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Proceedings Paper

Gel electrophoretic studies of photochemical cross-linking of type I collagen with brominated 1,8-naphthalimide dyes and visible light
Author(s): Millard M. Judy; L. Fuh; James Lester Matthews; David E. Lewis; Ronald E. Utecht
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Paper Abstract

Insoluble Type I collagen from bovine Achilles tendon (Sigma C9879) was suspended in a 3 mM solution of the dye diEd66Br dissolved in Cremophor ELR (BASF) to give a molecular concentration ratio. Fifty-microliter aliquots in 5-mm-diameter wells were exposed to 458 J/cm2 (225 mW/cm2, 1800 sec) of 457.9-nm light from an argon ion laser; similar aliquots with and without dye were kept in the dark to serve as controls. Following pelleting of the collagen by centrifugation and 3x washing in phosphate-buffered saline, aliquots of light-treated and control sample pellets were (1) digested in collagenase (Sigma C9891) or (2) extracted in 0.5 M acetic acid, followed by centrifugative ultrafiltration (10-kd cutoff) in 0.01 M acetic acid. Aliquots of the supernatant of the acid-extracted collagen also were digested in pepsin. Electrophoretic protein migration in 8% to 25% gradient polyacrylamide gels following SDS solubilization disclosed numerous, densely packed, essentially contiguous protein bands. These studies indicate that the dye and light treatment of insoluble Type I collagen (1) results in cross-linking of collagen molecules and (2) does not denature the trimer conformation sufficiently to enable significant digestion by pepsin.

Paper Details

Date Published: 7 September 1994
PDF: 4 pages
Proc. SPIE 2128, Laser Surgery: Advanced Characterization, Therapeutics, and Systems IV, (7 September 1994); doi: 10.1117/12.184876
Show Author Affiliations
Millard M. Judy, Baylor Research Institute and MicroBioMed Corp. (United States)
L. Fuh, Baylor Research Institute (United States)
James Lester Matthews, Baylor Research Institute and MicroBioMed Corp. (United States)
David E. Lewis, South Dakota State Univ. and MicroBioMed Corp. (United States)
Ronald E. Utecht, South Dakota State Univ. and MicroBioMed Corp. (United States)


Published in SPIE Proceedings Vol. 2128:
Laser Surgery: Advanced Characterization, Therapeutics, and Systems IV
R. Rox Anderson; R. Rox Anderson, Editor(s)

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