Two-dimensional (2D) graphene oxide (GO) and molybdenum disulfide (MoS2) nanosheets (NSs) have been widely used as photothermal agents and as potential carriers of antitumor drugs. Their spatial thermal effects have been extensively explored for use at physiological and hyperthermic temperatures (37 to 46 °C). Furthermore, the modulation of the spatial thermal distributions with these NSs may have even more profound applications in the microstructural control of biomaterials at cryogenic temperatures (-196 to 37 °C). These applications include bioinspired microfabrication via freezing, food and drug freeze-drying, and biomaterial cryopreservation. However, such thermal effects of NSs and their applications at cryogenic temperatures had never been fully explored. Therefore, in this study, we have utilized the near-infrared laser induced photothermal effects of GO and MoS2 NSs to suppress the ice nucleation and ice crystal growth during warming of the biosamples. Using this approach, biological cells subjected to fast cooling to a deeply frozen state (-196 °C) were successfully recovered with high survival rates and full biological functionality. Thus, we provide a NS based effective approach to control the crystallization behaviors of water during warming at cryogenic temperatures, as NSs may have wide applications in both materials science and bioengineering.

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Vitrification is considered as an important alternative approach to traditional slow freezing method for cryopreservation of cells. A typical cell vitrification procedure involves a non-equilibrium cooling process commonly accomplished in liquid nitrogen, while in which film boiling is believed to greatly hinder heat transfer surrounding the sample, resulting in incomplete vitrification or a much higher critical concentration. In this study, we developed a simple while effective approach, wrapping traditional French-type straw with medical gauze, to greatly enhance convective heat transfer during cooling by suppress film boiling. We further established a coupled heat transfer model for cooling and warming of cell suspensions to investigate the inherent thermodynamic mechanism in this approach. The model describes both the macroscale thermal distributions in extracellular solution and the microscale ice crystallization inside the cells. The simulation indicated that straws wrapped with medical gauze would increase cell survival subject to vitrification cryopreservation by significantly increasing the cooling rate to inhibit intracellular ice formation (IIF). Our experiments on human umbilical vein endothelial cells (HUVECs) further confirmed the predictions in that the cell survival rate was significantly increased by wrapping straws with medical gauze.

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