# Osteoimmunology of Retarded Bone Regeneration in Periodontitis

> **NIH NIH R01** · NOVA SOUTHEASTERN UNIVERSITY · 2021 · $74,336

## Abstract

This application for Diversity Administrative Supplement (PA-21-071) is written to request for the one-year
support for Hispanic Post Bachelorette Trainee, Ms. Elizabeth Leon, under the parent RO1 grant, titled,
“Osteoimmunology of regarded bone regeneration in periodontitis.” She will perform the basic research to
elucidate the mechanism underlying the pathogenic bone resorption in periodontitis. Through the proposed
training, she will not only become familiar with the operational approach to perform basic research related to
the oral diseases, but also master the conceptual approach to find the knowledge gap and the way to solve the
existing problem. The fundamentals for increasing the rigor and reproducibility, as well as the principle for
responsible research conduct will be also learned. Parallelly, she will be working toward acceptance in the
program of dentistry. The goal of this training is to support the career development of a future clinician/scientist
in the basic and translational research in the dental sciences.
Candidate’s research project background: Our unpublished data demonstrated that P. gingivalis’ vesicles
can promote the osteoclastogenesis which was significantly attenuated by the vesicles isolated from PAD-KO
P. gingivalis. Furthermore, P. gingivalis’ vesicles up-regulated the expression of OC-STAMP in the RANKL-
stimulated RAW264.7 osteoclast precursor cells.
Hypothesis: We hypothesized that PAD expressed by vesicles of P. gingivalis can citrullinate the host derived
Vimentin which, in turn, acts on OC-STAMP expressed by osteoclasts and promotes osteoclastogenesis
Project Objectives for Candidate:
 Objective 1: To evaluate the role of PAD expressed by vesicles of P. gingivalis (Pg vesicle-PAD) in
citrullinating the host proteins, including Vimentin, in the co-culture of vesicles and osteoclasts.
Objective 2: To examine whether Pg vesicle-PAD-mediated citrullination of host vimentin can alter
 Vimentin’s effect on promoting RANKL-induced osteoclastogenesis.
The vesicles will be isolated from PAD-KO Pg W83 (gift from Dr. Potempa) and WT Pg W83 using a
ultracentrifuge machine. Those isolated vesicles, as well as whole bacterial cells, will be co-cultured with
osteoclast precursors. The level of citrullination of host proteins will be determined by W-blot and Mass-Spec.
The expression of PAD by vesicles will be monitored using a W-blot. Finally, possible engagement of OC-
STAMP expressed by osteoclasts which is the receptor for citrullinated Vimentin in osteoclastogenesis will be
determined by blocking with anti-OC-STAMP mAb or anti-citrullinated Vimentin mAb.
 After completing these two objectives, the candidate will have mastered the methods of cell culture, W-
 blotting, qPCR, flow cytometry, and osteoclastogenesis assays, as well as basic statistical analysis and
 interpretation of data which strengthen her credential to be a candidate for graduate program.

## Key facts

- **NIH application ID:** 10451355
- **Project number:** 3R01DE029709-02S1
- **Recipient organization:** NOVA SOUTHEASTERN UNIVERSITY
- **Principal Investigator:** TOSHIHISA KAWAI
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $74,336
- **Award type:** 3
- **Project period:** 2021-08-01 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10451355

## Citation

> US National Institutes of Health, RePORTER application 10451355, Osteoimmunology of Retarded Bone Regeneration in Periodontitis (3R01DE029709-02S1). Retrieved via AI Analytics 2026-06-01 from https://api.ai-analytics.org/grant/nih/10451355. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*