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      Evaluation of Cytotoxicity of Allium sativum (Garlic Extract) against Human Dental Pulp Fibroblasts

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          ABSTRACT

          Background

          Vital pulp therapy procedures in primary dentition focuses on preservation and maintenance of pulp tissue that has been compromised due to caries, trauma, etc. Several pulp dressing materials have been used in primary teeth and some natural materials from the field of traditional medicine have also been introduced as medicaments in vital pulp therapy. The understanding of biologic and cytotoxic properties of newer materials is important for safe clinical usage. The biologic compatibility of these newer materials is imperative to limit or avoid tissue irritation or degeneration.

          Aim

          To evaluate the cytotoxic effects of Allium sativum on cultured human primary dental pulp fibroblasts.

          Materials and methods

          Primary pulp fibroblasts were cultured from the pulp tissue obtained from extracted deciduous primary canines and central incisor teeth. The freshly prepared concentrations of 1000, 500, 250, 125, and 62.5 µg/mL A. sativum extract were added to the 96-well plate in triplicates to which culture medium containing fourth passage cell suspension was added previously. Cells without treatment served as control, while cells treated with 5% dimethyl sulfoxide (DMSO) served as toxic control. After the addition of experimental and control agents, cells were incubated for 24 and 48 hours at 37°C in 5% CO 2 atmosphere. After the incubation period, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to determine the number of viable cells. Absorbance was read with a microplate reader at 570 nm wavelength and the relative viability of dental pulp fibroblasts at various concentrations was expressed as color intensity of the experimental wells relative to that of control. The percentage of cell viability was also calculated accordingly.

          Results

          The MTT assay results revealed that A. sativum extract, in all the concentrations tested at both the time intervals maintained a cell viability of greater than 90%. At 24 hours, the mean absorbance value of untreated control wells was recorded as 0.84400 ± 0.00916 with 100% cell viability. Among all the concentrations of garlic extract tested, highest mean absorbance value of 0.83933 ± 0.00550 with 99.44% cell viability was recorded for 62.5 µg/mL concentration. At 48 hours, the mean absorbance value of untreated control wells was recorded as 1.22767 ± 0.01106 with 100% cell viability, and the highest mean absorbance value of 1.22567 ± 0.01006 with 99.83% cell viability was recorded for 62.5 µg/mL concentration. The cell viability did not seem to be affected by the concentration of A. sativum extract at 24 hours. However, at 48 hours, the sensitivity of the cells was observed to be dependent on the concentration of A. sativum with a decrease in the viability of cells noted with the increase in concentration.

          Conclusion

          A. sativum extract is noncytotoxic in nature and preserves the vitality of cultured human primary dental pulp fibroblasts making it a suitable material for use in vital pulp therapy procedures of primary teeth.

          How to cite this article

          Devaraju R, Reddy D, Paul ST, et al. Evaluation of Cytotoxicity of Allium sativum (Garlic Extract) against Human Dental Pulp Fibroblasts. Int J Clin Pediatr Dent 2024;17(2):143–148.

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          Most cited references41

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          Guidelines for cell viability assays

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            Inhibitory effect of garlic extract on oral bacteria.

            Garlic (Allium sativum) has long been known to have antibacterial, antifungal and antiviral properties but there are few data on its effects against oral bacterial species particularly putative periodontal pathogens or their enzymes. Filter sterilised, aqueous extract of garlic was tested for ability to inhibit the growth of a range of oral species and to inhibit the trypsin-like and total protease activity Porphyromonas gingivalis. The garlic extract (57.1% (w/v), containing 220 microg/ml allicin) inhibited the growth and killed most of the organisms tested. In general, the minimal inhibitory and minimum bactericidal concentrations for the Gram-negative strains (garlic MIC range 35.7-1.1 mg/ml; allicin mean MIC 4.1 microg/ml; mean MBC 7.9 microg/ml) were lower than those for the Gram-positive strains tested (garlic MIC range 142.7-35.7 mg/ml; allicin mean MIC 27.5 microg/ml; mean MBC 91.9 microg/ml). Also, of the organisms tested, the putative periodontal pathogens had among the lowest MICs (17.8-1.1 mg/ml garlic) and MBCs (35.7-1.1 mg/ml garlic). Time-kill curves for Streptococcus mutans and P. ginigvalis, showed that killing of the latter started almost immediately, whereas there was a delay before S. mutans was killed. The garlic extract also inhibited the trypsin-like and total protease activity of P. gingivalis by 92.7% and 94.88%, respectively. These data indicate that garlic extract inhibits the growth of oral pathogens and certain proteases and so may have therapeutic value, particularly for periodontitis.
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              In-vitro antimicrobial activity of four diallyl sulphides occurring naturally in garlic and Chinese leek oils.

              S Tsao, M C Yin (2001)
              The in-vitro antimicrobial activity of garlic oil, Chinese leek oil and four diallyl sulphides occurring naturally in these oils against Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), three Candida spp. and three Aspergillus spp. (total of 276 clinical isolates) was studied. The magnitude of activity of the four diallyl sulphides followed the order diallyl tetrasulphide > diallyl trisulphide > diallyl disulphide > diallyl monosulphide. These results suggest that disulphide bonds are an important factor in determining the antimicrobial capabilities of these sulphides. The concentration of four diallyl sulphides in garlic and Chinese leek oils was in the range 41.7-52.7% of total sulphides. Garlic oil, with a higher concentration of four diallyl sulphides, showed greater antimicrobial activity than Chinese leek oil. Diallyl disulphide, diallyl trisulphide, diallyl tetrasulphide and the oils rich in these sulphides may have a role in the prevention or treatment of infections.
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                Author and article information

                Journal
                Int J Clin Pediatr Dent
                Int J Clin Pediatr Dent
                IJCPD
                International Journal of Clinical Pediatric Dentistry
                Jaypee Brothers Medical Publishers
                0974-7052
                0975-1904
                February 2024
                : 17
                : 2
                : 143-148
                Affiliations
                [1–4 ]Department of Paedodontics and Preventive Dentistry, Sri Rajiv Gandhi College of Dental Sciences and Hospital, Bengaluru, Karnataka, India
                [5 ]Department of Periodontology and Implantology, Priya Multispeciality Dental Clinic, Sainagar, Bengaluru, Karnataka, India
                [6 ]Department of Oral and Maxillofacial Pathology & Microbiology, Vista 32 Multispeciality Dental Clinic, Yelahanka New Town, Bengaluru, Karnataka, India
                Author notes
                Raghu Devaraju, Department of Paedodontics and Preventive Dentistry, Sri Rajiv Gandhi College of Dental Sciences and Hospital, Bengaluru, Karnataka, India, Phone: +91 9738586632, e-mail: rd51824@ 123456gmail.com
                Article
                10.5005/jp-journals-10005-2781
                11339490
                e181218d-0965-454a-8b04-26117ce09234
                Copyright © 2024; The Author(s).

                © The Author(s). 2024 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted use, distribution, and non-commercial reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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                Original Research

                allium sativum,cytotoxicity,garlic extract,human dental pulp fibroblasts

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