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

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.

Keywords

cartilage, articular - allografts - bone transplantation - tissue and organ harvesting - osteochondral lesion

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

• Referências

• 1 Zouzias IC, Bugbee WD. Osteochondral Allograft Transplantation in the Knee. Sports Med Arthrosc Rev 2016; 24 (02) 79-84
• 2 Dhollander A, Verdonk P, Tirico LE, Gomoll AH. Treatment of failed cartilage repair: State of the Art. In: Journal of ISAKOS: Joint Disorders & Orthopaedic Sports Medicine, p. jisakos. 2016-000057.
• 3 LaPrade RF, Botker J, Herzog M, Agel J. Refrigerated osteoarticular allografts to treat articular cartilage defects of the femoral condyles. A prospective outcomes study. J Bone Joint Surg Am 2009; 91 (04) 805-811
• 4 Gracitelli GC, Moraes VY, Franciozi CE, Luzo MV, Belloti JC. Surgical interventions (microfracture, drilling, mosaicplasty, and allograft transplantation) for treating isolated cartilage defects of the knee in adults. Cochrane Database Syst Rev 2016; 9: CD010675
• 5 De Caro F, Bisicchia S, Amendola A, Ding L. Large fresh osteochondral allografts of the knee: a systematic clinical and basic science review of the literature. Arthroscopy 2015; 31 (04) 757-765
• 6 Sherman SL, Garrity J, Bauer K, Cook J, Stannard J, Bugbee W. Fresh osteochondral allograft transplantation for the knee: current concepts. J Am Acad Orthop Surg 2014; 22 (02) 121-133
• 7 Pearsall IV AW, Tucker JA, Hester RB, Heitman RJ. Chondrocyte viability in refrigerated osteochondral allografts used for transplantation within the knee. Am J Sports Med 2004; 32 (01) 125-131
• 8 Mankin HJ, Dorfman H, Lippiello L, Zarins A. Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. II. Correlation of morphology with biochemical and metabolic data. J Bone Joint Surg Am 1971; 53 (03) 523-537
• 9 Pritzker KP, Gay S, Jimenez SA. , et al. Osteoarthritis cartilage histopathology: grading and staging. Osteoarthritis Cartilage 2006; 14 (01) 13-29
• 10 Richter DL, Schenck Jr RC, Wascher DC, Treme G. Knee articular cartilage repair and restoration techniques: a review of the literature. Sports Health 2016; 8 (02) 153-160
• 11 Raz G, Safir OA, Backstein DJ, Lee PT, Gross AE. Distal Femoral Fresh Osteochondral Allografts: Follow-up at a Mean of Twenty-two Years. J Bone Joint Surg Am 2014; 96 (13) 1101-1107
• 12 Williams 3rd RJ, Dreese JC, Chen CT. Chondrocyte survival and material properties of hypothermically stored cartilage: an evaluation of tissue used for osteochondral allograft transplantation. Am J Sports Med 2004; 32 (01) 132-139
• 13 Ball ST, Amiel D, Williams SK. , et al. The effects of storage on fresh human osteochondral allografts. Clin Orthop Relat Res 2004; (418) 246-252
• 14 Garrity JT, Stoker AM, Sims HJ, Cook JL. Improved osteochondral allograft preservation using serum-free media at body temperature. Am J Sports Med 2012; 40 (11) 2542-2548
• 15 Pallante AL, Bae WC, Chen AC, Görtz S, Bugbee WD, Sah RL. Chondrocyte viability is higher after prolonged storage at 37 degrees C than at 4 degrees C for osteochondral grafts. Am J Sports Med 2009; 37 (Suppl. 01) 24S-32S
• 16 Xia Z, Murray D, Hulley PA, Triffitt JT, Price AJ. The viability and proliferation of human chondrocytes following cryopreservation. J Bone Joint Surg Br 2008; 90 (09) 1245-1248
• 17 Cook JL, Stoker AM, Stannard JP. , et al. A novel system improves preservation of osteochondral allografts. Clin Orthop Relat Res 2014; 472 (11) 3404-3414
• 18 de Sousa EB, Aguiar DP, Barcelos JF, Duarte ME, Olej B. Approaches to preserve human osteochondral allografts. Cell Tissue Bank 2015; 16 (03) 425-431
• 19 Bian L, Stoker AM, Marberry KM, Ateshian GA, Cook JL, Hung CT. Effects of dexamethasone on the functional properties of cartilage explants during long-term culture. Am J Sports Med 2010; 38 (01) 78-85
• 20 Yamada T, Uchida K, Onuma K. , et al. Hyaluronic Acid (800 kDa) Supplementation of University of Wisconsin Solution Improves Viability of Osteochondral Grafts and Reduces Matrix Metalloproteinase Expression during Cold Preservation. ScientificWorldJournal 2015; 2015: 631369
• 21 Onuma K, Urabe K, Naruse K, Uchida K, Itoman M. Allogenic serum improves cold preservation of osteochondral allografts. Clin Orthop Relat Res 2012; 470 (10) 2905-2914
• 22 Linn MS, Chase DC, Healey RM, Harwood FL, Bugbee WD, Amiel D. Etanercept enhances preservation of osteochondral allograft viability. Am J Sports Med 2011; 39 (07) 1494-1499
• 23 McCulloch K, Litherland GJ, Rai TS. Cellular senescence in osteoarthritis pathology. Aging Cell 2017; 16 (02) 210-218