Simple Methods for Staining Chitosan in Biotechnological Applications

AbstractAbstractChitosan is a naturally abundant cationic polysacchaside with unique physicochemical properties resulting in its use in a vast array of biomedical applications, including wound repair, drug delivery, and tissue engineering. However, despite numerous and diverse studies investigating its use, there have been no detailed reports describing specific staining methods for cell culture and histopathological studies using chitosan based materials. Here, we describe methods for staining chitosan in both cell culture and within paraffin sections. Procedural models used included identifying chitosan within in vitro whole blood clots, and culture wells, and in vivo chitosan hydrogel implants in multiple anatomical sites. First, a staining process for identifying chitosan was optimized using 0.03% Cibacron Brilliant Red-3BA. A second sensitive and versatile method using 0.01% Fast Green in conjunction with the known Safranin-O Method was demonstrated and may prove to be a key advancement in the histological study of chitosan-based biotechnological products. The method described here is broadly applicable to histological in situ detection of chitosan scaffolds of multiple deacetylation level and of variable fabrication. (The J Histotechnol 27:31, 2004)Submitted: September 30, 2003; Accepted: December 10, 2003Keywords: chitinFast GreenSafranin-OCibacron Brilliant Red-3BAtissue repairdrug deliveryblood clots