Measurement of skeletal muscle collagen breakdown by microdialysis

TY - JOUR

T1 - Measurement of skeletal muscle collagen breakdown by microdialysis

AU - Miller, B F

AU - Ellis, D

AU - Robinson, M M

AU - Rivera, J D

AU - Kjaer, M

AU - Langberg, Henning

N1 - © 2010 John Wiley & Sons A/S.

PY - 2011/12

Y1 - 2011/12

N2 - Exercise increases the synthesis of collagen in the extracellular matrix of skeletal muscle. Breakdown of skeletal muscle collagen has not yet been determined because of technical limitations. The purpose of the present study was to use local sampling to determine skeletal muscle collagen breakdown. Microdialysis fibers were tested in vitro to predict bath hydroxyproline (OHP) concentrations. We used an N-methyl-N-[tert-butyldimethyl-silyl]trifluoroacetimide derivative to analyze OHP using gas chromatography-mass spectroscopy (GC-MS) and compared the results with a colorimetric OHP assay. Ten young, healthy male subjects performed a bout of resistance exercise with one leg, followed 17-21h later by in vivo skeletal muscle sampling by microdialysis in exercised (EX) and control (CON) legs. Microdialysis reliably predicted [OHP] in vitro (R 2=0.90). Analysis with GC-MS was strongly correlated to traditional analysis methods (CON: slope=1.03, R 2=0.896, and P<0.05, EX: slope=0.795, R 2=0.896, and P<0.05). We conclude that in vitro, microdialysis fibers were able to measure OHP concentrations and were sensitive to changes in concentrations, a strenuous bout of exercise did not increase skeletal muscle collagen breakdown 17-21h post-exercise, and our measurement of OHP using GC-MS was in agreement with traditional assays.

AB - Exercise increases the synthesis of collagen in the extracellular matrix of skeletal muscle. Breakdown of skeletal muscle collagen has not yet been determined because of technical limitations. The purpose of the present study was to use local sampling to determine skeletal muscle collagen breakdown. Microdialysis fibers were tested in vitro to predict bath hydroxyproline (OHP) concentrations. We used an N-methyl-N-[tert-butyldimethyl-silyl]trifluoroacetimide derivative to analyze OHP using gas chromatography-mass spectroscopy (GC-MS) and compared the results with a colorimetric OHP assay. Ten young, healthy male subjects performed a bout of resistance exercise with one leg, followed 17-21h later by in vivo skeletal muscle sampling by microdialysis in exercised (EX) and control (CON) legs. Microdialysis reliably predicted [OHP] in vitro (R 2=0.90). Analysis with GC-MS was strongly correlated to traditional analysis methods (CON: slope=1.03, R 2=0.896, and P<0.05, EX: slope=0.795, R 2=0.896, and P<0.05). We conclude that in vitro, microdialysis fibers were able to measure OHP concentrations and were sensitive to changes in concentrations, a strenuous bout of exercise did not increase skeletal muscle collagen breakdown 17-21h post-exercise, and our measurement of OHP using GC-MS was in agreement with traditional assays.

KW - Adolescent

KW - Adult

KW - Collagen

KW - Denmark

KW - Exercise Test

KW - Gas Chromatography-Mass Spectrometry

KW - Humans

KW - Hydroxyproline

KW - Male

KW - Microdialysis

KW - Muscle Fibers, Skeletal

KW - Young Adult

U2 - 10.1111/j.1600-0838.2010.01105.x

DO - 10.1111/j.1600-0838.2010.01105.x

M3 - Journal article

C2 - 20561272

SN - 0905-7188

VL - 21

SP - e1-e8

JO - Scandinavian Journal of Medicine & Science in Sports

JF - Scandinavian Journal of Medicine & Science in Sports

IS - 6

ER -