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.

Osteochondral tissue cultures: Between limits and sparks, the next step for advanced in vitro models

“…One particular work by Mueller-Rath et al ( 2007) stands out from the literature that describes a cultivation of autologous chondrocytes in a defect created in a OCT graft and then implanted in subcutaneous pockets of nude mice. The remaining five papers had cultivation times more homogeneous of 2, 3, and 4 weeks (Brecelj et al, 2007;Lyman et al, 2012;Gavénis et al, 2010;Secretan et al, 2010;de Sousa et al, 2015). In particular, the study Although all papers describe careful washing steps of the samples to remove as much as possible bone marrow from the subchondral bone the experimental set ups, on the other hand, vary greatly.…”

“…reported byde Sousa et al (2015) on a large number of patients (22) and a very large number of specimens (220 OCTs) analyzed the best culture conditions and compared specimen cultivated at 4 or 37°C for 1, 3, and 14 days. Major findings showed an increase in cell death at 14 days in comparison with shorter experimental times, and when plugs were cultured at 37°C in comparison with 4°C.…”

Harvest, Transport, and Storage of Fresh Humeral Head Osteochondral Allograft: Step-by-Step Protocol

“…11 Preservation of human osteochondral allografts has previously been studied for the treatment of knee lesions. 16,17 Regarding the shoulder, few studies have evaluated the use of osteochondral allografts for treating humeral head articular cartilage lesions. 7 The few existing reports have mainly described the use of frozen allografts, 7,18,19 whereas studies involving the use of fresh allografts are scarce.…”

Protocolo para captação, transporte e preservação de tecido osteocondral humano

“…In a preliminary study, performed with surgical disposal material, our group showed that refrigeration at 4°C offers better preservation capacity of osteochondral fragments than at 37°C for up to 14 days. 18 For this reason, in the current study with young CD tissues, we kept the option of cooling the fragments to 4°C. Our results confirm previous findings that the temperature of 4°C is adequate for allograft preservation.…”