title: Approaches to preserve human osteochondral allografts.
abstract: Osteochondral defects may progress to osteoarthritis. Many attempts have been developed to overcome this issue, including osteochondral autografts and allografts. The goal of this study was to develop a new protocol for storage of human osteochondral allografts. Osteochondral plugs were randomly allocated in the following groups: control, immediate freezing up to -70 °C, cooling at 4 °C, and storage at 37 °C. Samples from the cooling at 4 °C and storage at 37 °C groups were stored in tubes containing medium plus human albumin and analyzed after 1, 3, and 14 days. The frozen groups' samples were cryopreserved for 1 year in cryotubes containing medium only (FM), medium plus human albumin (FA), and medium plus human albumin and glucose (FG) and were then analyzed. Analysis involved histological study with hematoxylin-eosin and Safranin O and a modified Live/Dead assay. In samples stored both at 37 and 4 °C, analysis showed statistically significant higher cellular mortality at 14 days compared to 1 and 3 days, but mortality in the 4 °C group was lower. In the freezing protocols, the FA group showed less cellular mortality than the FM and FG groups. Cooling at 4 °C offers better preservation capacity than storage at 37 °C, but both offer the capacity for preservation for 14 days. Adding human albumin to the storage medium is useful in reducing cellular mortality in samples frozen for 1 year.
authors.affiliation: Research Division, National Institute of Traumatology and Orthopaedics, Avenida Brasil, 500 - 5° Andar, Rio de Janeiro, RJ, 20940-070, Brazil, eduardobsousa@globo.com.
authors.forename: Eduardo Branco
authors.initials: EB
authors.lastname: de Sousa
authors.forename: Diego Pinheiro
authors.initials: DP
authors.lastname: Aguiar
authors.forename: José Fernando Marques
authors.initials: JF
authors.lastname: Barcelos
authors.forename: Maria Eugênia Leite
authors.initials: ME
authors.lastname: Duarte
authors.forename: Beni
authors.initials: B
authors.lastname: Olej
mesh_terms: D064591:Allografts / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D002358:Cartilage, Articular / Q000737:chemistry / Q000166:cytology* / Q000502:physiology*; D015925:Cryopreservation / Q000379:methods*; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008875:Middle Aged; D009924:Organ Culture Techniques / Q000379:methods*; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000737:chemistry; D014022:Tissue Survival / Q000502:physiology
chemical_list: D003470:Culture Media; D019188:Organ Preservation Solutions