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Effect of Local Injections of Mesenchymal Stem Cells and Osteoblasts on Bone Regeneration

Grant number: 14/26864-4
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): June 01, 2016
Effective date (End): January 31, 2019
Field of knowledge:Health Sciences - Dentistry - Oral and Maxillofacial Surgery
Principal Investigator:Adalberto Luiz Rosa
Grantee:Gileade Pereira Freitas
Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated scholarship(s):16/23850-8 - Gene therapy for bone regeneration using CRISPR-Cas9 system, BE.EP.DR


Bone has a high regenerative capacity, but in some situations the lesion extension prevents the tissue repair. In this context, cell therapy has been the focus of several research groups as an alternative to the current treatments. However, there are experimental aspects that need to be deeply investigated in order to make cell therapy an effective treatment for regenerating bone defects. Then, the aim of this study will be evaluate the potential of mesenchymal stem cells (MSCs) derived from either bone marrow or adipose tissue, as well as, osteoblasts differentiated from these cells to regenerate bone tissue when directly injected (repeated injections) into rat calvarial bone defects. MSCs will be obtained from bone marrow and inguinal adipose tissue of rats, cultured in conditions to either keep MSCs features or to differentiate into osteoblasts. MSCs will be characterized 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 in vivo experiments will be carried out with the four groups of cells, (1) MSCs derived from bone marrow, (2) MSCs derived from adipose tissue, (3) osteoblasts differentiated from bone marrow MSCs and (4) osteoblasts differentiated from adipose tissue MSCs, expressing luciferase. This will allow tracking these cells after direct injection into calvarial bone defects to determine the number of injections and its frequency. The first cell injection will be carried out after 2 weeks of the bone defect creation and at the end of 8 weeks the new-formed bone will be evaluated by microtomography, histological analysis and elastic modulus and hardness analyses. The data will be submitted to adherence to normal distribution to determine the appropriate statistical test. The results of this study may stablish new parameters regarding cell injections to regenerate bone tissue as, up to now, there are no studies investigating the effect of repeated injections.

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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)
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, . (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, . (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., . (14/26864-4, 15/21439-6)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
FREITAS, Gileade Pereira. Effect of local injections of mesenchymal stem cells or osteoblasts on bone regeneration. 2019. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Odontologia de Ribeirão Preto (PCARP/BC) Ribeirão Preto.

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