As an ideal antioxidant and anti-apoptotic substance, hydrogen (H2) has protective effects on many isolated organs, such as the heart, lung and kidney. In this study, we explore whether H2 improves the preservation effect of osteochondral allograft by adding it to Dulbecco’s Modified Eagle Medium (DMEM) solution during the tissue culture stage. The osteochondral allograft apparatus was used to harvest 60 pieces of cylindrical allografts (l = 10 mm, d = 6 mm) cartilage in the lateral loading area of the femoral condyle from the pig knee joint in the aseptic condition, and the grafts were randomly divided into 4 groups: Control group (DMEM solution without hydrogen); H-1 group (DMEM solution with hydrogen concentration of 0.2 mmol/L); H-2 group (DMEM solution with hydrogen concentration of 0.4 mmol/L); and H-3 group (DMEM solution with hydrogen concentration of 0.8 mmol/L). The chondrocyte viability, histological changes (hematoxylin and eosin staining, Safranine O staining, and collagen type II immunohistochemistry staining) and biomechanical properties (Young’s modulus) of the osteochondral allograft were investigated after 28 days’ storage. The chondrocyte viability and proteoglycan and collagen type II contents in the H-3 and H-2 groups were higher than that in the Control and H-1 groups, and the H-3 group had the highest values. However, significant differences were not observed between the four groups based on Young’s modulus. Hydrogen as an additive to the DMEM solution improved the preservation effect of osteochondral allograft. The preservation effect of hydrogen occurred in a concentration-dependent manner.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Bian L, Stoker AM, Marberry KM, Ateshian GA, Cook JL, Hung CT (2010) Effects of dexamethasone on the functional properties of cartilage explants during long-term culture. Am J Sports Med 38:78–85
Buokwalter JA (2001) Basic science of orthopedics. People’s Medical Publishing House, Beijing
Cameron JI, Pulido PA, McCauley JC, Bugbee WD (2016) Osteochondral allograft transplantation of the femoral trochlea. Am J Sports Med 44:633–638
Campbell J, Filardo G, Bruce B, Bajaj S, Friel N, Hakimiyan A, Wood S, Grumet R, Shafikhani S, Chubinskaya S, Cole BJ (2014) Salvage of contaminated osteochondral allografts: the effects of chlorhexidine on human articular chondrocyte viability. Am J Sports Med 42:973–978
Chen K, Wang N, Diao Y, Dong W, Sun Y, Liu L, Wu X (2017) Hydrogen-rich saline attenuates brain injury induced by cardiopulmonary bypass and inhibits microvascular endothelial cell apoptosis via the PI3K/Akt/GSK3beta signaling pathway in rats. Cell Physiol Biochem 43:1634–1647
Cook JL, Stoker AM, Stannard JP, Kuroki K, Cook CR, Pfeiffer FM, Bozynski C, Hung CT (2014) A novel system improves preservation of osteochondral allografts. Clin Orthop Relat Res 472:3404–3414
Cook JL, Stannard JP, Stoker AM, Bozynski CC, Kuroki K, Cook CR, Pfeiffer FM (2016) Importance of donor chondrocyte viability for osteochondral allografts. Am J Sports Med 44:1260–1268
Dang Y, Liu T, Mei X, Meng X, Gou X, Deng B, Xu H, Xu L (2017) Hyperoxygenated hydrogen-rich solution suppresses shock- and resuscitation-induced liver injury. J Surg Res 220:363–371
Diao HJ, Fung HS, Yeung P, Lam KL, Yan CH, Chan BP (2017) Dynamic cyclic compression modulates the chondrogenic phenotype in human chondrocytes from late stage osteoarthritis. Biochem Biophys Res Commun 486:14–21
Forriol F, Longo UG, Alvarez E, Campi S, Ripalda P, Rabitti C, Maffulli N, Denaro V (2011) Scanty integration of osteochondral allografts cryopreserved at low temperatures with dimethyl sulfoxide. Knee Surg Sports Traumatol Arthrosc 19:1184–1191
Gao Y, Yang H, Chi J, Xu Q, Zhao L, Yang W, Liu W, Yang W (2017) Hydrogen gas attenuates myocardial ischemia reperfusion injury independent of postconditioning in rats by attenuating endoplasmic reticulum stress-induced autophagy. Cell Physiol Biochem 43:1503–1514
Ge L, Yang M, Yang NN, Yin XX, Song WG (2017) Molecular hydrogen: a preventive and therapeutic medical gas for various diseases. Oncotarget 8:102653–102673
Glenn RE Jr, McCarty EC, Potter HG, Juliao SF, Gordon JD, Spindler KP (2006) Comparison of fresh osteochondral autografts and allografts: a canine model. Am J Sports Med 34:1084–1093
Haaparanta AM, Jarvinen E, Cengiz IF, Ella V, Kokkonen HT, Kiviranta I, Kellomaki M (2014) Preparation and characterization of collagen/PLA, chitosan/PLA, and collagen/chitosan/PLA hybrid scaffolds for cartilage tissue engineering. J Mater Sci Mater Med 25:1129–1136
Hanaoka T, Kamimura N, Yokota T, Takai S, Ohta S (2011) Molecular hydrogen protects chondrocytes from oxidative stress and indirectly alters gene expressions through reducing peroxynitrite derived from nitric oxide. Med Gas Res 1:18
Henrotin Y, Kurz B, Aigner T (2005) Oxygen and reactive oxygen species in cartilage degradation: friends or foes? Osteoarthr Cartil 13:643–654
Krych AJ, Robertson CM, Williams RJ 3rd, Cartilage Study G (2012) Return to athletic activity after osteochondral allograft transplantation in the knee. Am J Sports Med 40:1053–1059
LaPrade RF, Bursch LS, Olson EJ, Havlas V, Carlson CS (2008) Histologic and immunohistochemical characteristics of failed articular cartilage resurfacing procedures for osteochondritis of the knee: a case series. Am J Sports Med 36:360–368
LaPrade RF, Botker J, Herzog M, Agel J (2009) Refrigerated osteoarticular allografts to treat articular cartilage defects of the femoral condyles: a prospective outcomes study. J Bone Joint Surg Am 91:805–811
Linn MS, Chase DC, Healey RM, Harwood FL, Bugbee WD, Amiel D (2011) Etanercept enhances preservation of osteochondral allograft viability. Am J Sports Med 39:1494–1499
Maher P, Salgado KF, Zivin JA, Lapchak PA (2007) A novel approach to screening for new neuroprotective compounds for the treatment of stroke. Brain Res 1173:117–125
Ohsawa I, Ishikawa M, Takahashi K, Watanabe M, Nishimaki K, Yamagata K, Katsura K, Katayama Y, Asoh S, Ohta S (2007) Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med 13:688–694
Ohta S (2011) Recent progress toward hydrogen medicine: potential of molecular hydrogen for preventive and therapeutic applications. Curr Pharm Des 17:2241–2252
Pennock A, Ferdinand W, Robertson C, Harwood F, Bugbee W (2006) Prolonged storage of osteochondral allografts: does the addition of fetal bovine serum improve chondrocyte viability? J Knee Surg 19:265–272
Poleni PE, Bianchi A, Etienne S, Koufany M, Sebillaud S, Netter P, Terlain B, Jouzeau JY (2007) Agonists of peroxisome proliferators-activated receptors (PPAR) alpha, beta/delta or gamma reduce transforming growth factor (TGF)-beta-induced proteoglycans’ production in chondrocytes. Osteoarthr Cartil 15:493–505
Qi J, Hu Z, Song H, Chen B, Xie D, Zhou L, Zhang Y (2016) Cartilage storage at 4 °C with regular culture medium replacement benefits chondrocyte viability of osteochondral grafts in vitro. Cell Tissue Bank 17:473–479
Salahudeen AK, Huang H, Patel P, Jenkins JK (2000) Mechanism and prevention of cold storage-induced human renal tubular cell injury. Transplantation 70:1424–1431
Tan M, Sun X, Guo L, Su C, Sun X, Xu Z (2013) Hydrogen as additive of HTK solution fortifies myocardial preservation in grafts with prolonged cold ischemia. Int J Cardiol 167:383–390
Teng MS, Yuen AS, Kim HT (2008) Enhancing osteochondral allograft viability: effects of storage media composition. Clin Orthop Relat Res 466:1804–1809
Yamada T, Uchida K, Onuma K, Kuzuno J, Ujihira M, Inoue G, Sato B, Kurokawa R, Sakai R, Takaso M (2014) Hydrogen supplementation of preservation solution improves viability of osteochondral grafts. Sci World J 2014:109876
Zhang X, Liu J, Jin K, Xu H, Wang C, Zhang Z, Kong M, Zhang Z, Wang Q, Wang F (2017) Subcutaneous injection of H2 is a novel effective treatment for type 2 diabetes. J Diabetes Investig 9:83–90
This work was supported by the Shandong Provincial Natural Science Foundation of China (Grant No. ZR2017LH018, ZR2013HM034), the Projects of Medical and Health Technology Development Program in Shandong Province (Grant No. 2015WS0109) and Tai'an Science and Technology Development Plan (Grant No. 2018NS0194).
Conflicts of interest
The authors declare they have no conflicts of interest to disclose.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Han, Y., Qu, P., Zhang, K. et al. Storage solution containing hydrogen improves the preservation effect of osteochondral allograft. Cell Tissue Bank 20, 201–208 (2019). https://doi.org/10.1007/s10561-019-09758-7
- Tissue preservation
- Chondrocyte viability
- Osteochondral allograft