Riboflavin Collagen Crosslinking
08/26/19
Crosslinking collagen with riboflavin is a widely used application for corneal treaments. Below is a simple protocol outlining a starting strategy for photocrosslinking collagen using riboflavin and UV light.
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Mix riboflavin at a 0.02% solution (20 mg/100 ml) in 1X PBS.
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Bathe your collagen construct or hydrogel in the riboflavin solution for 5 minutes.
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Expose to UV light (365nm) for 5 minutes.
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Wash out the riboflavin with cell culture media (ensure the riboflavin is fully exchanged out for media).
This should result in about a 2X increase in collagen gel stiffness. UV light intensity, crosslinking time, riboflavin concentration, collagen concentration and other variables will affect your results.
It should also be noted that prolonged UV light exposure can be damaging to cells and affect cell viability.
