Connective tissue growth stimulated by carrageenin. 3. The nature and amount of polysaccharide produced in normal and scorbutic guinea pigs and the metabolism of a chondroitin sulphuric acid fraction

Abstract

Growth of new connective tissue was stimulated by the injection of 50 mg of carrageenin beneath the abdominal skin of guinea pigs. Groups of guinea pigs were maintained on an ascorbic acid-deficient diet for 7 or 10 days before injection with carrageenin and killed at intervals from 6 to 12 days later. Corresponding groups of normal guinea pigs were given ascorbic acid supplements during the periods of granuloma production. The granulomata were dissected out and pooled from groups of normal and scorbutic animals at each time interval of killing. After digestion of the granulation tissues with papain, polysaccharide materials were extracted by a fractionation scheme utilizing cetylpyridinium chloride. By this means 3 groups of polysaccharide material were obtained: (1) total polysaccharides; (2) a chondroitin sulfate fraction; (3) a hyaluronic acid-like fraction. The granuloma tissue from the scorbutic animals showed a greatly increased production of total polysaccharides in comparison with that from normal animals. Evidence is presented that most of this increased polysaccharide production is due to greatly increased amounts of a polysaccharide fraction with properties similar to those of hyaluronic acid. Production of the chondroitin sulfate fraction was generally slightly reduced in the scorbutic granuloma in comparison with the normal animals. Although most of the polysaccharide material obtained could be accounted for in terms of the chondroitin sulfate and hyaluronic acid fractions the presence of other mucopolysaccharides is not excluded. Normal and scorbutic guinea pigs, 5 days after injection with carrageenin, were injected with 1 [mu]curie/g body weight of Na2S35O4 and killed at intervals of time from 1 to 7 days later. The specific activity of the chondroitin sulfate fraction from rib cartilage in the scorbutic animals was much less at 24 hours than that in corresponding normal animals. By contrast, incorporation of S35 sulfate into chondroitin sulfate from the granuloma of scorbutic guinea pigs at 24 hours was only a little less than into chondroitin sulfate from granuloma of normal animals. The rate of loss of specific activity in the scorbutic granuloma, however, was less rapid than in the normal granuloma and suggests some reduction in the rate of metabolism of chondroitin sulfate in scorbutic granulation tissue.