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Optical clearing agent perfusion enhancement via combination of microneedle poration, heating and pneumatic pressure.

Lasers in surgery and medicine (2014-05-28)
Yasaman Damestani, Bissrat Melakeberhan, Masaru P Rao, Guillermo Aguilar
ABSTRACT

Optical clearing agents (OCAs) have shown promise for increasing the penetration depth of biomedical lasers by temporarily decreasing optical scattering within the skin. However, their translation to the clinic has been constrained by lack of practical means for effectively perfusing OCA within target tissues in vivo. The objective of this study was to address this limitation through combination of a variety of techniques to enhance OCA perfusion, including heating of OCA, microneedling and/or application of pneumatic pressure over the skin surface being treated (vacuum and/or positive pressure). While some of these techniques have been explored by others independently, the current study represents the first to explore their use together. Propylene glycol (PG) OCA, either at room-temperature or heated to 45°C, was topically applied to hydrated, body temperature ex vivo porcine skin, in conjunction with various combinations of microneedling pre-treatment (0.2 mm length microneedles, performed prior to OCA application), vacuum pre-treatment (17-50 kPa, performed prior to OCA application), and positive pressure post-treatment (35-172 kPa, performed after OCA application). The effectiveness of OCA perfusion was characterized via measurements of transmittance, reduced scattering coefficient, and penetration depth at a number of medically-relevant laser wavelengths across the visible spectrum. Topical application of room-temperature (RT) PG led to an increase in transmittance across the visible spectrum of up to 21% relative to untreated skin. However, only modest increases were observed with addition of various combinations of microneedling pre-treatment, vacuum pre-treatment, and positive pressure post-treatment. Conversely, when heated PG was used in conjunction with these techniques, we observed significant increases in transmittance. Using an optimal PG perfusion enhancement protocol consisting of 45°C heated PG + microneedle pre-treatment + 35 kPa vacuum pre-treatment + 103 kPa positive pressure post-treatment, we observed up to 68% increase in transmittance relative to untreated skin, and up to 46% increase relative to topical RT PG application alone. Using the optimal PG perfusion enhancement protocol, we also observed up to 30% decrease in reduced scattering coefficient relative to untreated skin, and up to 20% decrease relative to topical RT PG alone. Finally, using the optimal protocol, we observed up to 25% increase in penetration depth relative to untreated skin, and up to 23% increase relative to topical RT PG alone. The combination of heated PG, microneedling pre-treatment, vacuum pre-treatment, and positive pressure-post treatment were observed to significantly enhance the perfusion of topically applied PG. Although further studies are required to evaluate the efficacy of combined perfusion enhancement techniques in vivo, the current results suggest promise for facilitating the translation of OCAs to the clinic.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Propylene glycol, FCC, FG
Supelco
Propylene glycol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Propylene Glycol, meets USP testing specifications
USP
Propylene glycol, United States Pharmacopeia (USP) Reference Standard
Propylene glycol, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
1,2-Propanediol, SAJ special grade, ≥99.0%
Sigma-Aldrich
1,2-Propanediol, SAJ first grade, ≥99.0%
Sigma-Aldrich
1,2-Propanediol, ACS reagent, ≥99.5%
Sigma-Aldrich
1,2-Propanediol, ReagentPlus®, 99%
Sigma-Aldrich
1,2-Propanediol, meets analytical specification of Ph. Eur., BP, USP, ≥99.5%
Sigma-Aldrich
1,2-Propanediol, puriss. p.a., ACS reagent, ≥99.5% (GC)
Sigma-Aldrich
1,2-Propanediol, tested according to Ph. Eur.
Supelco
1,2-Propanediol, analytical standard
Sigma-Aldrich
1,2-Propanediol, LR, ≥99%