sdj-10161

Evaluation of 5-Aminolevulinic acid, Methylene Blue, Toluidine Blue O Photosensitizers on Clinically Isolated Porphyromonas gingivalis and Prevotella intermedia: An in vitro Study

Kashan K. Najm* , Sarhang S. Gul*

 

* Department of Periodontics, College of Dentistry, University of Sulaimani, Sulaimani, Iraq.

 

 

 

Submitted: July 8, 2022, Accepted: August 27, 2022, Published: December 1, 2022.

 

DOI: https://doi.org/10.17656/sdj.10161

Objective: To evaluate the antibacterial efficacy of 5-aminolevulinic acid (5-ALA), methylene blue (MB), and toluidine blue O (TBO) photosensitizers against clinically isolated Porphyromonas gingivalis (P. gingivalis) and Prevotella intermedia (P. intermedia).

Methods: Patients with a periodontal pocket depth of ≥5 mm were selected for subgingival plaque sampling. Bacterial identification was carried out using colony morphology, gram staining and then validated using the PCR technique. The photosensitizers tested were 5-ALA, MB, and TBO. The antibacterial test was performed using an agar well, and disk diffusion methods and double-fold serial dilution was used for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC).     

Results: Glossy, smooth, and looked grey, light brown, or black-pigmented colonies of P. intermedia on Columbia agar plates were confirmed by gram staining and PCR tests. Contrary to MB and TBO, 5-ALA photosensitizers didn’t have antibacterial efficacy against P. gingivalis and P. intermedia. The antibacterial test showed an increased inhibition zone with increasing the photosensitizers’ concentration. The greatest inhibition zones were at 10mg/ml concentration for both MB and TBO. The MIC for MB was 2 mg/ml, and 1mg/ml, while for TBO, it was 1mg/ml and 0.5mg/ml for P. gingivalis and P. intermedia, respectively.    

Conclusions: MB and TBO have antibacterial properties against P. gingivalis and P. intermedia, and their efficacy increase with increasing concentration. TBO is more effective when compared to MB. Furthermore, P. intermedia is more sensitive to photoactivation in comparison to P. gingivalis.                                                                                     

Keywords: Antibacterial effect, Methylene blue, Porphyromonas gingivalis, Prevotella intermedia, Toluidine blue O, 5-aminolevulinic acid.         

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References:

1.Jakubovics NS, Goodman SD, Mashburn‐Warren L, Stafford GP, Cieplik F. The dental plaque biofilm matrix. Periodontol 2000. 2021;86(1):32- 56.

2. Zhang Z, Liu D, Liu S, Zhang S, Pan Y. The role of Porphyromonas gingivalis outer membrane vesicles in periodontal disease and related systemic diseases. Front Cell Infect Microbiol. 2020;10:585917:1-12.

3. Cheng WC, Van Asten SD, Burns LA, Evans HG, Walter GJ, Hashim A, et al. Periodontitis- associated pathogens P. gingivalis and A. actinomycetemcomitans activate human CD14(+) monocytes leading to enhanced Th17/IL-17 responses. Eur J Immunol. 2016;46(9):2211-21.

4. Naito M, Belvin BR, Shoji M, Gui Q, Lewis JP. Insertional inactivation of Prevotella intermedia OxyR results in reduced survival with oxidative stress and in the presence of host cells. Microorganisms. 2021;9(3):551.

5. Pan W, Wang Q, Chen Q. The cytokine network involved in the host immune response to periodontitis. Int J Oral Sci. 2019;11(3):1-13.

6. Colombo APV, Magalhães CB, Hartenbach FaRR, Do Souto RM, Da Silva-Boghossian CM. Periodontal-disease-associated biofilm: A reservoir for pathogens of medical importance. Microb Pathog. 2016;94(1):27-34.

7. Kuboniwa M, Houser JR, Hendrickson EL, Wang Q, Alghamdi SA, Sakanaka A. Metabolic crosstalk regulates Porphyromonas gingivalis colonization and virulence during oral polymicrobial infection. Nat Microbiol. 2017;2(11):1493-9.

8. Hajishengallis G, Lambris JD. Microbial manipulation of receptor crosstalk in innate immunity. Nat Rev Immunol. 2011;11(3):187- 200.

9. Xu W, Zhou W, Wang H, Liang S. Roles of Porphyromonas gingivalis and its virulence factors in periodontitis. Adv Protein Chem Struct Biol. 2020;120:45-84.

10. Könönen E, Gursoy M, Gursoy UK. Periodontitis: A multifaceted disease of tooth-supporting tissues. J Clin Med. 2019;8(8):1135.

11. Hartenbach F, Silva-Boghossian CM, Colombo APV. The effect of supragingival biofilm re- development on the subgingival microbiota in chronic periodontitis. Arch Oral Biol. 2018;85(1):51-7.

12. Tariq M, Iqbal Z, Ali J, Baboota S, Talegaonkar S, Ahmad Z, et al. Treatment modalities and evaluation models for periodontitis. Int J Pharm Investig. 2012;2(3):106-22.

13. Van Der Weijden G, Dekkers GJ, Slot DE. Success of non‐surgical periodontal therapy in adult periodontitis patients: A retrospective analysis. Int J Dent Hyg. 2019;17(4):309-17.

14. Barca E, Cifcibasi E, Cintan S. Adjunctive use of antibiotics in periodontal therapy. J Istanb Univ Fac Dent. 2015;49(3):55-62.

15. Heta S, Robo I. The side effects of the most commonly used group of antibiotics in periodontal treatments. Med Sci (Basel). 2018;6(1):1-6.

16. Liu Y, Qin R, Zaat SA, Breukink E, Heger M. Antibacterial photodynamic therapy: overview of a promising approach to fight antibiotic-resistant bacterial infections. J Clin Transl Res. 2015;1(3):140-67.

17. Liu C, Mo L, Niu Y, Li X, Zhou X, Xu X. The Role of Reactive Oxygen Species and Autophagy in Periodontitis and Their Potential Linkage. Front Physiol. 2017;8(1):439.

18. Sha AM, Garib BT. Antibacterial effect of curcumin against clinically isolated porphyromonas gingivalis and connective tissue reactions to curcumin gel in the subcutaneous tissue of rats. Biomed Res Int. 2019;2019:6810936:1-14.

19. Hovhannisyan GG. Fluorescence in situ hybridization in combination with the comet assay and micronucleus test in genetic toxicology. Molecular cytogenetics. 2010;3(1):17.

20. Myer PR, Kim M, Freetly HC, Smith TPL. Evaluation of 16S rRNA amplicon sequencing using two next-generation sequencing technologies for phylogenetic analysis of the rumen bacterial community in steers. J Microbiol Methods. 2016;127(1):132-40.

21. Lee PY, Costumbrado J, Hsu C-Y, Kim YH. Agarose gel electrophoresis for the separation of DNA fragments. JoVE. 2012;20(62):e3923:1-5.

22. Hasegawa GK, Morais JLD, Soares M, Freitas AMD. Photodynamic inactivation of Escherichia coli by methylene blue and malachite green under red LED light. Revista Ambiente & Água. 2015;10(1):318-26.

23. Nakonechny F, Barel M, David A, Koretz S, Litvak B, Ragozin E, et al. Dark Antibacterial Activity of Rose Bengal. Int J Mol Sci. 2019;20(13):3196.

24. Nitzan Y, Salmon-Divon M, Shporen E, Malik Z. ALA induced photodynamic effects on gram positive and negative bacteria. Photochem Photobiol Sci. 2004;3(5):430-5.

25. Harris F, Pierpoint L. Photodynamic therapy based on 5-aminolevulinic acid and its use as an antimicrobial agent. Med Res Rev. 2012;32(6):1292-327.

26. Fotinos N, Convert M, Piffaretti JC, Gurny R, Lange N. Effects on gram-negative and gram- positive bacteria mediated by 5-aminolevulinic Acid and 5-aminolevulinic acid derivatives. Antimicrob Agents Chemother. 2008;52(4):1366- 73.

27. Lauritano D, Moreo G, Palmieri A, Vella FD, Petruzzi M, Botticelli D, et al. Photodynamic Therapy Using 5-Aminolevulinic Acid (Ala) for the Treatment of Chronic Periodontitis: A Prospective Case Series. Appl Sci. 2022;12(6):3102:1-12.

28. Mccord JM, Edeas MA. SOD, oxidative stress and human pathologies: a brief history and a future vision. Biomed Pharmacother. 2005;59(4):139-42.

29. Zou P, Cao P, Liu J, Li P, Luan Q. Comparisons of the killing effect of direct current partially mediated by reactive oxygen species on Porphyromonas gingivalis and Prevotella intermedia in planktonic state and biofilm state - an in vitro study. J Dent Sci. 2022;17(1):459-67.

30. Floury J, El Mourdi I, Silva JV, Lortal S, Thierry A, Jeanson S. Diffusion of solutes inside bacterial colonies immobilized in model cheese depends on their physicochemical properties: a time-lapse microscopy study. Front Microbiol. 2015;6(1):366.

31. Usacheva MN, Teichert MC, Biel MA. Comparison of the methylene blue and toluidine blue photobactericidal efficacy against gram- positive and gram-negative microorganisms. Lasers Surg Med. 2001;29(2):165-73.

Abstract

 

 

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