The Extraction of Tropocollagen and Proteoglycans from Bovine Eyes and Investigation of Their Recombination Using a New Method to Quantify Fibrillogenesis.

摘要

Current methods for observing collagen fibrillogenesis lack the ability to dynamically examine fibril formation, sacrificing valuable information. The primary aim of this series of investigations was to develop and validate a new, robust system for observation of fibrillogenesis utilizing Differential Interference Contrast (DIC) microscopy with edge detection image analysis. A secondary goal was the investigation of the effect of telopeptides and bovine corneal and scleral proteoglycans on the fibrillogenesis of bovine scleral tropocollagen.;The DIC method developed offers a highly controllable system, offering augmented information and advantages over conventional methods. The presence of intact telopeptides on the collagen molecule alters the kinetics of fibrillogenesis both morphologically, and significantly in linear growth rates and characteristic turbidity curve times. Inclusion of scleral and corneal proteoglycans both exhibit significant effects on fibrillogenesis, though those extracted from cornea were shown to offer greater control over rates and density of final matrix via DIC imaging.