Nanowarming and ice-free cryopreservation of large sized, intact porcine articular cartilage

Abstract

Successful organ or tissue long-term preservation would revolutionize biomedicine. Cartilage cryopreservation enables prolonged shelf life of articular cartilage, posing the prospect to broaden the implementation of promising osteochondral allograft (OCA) transplantation for cartilage repair. However, cryopreserved large sized cartilage cannot be successfully warmed with the conventional convection warming approach due to its limited warming rate, blocking its clinical potential. Here, we develope a nanowarming and ice-free cryopreservation method for large sized, intact articular cartilage preservation. Our method achieves a heating rate of 76.8 °C min-1, over one order of magnitude higher than convection warming (4.8 °C min-1). Using systematic cell and tissue level tests, we demonstrate the superior performance of our method in preserving large cartilage. A depth-dependent preservation manner is also observed and recapitulated through magnetic resonance imaging and computational modeling. Finally, we show that the delivery of nanoparticles to the OCA bone side could be a feasible direction for further optimization of our method. This study pioneers the application of nanowarming and ice-free cryopreservation for large articular cartilage and provides valuable insights for future technique development, paving the way for clinical applications of cryopreserved cartilage.

Cigarette smoke extract exacerbates progression of osteoarthritic‐like changes in cartilage explant cultures

“…Fresh cartilage explants ( n = 4) and cartilage explants after 12 days in culture, under control ( n = 8), OA only ( n = 8), and CSE + OA ( n = 8) conditions, were stored frozen in PBS‐soaked gauze. The hydraulic permeability and equilibrium modulus of cartilage explants were determined using a microindentation procedure 33,34 . Specifically, indentation was performed using an Anton Paar UHNT 3 surface testing platform in conjunction with a ruby ball indenter probe with a radius of 0.5 mm.…”

“…procedure. 33,34 Specifically, indentation was performed using an Anton Paar UHNT 3 surface testing platform in conjunction with a ruby ball indenter probe with a radius of 0.5 mm. A preload of 200 uN was applied for 400 s so that the tissue reached equilibrium between 5 and 9 um.…”

Optimized Partial Freezing Protocol Enables 10-Day Storage of Rat Livers

“…Alternately, vitri cation can stop biological time altogether, reaching temperatures as low as − 196°C, enabling inde nite storage [20]. However, these approaches are limited, with supercooling requiring highly stable storage conditions to prevent ice nucleation, isochoric preservation requiring technologically advanced storage systems for maintenance of constant volume, and vitri cation requiring high-powered radiofrequency coils for rapid nanowarming to enable the warming rates necessary to avoid ice nucleation [23,24]. Partial freezing (PF) is an attractive alternative, taking advantage of subzero preservation with controlled ice formation.…”