Development of slow-release carriers of vitamin D3 and Sincalide for periodontal regeneration

• Grant number: 24/17290-6
• Support Opportunities: Regular Research Grants
• Start date: August 01, 2025
• End date: July 31, 2028
• Field of knowledge: Health Sciences - Dentistry - Periodontology
• Principal Investigator: Karina Gonzales Silvério Ruiz
• Grantee: Karina Gonzales Silvério Ruiz
• Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil
• Associated researchers: Ana Paula Piovezan Fugolin ; Catharina Marques Sacramento

Abstract

One of the main challenges of contemporary periodontal therapy has been the re-establishment of supporting periodontal tissues lost due to the progression of periodontal disease. Our research group has tested several molecules that activate cell signaling pathways, modulating the process of alveolar bone and dental cementum neoformation. Two molecules, Sincalide and vitamin D3, have shown the potential to stimulate the deposition of mineralized matrix in vitro. The aim of this study is to develop hydrogel-based carrier platforms functionalized with slow-release nanoparticles made up of two types of biomodulators, vitamin D3 and Sincalide, and to evaluate their osteogenic properties in in vitro and in vivo experimental models. For the in vitro experiments, undifferentiated human periodontal ligament mesenchymal cells with a low potential for differentiation into an osteo/cementogenic phenotype (LOP cells) will be used, which will be treated with the hydrogels according to the following experimental groups: a) Control: cells cultured in standard culture medium (DMEM supplemented with 10% fetal bovine serum and 1% antibiotic), b) OM: cells cultured in osteogenic induction medium (OM medium - DMEM supplemented with 10% fetal bovine serum, 1% antibiotic, 50 µg/mL ascorbic acid, 10mM ¿-glycerophosphate and 10-5 M dexamethasone), c) Hsinc (Hydrogel-Sincalide): cells cultured in OM associated with Sincalide-based hydrogel, d) HVitD (Hydrogel-Vitamin D3): cells cultured in OM medium associated with Vitamin D3-based hydrogel, e) HSinc-VitD (Hydrogel-Sincalide + Vitamin D3): cells cultured in OM medium associated with Sincalide + Vitamin D3-based hydrogel. After the treatments, the following tests will be carried out to assess the osteogenic properties of the biomaterials: 1) cell viability tests (MTT), 2) deposition of mineralized matrix (alizarin red test), 3) quantification of extracellular calcium and alkaline phosphatase and 4) expression of genes for ASPN, BMP2, RUNX2, ALP, OC, VDR, CCK, CCKRA, RGS2, FOS, JNK3 and MKK6 (RT-qPCR). For in vivo analysis, experimental periodontitis will be induced in 150 male rats by installing a cotton ligature on the lower first molar. After 14 days, the ligatures will be removed and the furcation defects treated with the hydrogels according to the experimental groups: a) Control: no periodontal treatment, b) RACR: coronal root scraping and smoothing, c) RACR + HSinc: coronal scraping and smoothing followed by the application of 1mL of Sincalide-based hydrogel, d) RACR + HVitD: coronal scraping and smoothing followed by the application of 1mL of Vitamin D3-based hydrogel, e) RACR + HSinc + VitD: coronal scraping and smoothing followed by the application of 1mL of Sincalide + Vitamin D3-based hydrogel. After 7, 15 and 30 days, the animals will be euthanized and the samples submitted to routine histological procedures for histometric evaluation and micro-computed tomography analysis. (AU)