RSUSSH 2020
IN20-045 Utilization of modified mRNA encoding bone morphogenetic protein-2 for periodontal regeneration: an in vitro study
Presenter: Promphakkon Kulthanaamondhita
Chulalongkorn University, Thailand
Abstract
Current modalities for periodontal regeneration provide modest success, however complete periodontal regeneration is still not achievable. Recently in vitro synthesized nucleoside-modified messenger RNA (mRNA) has emerged as a novel platform in regenerative medicine. The aims of this study are to investigate the ability of human periodontal ligament cells (PDLCs), clinically relevant target cells, to produce bone morphogenetic protein-2 (BMP-2), a significant protein for bone formation after transfected with modified mRNA that encodes this protein. We investigated the biological activity of the translated protein for enhancing PDLC proliferation. Isolated PDLCs from healthy periodontal tissue were transfected with N1-methylpseudouridine modified mRNA encoding BMP-2 (m1Ψ-BMP-2 mRNA) complexed with transfecting agent, Lipofectamine 2000. Cell lysates and supernatants were collected at 24, 48 and 72 hours (h) after transfection for protein production by ELISA and cell viability by AlamarBlue assay. High levels of BMP-2 production were detected intracellulary and extracellulary. Secreted BMP-2 gradually increased up to 72 h. Cell viability was maintained above 90% throughout the observation period. In conclusion, the transfection of PDLCs with N1-methylpseudouridine modified mRNA encoding BMP-2 in lipofectamine 2000 led to high levels of functional BMP-2 protein. Using the in vitro synthesized nucleoside-modified mRNA may allow future application as a novel therapeutics platform for periodontal regeneration, however, further researches are required.
Citation format:
Kulthanaamondhita, P., Mahanonda, R., Champaiboon, C., Sa-Ard-Iam, N., Rerkyen, P., & Chanamuangkon, T.. (2020). Utilization of modified mRNA encoding bone morphogenetic protein-2 for periodontal regeneration: an in vitro study. Proceeding in RSU International Research Conference, May 1, 2020. Pathum Thani, Thailand.QUESTIONS & ANSWERS
Very interesting research. Are there any other growth factors used in mRNA encoding techniqe and how was the result of those studies compared to BMP-2?
How would you design your future reseach in animal model? Thank you.
The experiment is vey novel and interesting for periodontology field. I would like to ask about your idea on applying this result for real clinical practice? How would you incorporate this finding in the clinical practice?
Dear ทพ.ปภาตพงศ์ ศิริคุรุรัตน์
Thank you for the questions. Until today there are several other growth factors being studied under the chemically modified mRNA technology such as vascular endothelial growth factor-A (VEGF-A), platelet-derived growth factor-BB (PDGF-BB) and bone morphogenetic protein-9 (BMP-9). All of these studies reported positive results with some are better and some are inferior to this BMP-2 study. Interestingly, VEGF-A mRNA study has recently been conducted as a randomized phase I clinical trial in human with aims to improve diabetic wound healing in diabetes melltius patient where subjects were injected intradermally with chemically mRNA encoding VEGF-A. Increase in VEGF-A production and skin blood flow without any severe side effects were reported.
For future experiment in animal model, calvarial defects in rat or critical size defect in dog seems to be appropriate since BMP-2 is most potent in bone formation. The chemically modified mRNA can be delivered to the defect using appropriate delivery system such as collagen sponge and observation in bone formation enhancement can be seen.
Dear Dr.Kraisorn Sappayatosok
Thank you for the question. The concept of this mRNA therapy is to use patient own cell to produce desired proteins that aids in tissue regeneration. In clinical setting, this chemically modified mRNA complex could be delivered using appropriate carrier such as collagen sponge to the site in need for regeneration such as in a periodontal defect. After that the mRNA complex are release, travel to the neighboring cells, transfect the target cell and release the mRNA into the cytoplasm. Then the translation process begins and the produced protein are released and function in enhancing regeneration. Nevertheless, in orderfor the application in clinical practice to be reached, furthur researches are required.
Thank you for the answer Dr, Promphakkon