Mesenchymal stem cells and osteoblasts: evaluation of molecular signatures and their potential for bone regeneration

• Grant number: 15/21439-6
• Support Opportunities: Regular Research Grants
• Start date: June 01, 2016
• End date: May 31, 2018
• Field of knowledge: Health Sciences - Dentistry - Oral and Maxillofacial Surgery
• Principal Investigator: Adalberto Luiz Rosa
• Grantee: Adalberto Luiz Rosa
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated researchers: José César Rosa ; Karina Fittipaldi Bombonato Prado ; Márcio Mateus Beloti ; Paulo Tambasco de Oliveira

Abstract

Bone has a high regenerative capacity, but in some cases the extension of the lesion prevents tissue repair. In this context, cell therapy has been the focus of several research groups as an alternative to the current treatments. However, several molecular, cellular and tissue aspects were not addressed by the literature and need to be investigated in order to make cell therapy an effective treatment for regenerating bone defects. Then, the aim of this study is to compare the molecular signatures and the potential of mesenchymal stem cells (MSCs) and osteoblasts to regenerate bone tissue. MSCs will be obtained from bone marrow (MSCs-BM) and adipose tissue (MSCs-AT) of rats, cultured in conditions to either keep MSCs features or to differentiate into osteoblasts (OBs-BM and OBs-AT, respectively). MSCs will be identified by the expression of CD29, CD31, CD34, CD45, CD90 and CD106 by flow cytometry. Gene expression of Runx2, alkaline phosphatase and osteocalcin will be evaluated by real-time PCR to confirm osteoblast differentiation. The molecular signatures of MSCs-BM, MSCs-AT, OBs-BM and OBs-AT will be evaluated by large-scale analyses of gene (microarray) and protein (proteomic) expression. For bone regeneration, cells will be directly injected (single injection x repeated injections) in bone defects created in rat calvarial two weeks before the first injection to mimic bone defect without self-regeneration capacity. Firstly, MSCs-BM, MSCs-AT, OBs-BM and OBs-AT transfected to express luciferase will be injected in bone defects to allow tracking cells to determine the interval between repeated injections. Then, the first cell injection will be carried out after two weeks of the bone defect creation and the additional injections will be done following the interval previously determined. Bone defects treated with a single cell injection and with the vehicle phosphate buffered saline without cells will also be evaluated. Six weeks after the first injection, the rats will be killed and the new-formed bone will be evaluated by microtomography, histological and real-time PCR analysis and elastic modulus and hardness analyses. The data will be submitted to normal distribution adherence to determine the appropriate statistical test. The results of this study may stablish new parameters regarding characteristics of candidate cells to be used in cell therapy and their injections to regenerate bone tissue as, up to now, there are no studies investigating the effect of repeated injections. (AU)

Scientific publications

(8)

(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)

FREITAS, GILEADE P.; LOPES, HELENA B.; SOUZA, ALANN T. P.; OLIVEIRA, PAULA G. F. P.; ALMEIDA, ADRIANA L. G.; COELHO, PAULO G.; FERREIRA, FERNANDA U.; COVAS, DIMAS T.; BELOTI, MARCIO M.; ROSA, ADALBERTO L.. Effect of cell therapy with osteoblasts differentiated from bone marrow or adipose tissue stromal cells on bone repair. REGENERATIVE MEDICINE, v. 14, n. 12, p. 1107-1119, DEC 2019. (14/26864-4, 15/21439-6, 09/54142-5)

FREITAS, GILEADE P.; LOPES, HELENA B.; SOUZA, ALANN T. P.; OLIVEIRA, PAULA G. F. P.; ALMEIDA, ADRIANA L. G.; SOUZA, LUCAS E. B.; COELHO, PAULO G.; BELOTI, MARCIO M.; ROSA, ADALBERTO L.. Cell Therapy: Effect of Locally Injected Mesenchymal Stromal Cells Derived from Bone Marrow or Adipose Tissue on Bone Regeneration of Rat Calvarial Defects. SCIENTIFIC REPORTS, v. 9, SEP 17 2019. (14/26864-4, 15/21439-6)

FREITAS, G. P.; LOPES, H. B.; ALMEIDA, A. L.; BELOTI, M. M.; ROSA, A. L.. Bone formation in calvarial defects treated with mesenchymal stem cells or osteoblasts derived from adipose tissue: microtomographic analysis.. MOLECULAR BIOLOGY OF THE CELL, v. 27, p. 2-pg., 2016-01-01. (14/26864-4, 15/21439-6)

GONCALVES ALMEIDA, ADRIANA LUISA; FREITAS, GILEADE PEREIRA; LOPES, HELENA BACHA; GIMENES, ROSSANO; SIESSERE, SELMA; SOUSA, LUIZ GUSTAVO; BELOTI, MARCIO MATEUS; ROSA, ADALBERTO LUIZ. Effect of stem cells combined with a polymer/ceramic membrane on osteoporotic bone repair. BRAZILIAN ORAL RESEARCH, v. 33, p. 11-pg., 2019-01-01. (15/21439-6)

ADRIANA LUISA GONCALVES ALMEIDA; GILEADE PEREIRA FREITAS; HELENA BACHA LOPES; ROSSANO GIMENES; SELMA SIESSERE; LUIZ GUSTAVO SOUSA; MARCIO MATEUS BELOTI; ADALBERTO LUIZ ROSA. Effect of stem cells combined with a polymer/ceramic membrane on osteoporotic bone repair. Brazilian Oral Research, v. 33, 2019-09-09. (15/21439-6)

SOUZA, ALANN THAFFARELL PORTILHO; LOPES, HELENA BACHA; FREITAS, GILEADE PEREIRA; FERRAZ, EMANUELA PRADO; OLIVEIRA, FABIOLA SINGARETTI; ALMEIDA, ADRIANA LUISA GONCALVES; WEFFORT, DENISE; BELOTI, MARCIO MATEUS; ROSA, ADALBERTO LUIZ. Role of embryonic origin on osteogenic potential and bone repair capacity of rat calvarial osteoblasts. JOURNAL OF BONE AND MINERAL METABOLISM, v. 38, n. 4, FEB 2020. (15/21439-6, 09/54142-5)

PORTILHO SOUZA, ALANN THAFFARELL; FREITAS, GILEADE PEREIRA; LOPES, HELENA BACHA; FERRAZ, EMANUELA PRADO; OLIVEIRA, FABIOLA SINGARETTI; BELOTI, MARCIO MATEUS; ROSA, ADALBERTO LUIZ. Effect of cell therapy with allogeneic osteoblasts on bone repair of rat calvaria defects. CYTOTHERAPY, v. 20, n. 10, p. 1267-1277, OCT 2018. (15/21439-6)

MOURA FIDELES, SIMONE ORTIZ; ORTIZ, ADRIANA CASSIA; ASSIS, AMANDA FREIRE; DUARTE, MAX JORDAN; OLIVEIRA, FABIOLA SINGARETTI; PASSOS, GERALDO ALEIXO; BELOTI, MARCIO MATEUS; ROSA, ADALBERTO LUIZ. Effect of cell source and osteoblast differentiation on gene expression profiles of mesenchymal stem cells derived from bone marrow or adipose tissue. Journal of Cellular Biochemistry, v. 120, n. 7, p. 11842-11852, JUL 2019. (15/21439-6)