Abstract

Rose Bengal (RB) is a potential photosuturing agent that may improve standard dermatologic surgical closure techniques. However, RB produces reactive oxygen species with photoactivation and its photomutagenic potential must be considered in clinical application. We investigated cytotoxicity, mutagenicity, and singlet oxygen (SO) production of RB on epithelial Chinese hamster ovary cell line. Cells were exposed to RB concentrations: 0.1%, 0.01%, 0.001%, 0.0001%, 0.00001%; irradiated for 400 s using a high-intensity visible wavelength lamp or maintained in the dark. Cell viability was assessed by XTT assay, mutagenicity by HPRT gene mutation assay, and SO production by Sensor Green reagent. RB > 0.001% was significantly cytotoxic. Viabilities were uninfluenced by ≤0.0001% RB controls, or 30-min incubation. 49% of irradiated cells died after 24-h in 0.0001% RB. At ≥0.001% RB, >90% of cells died. Irradiating 0.00001% - 0.001% RB increased SO; levels dropped significantly between 0.01% - 0.1%. Controls exhibited negligible SO production. HPRT suggested that RB was not mutagenic (0.0001%, 0.00001%); SO induction increased between 0.00001% - 0.001%, with reduced production at higher concentrations. Pilot studies suggested irradiated 0.0001% RB is mutagenic in vitro; current data suggest RB is not photomutagenic. The contribution of RB’s cytotoxicity on observed clinical improvement of scars and mutagenic potential remains unclear, necessitating further study.

Highlights

  • Wound healing and scar formation are researched extensively, and especially important to dermatological surgery

  • 45% of cells were lost after light irradiation and 24 hours of incubation with 0.0001% Rose Bengal (RB) (Figure 3)

  • No Light, 30 Minutes Incubation There was no difference between the control and the lowest dose (0.0001% RB); comparing these to the higher concentrations in this group, controls and 0.0001% RB were significantly different from all other concentrations (p < 0.01), with viabilities greatly reduced in the latter concentrations

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Summary

Introduction

Wound healing and scar formation are researched extensively, and especially important to dermatological surgery. Skin incisions and excisions are closed by a variety suture types, staples, wound closure strips, and adhesives. Recent research shows promise in improved epidermal wound closure and healing with a photochemical tissue bonding (PTB) agent, 0.1% Rose Bengal (RB;4,5,6,7-tetrachloro-2’,4’,5’,7’-tetraiodo-fluorescein) suspended in phosphate buffered saline (PBS), when applied to wound edges and activated by green laser light [1,2]. Rose Bengal may behave as a sort of “nanosuture,” as the dye may crosslink proteins between tissues surfaces thereby sealing skin wounds [2]. We investigated toxicity, mutagenicity, and singlet oxygen production of photoactivated Rose Bengal using epithelial cells in culture

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