title: Viability of chondrocytes seeded onto a collagen I/III membrane for matrix-induced autologous chondrocyte implantation.
abstract: Cell viability is crucial for effective cell-based cartilage repair. The aim of this study was to determine the effect of handling the membrane during matrix-induced autologous chondrocyte implantation surgery on the viability of implanted chondrocytes. Images were acquired under five conditions: (i) Pre-operative; (ii) Handled during surgery; (iii) Cut edge; (iv) Thumb pressure applied; (v) Heavily grasped with forceps. Live and dead cell stains were used. Images were obtained for cell counting and morphology. Mean cell density was 6.60 × 10(5) cells/cm(2) (5.74-7.11 × 10(5) ) in specimens that did not have significant trauma decreasing significantly in specimens that had been grasped with forceps (p < 0.001) or cut (p = 0.004). Cell viability on delivery grade membrane was 75.1%(72.4-77.8%). This dropped to 67.4%(64.1-69.7%) after handling (p = 0.002), 56.3%(51.5-61.6%) after being thumbed (p < 0.001) and 28.8%(24.7-31.2%) after crushing with forceps (p < 0.001). When cut with scissors there was a band of cell death approximately 275 µm in width where cell viability decreased to 13.7%(10.2-18.2%, p < 0.001). Higher magnification revealed cells without the typical rounded appearance of chondrocytes. We found that confocal laser-scanning microscope (CLSM) can be used to quantify and image the fine morphology of cells on a matrix-induced autologous chondrocyte implantation (MACI) membrane. Careful handling of the membrane is essential to minimise chondrocyte death during surgery.
authors.affiliation: Department of Trauma and Orthopaedic Surgery, The Royal Infirmary of Edinburgh, Edinburgh, The United Kingdom; Centre for Integrative Physiology, School of Biomedical Sciences, The University of Edinburgh, Edinburgh, Edinburgh, The United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Hindle
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Hall
authors.forename: Leela C
authors.initials: LC
authors.lastname: Biant
mesh_terms: D000328:Adult; D001706:Biopsy; D002358:Cartilage, Articular / Q000502:physiology / Q000601:surgery*; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000637:transplantation*; D024042:Collagen Type I / Q000378:metabolism*; D024061:Collagen Type III / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007717:Knee / Q000033:anatomy & histology; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D015999:Multivariate Analysis; D019637:Orthopedic Procedures / Q000379:methods
chemical_list: D024042:Collagen Type I; D024061:Collagen Type III; D005456:Fluorescent Dyes
keywords: MACI; cell viability; chondrocyte; confocal microscopy; knee