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Pharmacologic Alternatives to Riboflavin Photochemical Corneal Cross-Linking: A Comparison Study of Cell Toxicity Thresholds
摘要： PURPOSE. The efficacy of therapeutic cross-linking of the cornea using riboflavin photochemistry (commonly abbreviated as CXL) has caused its use to become widespread. Because there are known chemical agents that cross-link collagenous tissues, it may be possible to cross-link tissue pharmacologically. The present study was undertaken to compare the cell toxicity of such agents. METHODS. Nine topical cross-linking agents (five nitroalcohols, glyceraldehyde [GLYC], genipin [GP], paraformaldehyde [FA], and glutaraldehyde [GLUT]) were tested with four different cell lines (immortalized human corneal epithelial cells, human skin fibroblasts, primary bovine corneal endothelial cells, and immortalized human retinal pigment epithelial cells [ARPE-19]). The cells were grown in planar culture and exposed to each agent in a range of concentrations (0.001 mM to 10 mM) for 24 hours followed by a 48-hour recovery phase. Toxicity thresholds were determined by using the trypan blue exclusion method. RESULTS. A semiquantitative analysis using five categories of toxicity/fixation was carried out, based on plate attachment, uptake of trypan blue stain, and cellular fixation. The toxicity levels varied by a factor of 103 with the least toxic being mononitroalcohols and GLYC, intermediate toxicity for a nitrodiol and nitrotriol, and the most toxic being GLUT, FA, GP, and bronopol, a brominated nitrodiol. When comparing toxicity between different cell lines, the levels were generally in agreement. CONCLUSIONS. There are significant differences in cell toxicity among potential topical cross-linking compounds. The balance between cross-linking of tissue and cell toxicity should be borne in mind as compounds and strategies to improve mechanical tissue properties through therapeutic tissue cross-linking continue to develop.
关键词： protein cross-linking,cornea,cellular toxicity,keratoconus
更新于2025-09-23 15:23:52
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Application of Matrix-Assisted Laser Desorption/Ionization Mass Spectrometric Imaging in Analysis of Medicinal Plants
摘要： Whey proteins (WPs) modification can improve their functional properties. To assess the crosslinking characteristics of Bacillus subtilis transglutaminase (BTG), it was used to reacted with WPs (12%, w/v) (40 °C, pH 8.0) for 6 h and 12 h, respectively. Both particle size profile and sodium dodecyl sulfate polyacrylamide gel electrophoresis demonstrated that BTG treatment enhanced the molecular weight of WPs. Meanwhile, BTG crosslinking greatly increased (*P < 0.05) the foaming and emulsifying properties of WPs. Furthermore, the rheological evaluation indicated that the cross-linked WPs showed greater storage modulus and loss modulus in comparison to the native WPs, leading to the increased gel strength. Additionally, BTG-induced WPs gels exhibited significantly enhanced (*P < 0.05) textural properties and water holding capacity. These results suggested the potential of BTG to enhance the functional properties of many proteins in the food industry.
关键词： Functional property,Bacillus subtilis transglutaminase,Whey protein,Cross-linking
更新于2025-09-16 10:30:52