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OR13 - Non-Viral Nanoparticles Delivering Plasmid DNA Encoding miR-200c for Regenerative Endodontics

Wednesday, May 3, 2023

1:00 PM – 1:15 PM CT

Location: E353A (Level 3)

Presenter(s)

Tadkamol Krongbaramee, D.D.S., Ph.D.

Resident
University of Iowa
Iowa city, Iowa

Disclosure(s): NIDCR R01 DE026433: Grant/Research Support (Ongoing)

Co-Author(s)

JH

Jue Hu, Ph.D.

Postdoctoral fellowship
University of Iowa
Iowa city, Iowa

Disclosure(s): NIH R01DE029159.: Grant/Research Support (Ongoing)
MR

Matthew Remy, N/A

PhD candidate
University of Iowa
Iowa city, Iowa

Disclosure(s): NIDCR F31 DE031153: Grant/Research Support (Ongoing)
BC

Bruno Cavalcanti, D.D.S., Ph.D.

Clinical Associate Professor
University of Michigan School of Dentistry
Ann Arbor, Michigan

Disclosure information not submitted.
Brad Amendt, Ph.D.

Professor, Director, Craniofacial Anomalies Research Center
University of Iowa
Iowa city, Iowa

Disclosure(s): NIDCR R01 DE026433: Grant/Research Support (Ongoing); NIDCR R01 DE028527: Grant/Research Support (Ongoing)
EZ

Erliang Zeng, M.S., Ph.D.

Associate Professor
University of Iowa
Iowa city, Iowa

Disclosure information not submitted.
Hs

Hongli sun, Ph.D.

Associate Professor
University of Iowa
Iowa city, Iowa

Disclosure(s): NIH R01DE029159: Grant/Research Support (Ongoing)
Fabricio B. Teixeira, D.D.S., M.S., Ph.D.

Professor and Chair - Program Director
University of Iowa
Iowa City, Iowa

Disclosure information not submitted.
Liu Hong, M.S., Ph.D.

Professor
University of Iowa
Iowa city, Iowa

Disclosure(s): NIDCR R01 DE026433: Grant/Research Support (Ongoing)

Introduction: MicroRNA-200c (miR-200c) regulates tooth development, modulates inflammation, and promotes osteogenesis. We aim to characterize miR-200c in pulpitis and determine their potential to attenuate inflammation and promote odontogenesis using in vitro and in vivo models.

Methods: miR-200c and proinflammatory cytokine expression levels from human dental pulp diagnosed with irreversible pulpitis and healthy pulp were quantified using RNA sequencing (miRNA-seq, mRNA-seq) and PCR. Transfection efficiency of plasmid DNA (pDNA) encoding miR-200c to transfect human dental pulp stem cells (hDPSCs) delivered by CaCO₃-based nanoparticles was also assessed. IL-6 and IL-8 transcript and protein levels of hDPSCs with miR-200c overexpression were quantified after exposure to P.gingivalis lipopolysaccharide (Pg-LPS). Odontogenic differentiation markers in hDPSCs were additionally assessed after miR-200c overexpression via qPCR and western blotting. Finally, a rat pulp capping model was used to evaluate the function of miR-200c in vivo using histology. Statistical analyses included one-way ANOVA and Student T-Tests.

Results: We observed that miR-200c was downregulated in inflamed pulps, while IL-6 and IL-8 were significantly upregulated (n=12). CaCO₃ nanoparticles effectively improved the transfection efficiency of pDNA with minimal cytotoxicity in hDPSCs. Overexpression of miR-200c using the CaCO₃ delivery system significantly increased hDPSCs odontogenic differentiation markers including OCN,DMP₁ and DSPP. Additionally, miR-200c also effectively downregulated IL-6 and IL-8 under the Pg-LPS challenge in hDPSCs and mitigated inflammation in a rat model of pulpitis.

Conclusion: Our findings demonstrated that pDNA encoding miR-200c delivered by CaCO₃ nanoparticles might serves as a novel therapeutic agent for regenerative endodontics by attenuating inflammation and promoting odontogenic differentiation.