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Arrabidaea chica Verlot: in vitro evaluation of toxicity, antimicrobial and cytoprotective activities from standardized free crude extract combined with zoledronic acid, and microencapsulation studies

Grant number: 10/07781-0
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): October 01, 2010
Effective date (End): February 28, 2014
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Mary Ann Foglio
Grantee:Patrícia Maria Wiziack Zago
Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil
Associated scholarship(s):11/21006-1 - Arrabidaea chica Verlot ethanolic extract activity on fibroblasts, keratinocytes, osteoblasts and osteoclasts under a bisphosphonate treatment, BE.EP.DR


The bisphosphonate related osteonecrosis of the jaw (BRONJ) is a condition associated to bisphosphonates, medicines used to treat bone disorders, as zoledronic acid. In vitro studies showed epithelial cells viability and growth can be altered by bisphosphonates, and may cause a deficient wound healing in BRONJ. Arrabidaea chica (H&B.) Verlot is a Bignoniaceae with wound healing properties, however, anthocyanidins, the main compounds of the plant extract, can be easily degraded under environmental conditions. This in vitro study aimed to evaluate the antimicrobial, citotoxic and citoprotective actions of A. chica hydroalcoholic extract (AC) in the presence of ZA therapy. Stability studies at 40 °C and 75% humidity conditions, from microencapsulated into arabic gum plus maltodextrin (1:1) and free crude extract samples were conducted. Keratinocytes (HaCaT), fibroblasts (PT64-04HF) and osteoblasts (MC3T3-E1) were submitted to 2 treatment protocols: zoledronic acid 10 µM (ZA) therapy during 24 h and then AC during 48 h and vice-versa; or co-treatment with both. Parameters as viability, apoptosis and caspase 3/7 activation were evaluated. Cancer cells (C4-2B) viability was determined after AC treatment. Streptococcus mutans were submitted to co-treatment with ZA and AC. Microcapsules containing AC were obtained after spray dryer processing, aliquoted and stored into climatic chamber for stability study according to ANVISA protocol (RE 01/2005). ANOVA and T Student test, a=0,05, were applied for statistical analysis. After 48 h, ZA treatment decreased viability of keratinocytes, fibroblasts and osteoblasts repectively by 34%, 47% and 51%. Co-treatments ZA and AC 5 µg mL-1 (82%, 83% and 55%), and ZA and AC 10 µg mL-1 (69%, 70% and 64%) increased cell viability. No apoptosis or caspase activation were detected on fibroblasts, however, caspase 3/7 was active on osteoblasts after ZA 48 h treatment. Concentrations higher than 10 µg mL-1 of AC were toxic to cancer cells and showed a decrease by 40% in cell viability, while no effect was seen with AC 5 or 10 µg mL-1. Higher ZA concentrations (50 µM) seemed to induce S. mutans growth, while AC showed no effect on bacteria. Finally, the studied core matherial (arabic gum plus matodextrin 1:1) was ineffective on protecting AC extract against degradation, since anthocyanidins concentration was reduced after 15 days incubation. Therefore, lower AC concentrations (5 and 10 µg mL-1) were promising on treating BRONJ wounds, as they were effective in protect epithelial cells against harmfull effects of ZA, without affecting cancer cells or S. mutans growth. Different types of core matherials must be evaluated for improving AC chemical stability. More studies are required to prove AC beneficial effects on treating BRONJ. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
SERVAT-MEDINA, LEILA; GONZALEZ-GOMEZ, ALVARO; REYES-ORTEGA, FELISA; OLIVEIRA SOUSA, ILZA MARIA; ALMEIDA QUEIROZ, NUBIA DE CASSIA; WIZIACK ZAGO, PATRICIA MARIA; JORGE, MICHELLE PEDROSA; MONTEIRO, KARIN MAIA; DE CARVALHO, JOAO ERNESTO; SAN ROMAN, JULIO; et al. Chitosan-tripolyphosphate nanoparticles as Arrabidaea chica standardized extract carrier: synthesis, characterization, biocompatibility, and antiulcerogenic activity. INTERNATIONAL JOURNAL OF NANOMEDICINE, v. 10, p. 3897-3909, . (09/16484-1, 11/09127-8, 10/07781-0)
WIZIACK ZAGO, PATRICIA MARIA; OLIVEIRA SOUSA, ILZA MARIA; SERVA-MEDINA, LEILA; JORGE, MICHELLE PEDROZA; LIMA NETO, LIDIO GONCALVES; HASS, VIVIANE; LI, XIN; GOIS RUIZ, ANA LUCIA TASCA; SAXENA, DEEPAK; FOGLIO, MARY ANN. Standardized Arrabidaea chica Extract Shows Cytoprotective Effects in Zoledronic Acid-Treated Fibroblasts and Osteoblasts. CLINICAL COSMETIC AND INVESTIGATIONAL DENTISTRY, v. 12, p. 327-333, . (11/09127-8, 10/07781-0, 11/21006-1)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
ZAGO, Patrícia Maria Wiziack. Arrabidaea chica Verlot: in vitro evaluation of toxicity, antimicrobial and cytoprotective activities from standardized free crude extract combined with zoledronic acid, and microencapsulation studies. 2014. Doctoral Thesis - Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba Piracicaba, SP.

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Filed patent(s) as a result of this research project

COMPOSIÇÕES FARMACÊUTICAS COMPREENDENDO EXTRATO DE ARRABIDAEA CHICA, VERLOT EM SISTEMAS DE LIBERAÇÃO MICRO E NANOPARTICULADOS E LIPOSSOMAIS, PROCESSOS DE FABRICAÇÃO E USO DOS MESMOS PI1105312-7 - Universidade Estadual de Campinas (UNICAMP) . Marcos Nogueira Eberlin; Rodney Alexandre Ferreira Rodrigues; Mary Ann Foglio; Leila Servat; Michelle Pedroza Jorge; Ilza Maria Oliveira Souza; Joao Ernesto de Carvalho; Ana Lúcia Tasca Góis Ruiz; Patrícia Maria Wiziack Zago; Elaine Cristina Cabral Polcelli; Maria Helena Andrade Santana; Viviane Ferre de Souza; Glyn Mara Figueira - December 2011, 20

PHARMACEUTICAL COMPOSITIONS COMPRISING ARRABIDAEA CHICA EXTRACT IN CONTROLLED RELEASE SYSTEMS, PRODUCTION PROCESS AND USE THEREOF PCT/BR2012/000532 - Universidade Estadual de Campinas (UNICAMP) . Mary Ann Foglio; João Ernesto De Carvalho; Ana Lucia Tasca Góis Ruiz; Michelle Pedroza Jorge; Leila Servat; Patricia Maria Wiziack Zago; Marcos Nogueira Eberlin; Elaine Cristina Cabral Polcelli; Maria Helena Andrade Santana; Viviane Ferre De Souza; Glyn Mara Figueira; Ilza Maria Oliveira; Rodney Alexandre Ferreira Rodrigues - December 2012, 19