Research Article
Volume 10 Issue 7 - 2022
Permeation, Bio-Distribution, and Subcellular Localization of Azulene Photosensitizer in Normal and Inflammatory Peripheral Blood Mononuclear Cells
Teerasak Damrongrungruang1,3,4*, Nujjakorn Yuhun2, Arunroong Kuntiya2 and Patcharaporn Klaipithak2
1Division of Oral Diagnosis, Department of Oral Biomedical Sciences, Faculty of Dentistry, Khon Kaen University, Thailand
2Dental Student, Faculty of Dentistry, Khon Kaen University, Khon Kaen, Thailand
3Laser in Dentistry Research Group, Khon Kaen University, Thailand
4Melatonin Research Program, Khon Kaen University, Thailand
*Corresponding Author: Teerasak Damrongrungruang, Division of Oral Diagnosis, Department of Oral Biomedical Sciences, Faculty of Dentistry, Khon Kaen University, Amphur Muang, Khon Kaen, Thailand.
Received: May 19, 2022; Published: June 29, 2022


Permeation and subcellular localization of photosensitizer in photodynamic therapy leads to precisely explaining the biological effects in the clinic. We aim to study cell permeation and subcellular localization of azulene between inflamed and normal human peripheral blood mononuclear cells (hPBMCs). 1, 10, 100 and, 1,000 mM (in 1%v/v methanol+deionized water) of azulene were incubated into hPBMCs and observed under a confocal fluorescence microscope. After the optimum concentration was determined, the permeation of this concentration of azulene was observed at 5, 10, 15, 20, 25 and, 30 minutes in either inflamed or normal hPBMCs. The subcellular localization was conducted by staining with 500 nM Mitotracker Red CMXRos and Lysotracker Green DND-26 followed the manufacturer's instruction for localization determination in mitochondria and lysosome, respectively, compared with the position of azulene in hPBMCs. The result shows that the longer the incubation time, the higher the permeation. In inflamed cells, the maximum permeation was at 25 minutes which gave the emission intensity close to 30 minutes. In normal PBMCs, the maximum permeation of azulene was in 20 minutes and azulene signal could not be detected at 30 minutes. The permeation of azulene in inflamed hPBMCs is higher than in normal hPBMCs. Regarding subcellular localization, indifference between mitochondria and lysosome localization of azulene in both inflamed and normal hPBMCs was demonstrated. In conclusion, the permeation of azulene was higher in inflamed hPBMCs compared with normal hPBMCs. With the limitation of this study, azulene appears to localize at both mitochondria and lysosome.

Keywords: Azulene; Permeation; Subcellular Localization; Peripheral Blood Mononuclear Cells; Inflammation


  1. Allison RR., et al. “Photodynamic therapy for the treatment of nonmelanomatous cutaneous malignancie”. Seminars in Cutaneous Medicine and Surgery2 (1998): 153-163.
  2. Hamblin MR and Abrahamse A. “Factors affecting photodynamic therapy and anti-tumor immune response”. Anticancer Agents in Medical Chemistry2 (2021): 123-136.
  3. Andrejevic-Blant S., et al. “Photodynamic therapy of early squamous cell carcinoma with tetra(m-hydroxyphenyl) chlorin: optimal drug-light interval”. British Journal of Cancer8 (1997): 1021-1028.
  4. Morton CA., et al. “A randomized, multinational, noninferiority, phase III trial to evaluate the safety and efficacy of BF‐200 aminolaevulinic acid gel vs. methyl aminolaevulinate cream in the treatment of nonaggressive basal cell carcinoma with photodynamic therapy”. British Journal of Dermatology2 (2018): 309-319.
  5. Zhang Y., et al. “Clinical study on clinical operation and post-treatment reactions of HMME-PDT in treatment of PWS”. Photodiagnosis and Photodynamic Therapy 20 (2017): 253-256.
  6. Zhao Y., et al. “Hemoporfin photodynamic therapy for port-wine stain: a randomized controlled trial”. PLoS One5 (2016): e0156219.
  7. Miola AC., et al. “Effectiveness and safety of 0.5% colchicine cream vs. photodynamic therapy with methyl aminolaevulinate in the treatment of actinic keratosis and skin field cancerization of the forearms: a randomized controlled trial”. British Journal of Dermatology5 (2018): 1081-1087.
  8. Cai XJ., et al. “Photodynamic therapy for intractable bronchial lung cancer”. Photodiagnosis Photodynamic Therapy4 (2013): 672-676.
  9. Damrongrungruang T., et al. “Effect of different irradiation modes of azulene-mediated photodynamic therapy on singlet oxygen and PGE2 formation”. Photochemistry and Photobiology2 (2021): 427-434.
  10. Damrongrungruang T., et al. “Effects of photodynamic therapy with azulene on peripheral blood mononuclear cell viability and singlet oxygen formation”. Photodiagnosis and Photodynamic Therapy 24 (2018): 318-323.
  11. Wang L., et al. “Phototoxicity and DNA damage induced by the cosmetic ingredient chemical azulene in human Jurkat T-cells”. Mutation Research1-2 (2004): 143-150.
  12. Dummin H., et al. “Selective photosensitization of mitochondria in HeLa cells by cationic Zn (II) phthalocyanines with lipophilic side-chains”. Journal of Photochemistry and Photobiology B: Biology3 (1997): 219-229.
  13. Teiten M-H., et al. “Endoplasmic reticulum and Golgi apparatus are the preferential sites of Foscan® localization in tumor cells”. British Journal of Cancer1 (2003): 146-152.
  14. Hsieh YJ., et al. “Subcellular localization of Photofrin determines the death phenotype of human epidermoid carcinoma A431 cells triggered by photodynamic therapy: when plasma membranes are the main targets”. Journal of Cellular Physiology3 (2003): 363-375.
  15. Daugelaviciene N., et al. “Lysosome-targeted photodynamic treatment induces primary keratinocyte differentiation”. Journal of Photochemistry and Photobiology B 218 (2021): 112183.
  16. Wang YY., et al. “MAL-PDT inhibits oral precancerous cells and lesions via autophagic cell death”. Oral Diseases3 (2019): 758-771.
  18. Tang D., et al. “The molecular machinery of regulated cell death”. Cell Research5 (2019): 347-364.
  19. Andersen FA. “Final report on the safety assessment of azulene”. International Journal of Toxicology3 (1999): 27-32.
  20. Miller GG., et al. “Uptake kinetics and intracellular localization of hypocrellin photosensitizers for photodynamic therapy: a confocal microscopy study”. Photochemistry and Photobiology6 (1995): 632-638.
  21. Sedgwick JB., et al. “Effects of inflammatory cytokines on the permeability of human lung microvascular endothelial cell monolayers and differential eosinophil transmigration”. Journal of Allergy and Clinical Immunology5 (2002): 752-756.
  22. Bakun P., et al. “In vitro and in vivo biological activities of azulene derivatives with potential applications in medicine”. Medicinal Chemistry Research4 (2021): 834-846.
  23. Luo W., et al. “Subcellular location and photodynamic therapeutic effect of chlorin e6 in the human tongue squamous cell cancer Tca8113 cell line”. Oncology Letters2 (2015): 551-556.
  24. Sandberg S., et al. “Porphyrin-induced photodamage as related to the subcellular localization of the porphyrins”. Acta Dermato-Venereologica Supplement (Stockhol) 100 (1982): 75-80.
  25. Tynga IM., et al. “The primary subcellular localization of Zinc phthalocyanine and its cellular impact on viability, proliferation and structure of breast cancer cells (MCF-7)”. Journal of Photochemistry and Photobiology B 120 (2013): 171-176.
  26. Høgset A., et al. “Photochemical internalisation in drug and gene delivery”. Advanced Drug Delivery Reviews1 (2004): 95-115.
  27. Rodrigues GB., et al. “Chemical features of the photosensitizers new methylene blue N and S137 influence their subcellular localization and photoinactivation efficiency in Candida albicans”. Journal of Photochemistry and Photobiology B 209 (2020): 111942.
  28. Fan Z., et al. “ABT737 enhances cholangiocarcinoma sensitivity to cisplatin through regulation of mitochondrial dynamics”. Experimental Cell Research1 (2015): 68-81.
  29. Ali-Seyed M., et al. “Photolon™-photosensitization induces apoptosis via ROS-mediated cross-talk between mitochondria and lysosomes”. International Journal of Oncology4 (2011): 821-831.
  30. Trivedi NS., et al. “Quantitative analysis of Pc 4 localization in mouse lymphoma (LY‐R) cells via double‐label confocal fluorescence microscopy”. Photochemistry and Photobiology5 (2000): 634-639.
  31. Shapiro AB and Ling V. “Effect of quercetin on Hoechst 33342 transport by purified and reconstituted P-glycoprotein”. Biochemical Pharmacology4 (1997): 587-596.
  32. Ciano L., et al. “The dynamics of azulene S2 state characterised by femtosecond transient absorption spectroscopy”. Journal of Photochemistry and Photobiology A. Chemistry2-3 (1997): 129-134.
Citation: Teerasak Damrongrungruang., et al. “Permeation, Bio-Distribution, and Subcellular Localization of Azulene Photosensitizer in Normal and Inflammatory Peripheral Blood Mononuclear Cells”. EC Pharmacology and Toxicology 10.7 (2022): 39-49.

PubMed Indexed Article

EC Pharmacology and Toxicology
LC-UV-MS and MS/MS Characterize Glutathione Reactivity with Different Isomers (2,2' and 2,4' vs. 4,4') of Methylene Diphenyl-Diisocyanate.

PMID: 31143884 [PubMed]

PMCID: PMC6536005

EC Pharmacology and Toxicology
Alzheimer's Pathogenesis, Metal-Mediated Redox Stress, and Potential Nanotheranostics.

PMID: 31565701 [PubMed]

PMCID: PMC6764777

EC Neurology
Differences in Rate of Cognitive Decline and Caregiver Burden between Alzheimer's Disease and Vascular Dementia: a Retrospective Study.

PMID: 27747317 [PubMed]

PMCID: PMC5065347

EC Pharmacology and Toxicology
Will Blockchain Technology Transform Healthcare and Biomedical Sciences?

PMID: 31460519 [PubMed]

PMCID: PMC6711478

EC Pharmacology and Toxicology
Is it a Prime Time for AI-powered Virtual Drug Screening?

PMID: 30215059 [PubMed]

PMCID: PMC6133253

EC Psychology and Psychiatry
Analysis of Evidence for the Combination of Pro-dopamine Regulator (KB220PAM) and Naltrexone to Prevent Opioid Use Disorder Relapse.

PMID: 30417173 [PubMed]

PMCID: PMC6226033

EC Anaesthesia
Arrest Under Anesthesia - What was the Culprit? A Case Report.

PMID: 30264037 [PubMed]

PMCID: PMC6155992

EC Orthopaedics
Distraction Implantation. A New Technique in Total Joint Arthroplasty and Direct Skeletal Attachment.

PMID: 30198026 [PubMed]

PMCID: PMC6124505

EC Pulmonology and Respiratory Medicine
Prevalence and factors associated with self-reported chronic obstructive pulmonary disease among adults aged 40-79: the National Health and Nutrition Examination Survey (NHANES) 2007-2012.

PMID: 30294723 [PubMed]

PMCID: PMC6169793

EC Dental Science
Important Dental Fiber-Reinforced Composite Molding Compound Breakthroughs

PMID: 29285526 [PubMed]

PMCID: PMC5743211

EC Microbiology
Prevalence of Intestinal Parasites Among HIV Infected and HIV Uninfected Patients Treated at the 1o De Maio Health Centre in Maputo, Mozambique

PMID: 29911204 [PubMed]

PMCID: PMC5999047

EC Microbiology
Macrophages and the Viral Dissemination Super Highway

PMID: 26949751 [PubMed]

PMCID: PMC4774560

EC Microbiology
The Microbiome, Antibiotics, and Health of the Pediatric Population.

PMID: 27390782 [PubMed]

PMCID: PMC4933318

EC Microbiology
Reactive Oxygen Species in HIV Infection

PMID: 28580453 [PubMed]

PMCID: PMC5450819

EC Microbiology
A Review of the CD4 T Cell Contribution to Lung Infection, Inflammation and Repair with a Focus on Wheeze and Asthma in the Pediatric Population

PMID: 26280024 [PubMed]

PMCID: PMC4533840

EC Neurology
Identifying Key Symptoms Differentiating Myalgic Encephalomyelitis and Chronic Fatigue Syndrome from Multiple Sclerosis

PMID: 28066845 [PubMed]

PMCID: PMC5214344

EC Pharmacology and Toxicology
Paradigm Shift is the Normal State of Pharmacology

PMID: 28936490 [PubMed]

PMCID: PMC5604476

EC Neurology
Examining those Meeting IOM Criteria Versus IOM Plus Fibromyalgia

PMID: 28713879 [PubMed]

PMCID: PMC5510658

EC Neurology
Unilateral Frontosphenoid Craniosynostosis: Case Report and a Review of the Literature

PMID: 28133641 [PubMed]

PMCID: PMC5267489

EC Ophthalmology
OCT-Angiography for Non-Invasive Monitoring of Neuronal and Vascular Structure in Mouse Retina: Implication for Characterization of Retinal Neurovascular Coupling

PMID: 29333536 [PubMed]

PMCID: PMC5766278

EC Neurology
Longer Duration of Downslope Treadmill Walking Induces Depression of H-Reflexes Measured during Standing and Walking.

PMID: 31032493 [PubMed]

PMCID: PMC6483108

EC Microbiology
Onchocerciasis in Mozambique: An Unknown Condition for Health Professionals.

PMID: 30957099 [PubMed]

PMCID: PMC6448571

EC Nutrition
Food Insecurity among Households with and without Podoconiosis in East and West Gojjam, Ethiopia.

PMID: 30101228 [PubMed]

PMCID: PMC6086333

EC Ophthalmology
REVIEW. +2 to +3 D. Reading Glasses to Prevent Myopia.

PMID: 31080964 [PubMed]

PMCID: PMC6508883

EC Gynaecology
Biomechanical Mapping of the Female Pelvic Floor: Uterine Prolapse Versus Normal Conditions.

PMID: 31093608 [PubMed]

PMCID: PMC6513001

EC Dental Science
Fiber-Reinforced Composites: A Breakthrough in Practical Clinical Applications with Advanced Wear Resistance for Dental Materials.

PMID: 31552397 [PubMed]

PMCID: PMC6758937

EC Microbiology
Neurocysticercosis in Child Bearing Women: An Overlooked Condition in Mozambique and a Potentially Missed Diagnosis in Women Presenting with Eclampsia.

PMID: 31681909 [PubMed]

PMCID: PMC6824723

EC Microbiology
Molecular Detection of Leptospira spp. in Rodents Trapped in the Mozambique Island City, Nampula Province, Mozambique.

PMID: 31681910 [PubMed]

PMCID: PMC6824726

EC Neurology
Endoplasmic Reticulum-Mitochondrial Cross-Talk in Neurodegenerative and Eye Diseases.

PMID: 31528859 [PubMed]

PMCID: PMC6746603

EC Psychology and Psychiatry
Can Chronic Consumption of Caffeine by Increasing D2/D3 Receptors Offer Benefit to Carriers of the DRD2 A1 Allele in Cocaine Abuse?

PMID: 31276119 [PubMed]

PMCID: PMC6604646

EC Anaesthesia
Real Time Locating Systems and sustainability of Perioperative Efficiency of Anesthesiologists.

PMID: 31406965 [PubMed]

PMCID: PMC6690616

EC Pharmacology and Toxicology
A Pilot STEM Curriculum Designed to Teach High School Students Concepts in Biochemical Engineering and Pharmacology.

PMID: 31517314 [PubMed]

PMCID: PMC6741290

EC Pharmacology and Toxicology
Toxic Mechanisms Underlying Motor Activity Changes Induced by a Mixture of Lead, Arsenic and Manganese.

PMID: 31633124 [PubMed]

PMCID: PMC6800226

EC Neurology
Research Volunteers' Attitudes Toward Chronic Fatigue Syndrome and Myalgic Encephalomyelitis.

PMID: 29662969 [PubMed]

PMCID: PMC5898812

EC Pharmacology and Toxicology
Hyperbaric Oxygen Therapy for Alzheimer's Disease.

PMID: 30215058 [PubMed]

PMCID: PMC6133268

News and Events

March Issue Release

We always feel pleasure to share our updates with you all. Here, notifying you that we have successfully released the March issue of respective journals and the latest articles can be viewed on the current issue pages.

Submission Deadline for Upcoming Issue

ECronicon delightfully welcomes all the authors around the globe for effective collaboration with an article submission for the upcoming issue of respective journals. Submissions are accepted on/before April 03, 2023.

Certificate of Publication

ECronicon honors with a "Publication Certificate" to the corresponding author by including the names of co-authors as a token of appreciation for publishing the work with our respective journals.

Best Article of the Issue

Editors of respective journals will always be very much interested in electing one Best Article after each issue release. The authors of the selected article will be honored with a "Best Article of the Issue" certificate.

Certifying for Review

ECronicon certifies the Editors for their first review done towards the assigned article of the respective journals.

Latest Articles

The latest articles will be updated immediately on the articles in press page of the respective journals.