Ultrastructural studies of cartilage in genetic disorders of skeletal growth

Abstract

The human skeletal dysplasias are an heterogeneous group of heritable connective tissue disorders associated with abnormalities in the size and shape of the limbs, trunk and/or skull which frequently result in disproportionate short stature. In recent years it has become apparent that these comprise over 50 distinct conditions with a variety of subtypes distinguished on clinical and radiological grounds. We have investigated the pathogenesis of these conditions in over 100 patients by direct transmission electron microscopic examination of chondro-osseous tissue. Some of the ultrastructural studies have been previously reported.Small biopsies of chondro-osseous junction were collected for electron microscopy from the rib or iliac crest of patients with skeletal dysplasias or from normal controls at the time of surgery. These were cut into small blocks and fixed for one hour in either 5% glutaraldehyde in white's buffer or directly in 1% osmic acid in White's buffer or a modified Karnovsky's fixative, (2. 5% paraformaldehyde, 2. 5% glutaraldehyde, 2. 5mM calcium in cacodylate buffer). Subsequent processing included osmium fixation, block staining with uranyl acetate and embedding in Araldite or Spurr's low viscosity resin (firm composition). Sections were cut with glass knives or diamond knives. The latter produced sections which were much more even in thickness, permitting more consistent appraisal of matrix features.