Mineral trioxide aggregate (MTA) would influence healing of periapical tissues by modulating the production of growth factors and cytokines from PDL fibroblasts, however, the studies are insufficient. Therefore, the purpose of this study was to monitor the expression of transforming growth factor-beta1 (TGF-β1), fibroblast growth factor-2 (FGF-2), and interleukin-6 (IL-6) from PDL fibroblasts in the presence of MTA. The human PDL fibroblasts were seeded onto the set MTA or IRM at a level of 1 × 10⁵ cells per unit well, and further incubated for 6, 12, 24, and 48 hours. The levels of TGF-β1, FGF-2, and IL-6 from the supernatant were measured by enzyme-linked immunosorbent assay (ELISA). The data were analyzed using one-way ANOVA. The level of TGF-β1 was down-regulated when the cells were grown in the presence of MTA except at 6 hours. The levels of FGF-2 release were significantly suppressed when PDL fibroblasts were grown in the presence of MTA or IRM at all time intervals (p < 0.05). The expressions of IL-6 from MTA treated cells were comparable to those of untreated control cells throughout the observation periods. We presume that this material inhibits the stimulatory function of growth factors on granulation tissue formation and in turn, it promotes the healing process modulated by other bone-remodeling cells.

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• Osteo/odontogenic Differentiation of Human Mesenchymal Stem Cells with Platelet-rich Plasma and Mineral Trioxide Aggregate
  Shanthi Vanka, Amit Vanka, Sandeep Kumar Vishwakarma, Manohar K Bhat, Othman Wali, Aleem A Khan
  The Journal of Contemporary Dental Practice.2019; 20(10): 1171.     CrossRef
• The effect of several root-end filling materials on MG63 osteoblast-like cells
  Jeong-Ho Lee, Won-Jun Shon, WooCheol Lee, Seung-Ho Baek
  Journal of Korean Academy of Conservative Dentistry.2010; 35(3): 222.     CrossRef
• Biocompatibility of experimental mixture of mineral trioxide aggregate and glass ionomer cement
  Min-Jae Oh, Yu-Na Jeong, In-Ho Bae, So-Young Yang, Bum-Jun Park, Jeong-Tae Koh, Yun-Chan Hwang, In-Nam Hwang, Won-Mann Oh
  Journal of Korean Academy of Conservative Dentistry.2010; 35(5): 359.     CrossRef
• Biocompatibility of bioaggregate cement on human pulp and periodontal ligament (PDL) derived cells
  Choo-Ryung Chung, Euiseong Kim, Su-Jung Shin
  Journal of Korean Academy of Conservative Dentistry.2010; 35(6): 473.     CrossRef
• Effects of condensation techniques and canal sizes on the microleakage of orthograde MTA apical plug in simulated canals
  Deuk-Lim Nam, Jeong-Kil Park, Bock Hur, Hyeon-Cheol Kim
  Journal of Korean Academy of Conservative Dentistry.2009; 34(3): 208.     CrossRef

This in vitro study monitored MMP-8 production on PMN by stimulated with the following three groups; Sonicated extracts of E. faecalis (SEF), SEF treated with Ca(OH)₂ (12.5mg/ml) for 7 days, and lipopolysaccharides (LPS) of E. coli. The level of MMP-8 in each group was immediately measured by ELISA. The data were analyzed with Kruskal-Wallis test and Mann-Whitney U test.

In the SEF group, the level of production of MMP-8 was higher than the negative control group in low concentration (0.05µg/ml) of SEF (p < 0.05), but it decreased with an increase in the concentration of SEF (p < 0.05). In the case of SEF treated with Ca(OH)₂, all of the MMP levels were higher than negative control group (p < 0.05), but no statistical difference was found among the different SEF concentrations (p > 0.05). All of the levels in E. coli LPS were increased with increasing concentrations (p < 0.05).

According to this study we could summarize as follows:

1. MMP-8 was expressed at low level in untreated PMN group and the levels of MMP-8 were upregulated in PMN stimulated by E. coli LPS groups.

2. In the SEF groups, the level of production of MMP-8 decreased with an increase in the concentration of SEF (p < 0.05). So E. faecalis may have suppressive effect on the production of MMP-8 by PMN.

3. In the case of SEF treated with Ca(OH)₂, all of the MMP levels at different SEF concentrations were higher than untreated PMN group (p < 0.05), but no statistical difference was found among the different SEF concentrations (p > 0.05).

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• Anti-inflammatory effects of PPARγ on human dental pulp cells
  Jeong-Hee Kim
  Journal of Korean Academy of Conservative Dentistry.2006; 31(3): 203.     CrossRef

In order to examine the immunoresponse of host cells to Enterococcus faecalis, this in vitro study monitored the production of Interleukin-2 (IL-2), Interleukin-4 (IL-4) and Transforming growth factor-β1 (TGF-β1) in human lymphocytes. Lymphocytes were activated with PHA in the presence or abscence of sonicated extracts of E. Faecalis (SEF) and further incubated for 72 hours. The level of each cytokine was measured by ELISA. Data were analyzed with Kruskal-Wallis test and Mann-Whitney U test (P < 0.05). PHA-activated group did exhibit higher level of IL-2 and IL-4 than untreated control group. The levels of expression of both cytokines were significantly decreased following the treatment of high (25 µg/ml) and medium concentration (12.5 µg/ml) of SEF (P <0 .05) than those of PHA activated group. But low concentration (5 µg/ml) of SEF showed the similar level of IL-2 and IL-4 production as those of PHA activated group. TGF-β1 was unaffected by SEF treatment. These results suggested that E. faecalis may suppress IL-2 and IL-4 production by lymphocytes and this could be one of possible factors why E. faecalis are found frequently in the teeth with failed endodontic treatment.

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  Sepideh Sarfi, Ehsaneh Azaryan, Mohsen Naseri
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• Role(s) of cytokines in pulpitis: Latest evidence and therapeutic approaches
  Mohammad M.Y. Khorasani, Gholamhossein Hassanshahi, Aniela Brodzikowska, Hossein Khorramdelazad
  Cytokine.2020; 126: 154896.     CrossRef
• Educational needs of an integrated health and oral health project for community dental hygienists
  Su-kyung Park, Yang-Keum Han, Young-Kyung Kim, Hyun-Ju Lim, Yang-Ok Kown, Han-Mi Kim, Mag-Yup Oh, Nam-Hee Kim
  Journal of Korean Academy of Oral Health.2015; 39(2): 127.     CrossRef
• Microorganism penetration in dentinal tubules of instrumented and retreated root canal walls.In vitroSEM study
  Saad Al-Nazhan, Alaa Al-Sulaiman, Fellwa Al-Rasheed, Fatimah Alnajjar, Bander Al-Abdulwahab, Abdulhakeem Al-Badah
  Restorative Dentistry & Endodontics.2014; 39(4): 258.     CrossRef

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• Cytotoxicity Comparison of Sure-seal root and Adseal Sealers on mouse fibroblast Cells:Invitro study
  Azam haddadikohsar, Mohammad shokrzade, Marjan Fallah, Fatemeh Shakeri
  journal of research in dental sciences.2024; 21(1): 46.     CrossRef
• Effect of Different Sealers on the Cytocompatibility and Osteogenic Potential of Human Periodontal Ligament Stem Cells: An In Vitro Study
  Shehabeldin Saber, Shereen Raafat, Mohamed Elashiry, Ahmed El-Banna, Edgar Schäfer
  Journal of Clinical Medicine.2023; 12(6): 2344.     CrossRef
• In Vitro Comparison of Biocompatibility of Calcium Silicate-Based Root Canal Sealers
  Ju Kyung Lee, Sunil Kim, Sukjoon Lee, Hyeon-Cheol Kim, Euiseong Kim
  Materials.2019; 12(15): 2411.     CrossRef
• A comparative evaluation of cytotoxicity of root canal sealers: anin vitrostudy
  Gautam Pyarelal Badole, Manjusha Madhukar Warhadpande, Ganesh Kothiramji Meshram, Rakesh Namdeoraoji Bahadure, Shubha Gopal Tawani, Gopal Tawani, Shital Gautam Badole
  Restorative Dentistry & Endodontics.2013; 38(4): 204.     CrossRef
• The effect of restorative materials on the stress distribution of class V composite resin restorations - a 3D finite element investigation
  Hyoung-Ryoul Ahn, Hyeon-Cheol Kim, Bock Huh, Jeong-Kil Park
  Journal of Korean Academy of Conservative Dentistry.2006; 31(1): 20.     CrossRef
• Effects of occlusal load on the stress distribution of four cavity configurations of noncarious cervical lesions: a three-dimensional finite element analysis study
  Sang-Je Jeon, Jeong-Kil Park, Hyeon-Cheol Kim, Sung-Gwan Woo, Kwang-Hoon Kim, Kwon Son, Bock Hur
  Journal of Korean Academy of Conservative Dentistry.2006; 31(5): 359.     CrossRef
• Cytotoxicity of resin-based root canal sealer, adseal
  Hee-Jung Kim, Seung-Ho Baek, Woo-Cheol Lee, Han-Soo Park, Kwang-Shik Bae
  Journal of Korean Academy of Conservative Dentistry.2004; 29(6): 498.     CrossRef

Bacterial lipopolysaccharide (LPS) plays a major role in stimulating the synthesis and release of the principal osteoclast-activating cytokines, namely, interleukin 1 and tumor necrosis factor-α from immune cells. Although monocytes/macrophages are the main producers of these cytokines, recent evidence has indicated that polymorphonuclear leukocytes (PMN) have the ability to release IL-1 and TNF-α. Calcium hydroxide has been shown to be an effective medicament in root canal infections, reducing the microbial titre within the canal. It has been proposed that the therapeutic effect of Ca(OH)₂ may also be the result of direct inactivation of LPS. The purpose of this study was to investigate whether treatment of Porphyromonas endodontalis LPS with calcium hydroxide alters its biological action as measured by human PMN secretion of IL-1 and TNF-α, and it was compared with Escherichia coli LPS.

P. endodontalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted using the hot-phenol water extraction method and purified. Purchased E. coli LPS was also purified. 100 µg/ml of each LPS in pyrogen free water were incubated with 25mg/ml Ca(OH)₂ at 37℃ for 7 days. The supernatants were subjected to ultrafiltration, and the isolates were lyophilized and weighed. PMNs were obtained from peripheral blood by centrifugation layered over Lymphoprep. The cells were resuspended (4×10⁶ cells/ml) in RPMI 1640 followed by treatment with various concentrations of LPS (0, 0.1, 1, 10µg/ml) for 24 hours at 37℃ in 5% CO₂ incubator. The supernatants of cells were collected and the levels of IL-1α, IL-1β and TNF-α were measured by enzyme-linked immunosorbent assay.

The results were as follows;

1. The levels of IL-1α, IL-1β, TNF-α from PMN treated with each LPS were significantly higher than those released from unstimulated PMN of the control group (p<0.05).

2. The levels of all three cytokines released from PMN stimulated with each calcium hydroxide treated LPS were significantly lower than those released from PMN stimulated with each untreated LPS (p<0.05), while they were not significantly different from those released from unstimulated PMN of the control group (p>0.05).

3. The levels of secretion for all three cytokines were affected in a dose-dependent manner in PMN stimulated with each LPS (p<0.05), but not in PMN stimulated with each calcium hydroxide treated LPS (p>0.05).

4. The levels of all three cytokines released from PMN stimulated with P. endodontalis LPS were significantly lower than those released from PMN stimulated with E. coli LPS (p<0.05).

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• Lipopolysaccharide from Porphyromonas gingivalis, but Not from Porphyromonas endodontalis, Induces Macrophage M1 Profile
  Pablo Veloso, Alejandra Fernández, Jessica Astorga, David González-Quintanilla, Alfredo Castro, Alejandro Escobar, Anilei Hoare, Marcela Hernández
  International Journal of Molecular Sciences.2022; 23(17): 10011.     CrossRef

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• Role(s) of cytokines in pulpitis: Latest evidence and therapeutic approaches
  Mohammad M.Y. Khorasani, Gholamhossein Hassanshahi, Aniela Brodzikowska, Hossein Khorramdelazad
  Cytokine.2020; 126: 154896.     CrossRef

Immune responses associated with bacterial infection involve various inflammatory cells. Clinical symptoms and pathologic features are particularly influenced by the predominant cells. Among inflammatory cells, T cells have the heterogenity. T cells may develop into the mature cells expressing the cell surface markers with different functions and T helper cells are categorized into Th1 and Th2 cells based on their different patterns of cytokine production. The objective of this study was to investigate the change of expression of surface markers on T cells and the Th1/Th2 immune response in pulpal inflammation associated with specific bacteria.

We experimentally induced pulpal inflammation in rat incisors by drilling without coolant and innoculated with Streptococcus mutans (S.M. group), Porphyromonas endodontalis (P.E. group), or only sterile cotton (control group). After 1, 2, and 5 days, mandibular incisors were extracted and the pulp tissues were extirpated. The expressions of IL-2 recepters (CD25) and ICAM-1 (CD54) on CD4+ and CD8+ cells in the pulps were determined using a flow cytometer, and the concentration of IL-2, IFN-γ and IL-4 was measured by enzyme-linked immunosorbent assay.

The results were as follows;

1. In the S.M. group, CD4+ cells were more increased at 2nd day than 1st day and in the P.E. group, CD8+ cells were more increased at 2nd day than 1st day.

2. The percentages of CD4+, CD4+25+ and CD4+54+ cells were decreased in the pulp tissues at 5th day after irritation in all groups.

3. The ratios of CD4+/CD8+, CD4+/CD4+25+ and CD4+/CD4+54+ in the pulps at 2nd day after irritation by P. endodontalis were significantly lower than the other groups.

4. The higher concentrations of IFN-γ than IL-4 in the pulps at 2nd day after irritation by P. endodontalis showed that T helper 1 reaction were predominant in the early stage of the pulpal inflammation induced by P. endodontalis.

5. The higher concentrations of IL-4 than IFN-γ in the pulps at 1st day and 5th day after irritation by S. mutans were measured but the differences were not significant.