Efficacy of ultra short sub-30 minute incubation of 5-aminolevulinic acid photodynamic therapy in vitro

Eugene Koo, Evan Austin, Andrew Mamalis, Jared Jagdeo

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background and Objective: The estimated incidence of cutaneous squamous cell carcinoma (SCC) is 700,000 cases per year. In the US, SCC incidence is highest among fair skinned adults older than 50 years of age. Thus, as the population ages, the reported number of SCCs will likely increase in the future. Photodynamic therapy (PDT) is an FDA approved therapy for treatment of actinic keratoses (AKs), a precursor to cutaneous SCC lesions. The FDA approved incubation time of the photosensitizing agent 5-aminolevulinic acid (ALA) is 14-18 hours. Recent studies have investigated short ALA incubation times of 1-3 hours with ALA and PDT demonstrating treatment success. Therefore, the question exists whether ALA incubation periods of less than 30 minutes are efficacious. Herein, we evaluate the efficacy of short ALA incubation periods by measuring apoptosis after 10, 15, and 20 minutes of ALA incubation. Study Design/Materials and Methods: AG13145 normal human dermal fibroblasts HDFs were incubated with 10, 15, or 20 minute of ALA at various concentrations (0, 0.05, 0.075, 0.1, 0.25, 0.375, 0.5, 1, and 2mM). After ALA incubation, samples were treated with 1,000 seconds (16 minutes 40 seconds) of Blu-U fluorescent blue light (417±5nm) for a fluence of 10J/cm2. Immediately following treatment with blue light, samples were collected and stained for apoptosis and necrosis with annexin-V and 7-aminoactinomycin D (7-AAD), and then analyzed by flow cytometry. Results: HDFs incubated with ALA for 10minute at 36°C followed by 10J/cm2 of blue light had no statistically significant changes in apoptosis. HDFs incubated with ALA for 15 or 20 minutes at 36°C followed by 10J/cm2 of blue light had statistically significant increases in the percentages of cells positive for apoptosis in the 0.5, 1.0, and 2.0mM ALA doses (P<0.05). Conclusions: We found that incubation of ALA for at least 15 minutes followed by 10J/cm2 of blue light resulted in a statistically significant increase in apoptosis.

Original languageEnglish (US)
JournalLasers in Surgery and Medicine
DOIs
StateAccepted/In press - 2017

Fingerprint

Aminolevulinic Acid
Photochemotherapy
Apoptosis
Light
Squamous Cell Carcinoma
Skin
In Vitro Techniques
Actinic Keratosis
Photosensitizing Agents
Annexin A5
Incidence
Therapeutics
Flow Cytometry
Necrosis
Fibroblasts

Keywords

  • 5-aminolevulinic acid
  • Actinic keratosis
  • Blue light
  • Photodynamic therapy
  • Short incubation
  • Squamouscell carcinoma

ASJC Scopus subject areas

  • Surgery
  • Dermatology

Cite this

Efficacy of ultra short sub-30 minute incubation of 5-aminolevulinic acid photodynamic therapy in vitro. / Koo, Eugene; Austin, Evan; Mamalis, Andrew; Jagdeo, Jared.

In: Lasers in Surgery and Medicine, 2017.

Research output: Contribution to journalArticle

@article{5249b597ac244fe2a806319f7d6d4b13,
title = "Efficacy of ultra short sub-30 minute incubation of 5-aminolevulinic acid photodynamic therapy in vitro",
abstract = "Background and Objective: The estimated incidence of cutaneous squamous cell carcinoma (SCC) is 700,000 cases per year. In the US, SCC incidence is highest among fair skinned adults older than 50 years of age. Thus, as the population ages, the reported number of SCCs will likely increase in the future. Photodynamic therapy (PDT) is an FDA approved therapy for treatment of actinic keratoses (AKs), a precursor to cutaneous SCC lesions. The FDA approved incubation time of the photosensitizing agent 5-aminolevulinic acid (ALA) is 14-18 hours. Recent studies have investigated short ALA incubation times of 1-3 hours with ALA and PDT demonstrating treatment success. Therefore, the question exists whether ALA incubation periods of less than 30 minutes are efficacious. Herein, we evaluate the efficacy of short ALA incubation periods by measuring apoptosis after 10, 15, and 20 minutes of ALA incubation. Study Design/Materials and Methods: AG13145 normal human dermal fibroblasts HDFs were incubated with 10, 15, or 20 minute of ALA at various concentrations (0, 0.05, 0.075, 0.1, 0.25, 0.375, 0.5, 1, and 2mM). After ALA incubation, samples were treated with 1,000 seconds (16 minutes 40 seconds) of Blu-U fluorescent blue light (417±5nm) for a fluence of 10J/cm2. Immediately following treatment with blue light, samples were collected and stained for apoptosis and necrosis with annexin-V and 7-aminoactinomycin D (7-AAD), and then analyzed by flow cytometry. Results: HDFs incubated with ALA for 10minute at 36°C followed by 10J/cm2 of blue light had no statistically significant changes in apoptosis. HDFs incubated with ALA for 15 or 20 minutes at 36°C followed by 10J/cm2 of blue light had statistically significant increases in the percentages of cells positive for apoptosis in the 0.5, 1.0, and 2.0mM ALA doses (P<0.05). Conclusions: We found that incubation of ALA for at least 15 minutes followed by 10J/cm2 of blue light resulted in a statistically significant increase in apoptosis.",
keywords = "5-aminolevulinic acid, Actinic keratosis, Blue light, Photodynamic therapy, Short incubation, Squamouscell carcinoma",
author = "Eugene Koo and Evan Austin and Andrew Mamalis and Jared Jagdeo",
year = "2017",
doi = "10.1002/lsm.22648",
language = "English (US)",
journal = "Lasers in Surgery and Medicine",
issn = "0196-8092",
publisher = "Wiley-Liss Inc.",

}

TY - JOUR

T1 - Efficacy of ultra short sub-30 minute incubation of 5-aminolevulinic acid photodynamic therapy in vitro

AU - Koo, Eugene

AU - Austin, Evan

AU - Mamalis, Andrew

AU - Jagdeo, Jared

PY - 2017

Y1 - 2017

N2 - Background and Objective: The estimated incidence of cutaneous squamous cell carcinoma (SCC) is 700,000 cases per year. In the US, SCC incidence is highest among fair skinned adults older than 50 years of age. Thus, as the population ages, the reported number of SCCs will likely increase in the future. Photodynamic therapy (PDT) is an FDA approved therapy for treatment of actinic keratoses (AKs), a precursor to cutaneous SCC lesions. The FDA approved incubation time of the photosensitizing agent 5-aminolevulinic acid (ALA) is 14-18 hours. Recent studies have investigated short ALA incubation times of 1-3 hours with ALA and PDT demonstrating treatment success. Therefore, the question exists whether ALA incubation periods of less than 30 minutes are efficacious. Herein, we evaluate the efficacy of short ALA incubation periods by measuring apoptosis after 10, 15, and 20 minutes of ALA incubation. Study Design/Materials and Methods: AG13145 normal human dermal fibroblasts HDFs were incubated with 10, 15, or 20 minute of ALA at various concentrations (0, 0.05, 0.075, 0.1, 0.25, 0.375, 0.5, 1, and 2mM). After ALA incubation, samples were treated with 1,000 seconds (16 minutes 40 seconds) of Blu-U fluorescent blue light (417±5nm) for a fluence of 10J/cm2. Immediately following treatment with blue light, samples were collected and stained for apoptosis and necrosis with annexin-V and 7-aminoactinomycin D (7-AAD), and then analyzed by flow cytometry. Results: HDFs incubated with ALA for 10minute at 36°C followed by 10J/cm2 of blue light had no statistically significant changes in apoptosis. HDFs incubated with ALA for 15 or 20 minutes at 36°C followed by 10J/cm2 of blue light had statistically significant increases in the percentages of cells positive for apoptosis in the 0.5, 1.0, and 2.0mM ALA doses (P<0.05). Conclusions: We found that incubation of ALA for at least 15 minutes followed by 10J/cm2 of blue light resulted in a statistically significant increase in apoptosis.

AB - Background and Objective: The estimated incidence of cutaneous squamous cell carcinoma (SCC) is 700,000 cases per year. In the US, SCC incidence is highest among fair skinned adults older than 50 years of age. Thus, as the population ages, the reported number of SCCs will likely increase in the future. Photodynamic therapy (PDT) is an FDA approved therapy for treatment of actinic keratoses (AKs), a precursor to cutaneous SCC lesions. The FDA approved incubation time of the photosensitizing agent 5-aminolevulinic acid (ALA) is 14-18 hours. Recent studies have investigated short ALA incubation times of 1-3 hours with ALA and PDT demonstrating treatment success. Therefore, the question exists whether ALA incubation periods of less than 30 minutes are efficacious. Herein, we evaluate the efficacy of short ALA incubation periods by measuring apoptosis after 10, 15, and 20 minutes of ALA incubation. Study Design/Materials and Methods: AG13145 normal human dermal fibroblasts HDFs were incubated with 10, 15, or 20 minute of ALA at various concentrations (0, 0.05, 0.075, 0.1, 0.25, 0.375, 0.5, 1, and 2mM). After ALA incubation, samples were treated with 1,000 seconds (16 minutes 40 seconds) of Blu-U fluorescent blue light (417±5nm) for a fluence of 10J/cm2. Immediately following treatment with blue light, samples were collected and stained for apoptosis and necrosis with annexin-V and 7-aminoactinomycin D (7-AAD), and then analyzed by flow cytometry. Results: HDFs incubated with ALA for 10minute at 36°C followed by 10J/cm2 of blue light had no statistically significant changes in apoptosis. HDFs incubated with ALA for 15 or 20 minutes at 36°C followed by 10J/cm2 of blue light had statistically significant increases in the percentages of cells positive for apoptosis in the 0.5, 1.0, and 2.0mM ALA doses (P<0.05). Conclusions: We found that incubation of ALA for at least 15 minutes followed by 10J/cm2 of blue light resulted in a statistically significant increase in apoptosis.

KW - 5-aminolevulinic acid

KW - Actinic keratosis

KW - Blue light

KW - Photodynamic therapy

KW - Short incubation

KW - Squamouscell carcinoma

UR - http://www.scopus.com/inward/record.url?scp=85016766365&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85016766365&partnerID=8YFLogxK

U2 - 10.1002/lsm.22648

DO - 10.1002/lsm.22648

M3 - Article

C2 - 28370019

AN - SCOPUS:85016766365

JO - Lasers in Surgery and Medicine

JF - Lasers in Surgery and Medicine

SN - 0196-8092

ER -