Effect of dimethylsulfoxide on articular cartilage proteoglycan synthesis and degradation, chondrocyte viability, and matrix water content

Objective—To determine the effects of dimethylsulfoxide (DMSO) exposure on cartilage proteoglycan (PG) synthesis, PG degradation, chondrocyte viability, and matrix water content. Study Design—Using a cartilage explant culture system, PG synthesis, PG degradation, matrix water content, and chondrocyt...

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Veröffentlicht in:Veterinary surgery 1998-09, Vol.27 (5), p.438-444
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Morris, E.A
description Objective—To determine the effects of dimethylsulfoxide (DMSO) exposure on cartilage proteoglycan (PG) synthesis, PG degradation, chondrocyte viability, and matrix water content. Study Design—Using a cartilage explant culture system, PG synthesis, PG degradation, matrix water content, and chondrocyte viablity were determined for cartilage exposed to DMSO daily for selected periods of time. Animals or Sample Population—Juvenile bovine (calf) carpometacarpal joint cartilage ex‐plants. Methods—PG synthesis: Explants (n = 30/group) were separated into 10 groups based on the time of daily exposure to 10% DMSO. Exposure time was repeated daily for 3 days. The control group was incubated in basal medium alone for 3 days, with daily medium changes. Once all DMSO exposure times were complete for the third day, PG synthesis was determined by analysis of incorporation of radiolabelled sulfate. Cell viability: Explants (n = 3/group) were subjected to an identical DMSO exposure protocol, and examined histologically. The percentage of viable cells/high power field (hpf) was calculated for each group. PG degradation: Explants (n = 21/group) were preincubated with radiolabelled sulfate, then subjected to a similar DMSO exposure protocol. The medium was collected from all explants daily and assayed for PG content. After 3 days, the explants were digested and total labelled PG content determined. Percent of total explant labelled PG content released into the medium daily was determined for each group. Water content: Explants (n = 21/group) were separated into three treatment groups, one of which had no treatments performed, whereas the other two groups were incubated in basal medium for 72 hours, one with, and one without, 10% DMSO. Wet and dry weights were determined, and percent water calculated, for all three groups. Separate 1‐way ANOVA were performed, with appropriate post hoc tests (p < .05). Results—PG synthesis was significantly lower than control for all time periods of DMSO exposure except for 1 and 3 hours, and decreased in a time‐dependent manner after the 1‐hour exposure time. The mean percentage of viable cells/hpf was significantly lower than control for the 1‐, 3‐, 9‐, 12‐, and 24‐hour treatment groups. There was no significant difference in PG degradation for any group compared with control for the first 2 days of incubation. All groups except the 24‐hour group had a significantly higher degradation compared with control for the third day of incubation. Car
doi_str_mv 10.1111/j.1532-950X.1998.tb00153.x
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Study Design—Using a cartilage explant culture system, PG synthesis, PG degradation, matrix water content, and chondrocyte viablity were determined for cartilage exposed to DMSO daily for selected periods of time. Animals or Sample Population—Juvenile bovine (calf) carpometacarpal joint cartilage ex‐plants. Methods—PG synthesis: Explants (n = 30/group) were separated into 10 groups based on the time of daily exposure to 10% DMSO. Exposure time was repeated daily for 3 days. The control group was incubated in basal medium alone for 3 days, with daily medium changes. Once all DMSO exposure times were complete for the third day, PG synthesis was determined by analysis of incorporation of radiolabelled sulfate. Cell viability: Explants (n = 3/group) were subjected to an identical DMSO exposure protocol, and examined histologically. The percentage of viable cells/high power field (hpf) was calculated for each group. PG degradation: Explants (n = 21/group) were preincubated with radiolabelled sulfate, then subjected to a similar DMSO exposure protocol. The medium was collected from all explants daily and assayed for PG content. After 3 days, the explants were digested and total labelled PG content determined. Percent of total explant labelled PG content released into the medium daily was determined for each group. Water content: Explants (n = 21/group) were separated into three treatment groups, one of which had no treatments performed, whereas the other two groups were incubated in basal medium for 72 hours, one with, and one without, 10% DMSO. Wet and dry weights were determined, and percent water calculated, for all three groups. Separate 1‐way ANOVA were performed, with appropriate post hoc tests (p &lt; .05). Results—PG synthesis was significantly lower than control for all time periods of DMSO exposure except for 1 and 3 hours, and decreased in a time‐dependent manner after the 1‐hour exposure time. The mean percentage of viable cells/hpf was significantly lower than control for the 1‐, 3‐, 9‐, 12‐, and 24‐hour treatment groups. There was no significant difference in PG degradation for any group compared with control for the first 2 days of incubation. All groups except the 24‐hour group had a significantly higher degradation compared with control for the third day of incubation. Cartilage exposed to DMSO for 72 hours had a significantly lower water content, and cartilage incubated in basal medium alone for 72 hours had a significantly higher water content than cartilage that received no DMSO and no incubation. 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Study Design—Using a cartilage explant culture system, PG synthesis, PG degradation, matrix water content, and chondrocyte viablity were determined for cartilage exposed to DMSO daily for selected periods of time. Animals or Sample Population—Juvenile bovine (calf) carpometacarpal joint cartilage ex‐plants. Methods—PG synthesis: Explants (n = 30/group) were separated into 10 groups based on the time of daily exposure to 10% DMSO. Exposure time was repeated daily for 3 days. The control group was incubated in basal medium alone for 3 days, with daily medium changes. Once all DMSO exposure times were complete for the third day, PG synthesis was determined by analysis of incorporation of radiolabelled sulfate. Cell viability: Explants (n = 3/group) were subjected to an identical DMSO exposure protocol, and examined histologically. The percentage of viable cells/high power field (hpf) was calculated for each group. PG degradation: Explants (n = 21/group) were preincubated with radiolabelled sulfate, then subjected to a similar DMSO exposure protocol. The medium was collected from all explants daily and assayed for PG content. After 3 days, the explants were digested and total labelled PG content determined. Percent of total explant labelled PG content released into the medium daily was determined for each group. Water content: Explants (n = 21/group) were separated into three treatment groups, one of which had no treatments performed, whereas the other two groups were incubated in basal medium for 72 hours, one with, and one without, 10% DMSO. Wet and dry weights were determined, and percent water calculated, for all three groups. Separate 1‐way ANOVA were performed, with appropriate post hoc tests (p &lt; .05). Results—PG synthesis was significantly lower than control for all time periods of DMSO exposure except for 1 and 3 hours, and decreased in a time‐dependent manner after the 1‐hour exposure time. The mean percentage of viable cells/hpf was significantly lower than control for the 1‐, 3‐, 9‐, 12‐, and 24‐hour treatment groups. There was no significant difference in PG degradation for any group compared with control for the first 2 days of incubation. All groups except the 24‐hour group had a significantly higher degradation compared with control for the third day of incubation. Cartilage exposed to DMSO for 72 hours had a significantly lower water content, and cartilage incubated in basal medium alone for 72 hours had a significantly higher water content than cartilage that received no DMSO and no incubation. Conclusions—DMSO, in relatively low concentration, is detrimental to articular cartilage PG</description><subject>Administration, Topical</subject><subject>Animals</subject><subject>Anti-Inflammatory Agents - pharmacology</subject><subject>calves</subject><subject>cartilage</subject><subject>Cartilage, Articular - chemistry</subject><subject>Cartilage, Articular - cytology</subject><subject>Cartilage, Articular - drug effects</subject><subject>Cartilage, Articular - metabolism</subject><subject>Cattle</subject><subject>Cell Survival - drug effects</subject><subject>chondrocytes</subject><subject>Chondrocytes - cytology</subject><subject>Chondrocytes - drug effects</subject><subject>dimethyl sulfoxide</subject><subject>Dimethyl Sulfoxide - pharmacology</subject><subject>duration</subject><subject>In Vitro Techniques</subject><subject>metabolism</subject><subject>protein degradation</subject><subject>protein synthesis</subject><subject>proteoglycans</subject><subject>Proteoglycans - biosynthesis</subject><subject>Proteoglycans - metabolism</subject><subject>tissue culture</subject><subject>viability</subject><subject>Water - analysis</subject><subject>water content</subject><issn>0161-3499</issn><issn>1532-950X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkd9u0zAUxi0EGqXwCAiLC66W4r9JzMUkVG0DaQKkUdid5TgnrUsaD9vZklfgqUlp1Xt8Y8vnO7-j830IvaVkQafzfrugkrNMSXK3oEqVi1QRMn0thidodio9RTNCc5pxodRz9CLGLSFECcHP0JkqhJJUzNCfy6YBm7BvcO12kDZjG_u28YOrAfsOm5Cc7VsTsN0_W7MGfB98Ar9uR2s6HMcubSC6iE1X4xrWwdQmOd-dY7vxXR28HRPgB2cq17o0nv_T7UwKbsCPJsFE9l2CLr1EzxrTRnh1vOdodXX5ffkpu_l6_Xn58SazgtMyK4CrvJKSiZxAJSkt66ZhhhaW5bmtClLWzNiGkIoRabnlAmRlamAVAc4Y53P07sCd9vjdQ0x656KFtjUd-D7qgitRciIm4YeD0AYfY4BG3we3M2HUlOh9EHqr927rvdt6H4Q-BqGHqfn1cUpf7aA-tR6dn-oXh_qja2H8D7L-cbsSvJwA2QHgYoLhBDDhl84LXkj988u1lnfLgvLy27TUHL056BvjtVkHF_XqlhHKCVNccEL4X68etP8</recordid><startdate>199809</startdate><enddate>199809</enddate><creator>Matthews, G.L</creator><creator>Engler, S.J</creator><creator>Morris, E.A</creator><general>Blackwell Publishing Ltd</general><scope>FBQ</scope><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>199809</creationdate><title>Effect of dimethylsulfoxide on articular cartilage proteoglycan synthesis and degradation, chondrocyte viability, and matrix water content</title><author>Matthews, G.L ; Engler, S.J ; Morris, E.A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4318-7e396b552460eb5118dff2a17c266cb708d2acf00b205c3c34e5bade2b0e32233</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Administration, Topical</topic><topic>Animals</topic><topic>Anti-Inflammatory Agents - pharmacology</topic><topic>calves</topic><topic>cartilage</topic><topic>Cartilage, Articular - chemistry</topic><topic>Cartilage, Articular - cytology</topic><topic>Cartilage, Articular - drug effects</topic><topic>Cartilage, Articular - metabolism</topic><topic>Cattle</topic><topic>Cell Survival - drug effects</topic><topic>chondrocytes</topic><topic>Chondrocytes - cytology</topic><topic>Chondrocytes - drug effects</topic><topic>dimethyl sulfoxide</topic><topic>Dimethyl Sulfoxide - pharmacology</topic><topic>duration</topic><topic>In Vitro Techniques</topic><topic>metabolism</topic><topic>protein degradation</topic><topic>protein synthesis</topic><topic>proteoglycans</topic><topic>Proteoglycans - biosynthesis</topic><topic>Proteoglycans - metabolism</topic><topic>tissue culture</topic><topic>viability</topic><topic>Water - analysis</topic><topic>water content</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Matthews, G.L</creatorcontrib><creatorcontrib>Engler, S.J</creatorcontrib><creatorcontrib>Morris, E.A</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Veterinary surgery</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Matthews, G.L</au><au>Engler, S.J</au><au>Morris, E.A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Effect of dimethylsulfoxide on articular cartilage proteoglycan synthesis and degradation, chondrocyte viability, and matrix water content</atitle><jtitle>Veterinary surgery</jtitle><addtitle>Vet Surg</addtitle><date>1998-09</date><risdate>1998</risdate><volume>27</volume><issue>5</issue><spage>438</spage><epage>444</epage><pages>438-444</pages><issn>0161-3499</issn><eissn>1532-950X</eissn><abstract>Objective—To determine the effects of dimethylsulfoxide (DMSO) exposure on cartilage proteoglycan (PG) synthesis, PG degradation, chondrocyte viability, and matrix water content. Study Design—Using a cartilage explant culture system, PG synthesis, PG degradation, matrix water content, and chondrocyte viablity were determined for cartilage exposed to DMSO daily for selected periods of time. Animals or Sample Population—Juvenile bovine (calf) carpometacarpal joint cartilage ex‐plants. Methods—PG synthesis: Explants (n = 30/group) were separated into 10 groups based on the time of daily exposure to 10% DMSO. Exposure time was repeated daily for 3 days. The control group was incubated in basal medium alone for 3 days, with daily medium changes. Once all DMSO exposure times were complete for the third day, PG synthesis was determined by analysis of incorporation of radiolabelled sulfate. Cell viability: Explants (n = 3/group) were subjected to an identical DMSO exposure protocol, and examined histologically. The percentage of viable cells/high power field (hpf) was calculated for each group. PG degradation: Explants (n = 21/group) were preincubated with radiolabelled sulfate, then subjected to a similar DMSO exposure protocol. The medium was collected from all explants daily and assayed for PG content. After 3 days, the explants were digested and total labelled PG content determined. Percent of total explant labelled PG content released into the medium daily was determined for each group. Water content: Explants (n = 21/group) were separated into three treatment groups, one of which had no treatments performed, whereas the other two groups were incubated in basal medium for 72 hours, one with, and one without, 10% DMSO. Wet and dry weights were determined, and percent water calculated, for all three groups. Separate 1‐way ANOVA were performed, with appropriate post hoc tests (p &lt; .05). Results—PG synthesis was significantly lower than control for all time periods of DMSO exposure except for 1 and 3 hours, and decreased in a time‐dependent manner after the 1‐hour exposure time. The mean percentage of viable cells/hpf was significantly lower than control for the 1‐, 3‐, 9‐, 12‐, and 24‐hour treatment groups. There was no significant difference in PG degradation for any group compared with control for the first 2 days of incubation. All groups except the 24‐hour group had a significantly higher degradation compared with control for the third day of incubation. Cartilage exposed to DMSO for 72 hours had a significantly lower water content, and cartilage incubated in basal medium alone for 72 hours had a significantly higher water content than cartilage that received no DMSO and no incubation. Conclusions—DMSO, in relatively low concentration, is detrimental to articular cartilage PG</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>9749514</pmid><doi>10.1111/j.1532-950X.1998.tb00153.x</doi><tpages>7</tpages></addata></record>
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identifier ISSN: 0161-3499
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source MEDLINE; Wiley Online Library Journals Frontfile Complete
subjects Administration, Topical
Animals
Anti-Inflammatory Agents - pharmacology
calves
cartilage
Cartilage, Articular - chemistry
Cartilage, Articular - cytology
Cartilage, Articular - drug effects
Cartilage, Articular - metabolism
Cattle
Cell Survival - drug effects
chondrocytes
Chondrocytes - cytology
Chondrocytes - drug effects
dimethyl sulfoxide
Dimethyl Sulfoxide - pharmacology
duration
In Vitro Techniques
metabolism
protein degradation
protein synthesis
proteoglycans
Proteoglycans - biosynthesis
Proteoglycans - metabolism
tissue culture
viability
Water - analysis
water content
title Effect of dimethylsulfoxide on articular cartilage proteoglycan synthesis and degradation, chondrocyte viability, and matrix water content
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