In Vitro Photodynamic Treatment Modality for A375 Melanoma Cell Line Using a Sulphonated Aluminum Phthalocyanine Chloride-Photosensitizer-Gold Nanoparticle Conjugate
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Culture Conditions
2.2. Isolation of Stem Cells
2.3. Isolated A375 Melanoma Cancer Stem-Cell Characterization
2.3.1. CD133 and CD20 Flow Cytometry
2.3.2. CD133 and C20 Immunofluorescence (IF)
2.4. Subcellular Localization of AlPcS4Cl-AuNP in A375 Cells
2.5. Melanoma and Melanoma CSC AlPcS4Cl-AuNP PDT
2.6. Post-Irradiation Analyses
2.6.1. Light Microscopy
2.6.2. A375 CSC Post-Irradiation Lactate Dehydrogenase (LDH) Assay
2.6.3. A375 CSC Post-Irradiation Adenosine Triphosphate (ATP) Assay
2.6.4. Trypan Blue Dye Viability Exclusion Assay for A375 CSC Post-Irradiation
3. Results and Discussion
3.1. Identification and Analysis of A375 CSCs CD133 and CD20
3.2. Hoechst Nuclear Stain
3.3. Subcellular Localization of AlPcS4Cl-AuNP in A375 Cells
3.4. Post-Irradiation and Biochemical Assay Analysis
3.4.1. A375 CSC Morphology
3.4.2. A375 CSC Post-Irradiation Lactate Dehydrogenase (LDH) Assay
3.4.3. A375 CSC Post-Irradiation Adenosine Triphosphate (ATP) Assay
3.4.4. Trypan Blue Dye Viability Exclusion Assay for A375 CSC Post-Irradiation
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Groups (n = 6) |
---|
1. CSCs Only |
2. CSCs + Laser |
3. CSCs + AuNPs |
4. CSCs + 35 µm AlPcS4Cl |
5. A375 Cells + 35 µM AlPcS4Cl-AuNP |
6. CSCs + 35 µM AlPcS4Cl-AuNP |
7. CSCs + AuNPs + Laser |
8. CSCs + 35 µM AlPcS4Cl + Laser |
9. A375 Cells + 35 µM AlPcS4Cl-AuNP + Laser |
10. CSCs + 35 µM AlPcS4Cl-AuNP + Laser |
Name | Parameter |
---|---|
Laser Type | Semiconductor (Diode) |
Laser Average Output | 75 mW |
Wavelength | 673.2 nm |
Wave Emission | Continuous Wave (CW) |
Spectrum | Red (Visible) |
Radiant Exposure | 5 J/cm2 |
Photosensitizer (PS) | AlPcS4Cl-AuNP |
PS Concentrations | 35 µM (IC50 Standardized Dose) |
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Mkhobongo, B.; Chandran, R.; Abrahamse, H. In Vitro Photodynamic Treatment Modality for A375 Melanoma Cell Line Using a Sulphonated Aluminum Phthalocyanine Chloride-Photosensitizer-Gold Nanoparticle Conjugate. Pharmaceutics 2022, 14, 2474. https://doi.org/10.3390/pharmaceutics14112474
Mkhobongo B, Chandran R, Abrahamse H. In Vitro Photodynamic Treatment Modality for A375 Melanoma Cell Line Using a Sulphonated Aluminum Phthalocyanine Chloride-Photosensitizer-Gold Nanoparticle Conjugate. Pharmaceutics. 2022; 14(11):2474. https://doi.org/10.3390/pharmaceutics14112474
Chicago/Turabian StyleMkhobongo, Bridgette, Rahul Chandran, and Heidi Abrahamse. 2022. "In Vitro Photodynamic Treatment Modality for A375 Melanoma Cell Line Using a Sulphonated Aluminum Phthalocyanine Chloride-Photosensitizer-Gold Nanoparticle Conjugate" Pharmaceutics 14, no. 11: 2474. https://doi.org/10.3390/pharmaceutics14112474