2023 Fiscal Year Research-status Report
Study of the NLRP3 inflammasome-autophagy interplay in odontoblast differentiation using an extra-oral tooth transplantation model
Project/Area Number |
23K09227
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Research Institution | Aichi Gakuin University |
Principal Investigator |
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Co-Investigator(Kenkyū-buntansha) |
本田 雅規 愛知学院大学, 歯学部, 教授 (70361623)
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Project Period (FY) |
2023-04-01 – 2026-03-31
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Keywords | Autophagy / NLRP3 inflammasome / Dental pulp regeneration / Odontoblasts / NLRP3 inflammasome |
Outline of Annual Research Achievements |
Our study aims to analyze if by stopping or inhibiting the activity of the NLRP3 inflammasome or just "NLRP3", dental pulp regeneration in teeth transplanted from old mice into a host mouse is improved. Inhibiting NLRP3 would in turn activate autophagy, which has been described to have a role in dental pulp regeneration. We used a molecule called MCC950 which inhibits NLRP3. We first tested the dose and time of MCC950. We used 40 mg/kg of MCC950 orally and applied it daily for 2 weeks in 3-week-old mice and confirmed that NLRP3 activity was reduced by adding 25 mg/kg of LPS (which activates the inflammatory cytokine, IL-1b via NLRP3). Then we measured IL-1b release by ELISA. IL-1b was decreased in MCC950-treated mice compared to untreated controls. However, the difference was not statistically significant. We also performed extraoral transplantation using immature teeth (open apexes) in GFP host mice. With H&E staining we saw a healing pattern that resembled cementum filling the pulp cavity in the coronal portion and pulp-like tissue in the apical portion. These cells were nestin+ and had GFP expression near it, results that were similar to our previous study.
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Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Although the volume of data is not a lot yet, the initial techniques, such as the extraoral transplantation method and the efficiency of MCC950 have been carried out. The dose of MCC950 could be increased more to obtain a statistical difference in the reduction of IL-1b release. Initial histological data to confirm our previous study`s although more data is necessary for confirmation.
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Strategy for Future Research Activity |
In this fiscal year, we plan to apply the methodologies described above, which have been confirmed and then test if the inhibition of NLRP3 improves pulp regeneration in teeth with mature roots. We plan to prepare as many samples as necessary while in parallel histologically the samples that are finished as soon as possible. With this we can obtain a large amount of histological data and confirm if other measures are required to test our hypothesis.
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Causes of Carryover |
We plan to use this amount in the fiscal year for experiments which need to be replicated due to failure in the methodology, or complications in the animals health that lead to incomplete specimen analysis.
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