Efetividade de scaffolds de poli (butileno adipato-co-tereftalato) / nanohidroxiapatita obtidos por eletrofiação para aplicação biomédica: avaliação in vitro
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Data
2016-02-01
Autores
Santana-Melo, Gabriela de Fátima [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A necessidade da fabricação de novos biomateriais que possam, além de mimetizar o tecido ósseo, fornecer resistências mecânicas favoráveis próximas às do tecido ósseo natural têm despertado interesse de pesquisadores com o objetivo de melhorar a qualidade de vida de pessoas que sofreram algum tipo de lesão. Scaffolds de nanofibras poliméricas fabricados por eletrofiação apresentam características tridimensionais (3D) e poros interconectados que permitem a colonização de toda a superfície 3D por células com a consequente formação de tecidos. O scaffold de poli (butileno adipato-co-tereftalato) (PBAT) mostra-se um biomaterial promissor para regeneração óssea, porém tem sido pouco explorado até a data. Embora do uso da HA seja consagrado para uso biomédico, sua utilização em polímeros ainda é pouco estudada, principalmente em associação ao PBAT. Desta forma, o objetivo deste estudo foi avaliar a efetividade in vitro de scaffolds poliméricos (PBAT) com incorporação de nanopartículas de HA (nHAp) em diferentes concentrações, produzidos por eletrofiação, por meio da bioatividade celular e expressão gênica de osteoblast-like MG63. Células (MG63) foram cultivadas sobre scaffolds de PBAT; PBAT/3%nHAp e PBAT/5% nHAp e sem a presença dos mesmos (controle) e avaliadas pelos testes qualitativo (MEV) e quantitativo de adesão e proliferação celular (1 e 7 dias e aos 1, 3, 7, 14 e 21 dias, respectivamente), citotoxicidade celular (1, 3 e 7 dias), corante vermelho de alizarina e formação de mineralização (14 dias) e análise da expressão de genes relacionados à osteogênese por qRT-PCR aos 7, 14 e 21 dias de cultura celular. Os dados foram analisados estatisticamente por variância (ANOVA) e Tukey (p<0,05). Os scaffolds de PBAT e PBAT/nHAp não apresentaram efeito citotóxico e sua arquitetura tridimensional influenciou positivamente na adesão e proliferaçãocelular, formação de matriz mineralizada bem como em alguns períodos na expressão dos genes ALP, Col I, Runx2, OC e OPN em relação ao grupo controle. O efeito osteocondutor e osteoindutor da nHAp promoveu melhor resposta celular nos scaffolds de PBAT/nHAp, independente da concentração. Esses resultados demonstram a efetividade in vitro dos scaffolds de PBAT e PBAT/nHAp, apresentando grande potencial para aplicação biomédica.
The need for the manufacture of new biomaterials that may, in addition to mimic to bone tissue, providing favorable mechanical strength close to natural bone have aroused the interest of researchers in order to improve the quality of life of people who have suffered some kind of injury. Scaffolds polymer nanofibers fabricated by electrospinning have three dimensional features (3D) and interconnected pores that allow the colonization of the entire 3D surface of cells with the consequent formation of tissue. Poly (butylene adipate-co-terephthalate) (PABT) scaffold showed to be a promising biomaterial for bone regeneration, however, has been underexplored to date. Although the use of HA is consecrated to biomedical use, their use in polymers is not well known, especially in association with PBAT. The aim of this study was evaluating in vitro effectiveness of polymeric (PABT) scaffolds with incorporated HA (nHAp) nanoparticles, obtained by electrospinning, through cellular bioactivity and osteoblast-like MG63 gene expression. MG63 cells were grown on PABT; PABT/3%nHAp and PABT/5%nHAp scaffolds and without their presence (control), and evaluated by qualitative (MEV) and quantitative tests of cell adhesion and proliferation (1 and 7 days and at 1, 3, 7, 14 and 21 days, respectively), cell cytotoxicity (1, 3 and 7 days), alizarin red dye and mineralization formation (14 days) and expression of genes related to osteogenesis by qRT-PCR to 7, 14 and 21 days of cell culture. Data were statistically analyzed by variance (ANOVA) and Tukey test (p<0.05). The PBAT and PBAT/nHAp scaffolds showed no cytotoxic effect and its three-dimensional architecture influenced positively in the cell adhesion and proliferation, mineralized matrix formation as well as in some periods the expression of genes ALP, Col I, Runx2, OC and OPN in relation the control group. The osteoconductive and osteoinductive effect of nHAp promoted better cellular response in scaffolds of PABT/nHAp independent of concentration. These results demonstrate the in vitro effectiveness of PABT and PABT/nHAp scaffolds, presenting great potential for biomedical application.
The need for the manufacture of new biomaterials that may, in addition to mimic to bone tissue, providing favorable mechanical strength close to natural bone have aroused the interest of researchers in order to improve the quality of life of people who have suffered some kind of injury. Scaffolds polymer nanofibers fabricated by electrospinning have three dimensional features (3D) and interconnected pores that allow the colonization of the entire 3D surface of cells with the consequent formation of tissue. Poly (butylene adipate-co-terephthalate) (PABT) scaffold showed to be a promising biomaterial for bone regeneration, however, has been underexplored to date. Although the use of HA is consecrated to biomedical use, their use in polymers is not well known, especially in association with PBAT. The aim of this study was evaluating in vitro effectiveness of polymeric (PABT) scaffolds with incorporated HA (nHAp) nanoparticles, obtained by electrospinning, through cellular bioactivity and osteoblast-like MG63 gene expression. MG63 cells were grown on PABT; PABT/3%nHAp and PABT/5%nHAp scaffolds and without their presence (control), and evaluated by qualitative (MEV) and quantitative tests of cell adhesion and proliferation (1 and 7 days and at 1, 3, 7, 14 and 21 days, respectively), cell cytotoxicity (1, 3 and 7 days), alizarin red dye and mineralization formation (14 days) and expression of genes related to osteogenesis by qRT-PCR to 7, 14 and 21 days of cell culture. Data were statistically analyzed by variance (ANOVA) and Tukey test (p<0.05). The PBAT and PBAT/nHAp scaffolds showed no cytotoxic effect and its three-dimensional architecture influenced positively in the cell adhesion and proliferation, mineralized matrix formation as well as in some periods the expression of genes ALP, Col I, Runx2, OC and OPN in relation the control group. The osteoconductive and osteoinductive effect of nHAp promoted better cellular response in scaffolds of PABT/nHAp independent of concentration. These results demonstrate the in vitro effectiveness of PABT and PABT/nHAp scaffolds, presenting great potential for biomedical application.
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Palavras-chave
Eletrofiação, Nanofibras, PBAT, Nanohidroxiapatita, Células MG63, qRT-PCR, Regeneração óssea, Electrospinning, Nanofibers, Nanohydroxyapatite, MG63 cells, Bone regeneration