Caracterização e avaliação do potencial de reparo ósseo de um novo biomaterial substituto ósseo: estudos in vitro e in vivo

André Antonio Pelegrine
Divisão de Implantodontia, Faculdade São Leopoldo Mandic, Campinas, Brasil.
ORCID: 0000-0001-7935-1062

Paulo Sérgio Perri de Carvalho
Divisão de Implantodontia, Faculdade São Leopoldo Mandic, Campinas, Brasil.
ORCID: 0000-0003-1775-3108

Marcelo Henrique Napimoga
Laboratório de Imunologia e Biologia Molecular, Faculdade São Leopoldo Mandic, Campinas, Brazil.
ORCID: 0000-0003-4472-365X

DOI: https://doi.org/10.71440/2675-5610.10.5.25.666-673.art

RESUMO

Objetivo: Avaliar um novo biomaterial substituto ósseo composto pela matriz inorgânica de origem bovina (hidroxiapatita), tanto do ponto de vista estrutural e biológica, como também em cima de fatores que podem apresentar repercussão imunológica. Material e Métodos: Ensaios in vitro para avaliação da porosidade do material (microscopia eletronica de varredura – MEV), analises de proteína residual (BCA e gel SDS-PAGE) e de citotoxicidade (utilizando fibroblastos da linhagem celular L-929) foram executadas em um primeiro momento. Na sequência, foram realizados trabalhos in vivo por meio de analise pirogênica, de sensibilização dérmica, toxicidade sistêmica e histologia apos implantação, para avaliação do potencial reconstrutivo do biomaterial. Resultados: As analises em MEV mostraram uma superficie micro rugosa com muitas topografias macro e micro rugosas de forma semelhante a osso nativo. As porosidades para macroporos foram de 0,172 ± 0,049 mm e para microporos de 0,016 ± 0,010mm. Os cálculos de proteína residual não resultaram em qualquer remanescente que pudesse gerar uma resposta imunogênica, assim como os resultados de citotoxicidade. Os ensaios in vivo mostraram ausência de potenciais pirogênicos, ausência de sensibilização dérmica e de toxicidade sistêmica. Histologicamente, foi possível avaliar a performance de incorporação dos grânulos do biomaterial com osso neoformado. Conclusão: A hidroxiapatita de origem bovina avaliada neste estudo apresenta adequada porosidade, e altamente purificada, possibilita o reparo ósseo e não induz inflamação/imunogenicidade.

Palavras-chave: regeneração óssea; transplante ósseo; xenoenxerto.

Characterization and evaluation of the bone repair potential of a new bone substitute biomaterial: in vitro and in vivo studies

ABSTRACT

Objective: To evaluate a new bone substitute biomaterial composed of anorganic bovine matrix (hydroxyapatite), both from a structural and biological point of view, as well as factors that may have immunological repercussions. Material and Methods: In vitro tests to evaluate the material’s porosity (scanning electron microscopy – SEM), residual protein analysis (BCA and SDS-PAGE gel) and cytotoxicity (using fibroblasts of the L-929 cell line) were performed initially. Subsequently, in vivo studies were performed using pyrogenic analysis, dermal sensitization, systemic toxicity and histology after implantation, to evaluate the reconstructive potential of the biomaterial. Results: SEM analyses showed a microrough surface with many macro and microrough topographies similar to native bone. The porosities for macropores were 0.172 ± 0.049 mm and for micropores, 0.016 ± 0.010 mm. Residual protein calculations did not result in any remnant that could generate an immunogenic response, as did the cytotoxicity results. In vivo assays showed no pyrogenic potential, no dermal sensitization and no systemic toxicity. Histologically, it was possible to evaluate the incorporation performance of the biomaterial granules with newly formed bone. Conclusion: The bovine hydroxyapatite evaluated in this study presents adequate porosity, is highly purified, enables bone regeneration and does not induce inflammation/immunogenicity.

Keywords: bone regeneration; bone transplantation; xenograft.

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