Collagen banded fibril structure and the collagen-platelet reaction

Bovine hide collagen dispersions were swollen in the pH range 1.6-7.0, treated with glutaraldehyde, and dialyzed to neutral pH. The intensity with which these collagens reacted with human platelets in plasma was studied by aggregometry and scanning electron microscopy. Collagen swollen at a pH below 4.25 +/- 0.30 and treated with glutaraldehyde exhibited greatly reduced platelet aggregating ability after restoration of neutral pH. In addition, the state of supramolecular order in these collagens was investigated by transmission electron microscopy and infrared spectroscopy. Native, insoluble collagen fibrils were found to lose their banded structure, as observed by transmission electron microscopy, reversibly when exposed to low ionic strength aqueous solutions below pH 4.25 +/- 0.30. During the disorder transition, which occurred by time dependent swelling of fibrils, but without their disaggregation, the packing order in the fibrils was largely abolished while the triple helical structure of individual collagen molecules was retained. Chemical modification of collagen by glutaraldehyde treatment was found to prevent recrystallization of collagen during dialysis to neutral pH but did not otherwise affect the collagen-platelet reaction. The results of altering collagen mass dose (concentration) demonstrated the critical importance of traces of banded fibrils which resisted disordering below pH 4.25. The data suggest that collagen preparations which are free of significant traces of banded fibrils, but which are made up of collagen molecules possessing triple helical structure do not induce platelet aggregation, irrespective of dose.