Chondrocyte necrosis and apoptosis in impact damaged articular cartilage
A decrease in chondrocyte numbers is one characteristic of osteoarthritic cartilage. This decrease may be the result of apoptosis or other forms of cell death induced by mechanical damage. Furthermore, cell death may contribute to the structural and metabolic changes found in osteoarthritic cartilag...
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Published in | Journal of orthopaedic research Vol. 19; no. 4; pp. 703 - 711 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | English |
Published |
Hoboken
Elsevier Ltd
01.07.2001
Wiley Subscription Services, Inc., A Wiley Company Blackwell Publishing Ltd |
Subjects | |
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Abstract | A decrease in chondrocyte numbers is one characteristic of osteoarthritic cartilage. This decrease may be the result of apoptosis or other forms of cell death induced by mechanical damage. Furthermore, cell death may contribute to the structural and metabolic changes found in osteoarthritic cartilage. Therefore, we investigated cell viability and the mode of cell death in cartilage subjected to an increasing severity of impact loads expected to cause compositional damage and osteoarthritic-like metabolic alterations. Canine cartilage explants were subjected to cyclic indentation impacts of 5 megapascals at 0.3 Hz for 0, 2, 20, and 120 min and then kept in culture for 2, 4, 48, and 144 h. Cell death was assessed by the TUNEL assay and by uptake of propidium iodide. Viable cells were detected by the ability to metabolize fluorescein diacetate. Nuclear morphology and ultrastructure of the cell were examined using Hoechst 33342 fluorescent staining and transmission electron microscopy (TEM). As controls for necrosis and apoptosis, cartilage was, respectively, frozen and thawed or incubated with mitomycin-C, an apoptosis inducer. In cartilage that had been loaded for 2 h, 32% of the chondrocytes in the loaded core took up propidium iodide within 2 h after loading. Most of these were in the middle to superficial zones and reflected leaky cell membranes usually characteristic of necrosis. Less than 1% of these chondrocytes were positive in the TUNEL assay after 4 h. After additional culture for 2 days, however, the proportion of chondrocytes which were positive in the TUNEL assay reached 73%. A dose dependent response to duration of loading was detected with the TUNEL assay at this time. The TUNEL assay was not specific for apoptosis since 92% of chondrocytes in freeze/thawed cartilage were TUNEL positive. However, some cells with apoptotic bodies and chromatin condensation characteristic of apoptosis were found in the transition zone between necrotic and normal chondrocytes, but not in the superficial and upper zones, in impact damaged cartilage. We concluded that in this study, necrosis occurred first, followed by apoptosis. |
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AbstractList | A decrease in chondrocyte numbers is one characteristic of osteoarthritic cartilage. This decrease may be the result of apoptosis or other forms of cell death induced by mechanical damage. Furthermore, cell death may contribute to the structural and metabolic changes found in osteoarthritic cartilage. Therefore, we investigated cell viability and the mode of cell death in cartilage subjected to an increasing severity of impact loads expected to cause compositional damage and osteoarthritic-like metabolic alterations. Canine cartilage explants were subjected to cyclic indentation impacts of 5 megapascals at 0.3 Hz for 0, 2, 20, and 120 min and then kept in culture for 2, 4, 48, and 144 h. Cell death was assessed by the TUNEL assay and by uptake of propidium iodide. Viable cells were detected by the ability to metabolize fluorescein diacetate. Nuclear morphology and ultrastructure of the cell were examined using Hoechst 33342 fluorescent staining and transmission electron microscopy (TEM). As controls for necrosis and apoptosis, cartilage was, respectively, frozen and thawed or incubated with mitomycin-C, an apoptosis inducer. In cartilage that had been loaded for 2 h, 32% of the chondrocytes in the loaded core took up propidium iodide within 2 h after loading. Most of these were in the middle to superficial zones and reflected leaky cell membranes usually characteristic of necrosis. Less than 1% of these chondrocytes were positive in the TUNEL assay after 4 h. After additional culture for 2 days, however, the proportion of chondrocytes which were positive in the TUNEL assay reached 73%. A dose dependent response to duration of loading was detected with the TUNEL assay at this time. The TUNEL assay was not specific for apoptosis since 92% of chondrocytes in freeze/thawed cartilage were TUNEL positive. However, some cells with apoptotic bodies and chromatin condensation characteristic of apoptosis were found in the transition zone between necrotic and normal chondrocytes, but not in the superficial and upper zones, in impact damaged cartilage. We concluded that in this study, necrosis occurred first, followed by apoptosis. Abstract A decrease in chondrocyte numbers is one characteristic of osteoarthritic cartilage. This decrease may be the result of apoptosis or other forms of cell death induced by mechanical damage. Furthermore, cell death may contribute to the structural and metabolic changes found in osteoarthritic cartilage. Therefore, we investigated cell viability and the mode of cell death in cartilage subjected to an increasing severity of impact loads expected to cause compositional damage and osteoarthritic‐like metabolic alterations. Canine cartilage explants were subjected to cyclic indentation impacts of 5 megapascals at 0.3 Hz for 0, 2, 20, and 120 min and then kept in culture for 2, 4, 48, and 144 h. Cell death was assessed by the TUNEL assay and by uptake of propidium iodide. Viable cells were detected by the ability to metabolize fluorescein diacetate. Nuclear morphology and ultrastructure of the cell were examined using Hoechst 33342 fluorescent staining and transmission electron microscopy (TEM). As controls for necrosis and apoptosis, cartilage was, respectively, frozen and thawed or incubated with mitomycin‐C, an apoptosis inducer. In cartilage that had been loaded for 2 h, 32% of the chondrocytes in the loaded core took up propidium iodide within 2 h after loading. Most of these were in the middle to superficial zones and reflected leaky cell membranes usually characteristic of necrosis. Less than 1% of these chondrocytes were positive in the TUNEL assay after 4 h. After additional culture for 2 days, however, the proportion of chondrocytes which were positive in the TUNEL assay reached 73%. A dose dependent response to duration of loading was detected with the TUNEL assay at this time. The TUNEL assay was not specific for apoptosis since 92% of chondrocytes in freeze/thawed cartilage were TUNEL positive. However, some cells with apoptotic bodies and chromatin condensation characteristic of apoptosis were found in the transition zone between necrotic and normal chondrocytes, but not in the superficial and upper zones, in impact damaged cartilage. We concluded that in this study, necrosis occurred first, followed by apoptosis. © 2001 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved. A decrease in chondrocyte numbers is one characteristic of osteoarthritic cartilage. This decrease may be the result of apoptosis or other forms of cell death induced by mechanical damage. Furthermore, cell death may contribute to the structural and metabolic changes found in osteoarthritic cartilage. Therefore, we investigated cell viability and the mode of cell death in cartilage subjected to an increasing severity of impact loads expected to cause compositional damage and osteoarthritic‐like metabolic alterations. Canine cartilage explants were subjected to cyclic indentation impacts of 5 megapascals at 0.3 Hz for 0, 2, 20, and 120 min and then kept in culture for 2, 4, 48, and 144 h. Cell death was assessed by the TUNEL assay and by uptake of propidium iodide. Viable cells were detected by the ability to metabolize fluorescein diacetate. Nuclear morphology and ultrastructure of the cell were examined using Hoechst 33342 fluorescent staining and transmission electron microscopy (TEM). As controls for necrosis and apoptosis, cartilage was, respectively, frozen and thawed or incubated with mitomycin‐C, an apoptosis inducer. In cartilage that had been loaded for 2 h, 32% of the chondrocytes in the loaded core took up propidium iodide within 2 h after loading. Most of these were in the middle to superficial zones and reflected leaky cell membranes usually characteristic of necrosis. Less than 1% of these chondrocytes were positive in the TUNEL assay after 4 h. After additional culture for 2 days, however, the proportion of chondrocytes which were positive in the TUNEL assay reached 73%. A dose dependent response to duration of loading was detected with the TUNEL assay at this time. The TUNEL assay was not specific for apoptosis since 92% of chondrocytes in freeze/thawed cartilage were TUNEL positive. However, some cells with apoptotic bodies and chromatin condensation characteristic of apoptosis were found in the transition zone between necrotic and normal chondrocytes, but not in the superficial and upper zones, in impact damaged cartilage. We concluded that in this study, necrosis occurred first, followed by apoptosis. © 2001 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved. |
Author | Borden, Caroline Chen, Chih-Tung Lust, George Hueffer, Karsten Burton-Wurster, Nancy Bloom, Stephen E |
Author_xml | – sequence: 1 givenname: Chih-Tung surname: Chen fullname: Chen, Chih-Tung organization: James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA – sequence: 2 givenname: Nancy surname: Burton-Wurster fullname: Burton-Wurster, Nancy email: niw1@cornell.edu organization: James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA – sequence: 3 givenname: Caroline surname: Borden fullname: Borden, Caroline organization: James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA – sequence: 4 givenname: Karsten surname: Hueffer fullname: Hueffer, Karsten organization: James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA – sequence: 5 givenname: Stephen E surname: Bloom fullname: Bloom, Stephen E organization: Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA – sequence: 6 givenname: George surname: Lust fullname: Lust, George organization: James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/11518282$$D View this record in MEDLINE/PubMed |
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References_xml | – volume: 14 start-page: 417 year: 1996 end-page: 423 ident: BIB11 article-title: Swelling and fibronectin accumulation in articular cartilage explants after cyclical impact publication-title: J. Orthop. Res. contributor: fullname: Lust – volume: 9 start-page: 247 year: 1982 end-page: 248 ident: BIB10 article-title: A direct spectrophotometric microassay for sulfated glycosaminoglycans in cartilage cultures publication-title: Connect Tissue Res. contributor: fullname: Barrett – volume: 800 start-page: 52 year: 1984 end-page: 58 ident: BIB26 article-title: Synthesis of fibronectin in normal and osteoarthritic articular cartilage publication-title: Biochim. Biophys. Acta. contributor: fullname: Lust – volume: 6 start-page: 393 year: 1998 end-page: 399 ident: BIB22 article-title: Osteoarthritic cartilage fibrillation is associated with a decrease in chondrocyte adhesion to fibronectin publication-title: Osteoarth. 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1999 article-title: Injurious compression of bovine articular cartilage induced chondrocyte apoptosis before detectable mechanical damage publication-title: Proc 45th Ann Mtg Orthop Res Soc – volume: 14 start-page: 417 year: 1996 end-page: 23 article-title: Swelling and fibronectin accumulation in articular cartilage explants after cyclical impact publication-title: J Orthop Res – volume: 11 start-page: 319 year: 1997 end-page: 26 article-title: Effect of impact load on articular cartilage: development of an intra‐articular fracture model publication-title: J Orthop Trauma – volume: 146 start-page: 75 year: 1995 end-page: 85 article-title: Chondrocyte apoptosis induced by nitric oxide publication-title: Am J Pathol – volume: 58 start-page: 181 year: 1995 end-page: 90 article-title: Cell death: current difficulties in discriminating apoptosis from necrosis in the context of pathological processes in vivo publication-title: J Cell Biochem – volume: 800 start-page: 52 year: 1984 end-page: 8 article-title: Synthesis of fibronectin in normal and osteoarthritic articular cartilage publication-title: Biochim Biophys Acta – volume: 6 start-page: 393 year: 1998 end-page: 9 article-title: Osteoarthritic cartilage fibrillation is associated with a decrease in chondrocyte adhesion to fibronectin publication-title: Osteoarth Cart – volume: 9 start-page: 247 year: 1982 end-page: 8 article-title: A direct spectrophotometric microassay for sulfated glycosaminoglycans in cartilage cultures publication-title: Connect Tissue Res – start-page: 17 year: 1999 end-page: 19 – volume: 24 start-page: 672 year: 1999 article-title: Cell death following repetitive loading of cartilage explants publication-title: Proc 45th Ann Mtg Orthop Res Soc – volume: 16 start-page: 490 year: 1998 end-page: 9 article-title: Effects of injurious compression on matrix turnover around individual cells in calf articular cartilage explants publication-title: J Orthop Res – start-page: 317 year: 1993 end-page: 84 – 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and clinical aspects year: 1993 ident: e_1_2_1_7_2 contributor: fullname: Burton‐Wurster N – ident: e_1_2_1_13_2 doi: 10.1016/S0306-4522(99)00345-0 – ident: e_1_2_1_16_2 doi: 10.1002/1529-0131(199807)41:7<1266::AID-ART18>3.0.CO;2-Y – volume: 24 start-page: 42 year: 1999 ident: e_1_2_1_19_2 article-title: Injurious compression of bovine articular cartilage induced chondrocyte apoptosis before detectable mechanical damage publication-title: Proc 45th Ann Mtg Orthop Res Soc contributor: fullname: Loening AM – volume: 21 start-page: 1465 year: 1995 ident: e_1_2_1_15_2 article-title: In situ detection of fragmented DNA (TUNEL assay) fails to discriminate among apoptosis, necrosis, and autolytic cell death: a cautionary note publication-title: Hepatology doi: 10.1016/0270-9139(95)90071-3 contributor: fullname: Grasl‐Kraupp B – ident: e_1_2_1_18_2 doi: 10.1016/0003-2697(88)90532-5 – ident: e_1_2_1_23_2 doi: 10.1053/joca.1998.0138 – ident: e_1_2_1_24_2 doi: 10.1002/jor.1100160415 – ident: 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Snippet | A decrease in chondrocyte numbers is one characteristic of osteoarthritic cartilage. This decrease may be the result of apoptosis or other forms of cell death... Abstract A decrease in chondrocyte numbers is one characteristic of osteoarthritic cartilage. This decrease may be the result of apoptosis or other forms of... |
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SubjectTerms | Animals Apoptosis Cartilage, Articular - injuries Cartilage, Articular - pathology Cell Nucleus - ultrastructure Cell Survival Cells, Cultured Chondrocytes - pathology Chondrocytes - ultrastructure Dogs In Situ Nick-End Labeling Microscopy, Electron Necrosis |
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Title | Chondrocyte necrosis and apoptosis in impact damaged articular cartilage |
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