Dec 26;13(12):1881-1904.
- Marques-Smith P, Kallerud AS, Johansen GM, Boysen P, Jacobsen AM, Reitan KM, Henriksen MM, Löfgren M, Fjordbakk CT. Is clinical effect of autologous conditioned serum in spontaneously occurring equine articular lameness related to ACS cytokine profile?. BMC Vet Res 2020 Jun 8;16(1):181.
- Müller S, Lindemann S, Gigout A. Effects of Sprifermin, IGF1, IGF2, BMP7, or CNP on Bovine Chondrocytes in Monolayer and 3D Culture. J Orthop Res 2020 Mar;38(3):653-662.
- Chen T, McCarthy MM, Guo H, Warren R, Maher SA. The Scaffold-Articular Cartilage Interface: A Combined In Vitro and In Silico Analysis Under Controlled Loading Conditions. J Biomech Eng 2018 Sep 1;140(9):0910021-7.
- Lang HM, Schnabel LV, Cassano JM, Fortier LA. Effect of needle diameter on the viability of equine bone marrow derived mesenchymal stem cells. Vet Surg 2017 Jul;46(5):731-737.
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- Hurtig MB, Buschmann MD, Fortier LA, Hoemann CD, Hunziker EB, Jurvelin JS, Mainil-Varlet P, McIlwraith CW, Sah RL, Whiteside RA. Preclinical Studies for Cartilage Repair: Recommendations from the International Cartilage Repair Society. Cartilage 2011 Apr;2(2):137-52.
- Mullen LM, Best SM, Ghose S, Wardale J, Rushton N, Cameron RE. Bioactive IGF-1 release from collagen-GAG scaffold to enhance cartilage repair in vitro. J Mater Sci Mater Med 2015 Jan;26(1):5325.
- Ortved KF, Begum L, Mohammed HO, Nixon AJ. Implantation of rAAV5-IGF-I transduced autologous chondrocytes improves cartilage repair in full-thickness defects in the equine model. Mol Ther 2015 Feb;23(2):363-73.
- Mullen LM, Best SM, Brooks RA, Ghose S, Gwynne JH, Wardale J, Rushton N, Cameron RE. Binding and release characteristics of insulin-like growth factor-1 from a collagen-glycosaminoglycan scaffold. Tissue Eng Part C Methods 2010 Dec;16(6):1439-48.
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