Subcytotoxic transepidermal delivery using low intensity cold atmospheric plasma
Abstract Cold atmospheric plasma (CAP) has been utilized in various medical devices using its oxidative nature. Recent studies have provided evidence that CAP can facilitate the delivery of large, hydrophilic molecules through the epidermis to the dermis. On the other hand, a new approach called low...
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2025-01-01
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| author | Ga Ram Ahn Hyung-Joon Park Yu Jin Kim Min Gyo Song Hye Sung Han Woo Geon Lee Hyuck Ki Hong Kwang Ho Yoo Joon Seok Kyu Back Lee Beom Joon Kim |
| author_facet | Ga Ram Ahn Hyung-Joon Park Yu Jin Kim Min Gyo Song Hye Sung Han Woo Geon Lee Hyuck Ki Hong Kwang Ho Yoo Joon Seok Kyu Back Lee Beom Joon Kim |
| author_sort | Ga Ram Ahn |
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| description | Abstract Cold atmospheric plasma (CAP) has been utilized in various medical devices using its oxidative nature. Recent studies have provided evidence that CAP can facilitate the delivery of large, hydrophilic molecules through the epidermis to the dermis. On the other hand, a new approach called low-intensity CAP (LICAP) has been developed, allowing the plasma level to be controlled within a subtoxic range, thereby demonstrating various biological benefits without tissue damage. However, the ability of LICAP to enhance transepidermal delivery in sub-cytotoxic conditions has not been fully investigated. This study aims to determine the sub-cytotoxic range of exposure time for LICAP and, within the range, to investigate the effects of LICAP treatment on transepidermal drug delivery (TED) and mechanisms using human keratinocytes and a mouse model. For the in vitro studies, LICAP treatment was evaluated in human keratinocyte (HaCaT) cells by assessing reactive species production, DNA damage, and cytotoxicity profiles. Within the determined safety range, mechanistic analyses were conducted to examine LICAP-enhanced delivery pathways. mRNA expression and protein levels of tight and adherens junction genes were quantified, and changes in ultramicroscopic morphology of HaCaT monolayers were investigated. Intracellular delivery of fluorescein isothiocyanate (FITC)-dextran was also assessed. For the in vivo studies, E-cadherin expression and the transepidermal delivery (TED) of human epidermal growth factor (hEGF) were analyzed in LICAP-treated mouse dorsal skin. The upper safety range of LICAP exposure time, reducing cell viability by 70% (IC70 or LD30), was estimated at 34.3 s. Within the safety range, LICAP treatment downregulated multiple tight and adherens junction genes in HaCaT cells. Consistent with the in vitro results, the epidermal E-cadherin expression was reduced, and human epidermal growth factor (hEGF) was infiltrated in the dermis of the LICAP-treated mouse skin. Intercellular clefts were detected in the HaCaT cell monolayer immediately following LICAP treatment and intracellular delivery of FITC-dextran was confirmed after LICAP exposure. This study demonstrated that LICAP treatment enhances transepidermal permeation of hEGF, apparently via both paracellular and transcellular routes. Under our study conditions, LICAP treatment seems to be a novel approach to facilitate TED with low safety concerns in vitro. Further translational studies are needed for clinical evaluation. |
| format | Article |
| id | doaj-art-6c19f08a36b843c4aaed149f43b66a11 |
| institution | Kabale University |
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| publishDate | 2025-01-01 |
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| spelling | doaj-art-6c19f08a36b843c4aaed149f43b66a112025-01-19T12:19:57ZengNature PortfolioScientific Reports2045-23222025-01-0115111410.1038/s41598-024-83201-0Subcytotoxic transepidermal delivery using low intensity cold atmospheric plasmaGa Ram Ahn0Hyung-Joon Park1Yu Jin Kim2Min Gyo Song3Hye Sung Han4Woo Geon Lee5Hyuck Ki Hong6Kwang Ho Yoo7Joon Seok8Kyu Back Lee9Beom Joon Kim10Department of Dermatology, Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical SchoolDepartment of Interdisciplinary Bio/Micro System Technology, College of Engineering, Korea UniversityCollege of Medicine, Chung-Ang UniversityDepartment of Biomedical Engineering, College of Health Science, Korea UniversityCollege of Medicine, Chung-Ang UniversityCollege of Medicine, Chung-Ang UniversityHuman IT Convergence System R&D Division, Korea Electronics Technology InstituteCollege of Medicine, Chung-Ang UniversityDepartment of Dermatology, Chung-Ang University HospitalDepartment of Biomedical Engineering, College of Health Science, Korea UniversityDepartment of Dermatology, Chung-Ang University HospitalAbstract Cold atmospheric plasma (CAP) has been utilized in various medical devices using its oxidative nature. Recent studies have provided evidence that CAP can facilitate the delivery of large, hydrophilic molecules through the epidermis to the dermis. On the other hand, a new approach called low-intensity CAP (LICAP) has been developed, allowing the plasma level to be controlled within a subtoxic range, thereby demonstrating various biological benefits without tissue damage. However, the ability of LICAP to enhance transepidermal delivery in sub-cytotoxic conditions has not been fully investigated. This study aims to determine the sub-cytotoxic range of exposure time for LICAP and, within the range, to investigate the effects of LICAP treatment on transepidermal drug delivery (TED) and mechanisms using human keratinocytes and a mouse model. For the in vitro studies, LICAP treatment was evaluated in human keratinocyte (HaCaT) cells by assessing reactive species production, DNA damage, and cytotoxicity profiles. Within the determined safety range, mechanistic analyses were conducted to examine LICAP-enhanced delivery pathways. mRNA expression and protein levels of tight and adherens junction genes were quantified, and changes in ultramicroscopic morphology of HaCaT monolayers were investigated. Intracellular delivery of fluorescein isothiocyanate (FITC)-dextran was also assessed. For the in vivo studies, E-cadherin expression and the transepidermal delivery (TED) of human epidermal growth factor (hEGF) were analyzed in LICAP-treated mouse dorsal skin. The upper safety range of LICAP exposure time, reducing cell viability by 70% (IC70 or LD30), was estimated at 34.3 s. Within the safety range, LICAP treatment downregulated multiple tight and adherens junction genes in HaCaT cells. Consistent with the in vitro results, the epidermal E-cadherin expression was reduced, and human epidermal growth factor (hEGF) was infiltrated in the dermis of the LICAP-treated mouse skin. Intercellular clefts were detected in the HaCaT cell monolayer immediately following LICAP treatment and intracellular delivery of FITC-dextran was confirmed after LICAP exposure. This study demonstrated that LICAP treatment enhances transepidermal permeation of hEGF, apparently via both paracellular and transcellular routes. Under our study conditions, LICAP treatment seems to be a novel approach to facilitate TED with low safety concerns in vitro. Further translational studies are needed for clinical evaluation.https://doi.org/10.1038/s41598-024-83201-0Cold atmospheric plasmaCytotoxicityHormesisLow intensityLICAPPlasma jet |
| spellingShingle | Ga Ram Ahn Hyung-Joon Park Yu Jin Kim Min Gyo Song Hye Sung Han Woo Geon Lee Hyuck Ki Hong Kwang Ho Yoo Joon Seok Kyu Back Lee Beom Joon Kim Subcytotoxic transepidermal delivery using low intensity cold atmospheric plasma Scientific Reports Cold atmospheric plasma Cytotoxicity Hormesis Low intensity LICAP Plasma jet |
| title | Subcytotoxic transepidermal delivery using low intensity cold atmospheric plasma |
| title_full | Subcytotoxic transepidermal delivery using low intensity cold atmospheric plasma |
| title_fullStr | Subcytotoxic transepidermal delivery using low intensity cold atmospheric plasma |
| title_full_unstemmed | Subcytotoxic transepidermal delivery using low intensity cold atmospheric plasma |
| title_short | Subcytotoxic transepidermal delivery using low intensity cold atmospheric plasma |
| title_sort | subcytotoxic transepidermal delivery using low intensity cold atmospheric plasma |
| topic | Cold atmospheric plasma Cytotoxicity Hormesis Low intensity LICAP Plasma jet |
| url | https://doi.org/10.1038/s41598-024-83201-0 |
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