Pathological Features of Bovine Pericardium Implanted Into Mice Abdominal Wall after Preservation with Glutaraldehyde or Glycerin

Cláudia Borges Costa, Marta Fernanda Albuquerque da Silva, Gisele Braziliano de Andrade

Abstract


Background: Lately, the use of biological materials has been widely indicated in surgical procedures to restore structure and function of injured tissues. Bioimplants require different conservation techniques; among these, glutaraldehyde preservation stands out owing to its higher antimicrobial efficiency as compared to glycerin. In view of the need to determine a concentration of glutaraldehyde that can act as a biocide but do not cause undesirable tissue reactions, this study aimed to identify and quantify gross and microscopic tissue alterations after implantation of bovine pericardium, which was preserved in various concentrations of glutaraldehyde, in the abdominal wall of mice.

Materials, Methods & Results: Fresh pericardia from 18 bovines were fractioned into 1cm2 samples and treated with a 98% glycerin solution for 30 days (control group), or 0.625%, 1%, and 1.5% glutaraldehyde solution for 18 days (experimental groups). An abdominal muscle fragment was excised from each mouse, and a 1-cm2 fragment of preserved pericardium was implanted in the area. Sixty mice (n = 15 per treatment) divided into groups were observed for 7, 14, and 30 days, and five animals from each group were euthanized at each time point for gross and microscopic examination. Fragments of the implants and adjacent skin lesions were harvested, fixed in formalin, and processed for routine histology and microscopic analysis. Both the type of inflammatory infiltrate and the repair process of the tissue response were similar between the groups that received glycerin-preserved pericardium and those that were subjected to pericardium preserved with 0.625% glutaraldehyde. Animals that received 1% glutaraldehyde-preserved implants and were examined 30 days thereafter exhibited a chronic, intense reaction with fibrosis and necrosis of the abdominal wall muscles, as well as calcification and presence of giant cells, when compared to the animals examined at 7 and 14 days in the same treatment group. These changes were also present and more intense in animals that received 1.5% glutaraldehyde-preserved pericardium examined at 14 and 30 days later, with tissue destruction and impaired incorporation of the implant into the adjacent muscle tissue.

Discussion: The continuous cell destruction observed in animals treated with implants preserved with 1% or 1.5% glutaraldehyde is a hallmark of chronic inflammation, since several inflammatory cell molecules contribute to this lesion. A cycle is created: continuous degradation sustains inflammation, and inflammatory molecules contribute to the process of cell destruction. Consequently, we conclude that the use of glutaraldehyde at concentrations of 1 or 1.5% is not feasible for preservation of biological materials. Tissue repair was chronologically more effective in the group treated with glycerinpreserved implants, since animals treated with glutaraldehyde-preserved implants needed a longer period to restore due to presence of a persistent inflammatory response, immunogenicity, calcification, and deficient remodeling.  The ideal preservative for biological materials should not cause chronic and/or intense inflammatory reaction in order to preserve the implant’s structure and allow its perfect incorporation into the tissue, even if the chosen preservative is flexible and exhibits disinfectant properties. Therefore, we conclude that glutaraldehyde at concentrations of 0.625% to 1% is suitable as a preservative for biomaterials because the tissue reaction it causes is tolerable; additionally, glutaraldehyde at concentrations close to 1% has been described to have sterilizing properties.


Keywords


bioimplant; inflammatory reaction; chemical preservation; histopathology.

Full Text:

PDF

References


Baucia J.A., Leal Neto R.M., Rogero J.R. & Nascimento N. 2006. Tratamentos anticalcificantes do pericárdio bovino fixado com glutaraldeído: comparação e avaliação de possíveis efeitos sinérgicos. Brazilian Journal of Cardiovascular Surgery. 21(2): 180-187.

Braga F.A. & Pippi N.L. 2009. Uso de pericárdio heterólogo conservado em glicerina na reparação de hérnia abdominal traumática em cão. Redevet. Disponível em: < http://www.redevet.com.br/artigos/pericardio.htm# >. [Acessed October 2015].

Brun M.V., Pippi N.L., Driemeier D., Contesini E.A., Beck C.A.C., Cunha O., Filho S.T.L.P., Roehsig C., Stedile R. & Silva T.F. 2004. Solução Hipersaturada de sal ou de glicerina a 98% como conservantes de centro frênicos caninos utilizados na reparação de defeitos musculares em ratos wistar. Ciencia Rural. 34(1): 147-153.

Christian A.J., Alferiev I.S., Connolly J.M., Ischiropoulos H. & Levy R.J. 2015. The effects of the covalent attachment of 3-(4-hydroxy-3, 5-di-tert-butylphenyl) propyl amine to glutaraldehy depretreated bovine pericardium on structural degeneration, oxidative modification, and calcification of rat subdermal implants. Journal of Biomedical Materials Research. Part A. 103(7): 2441-2448.

Costa J.N.L., Pomerantzeff P.M.A., Braile D.M., Ramirez V.A., Goissis G. & Stolf N.A.G. 2005. Comparação entre pericárdio bovino decelularizado e pericárdio bovino convencional utilizado na confecção de biopróteses valvares cardíacas. Revista Brasileira de Cirurgia Cardiovascular. 20(1): 14-22.

Gamba P.G., Conconi M.T., Lo Piccolo R., Zara G., Spinazzi R. & Parnigotto P.P. 2002. Experimental abdominal wall defect repaired with acellular matrix. Pediatric Surgery International. 18(5-6): 327-331.

Greca F.H., Noronha L., Costa F.D.A., Souza Filho Z.A.S., Soccol A.T., Feres A.N., Duda J.R. & Adams E. 2005. Estudo Comparativo da biocompatibilidade da mucosa intestinal porcina e pericárdio bovino usados como enxertos na veia cava de cães. Acta Cirúrgica Brasileira. 20(4): 317-322.

Goissis G., Braile D.M. & Giglioti A.F. 1999. Desenvolvimento de processo automático para a reticulação progressiva de matrizes de colágeno com glutaraldeído. Polímeros. 9(4): 92-97.

Haddad Filho D., Marques A., Kafejian-Haddad A.P. & Zveibel D.K. 2004. Estudo Comparativo das reações teciduais ao implante de pericárdio bovino e a inclusão de politetrafluoroetileno expandido em ratos. Acta Cirúrgica Brasileira. 19(2): 131-135.

Hilbert S.L., Ferrans V.J. & Jones M. 1988-1989. Tissue-derived biomaterials and their use in cardiovascular prosthetic devices. Medical Progress Through Technology. 14(3-4): 115-163.

Jayakrishnan A. & Jameela S.R. 1996. Glutaraldehyde as a fixative in bioprostheses and drug delivery matrices. Biomaterials. 17(5): 471-484.

Lim H.G., Kim G.B., Jeong S. & Kim Y.J. 2014. Valved conduit with glutaraldehyde-fixed bovine pericardium treated by anticalcification protocol. The Korean Journal of Thoracic and Cardiovascular Surgery. 47(4): 333-343.

Lim H.G., Choi S.Y., Jeong S., Shin J.S., Park C.G. & Kim Y.J. 2015. In vivo efficacy for novel combined anticalcification treatment of glutaraldehyde-fixed cardiac xenograft using humanized mice. Journal of Biomaterials Application. 29(7): 929-940.

Maizato M.J., Pires M.D., Canzian M., Higa O.Z., Pitombo R.N. & Leirner A.A. 2008. Histological evaluation of biocompatibility of lyophilized bovine pericardium implanted subcutaneously in rats. Artificial Organs. 32(4): 268-271.

Paiva F.P., Maffili V.V. & Santos A.C.S. 2005. Curso de Manipulação de Animais de Laboratório. Salvador: Fundação Oswaldo Cruz - Centro de Pesquisas Gonçalo Moniz, 28p.

Pigossi N. 1964. Implantação de dura-máter homógena conservada em glicerina: estudo experimental em cães. Arquivos de Cirurgia e Clínica Experimental, São Paulo. 27: 213-247.

Plunkett S.J. 2006. Procedimentos de Emergência em Pequenos Animais. 2.ed. Rio de Janeiro: Revinter, 521p.

Quitzan J.G., Rahal S.C., Rocha N.S. & Crocci A.J. 2003. Comparação entre pericárdio bovino preservado em glicerina e malha de poliéster no reparo de falhas da parede abdominal em ratos. Acta Cirúrgica Brasileira. 18(4): 297-301.

Santillán-Doherty P., Jasso-Victoria R., Sotres-Vega A., Olmos R., Arreola J.L., Garc-ia D., Vanda B. & Gaxiola M. 1995. Repair of thoracoabdominal wall defects in dogs using a bovine pericardial bioprothesis. Revista de Investigacion Clinica; organo del Hospital de Enfermidades de la Nutrición. 47(6): 439-446.

Schoen F.J., Tsao J.W. & Levy R.J. 1986. Calcification of bovine pericardium used in cardiac valve bioprostheses. Implications for the mechanisms of bioprosthetic tissue mineralization. The American Journal of Pathology. 123(1): 134-145.

Tang L. & Eaton J.W. 1995. Inflammatory Responses to Biomaterials. American Journal of Clinical Pathology. 103(4): 466-471.

Thampi P., Nair D.R.L.N.V., Venugopal S. & Ramachandra U. 2013. Pathological effects of processed bovine pericardial scaffolds--a comparative in vivo evaluation. Artificial Organs. 37(7): 600-605.

Trani R.A.S. 2006. Eficácia das soluções de Glicerina 98% e Glutaraldeído 0,625% na desinfecção de pericárdio de camundongos (Mus musculus) experimentalmente inoculados com vírus da Raiva. 68f. Seropédica, RJ. Dissertação (Mestrado em Medicina Veterinária, Patologia Animal) - Program de Pós-graduação em Medicina Veterinária, Patologia Animal, Universidade Federal Rural do Rio de Janeiro.

Umashankar P.R., Mohanan P.V. & Kumari T.V. 2012. Glutaraldehyde treatment elicits toxic response compared to decellularization in bovine pericardium. Toxicology Internatiol. 19(1): 51-58.

Vincentelli A., Latrémouille C., Zegdi R., Shen M., Lajos P.S., Chachques J.C. & Fabiani J.N. 1998. Does glutaraldehyde induce calcification of bioprosthetic tissues? The Annals of Thoracic Surgery. 66(Suppl 6): S255-258.




DOI: https://doi.org/10.22456/1679-9216.81104

Copyright (c) 2018 Cláudia Borges Costa, Marta Fernanda Albuquerque da Silva, Gisele Braziliano de Andrade

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.