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Klorheksidinin İnsan Dişeti Fibroblastları Üzerindeki Hücre Canlılığı ve Sitotoksisite Etkinliğinin İn vitro Koşullarda Değerlendirilmesi

Year 2023, , 27 - 32, 30.04.2023
https://doi.org/10.29228/erd.40

Abstract

Amaç: Bu in vitro çalışmada, % 0,2’lik klorheksidin (CHX) solüsyonunun, insan dişeti fibroblastları (HGF) üzerindeki hücre canlılığı ve sitotoksisite etkinliklerinin değerlendirilmesi amaçlanmıştır.
Gereç ve Yöntemler: 30 saniye ve 2 dakikalık zaman aralıklarında, nötr pH değerindeki % 0,2’lik CHX solüsyonu kullanılarak çalışılmıştır. Hücre canlılığı ve sitotoksisitesi, HGF’ye karşı 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolyum bromür (MTT) testi ve laktat dehidrogenaz (LDH) testi kullanılarak değerlendirilmiştir. Sonuçlar ortalama ve standart sapma değerleri kullanılarak two way ANOVA testiyle istatistiksel olarak değerlendirilmiştir (p<0,05).
Bulgular: MTT testi sonuçlarına göre, % 0,2 CHX solüsyonunun zamanla hücre canlılığını azalttığı gözlendi. LDH testi sonuçlarına göre ise % 0,2 CHX’in sitotoksik etkinliğinin kontrol grubuna kıyasla arttığı ancak zaman içinde istatistiksel olarak anlamlı bir fark göstermediği gözlendi.
Sonuç: Bu in vitro çalışmanın sınırları dahilinde, % 0,2 CHX solüsyonu, 30 sn ve 2 dk’lık kısa maruz kalma sürelerinde, HGF hücre canlılığını azalttı ve HGF üzerine sitotoksik etkinlik gösterdi.

Supporting Institution

Marmara Üniversitesi Bilimsel Araştırma Projeleri Komisyonu

Project Number

TDK-2021-10281

References

  • Babgi W, Alhajaji M, Al-Mehmadi L, Elbaqli R, Khayat N, Aldahlawi S, Youssef AR. Effect of root conditioning agents hyaluronic acid, EDTA and chlorhexidine on the attachment of human gingival fibroblasts to healthy root surface. Saudi Dent J. 2021;33(6): 342-347.
  • Babich H, Wurzburger BJ, Rubin YL, Sinensky MC, Blau L. An in vitro study on the cytotoxicity of chlorhexidine digluconate to human gingival cells. Cell Biol Toxicol. 1995;11(2): 79-88.
  • Batra C, Alalshaikh M, Gregory RL, Windsor LJ, Blanchard SB, Hamada Y. An in vitro comparison of four antibacterial agents with and without nicotine and their effects on human gingival fibroblasts. J Periodontol. 2022;93(2): e24-e33.
  • Becerik S, Turkoglu O, Emingil G, Vural C, Ozdemir G, Atilla G. Antimicrobial effect of adjunctive use of chlorhexidine mouthrinse in untreated gingivitis: a randomized, placebo-controlled study. APMIS. 2011;119(6): 364-372.
  • Bidar M, Naderinasab M, Talati A, Ghazvini K, Asgari S, Hadizadeh B, Gharechahi M, Mashadi NA. The effects of different concentrations of chlorhexidine gluconate on the antimicrobial properties of mineral trioxide aggregate and calcium enrich mixture. Dent Res J (Isfahan). 2012;9(4): 466-471.
  • Castillo DM, Castillo Y, Delgadillo NA, Neuta Y, Jola J, Calderon JL, Lafaurie GI. Viability and Effects on Bacterial Proteins by Oral Rinses with Hypochlorous Acid as Active Ingredient. Braz Dent J. 2015;26(5): 519-524.
  • Coelho AS, Laranjo M, Goncalves AC, Paula A, Paulo S, Abrantes AM, Caramelo F, Ferreira MM, Silva MJ, Carrilho E, Botelho MF. Cytotoxic effects of a chlorhexidine mouthwash and of an enzymatic mouthwash on human gingival fibroblasts. Odontology. 2020;108(2): 260-270.
  • Dadpe MV, Dhore SV, Dahake PT, Kale YJ, Kendre SB, Siddiqui AG. Evaluation of antimicrobial efficacy of Trachyspermum ammi (Ajwain) oil and chlorhexidine against oral bacteria: An in vitro study. J Indian Soc Pedod Prev Dent. 2018;36(4): 357-363.
  • Decker EM, Bartha V, Kopunic A, von Ohle C. Antimicrobial efficiency of mouthrinses versus and in combination with different photodynamic therapies on periodontal pathogens in an experimental study. J Periodontal Res. 2017;52(2): 162-175.
  • Drisko CH. Nonsurgical periodontal therapy. Periodontol 2000. 2001;25: 77-88.
  • Etemadi A, Eftekhari Bayati S, Pourhajibagher M, Chiniforush N. In vitro effect of antimicrobial photodynamic therapy with phycocyanin on Aggregatibacter actinomycetemcomitans biofilm on SLA titanium discs. Photodiagnosis Photodyn Ther. 2020;32: 102062.
  • Faria G, Cardoso CR, Larson RE, Silva JS, Rossi MA. Chlorhexidine-induced apoptosis or necrosis in L929 fibroblasts: A role for endoplasmic reticulum stress. Toxicol Appl Pharmacol. 2009;234(2): 256-265.
  • Figuero E, Nobrega DF, Garcia-Gargallo M, Tenuta LM, Herrera D, Carvalho JC. Mechanical and chemical plaque control in the simultaneous management of gingivitis and caries: a systematic review. J Clin Periodontol. 2017;44 Suppl 18: S116-S134.
  • Giannelli M, Chellini F, Margheri M, Tonelli P, Tani A. Effect of chlorhexidine digluconate on different cell types: a molecular and ultrastructural investigation. Toxicol In Vitro. 2008;22(2): 308-317.
  • Hennessey TS. Some antibacterial properties of chlorhexidine. J Periodontal Res Suppl. 1973;12: 61-67.
  • Jalaluddin M, Jayanti I, Gowdar IM, Roshan R, Varkey RR, Thirutheri A. Antimicrobial Activity of Curcuma longa L. Extract on Periodontal Pathogens. J Pharm Bioallied Sci. 2019;11(Suppl 2): S203-S207.
  • Karpinski TM, Szkaradkiewicz AK. Chlorhexidine--pharmaco-biological activity and application. Eur Rev Med Pharmacol Sci. 2015;19(7): 1321-1326.
  • Koychev S, Dommisch H, Chen H, Pischon N. Antimicrobial Effects of Mastic Extract Against Oral and Periodontal Pathogens. J Periodontol. 2017;88(5): 511-517.
  • Kshitish D, Laxman VK. The use of ozonated water and 0.2% chlorhexidine in the treatment of periodontitis patients: A clinical and microbiologic study. Indian J Dent Res. 2010;21: 341-348.
  • Mima EG, Pavarina AC, Vargas Fda S, Giampaolo ET, Machado AL, Vergani CE. Effectiveness of chlorhexidine on the disinfection of complete dentures colonised with fluconazole-resistant Candida albicans: in vitro study. Mycoses. 2011;54(5): e506-512.
  • Muller HD, Eick S, Moritz A, Lussi A, Gruber R. Cytotoxicity and Antimicrobial Activity of Oral Rinses In Vitro. Biomed Res Int. 2017;2017: 4019723.
  • Nawrot-Hadzik I, Matkowski A, Pitulaj A, Sterczala B, Olchowy C, Szewczyk A, Choromanska A. In Vitro Gingival Wound Healing Activity of Extracts from Reynoutria japonica Houtt Rhizomes. Pharmaceutics. 2021;13(11).
  • Neely AL. Essential oil mouthwash (EOMW) may be equivalent to chlorhexidine (CHX) for long-term control of gingival inflammation but CHX appears to perform better than EOMW in plaque control. J Evid Based Dent Pract. 2012;12(3 Suppl): 69-72.
  • Petersilka GJ, Ehmke B, Flemmig TF. Antimicrobial effects of mechanical debridement. Periodontol 2000. 2002;28: 56-71.
  • Quirynen M, Teughels W, De Soete M, van Steenberghe D. Topical antiseptics and antibiotics in the initial therapy of chronic adult periodontitis: microbiological aspects. Periodontol 2000. 2002;28: 72-90.
  • Russell AD. Bacterial spores and chemical sporicidal agents. Clin Microbiol Rev. 1990;3(2): 99-119.
  • Schmidt J, Zyba V, Jung K, Rinke S, Haak R, Mausberg RF, Ziebolz D. Cytotoxic effects of octenidine mouth rinse on human fibroblasts and epithelial cells - an in vitro study. Drug Chem Toxicol. 2016;39(3): 322-330.
  • Schmidt J, Zyba V, Jung K, Rinke S, Haak R, Mausberg RF, Ziebolz D. Effects of octenidine mouth rinse on apoptosis and necrosis of human fibroblasts and epithelial cells - an in vitro study. Drug Chem Toxicol. 2018;41(2): 182-187.
  • Sogut MU. Jermisid ajanlardan katyonik deterjanlar. J Exp Clin Med. 2013;30: 75-79.
  • Sritrairat N, Nukul N, Inthasame P, Sansuk A, Prasirt J, Leewatthanakorn T, Piamsawad U, Dejrudee A, Panichayupakaranant P, Pangsomboon K, Chanowanna N, Hintao J, Teanpaisan R, Chaethong W, Yongstar P, Pruphetkaew N, Chongsuvivatwong V, Nittayananta W. Antifungal activity of lawsone methyl ether in comparison with chlorhexidine. J Oral Pathol Med. 2011;40(1): 90-96.
  • Sukumaran SK, Vadakkekuttical RJ, Kanakath H. Comparative evaluation of the effect of curcumin and chlorhexidine on human fibroblast viability and migration: An in vitro study. J Indian Soc Periodontol. 2020;24(2): 109-116.
  • Tirali RE, Bodur H, Sipahi B, Sungurtekin E. Evaluation of the antimicrobial activities of chlorhexidine gluconate, sodium hypochlorite and octenidine hydrochloride in vitro. Aust Endod J. 2013;39(1): 15-18.
  • Utami WS, Anggani HS, Purbiati M. Cytotoxicity effect of orthodontic miniscrew-implant in different types of mouthwash: An in-vitro study. J Orthod Sci. 2022;11: 5.
  • Vahabi S, Hakemi-Vala M, Gholami S. In vitro Antibacterial Effect of Hydroalcoholic Extract of Lawsonia inermis, Malva sylvestris, and Boswellia serrata on Aggregatibacter actinomycetemcomitans. Adv Biomed Res. 2019;8: 22.
  • Vitt A, Babenka A, Bostrom EA, Gustafsson A, Lira Junior R, Slizen V, Sorsa T, Tervahartiala T, Buhlin K. Adjunctive Antiseptic Irrigation of Periodontal Pockets: Effects on Microbial and Cytokine Profiles. Dent J (Basel). 2020;8(4).
  • Von Maltzahn NF, Stumpp NS, Stiesch M. Antibacterial Effect of Cupral((R)) on Oral Biofilms - An In-Vitro Study. Eur Endod J. 2020;5(1): 40-45.
  • Voos AC, Kranz S, Tonndorf-Martini S, Voelpel A, Sigusch H, Staudte H, Albrecht V, Sigusch BW. Photodynamic antimicrobial effect of safranine O on an ex vivo periodontal biofilm. Lasers Surg Med. 2014;46(3): 235-243.

In vitro Evaluation of Cell Viability and Cytotoxicity Effects of Chlorhexidine on Human Gingival Fibroblasts

Year 2023, , 27 - 32, 30.04.2023
https://doi.org/10.29228/erd.40

Abstract

Objectives: The aim of this study was to evaluate the cell viability and cytotoxicity effects of a 0.2 % chlorhexidine (CHX) solution on human gingival fibroblasts (HGF) in vitro.
Materials and Methods: A 0.2 % CHX solution with a neutral pH was tested at 30 second and 2 minute intervals. Cell viability and cytotoxicity against HGF were determined using the 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test and the lactate dehydrogenase (LDH) test. Results were evaluated by calculating means and standard deviations using the two-way ANOVA test (p<0.05).
Results: According to the MTT test results, it was observed that a 0.2 % CHX solution decreased cell viability over time. According to the LDH test results, the cytotoxic efficacy of 0.2 % CHX was higher compared to the control group, but did not show a statistically significant difference over time.
Conclusions: Within the limits of this in vitro study, 0.2 % CHX solution reduced HGF cell viability and showed cytotoxic activity on HGF during short exposure times of 30 sec and 2 min.

Project Number

TDK-2021-10281

References

  • Babgi W, Alhajaji M, Al-Mehmadi L, Elbaqli R, Khayat N, Aldahlawi S, Youssef AR. Effect of root conditioning agents hyaluronic acid, EDTA and chlorhexidine on the attachment of human gingival fibroblasts to healthy root surface. Saudi Dent J. 2021;33(6): 342-347.
  • Babich H, Wurzburger BJ, Rubin YL, Sinensky MC, Blau L. An in vitro study on the cytotoxicity of chlorhexidine digluconate to human gingival cells. Cell Biol Toxicol. 1995;11(2): 79-88.
  • Batra C, Alalshaikh M, Gregory RL, Windsor LJ, Blanchard SB, Hamada Y. An in vitro comparison of four antibacterial agents with and without nicotine and their effects on human gingival fibroblasts. J Periodontol. 2022;93(2): e24-e33.
  • Becerik S, Turkoglu O, Emingil G, Vural C, Ozdemir G, Atilla G. Antimicrobial effect of adjunctive use of chlorhexidine mouthrinse in untreated gingivitis: a randomized, placebo-controlled study. APMIS. 2011;119(6): 364-372.
  • Bidar M, Naderinasab M, Talati A, Ghazvini K, Asgari S, Hadizadeh B, Gharechahi M, Mashadi NA. The effects of different concentrations of chlorhexidine gluconate on the antimicrobial properties of mineral trioxide aggregate and calcium enrich mixture. Dent Res J (Isfahan). 2012;9(4): 466-471.
  • Castillo DM, Castillo Y, Delgadillo NA, Neuta Y, Jola J, Calderon JL, Lafaurie GI. Viability and Effects on Bacterial Proteins by Oral Rinses with Hypochlorous Acid as Active Ingredient. Braz Dent J. 2015;26(5): 519-524.
  • Coelho AS, Laranjo M, Goncalves AC, Paula A, Paulo S, Abrantes AM, Caramelo F, Ferreira MM, Silva MJ, Carrilho E, Botelho MF. Cytotoxic effects of a chlorhexidine mouthwash and of an enzymatic mouthwash on human gingival fibroblasts. Odontology. 2020;108(2): 260-270.
  • Dadpe MV, Dhore SV, Dahake PT, Kale YJ, Kendre SB, Siddiqui AG. Evaluation of antimicrobial efficacy of Trachyspermum ammi (Ajwain) oil and chlorhexidine against oral bacteria: An in vitro study. J Indian Soc Pedod Prev Dent. 2018;36(4): 357-363.
  • Decker EM, Bartha V, Kopunic A, von Ohle C. Antimicrobial efficiency of mouthrinses versus and in combination with different photodynamic therapies on periodontal pathogens in an experimental study. J Periodontal Res. 2017;52(2): 162-175.
  • Drisko CH. Nonsurgical periodontal therapy. Periodontol 2000. 2001;25: 77-88.
  • Etemadi A, Eftekhari Bayati S, Pourhajibagher M, Chiniforush N. In vitro effect of antimicrobial photodynamic therapy with phycocyanin on Aggregatibacter actinomycetemcomitans biofilm on SLA titanium discs. Photodiagnosis Photodyn Ther. 2020;32: 102062.
  • Faria G, Cardoso CR, Larson RE, Silva JS, Rossi MA. Chlorhexidine-induced apoptosis or necrosis in L929 fibroblasts: A role for endoplasmic reticulum stress. Toxicol Appl Pharmacol. 2009;234(2): 256-265.
  • Figuero E, Nobrega DF, Garcia-Gargallo M, Tenuta LM, Herrera D, Carvalho JC. Mechanical and chemical plaque control in the simultaneous management of gingivitis and caries: a systematic review. J Clin Periodontol. 2017;44 Suppl 18: S116-S134.
  • Giannelli M, Chellini F, Margheri M, Tonelli P, Tani A. Effect of chlorhexidine digluconate on different cell types: a molecular and ultrastructural investigation. Toxicol In Vitro. 2008;22(2): 308-317.
  • Hennessey TS. Some antibacterial properties of chlorhexidine. J Periodontal Res Suppl. 1973;12: 61-67.
  • Jalaluddin M, Jayanti I, Gowdar IM, Roshan R, Varkey RR, Thirutheri A. Antimicrobial Activity of Curcuma longa L. Extract on Periodontal Pathogens. J Pharm Bioallied Sci. 2019;11(Suppl 2): S203-S207.
  • Karpinski TM, Szkaradkiewicz AK. Chlorhexidine--pharmaco-biological activity and application. Eur Rev Med Pharmacol Sci. 2015;19(7): 1321-1326.
  • Koychev S, Dommisch H, Chen H, Pischon N. Antimicrobial Effects of Mastic Extract Against Oral and Periodontal Pathogens. J Periodontol. 2017;88(5): 511-517.
  • Kshitish D, Laxman VK. The use of ozonated water and 0.2% chlorhexidine in the treatment of periodontitis patients: A clinical and microbiologic study. Indian J Dent Res. 2010;21: 341-348.
  • Mima EG, Pavarina AC, Vargas Fda S, Giampaolo ET, Machado AL, Vergani CE. Effectiveness of chlorhexidine on the disinfection of complete dentures colonised with fluconazole-resistant Candida albicans: in vitro study. Mycoses. 2011;54(5): e506-512.
  • Muller HD, Eick S, Moritz A, Lussi A, Gruber R. Cytotoxicity and Antimicrobial Activity of Oral Rinses In Vitro. Biomed Res Int. 2017;2017: 4019723.
  • Nawrot-Hadzik I, Matkowski A, Pitulaj A, Sterczala B, Olchowy C, Szewczyk A, Choromanska A. In Vitro Gingival Wound Healing Activity of Extracts from Reynoutria japonica Houtt Rhizomes. Pharmaceutics. 2021;13(11).
  • Neely AL. Essential oil mouthwash (EOMW) may be equivalent to chlorhexidine (CHX) for long-term control of gingival inflammation but CHX appears to perform better than EOMW in plaque control. J Evid Based Dent Pract. 2012;12(3 Suppl): 69-72.
  • Petersilka GJ, Ehmke B, Flemmig TF. Antimicrobial effects of mechanical debridement. Periodontol 2000. 2002;28: 56-71.
  • Quirynen M, Teughels W, De Soete M, van Steenberghe D. Topical antiseptics and antibiotics in the initial therapy of chronic adult periodontitis: microbiological aspects. Periodontol 2000. 2002;28: 72-90.
  • Russell AD. Bacterial spores and chemical sporicidal agents. Clin Microbiol Rev. 1990;3(2): 99-119.
  • Schmidt J, Zyba V, Jung K, Rinke S, Haak R, Mausberg RF, Ziebolz D. Cytotoxic effects of octenidine mouth rinse on human fibroblasts and epithelial cells - an in vitro study. Drug Chem Toxicol. 2016;39(3): 322-330.
  • Schmidt J, Zyba V, Jung K, Rinke S, Haak R, Mausberg RF, Ziebolz D. Effects of octenidine mouth rinse on apoptosis and necrosis of human fibroblasts and epithelial cells - an in vitro study. Drug Chem Toxicol. 2018;41(2): 182-187.
  • Sogut MU. Jermisid ajanlardan katyonik deterjanlar. J Exp Clin Med. 2013;30: 75-79.
  • Sritrairat N, Nukul N, Inthasame P, Sansuk A, Prasirt J, Leewatthanakorn T, Piamsawad U, Dejrudee A, Panichayupakaranant P, Pangsomboon K, Chanowanna N, Hintao J, Teanpaisan R, Chaethong W, Yongstar P, Pruphetkaew N, Chongsuvivatwong V, Nittayananta W. Antifungal activity of lawsone methyl ether in comparison with chlorhexidine. J Oral Pathol Med. 2011;40(1): 90-96.
  • Sukumaran SK, Vadakkekuttical RJ, Kanakath H. Comparative evaluation of the effect of curcumin and chlorhexidine on human fibroblast viability and migration: An in vitro study. J Indian Soc Periodontol. 2020;24(2): 109-116.
  • Tirali RE, Bodur H, Sipahi B, Sungurtekin E. Evaluation of the antimicrobial activities of chlorhexidine gluconate, sodium hypochlorite and octenidine hydrochloride in vitro. Aust Endod J. 2013;39(1): 15-18.
  • Utami WS, Anggani HS, Purbiati M. Cytotoxicity effect of orthodontic miniscrew-implant in different types of mouthwash: An in-vitro study. J Orthod Sci. 2022;11: 5.
  • Vahabi S, Hakemi-Vala M, Gholami S. In vitro Antibacterial Effect of Hydroalcoholic Extract of Lawsonia inermis, Malva sylvestris, and Boswellia serrata on Aggregatibacter actinomycetemcomitans. Adv Biomed Res. 2019;8: 22.
  • Vitt A, Babenka A, Bostrom EA, Gustafsson A, Lira Junior R, Slizen V, Sorsa T, Tervahartiala T, Buhlin K. Adjunctive Antiseptic Irrigation of Periodontal Pockets: Effects on Microbial and Cytokine Profiles. Dent J (Basel). 2020;8(4).
  • Von Maltzahn NF, Stumpp NS, Stiesch M. Antibacterial Effect of Cupral((R)) on Oral Biofilms - An In-Vitro Study. Eur Endod J. 2020;5(1): 40-45.
  • Voos AC, Kranz S, Tonndorf-Martini S, Voelpel A, Sigusch H, Staudte H, Albrecht V, Sigusch BW. Photodynamic antimicrobial effect of safranine O on an ex vivo periodontal biofilm. Lasers Surg Med. 2014;46(3): 235-243.
There are 37 citations in total.

Details

Primary Language Turkish
Subjects Dentistry
Journal Section Original Articles
Authors

Gözdem Bayraktar 0000-0002-0672-3854

Ayse Yilmaz 0000-0003-0802-523X

Hafize Öztürk Özener 0000-0002-2156-584X

Project Number TDK-2021-10281
Early Pub Date April 29, 2023
Publication Date April 30, 2023
Published in Issue Year 2023

Cite

APA Bayraktar, G., Yilmaz, A., & Öztürk Özener, H. (2023). Klorheksidinin İnsan Dişeti Fibroblastları Üzerindeki Hücre Canlılığı ve Sitotoksisite Etkinliğinin İn vitro Koşullarda Değerlendirilmesi. European Journal of Research in Dentistry, 7(1), 27-32. https://doi.org/10.29228/erd.40