CC BY-NC-ND 4.0 · Rev Bras Ortop (Sao Paulo) 2020; 55(02): 163-169
DOI: 10.1055/s-0039-3400522
Artigo Original
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Sociedade Brasileira de Ortopedia e Traumatologia. Published by Thieme Revinter Publicações Ltda Rio de Janeiro, Brazil

Protocol for Harvest, Transport and Storage of Human Osteochondral Tissue[]

Article in several languages: português | English
1  Divisão de Pesquisa, Instituto Nacional de Traumatologia e Ortopedia Jamil Haddad, Rio de Janeiro, RJ, Brasil
,
Rafael Dantas Prinz
1  Divisão de Pesquisa, Instituto Nacional de Traumatologia e Ortopedia Jamil Haddad, Rio de Janeiro, RJ, Brasil
,
Naasson Cavanellas
1  Divisão de Pesquisa, Instituto Nacional de Traumatologia e Ortopedia Jamil Haddad, Rio de Janeiro, RJ, Brasil
,
João Maurício Barretto
1  Divisão de Pesquisa, Instituto Nacional de Traumatologia e Ortopedia Jamil Haddad, Rio de Janeiro, RJ, Brasil
,
Eduardo Branco de Sousa
1  Divisão de Pesquisa, Instituto Nacional de Traumatologia e Ortopedia Jamil Haddad, Rio de Janeiro, RJ, Brasil
,
Diego Pinheiro Aguiar
1  Divisão de Pesquisa, Instituto Nacional de Traumatologia e Ortopedia Jamil Haddad, Rio de Janeiro, RJ, Brasil
› Author Affiliations
Further Information

Publication History

27 August 2018

22 January 2019

Publication Date:
28 January 2020 (online)

Abstract

Objective To elaborate a protocol for the harvest, transport, and preservation of human osteochondral tissue for use in tissue banks (TBs).

Methods Osteochondral fragments measuring 2 cm3 of 5 corpse donors aged between 15 and 45 years old were analyzed. The samples were stored in cell preservation medium containing: human albumin, Iscove's and vancomycin preserved at 4°C. The concentration of proteoglycans in the extracellular medium was quantified by the use of Safranin-O, while tissue structural analysis was assessed by histological study with hematoxylin-eosin stained slides. The images obtained were analyzed according to the histological scores of Mankin and the score proposed by the OsteoArthritis Research Society International. The samples were analyzed with 0, 15, 30 and 45 days of preservation.

Results The osteochondral fragments studied showed a progressive decrease in proteoglycan concentration with increased preservation time. After 30 days of preservation, structural changes were identified with discontinuity of the cartilage surface layer. According to the results obtained by the Mankin score, there was a statistically significant difference between 15 and 30 days of tissue preservation.

Conclusion The protocol described defined knee transport immersed in Lactated Ringer at a controlled temperature of 10° C until its arrival at the TB. After processing, the preservation solution was composed of Iscove's serum-free cell culture medium supplemented with 10% human albumin and 100 μg/ml vancomycin. The tissue was preserved at a temperature of 4°C until the moment of transplantation characterizing the fresh preservation.

Study conducted at the Instituto Nacional de Traumatologia e Ortopedia Jamil Haddad, Rio de Janeiro, RJ, Brazil.