Janus Vitrification of Droplet via Cold Leidenfrost Phenomenon.

Janus particles with asymmetric crystals show great importance in optoelectronics and photocatalysis, but their synthesis usually requires complicated procedures. Here, an unexpected Janus vitrification phenomenon is observed in a droplet caused by the Leidenfrost effect at a cryogenic temperature, which is commonly regarded as symmetric. The Leidenfrost phenomenon levitates the droplet when it comes in contact with liquid nitrogen causing different cooling conditions on the droplet's top and bottom surfaces. It induces asymmetric crystallization in the droplet, forming a Janus vitrified particle with an asymmetric crystallization borderline after cooling, as further evidenced by cryotransmission electron microscopy (cryo-TEM) experiments. Theoretical analysis and experimental study indicate that the position of the asymmetric crystallization borderline is determined by the droplet radius and density, and the observation window of asymmetric crystallization borderline is determined by the chemical concentration. The finding reveals the asymmetric crystallization phenomenon in droplet vitrification for the first time, and provides a new insight for creating Janus particles through the Leidenfrost phenomenon.

Differential Responses of Primary Astrocytes Under Hyperthermic Temperatures.

Astrocyte is the most abundant cells in brain and plays critical roles in brain homeostasis and functions. Although hyperthermia (or fever) is a common symptom in patients, its influence on astrocyte viability, morphology, and functions remains elusive. Here we developed an in vitro astrocyte culture system capable of precisely controlling culture temperature to study astrocyte responses under clinically-relevant hyperthermic temperatures (38 ∼ 41 °C). We found that hyperthermia in this temperature range does not alter cell morphology, but significantly affects cell viability, activation and functions. Specifically, high-hyperthermia (40 °C and 41 °C) causes irreversible and permanent damages to astrocytes and compromises their normal viability and functionalities repairing damaged neural tissue, recycling neurotransmitters, and promoting brain development, while mild-hyperthermia (38 °C and 39 °C) induces astrocyte activation and cytokine secretion without significant decreases in cell viability. This study sheds new insights into our understanding of various fever-associated symptoms, enabling the future development of astrocyte-targeted therapy to treat brain diseases via hyperthermia.

Thermo-Chemical Resistance to Combination Therapy of Glioma Depends on Cellular Energy Level.

Thermal therapy has been widely used in clinical tumor treatment and more recently in combination with chemotherapy, where the key challenge is the treatment resistance. The mechanism at the cellular level underlying the resistance to thermo-chemical combination therapy remains elusive. In this study, we constructed 3D culture models for glioma cells (i.e., 3D glioma spheres) as the model system to recapitulate the native tumor microenvironment and systematically investigated the thermal response of 3D glioma spheres at different hyperthermic temperatures. We found that 3D glioma spheres show high viability under hyperthermia, especially under high hyperthermic temperatures (42 °C). Further study revealed that the main mechanism lies in the high energy level of cells in 3D glioma spheres under hyperthermia, which enables the cells to respond promptly to thermal stimulation and maintain cellular viability by upregulating the chaperon protein Hsp70 and the anti-apoptotic pathway AKT. Besides, we also demonstrated that 3D glioma spheres show strong drug resistance to the thermo-chemical combination therapy. This study provides a new perspective on understanding the thermal response of combination therapy for tumor treatment.

In Situ and Quantitatively Imaging of Heat-Induced Oxidative State and Oxidative Damage of Living Neurons Using Scanning Electrochemical Microscopy.

Central nervous system is sensitive and vulnerable to heat. Oxidative state and oxidative damage of neurons under heat stress are vital for understanding early consequences and mechanisms of heat-related neuronal injury, which remains elusive partly due to the technical challenge of in situ and quantitative monitoring methods. Herein, a temperature-controlled scanning electrochemical microscopy (SECM) platform with programmable pulse potential and depth scan modes is developed for in situ and quantitatively monitoring of oxygen consumption, extracellular hydrogen peroxide level, and cell membrane permeability of neurons under thermal microenvironment of 37-42 °C. The SECM results show that neuronal oxygen consumption reaches a maximum at 40 °C and then decreases, extracellular H2 O2 level increases from 39 °C, and membrane permeability increases from 2.0 ± 0.6 × 10-5 to 7.2 ± 0.8 × 10-5 m s-1 from 39 to 42 °C. The therapeutic effect on oxidative damage of neurons under hyperthermia conditions (40-42 °C) is further evaluated by SECM and fluorescence methods, which can be partially alleviated by the potent antioxidant edaravone. This work realizes in situ and quantitatively observing the heat-induced oxidative state and oxidative damage of living neurons using SECM for the first time, which results can contribute to a better understanding of the heat-related cellular injury mechanism.

A mechanoelectrical coupling model of neurons under stretching.

Neurons are situated in a microenvironment composed of various mechanical cues, where stretching is thought to have a major impact on neurons, resulting in microstructural changes in neural tissue and further leading to abnormal electrophysiological function. In spite of significant experimental efforts, the underlying mechanism remains elusive, more works are needed to provide a detailed description of the process that leads to the observed phenomena. Here, we developed a mechanoelectrical coupling model of central neurons under stretching and specially considered the plastic deformation of neurons. With the model, we showed that the increasing axial strain induces a decreased membrane action potential and a more frequent neuronal firing, which agree well with experimental observations reported in the literature. The simulation results also showed a faster electrophysiological signal conduction. Our model provides a reference for the prediction and regulation of neuronal function under simplified conditions of mechanical loadings.

Ultrarapid Inductive Rewarming of Vitrified Biomaterials with Thin Metal Forms.

Arteries with 1-mm thick walls can be successfully vitrified by loading cryoprotective agents (CPAs) such as VS55 (8.4 M) or less concentrated DP6 (6 M) and cooling at or beyond their critical cooling rates of 2.5 and 40 °C/min, respectively. Successful warming from this vitrified state, however, can be challenging. For example, convective warming by simple warm-bath immersion achieves 70 °C/min, which is faster than VS55's critical warming rate of 55 °C/min, but remains far below that of DP6 (185 °C/min). Here we present a new method that can dramatically increase the warming rates within either a solution or tissue by inductively warming commercially available metal components placed within solutions or in proximity to tissues with non-invasive radiofrequency fields (360 kHz, 20 kA/m). Directly measured warming rates within solutions exceeded 1000 °C/min with specific absorption rates (W/g) of 100, 450 and 1000 for copper foam, aluminum foil, and nitinol mesh, respectively. As proof of principle, a carotid artery diffusively loaded with VS55 and DP6 CPA was successfully warmed with high viability using aluminum foil, while standard convection failed for the DP6 loaded tissue. Modeling suggests this approach can improve warming in tissues up to 4-mm thick where diffusive loading of CPA may be incomplete. Finally, this technology is not dependent on the size of the system and should therefore scale up where convection cannot.