Mucopolysaccharide localization in gingival epithelium.

Abstract

The effect of extraneous hyaluronidase, trypsin, hyaluronic acid, chrondroitin sulphate, and commercial heparin on the synthesis and secretion of proteoglycans (mucopolysaccharides) by gingival epithelium in short term incubations was investigated by autoradiography. Small pieces of human gingivae were incubated at 37°C in tissue culture medium (T.C. 199) for 75 min. The first 15 min incubation included a “pulse” of (35S)‐sulphate, after which the radioactive incorporation was “chased” in radioactive free medium. Cryostat sections of these pieces were cut, air dried, slide fixed with cetylpyridinium chloride, and prepared for autoradiography. The effect of the various additives in the “pulse” or the “chase” incubations on the autoradiographic localization of incorporated (35S)‐sulphate in the epithelium was noted.The responses by the epithelium to the enzymes hyaluronidase and trypsin were different. The former caused marked tissue disruption, probably due in part to degradation of the ground substance. Synthesis and secretion of sulphated macromolecules were evident when the lower concentrations of hyaluronidase were used. Tissue disruption appeared to be less severe. Trypsin in the medium of the gingival incubations appeared to cause both metabolic and secretory disruption. Both the sulphated polyanions, chrondroitin sulphate and heparin, when added to gingival incubations, showed inhibition of localization of (35S)‐sulphate label inlercellularly. Chrondroitin sulphate included in the “chase” incubations alone resulted in the inhibition, while only low concentrations of heparin caused any inhibition. These latter data were curious. The interpretations of a comparison of the data from “pulse” and “chase” additions of the non‐sulphated hyaluronic acid imply that there was an initial inhibitory effect on secretion which in turn resulted in a subsequent regulation of synthesis of the intercellular sulphated macromolecules. These data and those derived from other tissues such as endothelium, cartilage, and chondrocyte cultures are briefly contrasted.