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| Author(s): |
Luciana Gonçalves Sicchieri
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Ribeirão Preto. |
| Institution: | Universidade de São Paulo (USP). Faculdade de Odontologia de Ribeirão Preto (PCARP/BC) |
| Defense date: | 2010-09-03 |
| Examining board members: |
Adalberto Luiz Rosa;
Luciana Asprino;
Cristina Antoniali Silva
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| Advisor: | Adalberto Luiz Rosa |
| Abstract | |
It has been suggested that an adequate bone repair can be obtained by hybrid biomaterials, produced by combining osteoprogenitor cells and macroporous bone substitutes. The aim of this study was to evaluate the application of hybrid biomaterial formed by PLGA/CaP scaffold and bone marrow-derived mesenchymal stem cells and osteoblasts in bone tissue engineering. The effect of scaffold pore size was evaluated in vitro assessing cell growth, alkaline phosphatase (ALP) activity, and quantitative gene expression of osteoblastic phenotype markers in osteoblasts cultured on scaffolds; and in vivo assesseing bone formation after implantation of scaffolds in critical size rat calvaria defects. It was also evaluated the effect of pore size on cell seeding by centrifugal force. To evaluate the effect of fetal calf serum used to supplement the cell culture medium on in vivo tissue response, scaffolds exposed to serum were implanted in critical size rat calvaria defects. The effect of withdrawal of fetal calf serum in the culture medium on osteoblasts was analyzed by cell growth, ALP activity, total protein content and mineralization. To evaluate the effect of cell differentiation stage on bone repair, cells either undifferentiated or in different stages of osteoblast differentiation associated with the PLGA/CaP were implanted autologously in critical size rat calvaria. Scaffolds with pore sizes of around 1000 µm would favor the osteoblast differentiation and display a better cell seeding, while the scaffolds with pore sizes of around 500 µm would favor increased bone and vascular formation. The fetal calf serum influenced negatively the in vivo tissue response and bone formation. The withdrawal of fetal calf serum in the culture medium reduced cell growth and ALP activity without affecting the total protein content and the formation of mineralized matrix Mesenchymal stem cells and osteoblats at the early stage of differentiation (7 days) promoted greater bone formation, therefore they would allow the obtention of a hybrid biomaterial with higher osteogenic potential. (AU) | |
| FAPESP's process: | 07/05757-1 - Bone tissue engineering: in vitro and in vivo evaluation of the hybrid biomaterial calcium phosphate/poly(lactide-co-glycolide) and osteoblastic cells derived from stem cells |
| Grantee: | Luciana Gonçalves Sicchieri |
| Support Opportunities: | Scholarships in Brazil - Master |