ESTUDO DE BIOCOMPATIBILIDADE

ESTUDO COMPARATIVO DA AVALIAÇÃO DA BIOCOMPATIBILIDADE DOS ABUTMENTS

FABRICADOS EM TITÂNIO ASTM F136 BASEADO NA NORMA ISO 10993-1

Resumo
Neste trabalho é apresentado o estudo comparativo da biocompatibilidade dos abutments perante
aos requisitos da normativa ISO 10993-1, com base na tabela A1 podemos analisar o grau de
contato do dispositivo medico para saúde com partes do corpo humano. A metodologia
empregada nesta pesquisa, foi desenvolvida, foi de caráter exploratório, com base em estudos e
artigos científicos publicados. Foi comparado a compatibilidade do produto com base no anexo da
norma ISO 10993-1. Os resultados obtidos pelo método empregado podemos concluir que os
abutments encontra-se aprovado demonstrando ser um produto biocompatível devido matéria
prima o titânio ASTM F136 ser um material bioinerte ou seja biocompatível.

Introdução
A avaliação de biocompatibilidade de produtos para saúde é realizada com o objetivo principal de
determinar a toxicidade potencial resultante do contato entre o produto e o corpo humano. Os
materiais utilizados na fabricação de produtos para a saúde não devem, direta ou indiretamente,
liberar constituintes, produzir reações adversas locais, efeitos sistêmicos ou ser carcinogênicos.
No entanto, a avaliação de qualquer produto indicado para uso em seres humanos requer dados
extraídos de testes usando metodologias que possam garantir que os benefícios obtidos com o
uso do produto superem quaisquer riscos potenciais causados pelo mesmo. As interações
biológicas dos Abutments foram avaliadas pela condução de ensaios de biocompatibilidade
conforme norma ISO 10993-1.

Avaliação

Um dos primeiros passos durante o desenvolvimento de um produto é a seleção de materiais

biologicamente compatíveis com a aplicação. Esta etapa é essencial pois os testes necessários
para avaliar um produto dependem da natureza física e química dos materiais utilizados em sua
fabricação e da natureza e tempo de exposição ao corpo humano. A Organização Internacional
de Normalização (ISO), a fim de harmonizar os testes de avaliação de biocompatibilidade,
desenvolveu um padrão para a avaliação biológica de dispositivos médicos (ISO 10993). O
mesmo documento foi adaptado pela ABNT para atender às regulamentações nacionais. A
primeira parte da norma, intitulada “ABNT NBR ISO 10993-1 - Avaliação biológica de produtos de
saúde - Parte 1: Avaliação e ensaio dentro de um processo de gerenciamento de risco”, fornece
 ESTUDO DE BIOCOMPATIBILIDADE
um guia para a seleção dos testes de avaliação de resposta biológica de um dispositivo médico. A
Tabela A.1 do Anexo A desta norma sugere uma estrutura para selecionar os testes de
biocompatibilidade a serem realizados dependendo da natureza do contato (produtos de
superfície, produtos de comunicação externos e produto de implante) e o tempo de exposição
(limitado - menor ou igual a 24h; prolongado - maior que 24h e menor que 30 dias; permanente -
maior que 30 dias).

Categorização conforme norma ISO 10993-1

Os Abutments fabricados pela xxx são classificados como “componente protético”, de

contato permanente com tecido gengival (mais de 30 dias), portanto, considerando esta
classificação, recomenda-se realizar os seguintes testes: Citotoxicidade, Sensitização e Irritação.

Natureza do contato com o corpo: Dispositivos de implante – tecido gengival Duração do contato:
contato permanente

Avaliação bibliográfica
Foram avaliados estudos científicos publicados referente ao emprego do titânio na fabricação de
dispositivo medico para saúde, demonstrando ser um material biocompatível atendendo altos
níveis de suas propriedades físico-químicas.

Titanium Alloys for Dental Implants: A Review John W. Nicholson Dental Materials Unit, Bart’s and
the London Institute of Dentistry, Queen Mary University of London, Mile End Road, London E1
4NS, UK and Bluefield Centre for Biomaterials, 67-68 Hatton Garden, London EC1N 8JY, UK;
john.nicholson@bluefieldcentre.co.uk or j.nicholson@qmul.ac.uk Received: 14 May 2020;
Accepted: 10 June 2020; Published: 15 June 2020

Abstract: The topic of titanium alloys for dental implants has been reviewed. The basis of the
review was a search using PubMed, with the large number of references identified being reduced
to a manageable number by concentrating on more recent articles and reports of biocompatibility
and of implant durability. Implants made mainly from titanium have been used for the fabrication of
dental implants since around 1981. The main alloys are so-called commercially pure titanium
(cpTi) and Ti-6Al-4V, both of which give clinical success rates of up to 99% at 10 years. Both
alloys are biocompatible in contact with bone and the gingival tissues, and are capable of
undergoing osseointegration. Investigations of novel titanium alloys developed for orthopaedics
show that they offer few advantages as dental implants. The main findings of this review are that
the alloys cpTi and Ti-6Al-4V are highly satisfactory materials, and that there is little scope for
improvement as far as dentistry is concerned. The conclusion is that these materials will continue
to be used for dental implants well into the foreseeable future.
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Mechanical, chemical and biological aspects of titanium and titanium alloys in implant dentistry

Implant dentistry has become a popular restorative option in clinical practice. Titanium and
titanium alloys are the gold standard for endo-osseus dental implants production, thanks to their
biocompatibility, resistance to corrosion and mechanical properties. The characteristics of the
titanium implant surface seem to be particularly relevant in the early phase of osseointegration.
Furthermore, the microstructure of implant surface can largely influence the bone remodelling at
the level of the bone-implant surface. Recently, research has stated on the long-term of both
survival and success rates of osseointegrated implants and mainly on biomechanical aspects,
such as load distribution and biochemical and histological processes at the bone-implant interface.
This short review reports recent knowledge on chemical and mechanical properties, biological
aspects, innovations in preventing peri-implantitis, describing clinical applications and recent
improvements of titanium dental implants. In addition, it highlights current knowledge about a new
implant coating that has been demonstrated to reduce the number of initially adhering bacteria
and peri-implantitis.

Biocompatibility of Advanced Manufactured Titanium Implants—A Review

Titanium (Ti) and its alloys may be processed via advanced powder manufacturing routes such as
additive layer manufacturing (or 3D printing) or metal injection moulding. This field is receiving
increased attention from various manufacturing sectors including the medical devices sector. It is
possible that advanced manufacturing techniques could replace the machining or casting of metal
alloys in the manufacture of devices because of associated advantages that include design
flexibility, reduced processing costs, reduced waste, and the opportunity to more easily
manufacture complex or custom-shaped implants. The emerging advanced manufacturing
approaches of metal injection moulding and additive layer manufacturing are receiving particular
attention from the implant fabrication industry because they could overcome some of the
difficulties associated with traditional implant fabrication techniques such as titanium casting.
Using advanced manufacturing, it is also possible to produce more complex porous structures
with improved mechanical performance, potentially matching the modulus of elasticity of local
bone. While the economic and engineering potential of advanced manufacturing for the
manufacture of musculo-skeletal implants is therefore clear, the impact on the biocompatibility of
the materials has been less investigated. In this review, the capabilities of advanced powder
manufacturing routes in producing components that are suitable for biomedical implant
applications are assessed with emphasis placed on surface finishes and porous structures. Given
that biocompatibility and host bone response are critical determinants of clinical performance,
published studies of in vitro and in vivo research have been considered carefully. The review
concludes with a future outlook on advanced Ti production for biomedical implants using powder
metallurgy.

In vitro evaluation of cytotoxicity and corrosion behavior of commercially pure titanium and Ti-
6Al- 4V alloy for dental implants
Taking into consideration the limitations related to the present study, it can be concluded that both
Ti 12 and Ti 31 are suitable for dental and medical applications. The commercially pure Ti showed
better cell viability compared to Ti 31

Study of Biocompatibility of Titanium Alloys for Biomedical Application. Objective: to evaluate the
biocompatibility of porous samples of five kinds of titanium alloys on osteoblast activity. The
modification in composition and porosity confection in samples aimed at reducing the elasticity,
rendering it closer to the bone elasticity modulus.

Method: Five groups of porous samples were manufactured by powder metallurgy: 1) control -
commercially pure titanium (TiCP); 2) Ti-6Al-4V alloy (titanium-aluminium-vanadium); 3) Ti-13Nb-
 ESTUDO DE BIOCOMPATIBILIDADE
13Zr alloy (titanium-niobium-zirconium); 4) Ti-35Nb alloy (niobium); 5) Ti-35Nb-7Zr-5Ta alloy
titanium-niobium-zirconium-tantalum). Samples were characterized by scanning electron
microscopy (SEM) and energy dispersion spectrophotometry (EDS). They were evaluated for
cytotoxicity on rat calvaria osteogenic cells after 3 and 10 days, and nodules formation and
quantification of mineralization after 14 days. Cells were cultivated on 18 samples of each group.
Data were analyzed by ANOVA one way, ANOVA two ways and Tukey tests (p<0.05).

Result: EDS detected presence of specific ions for each alloy and MEV showed an interconnected
porous structure, demonstrating the effectiveness of the powder metallurgy technique. In vitro
tests revealed similar biocompatibility among different alloys. Alizarin red analysis showed
mineralized matrix formation similar in TiCP, Ti-35Nb-7Zr-5Ta, Ti-35Nb, and Ti-13Nb-13Zr. Ti-6Al-
4V showed the significant highest value in this analysis. MTT assay demonstrated significant
interaction between time and alloy by ANOVA two ways. Tukey test revealed that Ti-6Al-4V, at 10
days, showed lower cell viability when compared to the other groups at 3 days and similar viability
when compared to other groups at 10 days. The results indicate that the alloys did not show
cytotoxicity under static culture conditions, comparing to the standard alloy Ti-6Al-4V. This
cytocompatibility was correlated with nodules of mineralization in all groups.

Conclusion: porous TiCP, Ti-35Nb-7Zr-5Ta, Ti-35Nb, Ti-13Nb-13 Zr and Ti-6Al-4V alloys

specimens induce favorable cellular response with mineralized matrix formation. These alloys are
biocompatible and have great potential for employment in biomedical implants.

Immunotoxicological aspects of biocompatibility of titanium

Kodama, Tsuyoshi. (1989). Study on biocompatibility of titanium alloys. Kōkūbyō Gakkai zasshi.
The Journal of the Stomatological Society, Japan. 56. 263-88. 10.5357/koubyou.56.263. The
biocompatibility of two different titanium alloys, Ti-6Al-4V ELI and Ti-5Al-2, 5Fe, and pure titanium
were evaluated. The results were as follows: 1) Titanium alloys were implanted into the dorsal
subcutaneous tissues of the Hartley guinea-pig for 12 weeks, immersed in calf serum or in
Ringer's solution for 8 weeks. The surface changes of the titanium alloys were observed by SEM
and the chemical composition was analyzed by XMA. No evident surface changes were found. 2)
Three hundred mg, 200 mg and 100 mg of the powders of the tested materials were immersed in
2ml of Eagle's MEM, incubated for 1-7 days, 8-21 days and 22-70 days at 37 C degrees. The
amount of metallic elements dissolved in the solutions was measured by ICP and AAS. The
detected corrosion rates of V and Al contained in the solution, in which Ti-6Al-4V ELI 100 mg was
immersed for 1-7 days, were 194.3 +/- 17.6 and 73.0 +/- 28, 1 pg/mg alloy/day, respectively. V
was released more than Al. The amount of Ti was below the detectable limit. The solution Ti-5Al-
2.5 Fe 100 mg immersed for 1-7 days contained 31.9 +/- 34.4 pg/mg alloy/day Fe and 25.7 +/- 6.3
pg/mg alloy/day Al. Only in the solution 300 mg immersed for 1-7 days was Ti detected at 1.4
pg/mg alloy/day. 3) By the bacterial mutation assay of Salmonella typhimurium TA 98, Salmonella
typhimurium TA 100 and Escherichia coli WP2 uvrA, the solutions, in which the tested materials
were immersed, were not found to be mutagenic. 4) By the UDS assay, the grain counts on
autoradiography with the solutions, in which the tested materials were immersed, were not greater
than the negative control. The results suggest an excellent corrosion resistance of the titanium
alloys. Mutagenicity was negative by these mutation assays, indicating that the tested alloys and
pure titanium are safe for humans and animals.

Biocompatibility of titanium and its alloys used in dentistry

Makuch, Krzysztof & Koczorowski, Ryszard. (2010). Biocompatibility of titanium and its alloys
used in dentistry. Dental and Medical Problems. 47. 81-88. The widely used biomaterials,
including titanium and its alloys, manifest a range of physicochemical properties which
determine the way
 ESTUDO DE BIOCOMPATIBILIDADE
they are exploited. Titanium, known for its great biocompatibility, resistance to corrosion and low
allergic action, occurs in several forms (four classes of pure titanium and compounds with other
metals) of varied characteristics. The knowledge of the properties of pure titanium and its alloys,
new data regarding corrosion as well as more and more frequent reports on allergy cases
resulting from the common use of titanium implants allow clinicists to be aware of the different
reactions of patients to this alloplastic material generally considered ideal. The forms of pure
titanium are characterized by greater biocompatibility and less resistance and elasticity, while in
the case of alloys the situation is reverse. The improvement of mechanical properties of pure
titanium seems to be a dominating trend defining commercial materials used in implantology. A
correlation of implantological treatment failure with the quantity and quality of the compounds
released as a result of corrosion has been demonstrated, on the basis of the literature review, in
addition to cases of oversensitivity to type four titanium whose reaction mechanism is similar to
nickel allergy. The article emphasizes the relevance of immunological tests, such as the MELISA
test, to dental prosthetics as an effective way of determining sensitivity to titanium, proving that the
skin tests commonly used in cases of allergy to other metals are not a reliable detection method.

Mechanical properties and biocompatibility of titanium with a high oxygen concentration for
dental implants

In order to improve the strength of commercially pure Ti (CP-Ti) for oral implants, the high oxygen
content Ti (HOC-Ti) was prepared via powder metallurgy. Its composition and mechanical
properties were then characterized. After surface treatment by sandblasting and acid etching
(SLA), the surface morphology, wettability and roughness of the HOC-Ti and CP-Ti sample were
examined. In an in vitro test that followed an evaluation of the protein adsorption capacity of HOC-
Ti, the mouse preosteoblast cells were inoculated onto the specimens to evaluate their
biocompatibility, in comparison with those of CP-Ti. The oxygen concentration of the HOC-Ti
increased to 0.62 wt%, which is higher than the 0.26 wt% of the CP-Ti, while their compositions
and microstructures were very similar. The tensile and compressive yield strength of the HOC-Ti
(800 MPa) was improved significantly in comparison to that of the CP-Ti (530 MPa). After surface
treatment, a unique structure of micropores with a diameter of 380 nm was observed on the entire
surface of the HOC-Ti that facilitates cell adhesion and proliferation. The wettability of the HOC-Ti
was obviously superior (p < 0.05). The in vitro study showed that the MC3T3-E1 cells inoculated
on the surface of HOC-Ti exhibited a homogeneous microstructure, and the viability was higher
than that of the control group on days 4 and 7 (p < 0.05). In addition, the number and
differentiation activity of cells that adhered to the surface of the HOC-Ti increased significantly on
day 7 (p < 0.05). The experimental results showed that, in view of its mechanical properties and
biocompatibility, HOC-Ti is superior to CP-Ti and is promising for oral implant applications.

In vitro evaluation of biocompatibility of experimental titanium alloys for dental restorations

The results of Ames test showed that none of the experimental titanium alloys, pure titanium, or
nickel-chromium samples were mutagenic. There were no leachable components to cause cellular
lysis or decolorization in the agar diffusion assay from the 4 group metals. The results of the cell
attachment assay demonstrated that there was no significant difference in the numbers of cells
attached to the 4 group metals. However, the mean number of cells attached to nickel-chromium
samples was significantly lower than that of the control group.

Application of titanium and titanium alloys to fixed dental prostheses

Titanium and its alloys can be the substitute materials for Ag–Pd–Au alloy, and should be
considered worthy of qualifying for the dental insurance coverage.
 ESTUDO DE BIOCOMPATIBILIDADE

Mechanical properties, surface morphology and stability of a modified commercially pure high
strength titanium alloy for dental implants
Objective. Commercially pure titanium (cp Ti) and Ti–6Al–4V (Ti G5) alloy have limitations for
biomedical application, due to lower mechanical strength and the possibility of ion release,
respectively. The purpose of this work was to compare the properties of a modified cp Ti grade 4
(Ti G4 Hard) with those of available cp Ti and Ti G5 alloys. Methods. Bars, discs and dental
implants made with Ti G2, G4, G5 and G4 Hard were used. Mechanical tests (tension,
compression, hardness and torque) and roughness measurements were performed. Clinical trials
were used to evaluate the biological behavior of dental implants made with Ti G4 Hard and Ti G4.
Results. The results of the mechanical tests showed that the mechanical strength of modified Ti
G4 is higher than that of Ti G2, G4 and G5. Scanning electron microscopy analysis showed that
modified Ti G4 after etching has better surface morphological features than conventional cp Ti and
Ti G5. The clinical performances of Ti G4 and Ti G4 Hard were similar. Significance. The
improvement of the mechanical properties of modified Ti G4 means that Ti G5 can be safely
replaced by Ti G4 Hard without compromising the fracture resistance, with the advantage of not
releasing toxic ions.

Conclusão
As ligas de titânio são materiais metálicos mais atraentes para aplicações biomédicas. Na
medicina, eles são usados para dispositivos de implante que substituem tecidos rígidos
danificados. Os exemplos incluem articulações artificiais do quadril, articulações artificiais do
joelho, placas ósseas, parafusos para fixação de fraturas, próteses valvares cardíacas, marca-
passos e corações artificiais dentre outros dispositivos medicos. O Ti-6Al-4V é há muito tempo
uma das principais ligas de titânio para uso médico. No entanto, para aplicações de implantes
permanentes, a liga tem um possível efeito tóxico resultante da liberação de vanádio e alumínio.
Por esse motivo, ligas livres de vanádio e alumínio foram introduzidas para aplicações de
implantes, com base nos implantes Ti-6Al-4V.
Um grande número de experimentos de titânio in vivo e in vitro foram realizados em universidades
e indústrias em todo o mundo nos últimos 50 anos. Esses experimentos descobriram que a
excelente biocompatibilidade do titânio está associada aos seus óxidos. C.B. Johansson
demonstrou estudos de modelo animal in vivo que o óxido de titânio pode diferir de biomateriais
metálicos, como Ti-6Al-4V, ligas de CoCr e aço inoxidável 316 LVM. A interface entre o implante
de titânio e o osso é uma fina camada de proteoglicanos.
O titânio comercialmente puro (Ti Cp) é considerado o melhor material metálico biocompatível
porque suas propriedades de superfície resultam na formação espontânea de uma camada
oxidada estável e inerte. As principais propriedades físicas do titânio responsáveis pela
biocompatibilidade são: baixo nível de condutividade eletrônica, alta resistência à corrosão,
estado termodinâmico em valores de pH fisiológicos, baixa tendência à formação de íons em
ambientes aquosos e um ponto isoelétrico do óxido de 5-6. Além disso, a superfície coberta por
 ESTUDO DE BIOCOMPATIBILIDADE
filme passivo é apenas ligeiramente carregada negativamente em pH fisiológico, e o titânio tem
uma constante dielétrica comparável à da água, com a consequência de que a interação de
Coulomb de espécies carregadas é semelhante à da água.

Os materiais usados para aplicações biomédicas cobrem um amplo espectro e devem exibir
propriedades específicas. A propriedade mais importante dos materiais utilizados na fabricação
de implantes é a biocompatibilidade, seguida da resistência à corrosão. Os principais biomateriais
metálicos são os aços inoxidáveis, ligas de cobalto e titânio e ligas de titânio. O aço inoxidável foi
o primeiro biomaterial metálico usado com sucesso como implante. Em 1932, a liga à base de
cobalto chamada Vitallium foi desenvolvida para aplicações médicas. O titânio é o mais novo
biomaterial metálico. Nos campos médico e odontológico, o titânio e suas ligas têm demonstrado
sucesso como dispositivos biomédicos.
De acordo com o anexo da ASTM F136 - Standard Specification for Wrought Titanium-
6Aluminum-4Vanadium ELI (Extra Low Interstitial) Alloy for Surgical Implant Applications (UNS
R56401) A composição da liga de titânio coberta por esta especificação tem sido empregado com
sucesso em aplicações de implantes humanos em contato com tecidos moles e ossos por mais
de uma década. Devido o nível bem caracterizado de resposta biológica exibida por esta liga, tem
sido usada como um material de controle na prática.
Nenhum material de implante cirúrgico conhecido jamais foi mostrou ser completamente livre de
reações adversas no corpo humano. Experiência clínica de longo prazo do uso do material
referido nesta especificação, no entanto, mostrou que um nível aceitável de resposta biológica
pode ser esperado, se o material for usado em aplicações apropriadas.
Com base nesta avaliação de biocompatibilidade de caráter exploratório experimental,
concluímos que os Abutments fabricados em titânio conforme ASTM F136 encontra-se aprovados
perante a ISO 10993-1.
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Referências
Koizumi, H. et al. Application of titanium and titanium alloys to fixed dental prostheses. Journal of
prosthodontic research 63 (2019) 266–270.

Chandar, S. et al. In vitro evaluation of cytotoxicity and corrosion behavior of commercially pure
titanium and Ti-6Al-4V alloy for dental implants. J Indian Prosthodont Soc 2017;17:35-40.

Carvalho, Y. et al. Study of Biocompatibility of Titanium Alloys for Biomedical Application. Annual
Meeting of the IADR Continental European Division 2013.

Wang, R and Li, Y. In vitro evaluation of biocompatibility of experimental titanium alloys for dental
restorations. The Journal of Prosthetic Dentistry. Volume 80, Issue 4, October 1998, Pages 495-
500.

Luo, H. et al. Mechanical properties and biocompatibility of titanium with a high oxygen
concentration for dental implants. Materials Science and Engineering: C. Volume 117, December
2020, 111306.

Makuch, K. and Koczorowski, R. Biocompatibility of titanium and its alloys used in dentistry.Dental
and Medical Problems 47(1):81-88.

Lyapina, M. et al. Immunotoxicological aspects of biocompatibility of titanium. Journal of IMAB -

Annual Proceeding (Scientific Papers) 23(2):1550-1559.

Sidambe, A. Biocompatibility of Advanced Manufactured Titanium Implants—A Review. Materials

(Basel). 2014 Dec 19;7(12):8168-8188. doi: 10.3390/ma7128168.

Ottria, L. et al. Mechanical, chemical and biological aspects of titanium and titanium alloys in
implant dentistry. J Biol Regul Homeost Agents. Jan-Feb 2018;32(2 Suppl. 1):81-90.

Nicholson, J. Titanium Alloys for Dental Implants. Prosthesis. 2020; 2(2):100-116.

ISO 10993-1: Biological evaluation of medical devices — Part 1: Evaluation and testing within a
risk management process

ASTM F136 - Standard Specification for Wrought Titanium-6Aluminum-4Vanadium ELI (Extra Low
Interstitial) Alloy for Surgical Implant Applications (UNS R56401)