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Research Articles
Antimicrobial and cytotoxic properties of calcium-enriched mixture cement, Iranian propolis, and propolis with herbal extracts in primary dental pulp stem cells
Mohammad Esmaeilzadeh, Shirin Moradkhani, Fahimeh Daneshyar, Mohammad Reza Arabestani, Sara Soleimani Asl, Soudeh Tayebi, Maryam Farhadian
Restor Dent Endod 2023;48(1):e2.   Published online December 1, 2022
DOI: https://doi.org/10.5395/rde.2023.48.e2
AbstractAbstract PDFPubReaderePub
Objectives

In this study, natural substances were introduced as primary dental pulp caps for use in pulp therapy, and the antimicrobial and cytotoxic properties of these substances were investigated.

Materials and Methods

In this in vitro study, the antimicrobial properties of calcium-enriched mixture (CEM) cement, propolis, and propolis individually combined with the extracts of several medicinal plants were investigated against Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Then, the cytotoxicity of each substance or mixture against pulp stem cells extracted from 30 primary healthy teeth was evaluated at 4 concentrations. Data were gathered via observation, and optical density values were obtained using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) test and recorded. SPSS software version 23 was used to analyze the data. Data were evaluated using 2-way analysis of variance and the Tukey test.

Results

Regarding antimicrobial properties, thyme alone and thyme + propolis had the lowest minimum inhibitory concentrations (MICs) against the growth of S. aureus, E. coli, and P. aeruginosa bacteria. For E. faecalis, thyme + propolis had the lowest MIC, followed by thyme alone. At 24 and 72 hours, thyme + propolis, CEM cement, and propolis had the greatest bioviability in the primary dental pulp stem cells, and lavender + propolis had the lowest bioviability.

Conclusions

Of the studied materials, thyme + propolis showed the best results in the measures of practical performance as a dental pulp cap.

Citations

Citations to this article as recorded by  
  • Comprehensive review of composition, properties, clinical applications, and future perspectives of calcium-enriched mixture (CEM) cement: a systematic analysis
    Saeed Asgary, Mahtab Aram, Mahta Fazlyab
    BioMedical Engineering OnLine.2024;[Epub]     CrossRef
  • Effects of aqueous and ethanolic extracts of Chinese propolis on dental pulp stem cell viability, migration and cytokine expression
    Ha Bin Park, Yen Dinh, Pilar Yesares Rubi, Jennifer L. Gibbs, Benoit Michot
    PeerJ.2024; 12: e18742.     CrossRef
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  • 13 Download
  • 2 Web of Science
  • 2 Crossref
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Antifungal effects of synthetic human β-defensin 3-C15 peptide
Sang-Min Lim, Ki-Bum Ahn, Christine Kim, Jong-Won Kum, Hiran Perinpanayagam, Yu Gu, Yeon-Jee Yoo, Seok Woo Chang, Seung Hyun Han, Won-Jun Shon, Woocheol Lee, Seung-Ho Baek, Qiang Zhu, Kee-Yeon Kum
Restor Dent Endod 2016;41(2):91-97.   Published online March 17, 2016
DOI: https://doi.org/10.5395/rde.2016.41.2.91
AbstractAbstract PDFPubReaderePub
Objectives

The purpose of this ex vivo study was to compare the antifungal activity of a synthetic peptide consisting of 15 amino acids at the C-terminus of human β-defensin 3 (HBD3-C15) with calcium hydroxide (CH) and Nystatin (Nys) against Candida albicans (C. albicans) biofilm.

Materials and Methods

C. albicans were grown on cover glass bottom dishes or human dentin disks for 48 hr, and then treated with HBD3-C15 (0, 12.5, 25, 50, 100, 150, 200, and 300 µg/mL), CH (100 µg/mL), and Nys (20 µg/mL) for 7 days at 37℃. On cover glass, live and dead cells in the biomass were measured by the FilmTracer Biofilm viability assay, and observed by confocal laser scanning microscopy (CLSM). On dentin, normal, diminished and ruptured cells were observed by field-emission scanning electron microscopy (FE-SEM). The results were subjected to a two-tailed t-test, a one way analysis variance and a post hoc test at a significance level of p = 0.05.

Results

C. albicans survival on dentin was inhibited by HBD3-C15 in a dose-dependent manner. There were fewer aggregations of C. albicans in the groups of Nys and HBD3-C15 (≥ 100 µg/mL). CLSM showed C. albicans survival was reduced by HBD3-C15 in a dose dependent manner. Nys and HBD3-C15 (≥ 100 µg/mL) showed significant fungicidal activity compared to CH group (p < 0.05).

Conclusions

Synthetic HBD3-C15 peptide (≥ 100 µg/mL) and Nys exhibited significantly higher antifungal activity than CH against C. albicans by inhibiting cell survival and biofilm.

Citations

Citations to this article as recorded by  
  • Current status of antimicrobial peptides databases and computational tools for optimization
    Madhulika Jha, Akash Nautiyal, Kumud Pant, Navin Kumar
    Environment Conservation Journal.2025; 26(1): 281.     CrossRef
  • Anti-fungal peptides: an emerging category with enthralling therapeutic prospects in the treatment of candidiasis
    Jyoti Sankar Prusty, Ashwini Kumar, Awanish Kumar
    Critical Reviews in Microbiology.2024; : 1.     CrossRef
  • Harnessing antimicrobial peptides in endodontics
    Xinzi Kong, Vijetha Vishwanath, Prasanna Neelakantan, Zhou Ye
    International Endodontic Journal.2024; 57(7): 815.     CrossRef
  • Human β-defensins and their synthetic analogs: Natural defenders and prospective new drugs of oral health
    Mumian Chen, Zihe Hu, Jue Shi, Zhijian Xie
    Life Sciences.2024; 346: 122591.     CrossRef
  • Candida albicans Virulence Factors and Pathogenicity for Endodontic Infections
    Yeon-Jee Yoo, A Reum Kim, Hiran Perinpanayagam, Seung Hyun Han, Kee-Yeon Kum
    Microorganisms.2020; 8(9): 1300.     CrossRef
  • Innate Inspiration: Antifungal Peptides and Other Immunotherapeutics From the Host Immune Response
    Derry K. Mercer, Deborah A. O'Neil
    Frontiers in Immunology.2020;[Epub]     CrossRef
  • Human salivary proteins and their peptidomimetics: Values of function, early diagnosis, and therapeutic potential in combating dental caries
    Kun Wang, Xuedong Zhou, Wei Li, Linglin Zhang
    Archives of Oral Biology.2019; 99: 31.     CrossRef
  • Endodontic biofilms: contemporary and future treatment options
    Yeon-Jee Yoo, Hiran Perinpanayagam, Soram Oh, A-Reum Kim, Seung-Hyun Han, Kee-Yeon Kum
    Restorative Dentistry & Endodontics.2019;[Epub]     CrossRef
  • Bioactive Peptides Against Fungal Biofilms
    Karen G. N. Oshiro, Gisele Rodrigues, Bruna Estéfani D. Monges, Marlon Henrique Cardoso, Octávio Luiz Franco
    Frontiers in Microbiology.2019;[Epub]     CrossRef
  • Anticandidal Potential of Stem Bark Extract from Schima superba and the Identification of Its Major Anticandidal Compound
    Chun Wu, Hong-Tan Wu, Qing Wang, Guey-Horng Wang, Xue Yi, Yu-Pei Chen, Guang-Xiong Zhou
    Molecules.2019; 24(8): 1587.     CrossRef
  • Synthetic Human β Defensin-3-C15 Peptide in Endodontics: Potential Therapeutic Agent in Streptococcus gordonii Lipoprotein-Stimulated Human Dental Pulp-Derived Cells
    Yeon-Jee Yoo, Hiran Perinpanayagam, Jue-Yeon Lee, Soram Oh, Yu Gu, A-Reum Kim, Seok-Woo Chang, Seung-Ho Baek, Kee-Yeon Kum
    International Journal of Molecular Sciences.2019; 21(1): 71.     CrossRef
  • Candida Infections and Therapeutic Strategies: Mechanisms of Action for Traditional and Alternative Agents
    Giselle C. de Oliveira Santos, Cleydlenne C. Vasconcelos, Alberto J. O. Lopes, Maria do S. de Sousa Cartágenes, Allan K. D. B. Filho, Flávia R. F. do Nascimento, Ricardo M. Ramos, Emygdia R. R. B. Pires, Marcelo S. de Andrade, Flaviane M. G. Rocha, Cristi
    Frontiers in Microbiology.2018;[Epub]     CrossRef
  • Perspectives for clinical use of engineered human host defense antimicrobial peptides
    María Eugenia Pachón-Ibáñez, Younes Smani, Jerónimo Pachón, Javier Sánchez-Céspedes
    FEMS Microbiology Reviews.2017; 41(3): 323.     CrossRef
  • The synthetic human beta-defensin-3 C15 peptide exhibits antimicrobial activity against Streptococcus mutans, both alone and in combination with dental disinfectants
    Ki Bum Ahn, A. Reum Kim, Kee-Yeon Kum, Cheol-Heui Yun, Seung Hyun Han
    Journal of Microbiology.2017; 55(10): 830.     CrossRef
  • Antibiofilm peptides against oral biofilms
    Zhejun Wang, Ya Shen, Markus Haapasalo
    Journal of Oral Microbiology.2017; 9(1): 1327308.     CrossRef
  • Humanβ-Defensin 3 Reduces TNF-α-Induced Inflammation and Monocyte Adhesion in Human Umbilical Vein Endothelial Cells
    Tianying Bian, Houxuan Li, Qian Zhou, Can Ni, Yangheng Zhang, Fuhua Yan
    Mediators of Inflammation.2017; 2017: 1.     CrossRef
  • Antifungal Effects of Synthetic Human Beta-defensin-3-C15 Peptide on Candida albicans –infected Root Dentin
    Yeon-Jee Yoo, Ikyung Kwon, So-Ram Oh, Hiran Perinpanayagam, Sang-Min Lim, Ki-Bum Ahn, Yoon Lee, Seung-Hyun Han, Seok-Woo Chang, Seung-Ho Baek, Qiang Zhu, Kee-Yeon Kum
    Journal of Endodontics.2017; 43(11): 1857.     CrossRef
  • A 15-amino acid C-terminal peptide of beta-defensin-3 inhibits bone resorption by inhibiting the osteoclast differentiation and disrupting podosome belt formation
    Ok-Jin Park, Jiseon Kim, Ki Bum Ahn, Jue Yeon Lee, Yoon-Jeong Park, Kee-Yeon Kum, Cheol-Heui Yun, Seung Hyun Han
    Journal of Molecular Medicine.2017; 95(12): 1315.     CrossRef
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  • 18 Crossref
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Basic Research
The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure
Jin-Ho Chung, Jin Kim, Seong-Ho Choi, Eui-Seong Kim, Jiyong Park, Seung-Jong Lee
J Korean Acad Conserv Dent 2010;35(4):285-294.   Published online July 31, 2010
DOI: https://doi.org/10.5395/JKACD.2010.35.4.285
AbstractAbstract PDFPubReaderePub

The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group.

Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at 0℃ under pressure of 2 MPa), group 8 (low-temperature preservation at 0℃ under no additional pressure), group 9 (low-temperature preservation at -5℃ under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in 37℃ water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization.

In both MTT and WST-1 assay, group 7 (0℃/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance).

By the results of this study, low-temperature preservation at 0℃ under pressure of 2 MPa suggest the possibility for long term preservation of teeth.

Citations

Citations to this article as recorded by  
  • Evaluation of the Viability of Rat Periodontal Ligament Cells after Storing at 0℃/2 MPa Condition up to One Week: In Vivo MTT Method
    Sun Mi Jang, Sin-Yeon Cho, Eui-Seong Kim, Il-Young Jung, Seung Jong Lee
    Journal of Korean Dental Science.2016; 9(1): 1.     CrossRef
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  • 1 Crossref
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Original Articles
THE EFFICACY OF PROGRAMMED CRYO-PRESERVATION UNDER PRESSURE IN RAT PERIODONTAL LIGAMENT CELLS
Young-Eun Lee, Eui-Seong Kim, Jin Kim, Seung-Hoon Han, Seung-Jong Lee
J Korean Acad Conserv Dent 2009;34(4):356-363.   Published online January 14, 2009
DOI: https://doi.org/10.5395/JKACD.2009.34.4.356
AbstractAbstract PDFPubReaderePub
Abstract

The purpose of this study was to evaluate the viability of periodontal ligament cells in rat teeth using slow cryo-preservation method under pressure by means of MTT assay and WST-1 assay. Eighteen teeth of Sprague-Dawley white female rats of 4 week-old were used for each group.

Both sides of the first and second maxillary molars were extracted as atraumatically as possible under Tiletamine anesthesia. The experimental groups were group 1 (Immediate control), group 2 (Cold preservation at 4°C for 1 week), group 3 (Slow freezing), group 4 (Slow freezing under pressure of 3 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in 37°C water bath, then MTT assay and WST-1 assay were processed. One way ANOVA and Tukey method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization.

In both MTT and WST-1 assay, group 4 showed significantly higher viability of periodontal ligament cells than group 2 and 3 (p < 0.05), but showed lower viability than immediate control group.

By the results of this study, slow cryo-preservation method under pressure suggests the possibility for long term cryo-preservation of the teeth.

Citations

Citations to this article as recorded by  
  • Effects of Slow Programmable Cryopreservation on Preserving Viability of the Cultured Periodontal Ligament Cells from Human Impacted Third Molar
    Jin-Woo Kim, Tae-Yi Kim, Ye-mi Kim, Eun-Kyoung Pang, Sun-Jong Kim
    Journal of Korean Dental Science.2015; 8(2): 57.     CrossRef
  • The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure
    Jin-Ho Chung, Jin Kim, Seong-Ho Choi, Eui-Seong Kim, Jiyong Park, Seung-Jong Lee
    Journal of Korean Academy of Conservative Dentistry.2010; 35(4): 285.     CrossRef
  • Comparison of viability of oral epithelial cells stored by different freezing methods
    Do-Young Baek, Seung-Jong Lee, Han-Sung Jung, EuiSeong Kim
    Journal of Korean Academy of Conservative Dentistry.2009; 34(6): 491.     CrossRef
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Evaluation of periodontal ligament cell viability in rat teeth according to various extra-oral dry storage times using MTT assay
In-Soo Jeon, Eui-Seong Kim, Jin Kim, Seung-Jong Lee
J Korean Acad Conserv Dent 2006;31(5):398-408.   Published online September 30, 2006
DOI: https://doi.org/10.5395/JKACD.2006.31.5.398
AbstractAbstract PDFPubReaderePub

The purpose of this study was to verify the usefulness of MTT analysis as a tool of measurement of the periodontal ligament cell viability from the extracted rat molar.

A total of 80 Sprague-Dawley white female rat of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted under Ketamine anesthesia. Twenty-four teeth of each group (divided as five groups depending upon the time-lapse after extraction such as immediate, 10, 20, 40 and 60 minutes) were immersed in 200 µl of MTT solution (0.5 mg/ml) and processed for optical density measurements . Another 10 teeth of each group were treated as same as above and sectioned at 10 µm for microscopic examination.

All measurements values were divided by the value of hematoxylin-eosin staining which represented the volume of each corresponding samples. Immediate and 10 minute groups showed highest MTT values followed by 20, 40, and 60 minutes consecutively. Statistical significance (p < 0.05) existed between all groups except in immediate versus 10 minute groups and 40 versus 60 minutes. Histological findings also showed similar findings with MTT results in crystal shape and crystal numbers between the experimental groups.

These data indicate that in vivo MTT analysis may be of value for evaluation of the periodontal ligament cell viability without time- consuming cell culturing processes.

Citations

Citations to this article as recorded by  
  • Evaluation of the Viability of Rat Periodontal Ligament Cells after Storing at 0℃/2 MPa Condition up to One Week: In Vivo MTT Method
    Sun Mi Jang, Sin-Yeon Cho, Eui-Seong Kim, Il-Young Jung, Seung Jong Lee
    Journal of Korean Dental Science.2016; 9(1): 1.     CrossRef
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  • 1 Crossref
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