Comparative Study of Cryopreserved Bone Tissue and Tissue Preserved in a 98% Glycerol Solution

Abstract

OBJECTIVE: To compare the bone graft cryopreservation method (at-80ºC) with a preservation method using a 98% glycerol solution at room temperature (10ºC-35ºC), by testing the antibacterial and fungal effects of 98% glycerol and comparatively analyzing the observed histological changes resulting from the use of both methods. METHOD: This study was of 30 samples of trabecular bone tissue from 10 patients undergoing total hip arthroplasty. Each femoral head provided 3 samples that were randomized into 3 groups, namely, the control group, the cryopreserved group, and the group preserved in a 98% glycerol at room temperature for 1 year. The samples were submitted to histomorphologic, cell feasibility, and microbiologic analyses. The results were statistically analyzed using the McNemar test, with a statistical significance index of 0.05. RESULTS: Values obtained using the McNemar test to compare probability distributions of histomorphologic variables (mature or lamellar bone, immature bone, and necrosis) and cell feasibility (osteoblasts and osteoclasts) indicated that there is no difference between the distributions of variables under the 3 experimental conditions. Microbiological analysis of the 98% glycerol solution and bone fragments from samples stored for 1 year at room temperature did not show bacterial or fungal growth. The histological and microbiological investigation were performed at 2 different time points: immediately after the sample processing and after 1 year. CONCLUSION: The method used to preserve bone grafts kept in 98% glycerol at room temperature (10ºC-35ºC) was similar to cryopreservation in terms of bone matrix preservation; no bacteria or fungi were found in the samples.

Caracterização físico-química do enxerto de osso bovino liofilizado

“…Among these are bone grafts of bovine origin that have been processed and lyophilized (7) . Bovine bone has practically unlimited availability and great physicochemical and structural similarity to human bone (8) .…”

Cryopreservation - A Viable Alternative in Preparation for Use of Allografts in Knee Ligament Reconstruction

“…Two studies carried out in Brazil address the biomechanical properties of patellar and calcaneus tendons of cadavers with the same preparation method as that used in our study, comparing fresh and cryopreserved allografts (Giovani et al, 2006;Reiff et al, 2007). They found no differences.…”

Evaluation of the presence of Leishmania infantum in bone stored in glycerol or sterilized by autoclave for cortical bone grafting

“…Glycerol may have a bacteriostatic activity, as demonstrated in studies by Del Carlo et al (1999) instead of a bactericidal activity, as mentioned by other authors (Gioso et al, 2002, Giovani et al, 2006.…”

“…It is important to mention, though, that both have limitations, such as the reduction of mechanical strength in autoclaved tissues and reduction of osteogenic capacity in both techniques. Thus, other techniques for processing bone should also be studied in the future for the elimination of Leishmania, given that each has its advantages and limitations that must be analyzed to choose the best technique to be applied in each case, as described in literature (Fitch et al, 1997;Boyce et al, 1999;Giovani et al, 2006). Thomson and Sinton (1921) observed growth of L. infantum in NNN medium, the same used in this study, with a sample of bone marrow from a corpse after 14 hours postmortem, claiming the high internal resistance of the parasite in tissues without specifying the mechanisms involved.…”