THE FORMATION OF FIBRILS FROM COLLAGEN SOLUTIONS : IV. Effect of Mucopolysaccharides and Nucleic Acids: An Electron Microscope Study

The kinetics of collagen reprecipitation from solutions of salt-extracted calf dermis in the presence of small amounts of mucopolysaccharide and nucleic acids (0.005 per cent in the final reaction mixture) has been reported by Wood (1960). The present paper is a parallel study using the same materials, and describes the electron microscopic (EM) morphology of the collagen precipitates replicated after 24 hours at room temperature. Satisfactory, uncontaminated EM preparations were obtained which showed that all the deposits were fibrous and bore the 640 A cross-banding characteristic of collagen except some narrow, background fibrils 200 to 1000 A wide precipitated in the presence of heparin. These exhibited fine striations about 220 A apart. Chondroitin sulfate greatly increased the rate of precipitation to give a deposit of low optical density consisting of narrow, rigid, discrete fibrils resembling fresh dermis. In contrast, heparin prevented macroscopic gelation, delayed precipitation, and only produced a scanty deposit of abnormal, short, wide, striated tactoids and compound fibers of varying length. The control preparations and the deposits formed in the presence of hyaluronic acid were intermediate between these two extremes. Delayed precipitation was associated with a coarser deposit and aggregation of the fibrils. A duplicate series of deposits precipitated in the presence of RNA and DNA, together with their controls, were examined after ½, 1, 1½, 3, 9, and 24 hours. One set employed an acetic extract of whole calf dermis and the other salt-extracted dermis. The presence of 0.005 per cent DNA in the reaction mixture markedly delayed collagen precipitation with the slow formation of abnormal, short, wide tactoids and compound fibers. RNA also interfered with the quantity and quality of the deposits which contained far less collagen resembling unfixed, normal, adult human dermis, than the controls at the corresponding time intervals. Comparison of the experiments employing whole calf dermis with those employing the salt-extracted material demonstrated that at every time interval in all the experiments the deposits were retarded when salt-extracted dermis was used. This suggests that the salt-soluble components of the dermis play a part in fiber formation.