Effects of freezing on bone histological morphology
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Effects of freezing on bone histological morphology Miguel Gustavo Setu´bal Andrade Æ Camila Neves Sa´ Æ Antoˆnio Ma´rcio Teixeira Marchionni Æ Thereza Cristina Bo´rio dos Santos Calmon de Bittencourt Æ Moyse´s Sadigursky
Received: 25 October 2007 / Accepted: 19 February 2008 / Published online: 5 March 2008 Ó Springer Science+Business Media B.V. 2008
Abstract Allograft bone has been widely used for reconstruction of different portions of the skeleton. The fragment of bone harvested must be kept under
M. G. S. Andrade A. M. T. Marchionni Maxillofacial Surgery Division, Bahia Foundation for Science Development, Salvador, BA, Brazil M. G. S. Andrade T. C. B. do. S. C. de Bittencourt M. Sadigursky Post-graduation Program in Immunology, Bahia Federal University, Salvador, BA, Brazil M. G. S. Andrade (&) Division of Maxillofacial Surgery, Fundac¸a˜o Bahiana para o Desenvolvimento das Cieˆncias, Avenida Silveira Martins, n. 3386. Cabula, Salvador, BA, Brazil CEP 41150-100 e-mail: [email protected] C. N. Sa´ Tutorial Educational Program. School of Dentistry, Bahia Federal University, Salvador, BA, Brazil A. M. T. Marchionni Post-graduation Program in Laser in Dentirtry, Bahia Federal University, Salvador, BA, Brazil T. C. B. do. S. C. de Bittencourt Veterinary School, Bahia Federal University, Salvador, BA, Brazil M. Sadigursky Edgard Santos Hospital, Bahia Federal University, Salvador, BA, Brazil
low temperatures. The cryopreservation also contributes to decrease the antigenic potential of the tissue. Although this technique is considered safe, there is little information about the morphological modifications that the medullary and cortical portions of bone suffer after freezing. Hence, the aim of this study was to investigate the morphology of bone tissue after freezing under different temperatures and periods. Twelve rabbits were used to analyze the effects of two temperatures, -20°C and -70°C, during four periods of time: 30, 60, 90, 120 days. Tissues were analyzed by HE, picro-sirius stains and also by Feulgen’s reaction, through qualitative and morphometric ways, considering the area occupied by cells and nuclei, medullary and cortical portions, as well as by collagen expression at cortical. The differences among the treatments were analyzed by Tukey´s test, at 5% significance level. Bone freezing increased cellular and nuclear areas at cancellous bone and diminished nuclear area at the cortical bone. Cortical bone collagen suffered denaturation proportionally to temperature decrease and to freezing duration. These alterations compromised the morphology of tissues after 90 or 120 days of freezing at the temperature of -70°C. Cells necrosed during freezing, contributing to reduce bone antigenicity. Keywords Bone transplantation Freezing Transplantation Immunology Necrosis
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Introduction Bone grafts are widely used for several surgical purposes. Autologous grafts are preferred however, in order to skirt a surgery in a second region, which functions as donor site (Wiltfang et al. 2004;
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