Quantitative analysis of repair tissue biopsies following chondrocyte implantation.
Outcome measures for cartilage repair techniques include clinical assessment of functional status, magnetic resonance imaging, mechanical indentation in situ and second-look biopsies, which are used for detailed ex vivo histological and immunohistochemical assessment. Biopsy analysis is considered an important outcome measure, despite being highly invasive, since it provides a visual record of the spatial organization of matrix proteins and cells. We propose that the value of second-look biopsies would be significantly enhanced if accurate quantification of cartilage matrix molecules could also be obtained. The goal of our work has been to develop a combined method for histological and biochemical analysis of a single biopsy. We have developed a method of cutting frozen sections of cartilage and recovering the uncut tissue for subsequent biochemical analysis. We have also developed a range of miniaturized assays that can be performed after cartilage digestion with trypsin. In this way we are now able to analyse biopsies with a wet weight as low as 5 mg using both histological and biochemical methods, so obtaining the maximum amount of information from the minimum volume of tissue. This new approach will allow a more accurate assessment of the quality of cartilage repair tissue than histological analysis alone.
Hollander
Anthony P
AP
University of Bristol Academic Rheumatology, Avon Orthopaedic Centre, Southmead Hospital, Bristol BS10 SNB, UK.
Dickinson
Sally C
SC
Sims
Trevor J
TJ
Soranzo
Carlo
C
Pavesio
Alessandra
A
D000367:Age Factors; D000818:Animals; D001672:Biocompatible Materials; D001706:Biopsy; D002358:Cartilage, Articular / Q000737:chemistry / Q000293:injuries / Q000473:pathology* / Q000601:surgery; D002417:Cattle; D002478:Cells, Cultured / Q000737:chemistry / Q000637:transplantation; D019902:Chondrocytes / Q000737:chemistry / Q000637:transplantation*; D024042:Collagen Type I / Q000032:analysis; D024043:Collagen Type II / Q000032:analysis; D010450:Endopeptidases / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005500:Follow-Up Studies; D005629:Frozen Sections; D006025:Glycosaminoglycans / Q000032:analysis; D006818:Hyalin / Q000737:chemistry; D006820:Hyaluronic Acid; D006909:Hydroxyproline / Q000032:analysis; D009929:Organ Size; D019736:Prostheses and Implants; D011817:Rabbits; D024162:Second-Look Surgery; D013526:Surgical Mesh; D023822:Tissue Engineering
D001672:Biocompatible Materials; D024042:Collagen Type I; D024043:Collagen Type II; D006025:Glycosaminoglycans; D006820:Hyaluronic Acid; D010450:Endopeptidases; D006909:Hydroxyproline