Comportamento de células MC3T3 e adesão bacteriana em liga β Ti-35Nb-7Zr- 5Ta caracterizada com íons Mg2+, Ca2+ e PO34-
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Data
2021-12-15
Autores
Reis, Bárbara Araújo dos
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Universidade Estadual Paulista (Unesp)
Resumo
Reabilitações com implantes dentais são procedimentos rotineiros na prática odontológica atual. Altas taxas de sobrevivência do implante foram alcançadas nas últimas décadas devido a modificações contínuas no design do implante e na topografia da superfície, no entanto ainda há uma busca contínua para o controle da perda óssea peri-implantar. São escassos os trabalhos que apresentem dados da atividade osteoblástica nas superfícies tratadas concomitante a resultados da proliferação de bactérias nas mesmas, que podem desencadear perda óssea ou peri-implantite, processo patológico que deve ser considerado em longo prazo. O objetivo do trabalho foi desenvolver ligas de Ti-35Nb-7Zr-5Ta (TNZT), realizar tratamentos de superfície com íons cálcio e fosfato e íons magnésio por deposição de plasma eletrolítico (PEO) e avaliar os distintos tratamentos quanto a morfologia, composição química, rugosidade, molhabilidade e adesão do filme. Avaliou-se em seguida a adesão e proliferação de células MC3T3 com teste de alamar blue, FEG/SEM, MTT e fosfatase alcalina. As espécies microbianas foram identificadas e quantificadas pelo método de hibridização DNA-DNA Checkerboard. Foram avaliados 3 grupos diferentes: grupo polido (sem anodização), grupo CaP (β-glicerofosfato de sódio 0,04mol/L+ acetato de cálcio 0,35 mol/L, 300V, 2,5A, 60 seg) e grupo Mg (0,1 mol/L cloreto de Mg, 200V, 60seg. O grupo CaP e Mg alteraram a morfologia, rugosidade, molhabilidade e cristalinidade da liga TNZT. A adesão dos filmes do grupo CaP e Mg resistiram a uma maior força de tração. Menor quantidade total de bactérias foi identificada no grupo Mg e diferenças relevantes foram encontradas no perfil microbiano associado aos diferentes tratamentos de superfície. As células MC3T3 conseguiram aderir e proliferar mais rapidamente nos grupos tratados. No teste alamar blue o grupo CaP apresentou maior atividade metabólica no dia 21. No teste de fosfatase alcalina, o grupo CaP induziu maior expressão de ALP no dia 10. As análises preliminares in vitro sugeriram possíveis vantagens dos tratamento de superfície, sendo que o grupo CaP foi mais benéfico para adesão e proliferação celular, enquanto o grupo Mg reduziu a adesão de bactérias.
Dental implant rehabilitations are routine procedures in current dental practice. High implant survival rates have been achieved in recent decades due to continual modifications in implant design and surface topography, however there is still an ongoing quest to control peri-implant bone loss. There are few studies that present data on osteoblastic activity on treated surfaces, concomitant with the results of bacterial proliferation on them, which can trigger bone loss or peri-implantitis, a pathological process that should be considered in the long term. The objective of this work was to develop Ti-35Nb-7Zr-5Ta (TNZT) alloys, perform surface treatments with calcium and phosphate ions and magnesium ions by electrolytic oxidized plasma (PEO) and evaluate the different treatments in terms of morphology, chemical composition, roughness, wettability and adhesion of the film. The adhesion and proliferation of MC3T3 cells were then evaluated with the alamar blue, FEG/SEM, MTT and alkaline phosphatase tests. The microbial species were identified and quantified by the DNA-DNA Checkerboard hybridization method. Three different groups were evaluated: polished group (without anodizing), CaP group (sodium βglycerophosphate 0.04mol/L+ calcium acetate 0.35mol/L, 300V, 2.5A, 60 sec) and Mg group (0 ,1 mol/L Mg chloride, 200V, 60sec. The CaP and Mg group changed the morphology, roughness, wettability and crystallinity of the TNZT alloy. The adhesion of the films of the CaP and Mg group resisted a higher tensile strength. Less amount total bacteria was identified in the Mg group and relevant differences were found in the microbial profile associated with the different surface treatments. MC3T3 cells were able to adhere and proliferate more quickly in the treated groups. In the alamar blue test, the CaP group showed greater metabolic activity on day 21 In the alkaline phosphatase test, the CaP group induced greater expression of ALP on day 10. Preliminary in vitro analyzes suggested possible advantages of surface treatments, with the CaP group being more beneficial for cell adhesion and proliferation, while the Mg group reduce the adhesion of bacteria on its surface
Dental implant rehabilitations are routine procedures in current dental practice. High implant survival rates have been achieved in recent decades due to continual modifications in implant design and surface topography, however there is still an ongoing quest to control peri-implant bone loss. There are few studies that present data on osteoblastic activity on treated surfaces, concomitant with the results of bacterial proliferation on them, which can trigger bone loss or peri-implantitis, a pathological process that should be considered in the long term. The objective of this work was to develop Ti-35Nb-7Zr-5Ta (TNZT) alloys, perform surface treatments with calcium and phosphate ions and magnesium ions by electrolytic oxidized plasma (PEO) and evaluate the different treatments in terms of morphology, chemical composition, roughness, wettability and adhesion of the film. The adhesion and proliferation of MC3T3 cells were then evaluated with the alamar blue, FEG/SEM, MTT and alkaline phosphatase tests. The microbial species were identified and quantified by the DNA-DNA Checkerboard hybridization method. Three different groups were evaluated: polished group (without anodizing), CaP group (sodium βglycerophosphate 0.04mol/L+ calcium acetate 0.35mol/L, 300V, 2.5A, 60 sec) and Mg group (0 ,1 mol/L Mg chloride, 200V, 60sec. The CaP and Mg group changed the morphology, roughness, wettability and crystallinity of the TNZT alloy. The adhesion of the films of the CaP and Mg group resisted a higher tensile strength. Less amount total bacteria was identified in the Mg group and relevant differences were found in the microbial profile associated with the different surface treatments. MC3T3 cells were able to adhere and proliferate more quickly in the treated groups. In the alamar blue test, the CaP group showed greater metabolic activity on day 21 In the alkaline phosphatase test, the CaP group induced greater expression of ALP on day 10. Preliminary in vitro analyzes suggested possible advantages of surface treatments, with the CaP group being more beneficial for cell adhesion and proliferation, while the Mg group reduce the adhesion of bacteria on its surface
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Biofilmes, Titânio, Ligas, Biologia celular, Osseointegração, Biofilms, Alloys, Titanium, Cell biology, Osseointegration