Reverse transcription-polymerase chain reaction detection of collagen transcripts in healing human wounds.
The aim of this study was to analyse the expression of COl1A1, COl1A2 and COl3A1 in 6 mm diameter punch biopsies obtained from human wounds. Total RNA was isolated from biopsies taken from Sacrococcygeal pilonidal sinus excision cavities at weekly intervals between surgery and clinical closure. cDNAs were generated from the RNA using reverse transcriptase and polymerase chain reaction (PCR) amplifications performed with oligonucleotide primer pairs specific for regions of the COl1A1, COl1A2 and COl3A1 genes. The expression of these three genes was demonstrated throughout the course of healing on 36 biopsies taken from nine patients between surgery and clinical closure. Amplification bands demonstrated on cDNAs generated from 6 mm diameter biopsies were comparable in intensity and specificity with those generated from 50 mg excised scar tissue and cultured fibroblasts. The RT-PCR technique described here allows the rapid 'routine' detection of specific gene expression in 6 mm biopsies obtained from healing wounds.
Hopkinson
I
I
Wound Healing Research Unit, University of Wales College of Medicine, Cardiff, UK.
Evans
W
W
Chant
D
D
Hiscox
S
S
Berry
D
D
Harding
K
K
D001706:Biopsy; D003094:Collagen / Q000096:biosynthesis*; D018076:DNA, Complementary; D006801:Humans; D010864:Pilonidal Sinus / Q000601:surgery*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011347:Procollagen / Q000096:biosynthesis*; D014158:Transcription, Genetic; D014945:Wound Healing / Q000502:physiology*
D018076:DNA, Complementary; D011347:Procollagen; D003094:Collagen