Cell therapy: potential of mesenchymal stem cells, VEGF-A and BMP-9 to regenerate bone tissue

• Grant number: 17/12622-7
• Support Opportunities: Research Projects - Thematic Grants
• Start date: August 01, 2018
• End date: July 31, 2024
• 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
• Pesquisadores principais: Márcio Mateus Beloti
• Associated researchers: Gary S. Stein ; Geraldo Aleixo da Silva Passos Júnior ; Jane Lian ; Janet L. Stein ; José César Rosa ; Karina Fittipaldi Bombonato Prado ; Marília Afonso Rabelo Buzalaf ; Paulo Tambasco de Oliveira
• Associated research grant(s): 23/01309-7 - Effect of triple-primed mesenchymal stem cells on bone regeneration, AP.R; 22/03820-8 - Micro- and nanotopography-based stem cell priming technology for cell therapy, AP.R; 19/05289-5 - Multi-User Equipment approved in grant 2017/21622-7: aparelho QuantStudio 7 Flex Real-Time PCR System, AP.EMU; 19/05291-0 - Multi-use equipment approved in grant 2017/12622-7:SkyScan 1276 high resolution desktop in-vivo microtomograph, AP.EMU
• Associated scholarship(s): 23/00538-2 - Effect of Engrailed-1 protein modulation in mesenchymal stem cells on osteoblastic differentiation and bone formation in defects created in mouse calvaria, BP.DR; 23/06099-0 - Effect of the secretome from mesenchymal stem cells overexpressing vascular endothelial growth factor A on bone repair, BP.IC; 23/02217-9 - Cell Therapy: Potential of Mesenchymal Stem Cells, VEGF-A and BMP-9 to Regenerate Bone Tissue, BP.PD; + associated scholarships 22/12618-8 - Cell therapy: potential of mesenchymal stem cells, VEGF-A and BMP-9 to regenerate bone tissue, BP.PD; 22/10128-3 - Evaluation of the protein expression of BMP-9 and/or VEGFA in bone marrow and adipose tissue mesenchymal stem cells genetically edited by CRISPR-Cas9, BP.IC; 21/04824-4 - Effect of agrin overexpression in adipose tissue-derived mesenchymal stem cells on osteoblastic and adipocytic differentiation and on bone regeneration, BP.DR; 21/03204-2 - Participation of agrin on osteoblast differentiation and bone tissue regeneration induced by mesenchymal stem cells in defects created in mouse calvaria, BP.DD; 20/06599-5 - Effect of conditioned medium by mesenchymal stem cells overexpressing bone morphogenetic protein 9 on osteoblast differentiation and bone regeneration, BP.MS; 20/03616-6 - Effect of local injection of mesenchymal stem cells combined with systemic injections of parathyroid hormone and pentoxifylline on bone repair, BP.DD; 19/18711-7 - Effect of diabete mellitus on osteoblastic differentiation of mesenchymal stem cells, BP.MS; 19/10076-0 - Effect of mesenchymal stem cells from periodontal ligament with high and low osteogenic potential on bone repair, BP.MS; 19/01344-1 - Evaluation of the osteogenic and bone tissue regeneration potentials of BM-MSCs and AT-MSCs associated with VEGF-A and BMP-9, BP.PD; 19/01346-4 - Evaluation of the angiogenic and bone regeneration potentials of BM-MSCs and AT-BMCs genetically edited by CRISP-Cas9 to overexpress VEGF-A and BMP-9, BP.PD; 18/13290-0 - Cell therapy: evaluation of the effect of mesenchymal stem cells to regenerate bone tissue of rats with Osteoporosis, Diabetes Mellitus or Arterial Hypertension, BP.DR; 18/19559-1 - Evaluation of the angiogenic and bone healing potential of BM-MSCs and AT-MSCs combined with VEGF-A and BMP-9, BP.IC; 18/16852-0 - Study of the angiogenic potential and of the capacity to regenerate bone tissue of BM-MSCs and AT-MSCs associated with VGFA-A and BMP-9, BP.IC - associated scholarships

Abstract

Bone tissue has high capacity of regeneration, but in several situations the extent of the injury overcomes its regenerative potential. In this scenario, therapies based on the use of mesenchymal stem cells (MSCs) have aroused the attention of many researchers for being a promising alternative compared with the available treatments. However, many molecular, cellular and tissue characteristics remain unmet in the literature to make cell therapy an effective treatment for bone repair both in healthy and systemic compromised patients by pathologies such as osteoporosis, diabetes and hypertension. In keeping with this, this research project consists of three subprojects that aim to: (1) evaluate the potential of MSCs harvested from either bone marrow (BM-MSCs) or adipose tissue (AT-MSCs) combined with vascular endothelial growth factor A (VEGF-A) and/or bone morphogenetic protein 9 (BMP-9) to repair bone defects; (2) evaluate the potential of these MSCs genetically edited to overexpress VEGF-A and/or BMP-9 to repair bone defects; and (3) evaluate the effect of BM-MSCs harvested from healthy rats on the osteoblastic differentiation of BM-MSCs harvested from osteoporotic, diabetic and hypertensive rats. To develop subprojects 1 and 2, BM-MSCs and AT-MSCs will be treated with VEGF-A and/or BMP-9 or genetically edited by clustered regularly interspaced short palindromic repeats (CRISPR-Cas9) to overexpress VEGF-A and/or BMP-9. Those cells will be evaluated in vitro to assess their angiogenic and osteogenic potentials as well as their large scale genomic and proteomic profile. For bone repair, BM-MSCs and AT-MSCs either combined with VEGF-A and/or BMP-9 or edited to overexpress these factors will be directly injected into rat bone calvarial defects. To evaluate the presence of cells in the defects, Luc-expressing cells will be tracked by bioluminescent imaging. Up to 4 weeks, vasculogenesis and bone formation will be evaluated by in vivo micro-CT. After 4 weeks, the animals will be euthanized and the harvested calvaria evaluated by histological analysis. In the subproject 3, the effect of BM-MSCs harvested from healthy rats on the osteoblastic differentiation of BM-MSCs from osteoporotic, diabetic and hypertensive rats will be evaluated using an indirect co-culture model. These studies are the first step aiming to apply these cell therapies to regenerate bone defects in the presence of such pathologies. (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)

ABUNA, RODRIGO P. F.; ALMEIDA, LUCIANA O.; SOUZA, ALANN T. P.; FERNANDES, ROGER R.; SVERZUT, THALES F. V.; ROSA, ADALBERTO L.; BELOTI, MARCIO M.. Osteoporosis and osteoblasts cocultured with adipocytes inhibit osteoblast differentiation by downregulating histone acetylation. Journal of Cellular Physiology, v. 236, n. 5, p. 3906-3917, MAY 2021. (18/17356-6, 18/13290-0, 16/14171-0, 17/12622-7, 16/14711-4)

WEFFORT, D.; ADOLPHO, L. F.; SOUZA, A. T. P.; FREITAS, G. P.; LOPES, H. B.; OLIVEIRA, F. S.; BIGHETTI-TREVISAN, R. L.; PITOL-PALIN, L.; MATSUSHITA, D. H.; OKAMOTO, R.; et al. Effect of Normoglycemia on the Disrupted Osteoblast Differentiation of Mesenchymal Stem Cells Induced by Type 1 and Type 2 Diabetes Mellitus. MOLECULAR BIOLOGY OF THE CELL, v. 34, n. 2, p. 2-pg., 2023-02-01. (19/18711-7, 17/12622-7)

SOUZA, ALANN THAFFARELL PORTILHO; FREITAS, GILEADE PEREIRA; LOPES, HELENA BACHA; TOTOLI, GABRIELA GUARALDO CAMPOS; TARONE, ADRIANA GADIOLI; MAROSTICA-JUNIOR, MARIO ROBERTO; ROSA, ADALBERTO LUIZ; BELOTI, MARCIO MATEUS. Jabuticaba peel extract modulates adipocyte and osteoblast differentiation of MSCs from healthy and osteoporotic rats. JOURNAL OF BONE AND MINERAL METABOLISM, v. 39, n. 2, SEP 2020. (15/13320-9, 17/20349-9, 18/11069-5, 15/50333-1, 17/12622-7)

FREITAS, GILEADE P.; SOUZA, ALANN T. P.; LOPES, HELENA B.; TREVISAN, RAYANA L. B.; OLIVEIRA, FABIOLA S.; FERNANDES, ROGER R.; FERREIRA, FERNANDA U.; ROSE, FELIPE A.; BELOTI, MARCIO M.; ROSA, ADALBERTO L.. Mesenchymal Stromal Cells Derived from Bone Marrow and Adipose Tissue: Isolation, Culture, Characterization and Differentiation. BIO-PROTOCOL, v. 10, n. 4, FEB 20 2020. (18/17356-6, 17/12622-7)

ADOLPHO, LETICIA FAUSTINO; LOPES, HELENA BACHA; FREITAS, GILEADE PEREIRA; WEFFORT, DENISE; CAMPOS TOTOLI, GABRIELA GUARALDO; LOYOLA BARBOSA, ANA CAROLINA; FREIRE ASSIS, RAHYZA INACIO; SILVERIO RUIZ, KARINA GONZALES; ANDIA, DENISE CARLETO; ROSA, ADALBERTO LUIZ; et al. Human periodontal ligament stem cells with distinct osteogenic potential induce bone formation in rat calvaria defects. REGENERATIVE MEDICINE, v. 17, n. 6, p. 13-pg., 2022-03-16. (19/10076-0, 17/12622-7, 13/09650-8)

TREVISAN, R. L. B.; FERRAZ, E. P.; GORDON, J. A. R.; TYE, C.; LIAN, J. B.; STEIN, G. S.; STEIN, J. L.; ROSA, A. L.; BELOTI, M. M.. Effects of Osteoclasts on Transcriptional Regulation of Non-Coding RNAs in Osteoblasts Grown on Titanium with Nanotopography. MOLECULAR BIOLOGY OF THE CELL, v. 34, n. 2, p. 2-pg., 2023-02-01. (17/12622-7, 18/17356-6, 19/09349-2, 17/23888-8)

WEFFORT, DENISE; ADOLPHO, LETICIA F.; SOUZA, ALANN T. P.; FREITAS, GILEADE P.; LOPES, HELENA B.; OLIVEIRA, FABIOLA S.; BIGHETTI-TREVISAN, RAYANA L.; PITOL-PALIN, LETICIA; MATSUSHITA, DORIS H.; OKAMOTO, ROBERTA; et al. Normoglycemia partially recovers the disrupted osteoblast differentiation of mesenchymal stem cells induced by type 1 but not type 2 diabetes mellitus. Journal of Cellular Biochemistry, v. 124, n. 7, p. 14-pg., 2023-06-09. (19/18711-7, 17/12622-7)

CALIXTO, ROBSON DIEGO; FREITAS, GILEADE PEREIRA; SOUZA, PAOLA GOMES; RAMOS, JAQUELINE ISADORA REIS; SANTOS, ISABELA CRISTINE; DE OLIVEIRA, FABIOLA SINGARETTI; ALMEIDA, ADRIANA LUISA GONCALVES; ROSA, ADALBERTO LUIZ; BELOTI, MARCIO MATEUS. Effect of the secretome of mesenchymal stem cells overexpressing BMP-9 on osteoblast differentiation and bone repair. Journal of Cellular Physiology, v. N/A, p. 13-pg., 2023-09-03. (17/12622-7, 20/06599-5, 19/01346-4)