Fixation-dependent immunolocalization shift and immunoreactivity of intracellular growth factors in cartilage |
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| Authors: | Bos P K van Osch G J van der Kwast T Verwoerd-Verhoef H L Verhaar J A |
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| Institution: | (1) Department of Orthopaedics, Erasmus University Medical Centre Rotterdam, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands;(2) Department of Otorhinolaryngology, Erasmus University Medical Centre Rotterdam, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands;(3) Department of Pathology, Erasmus University Medical Centre Rotterdam, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands |
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| Abstract: | The effects of fixation on immunolocalization and immunoreactivity in cartilage tissues were studied using monoclonal antibodies against peptides that can effectively stimulate chondrocytes in vitro and have been shown to play a role in musculoskeletal tissue regeneration: transforming growth factor  1, transforming growth factor 3, insulin-like growth factor I, insulin-like growth factor II and fibroblast growth factor 2. Paraffin sections fixed in buffered formalin, buffered paraformaldehyde, Carnoy and methacarn, as well as cryosections, were tested. A strong immunoreaction was observed in tissue fixed in formaldehyde-based fixatives, with a resemblance to that in cryopreserved tissues. Immunoreactivity was reduced in alcohol-fixed tissues. Furthermore, a striking intracellular immunolocalization shift from cytoplasm to nucleus was observed using alcohol-based fixatives as compared to cryopreserved or formaldehyde-based fixatives. We concluded that, for the detection and localization of growth factors in cartilage tissues, fixation in buffered formalin or paraformaldehyde is optimal. |
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