Stress is part of everyone's life and is exacerbated by traumatic events such as pandemics, disasters, violence, lifestyle changes, and health disorders. Chronic stress has many detrimental health effects and can even be life-threatening. Long-term stress monitoring outside of a hospital is often accomplished by measuring heart rate variability. While easy to measure, this digital biomarker has low specificity, greatly limiting its utility. To address this shortcoming, we report a non-invasive, wearable biomolecular sensor to monitor cortisol levels in sweat. Cortisol is a neuroendocrine hormone that regulates homeostasis as part of the stress pathway. Cortisol is detected using an electrochemical sensor functionalized with a pseudoknot-assisted aptamer and a flexible microfluidic sweat sampling system. The skin-worn microfluidic sampler provides rapid sweat collection while separating old and new sweat. The conformation-switching aptamer provides high specificity towards cortisol while being regenerable, allowing it to monitor temporal changes continuously. The aptamer was engineered to add a pseudoknot, restricting it to only two states, thus minimizing the background signal and enabling high sensitivity. An electrochemical pH sensor allows pH-corrected amperometric measurements. Device operation was demonstrated invitro with a broad linear dynamic range (1 pM - 1 μM) covering the physiological range and a sub-picomolar (0.2 pM) limit of detection in sweat. Real-time, on-body measurements were collected from human subjects using an induced stress protocol, demonstrating in-situ signal regeneration and the ability to detect dynamic cortisol fluctuations continuously for up to 90 min. The reported device has the potential to improve prognosis and enable personalized treatments.

Molecular monitoring in the dermal interstitial fluid (ISF) is an attractive approach to painlessly screen markers of health and disease status on the go. One promising strategy for accessing ISF involves the use of wearable patches containing microneedle sensor arrays. To date, such microneedle sensors have been fabricated via various manufacturing strategies based on injection molding, machining, and advanced lithography to name a few. Our groups previously reported 3D-printed microneedles as a convenient and scalable approach to sensor fabrication that, when combined with aptamer-based molecular measurements, can support continuous molecular monitoring in ISF. However, the original platform suffered from poor patch stability when deployed on the skin of rodents in vivo. We identified that this problem was due to the rheological properties of the rodent skin, which can contract post microneedle placement, physically pushing the microneedles out of the skin. This sensor retraction caused a loss of electrical contact between working and reference needles, irreversibly damaging the sensors. To address this problem, we report here an innovative approach that allows magnetic placement of microneedle sensor arrays on the skin of live rodents, affixing the patches under light pressure that prevents needle retraction. Using this strategy, we achieved sensor signaling baselines that drift at rates comparable to those seen with other in vivo deployments of electrochemical, aptamer-based sensors. We illustrate real-time pharmacokinetic measurements in live Sprague-Dawley rats using SLA-printed, aptamer-functionalized microneedles and demonstrate their ability to support drift correction via kinetic differential measurements. We also discuss future prospects and challenges.

Amidst the rising prevalence of respiratory diseases, the importance of effective lung treatment modalities is more critical than ever. However, current drug delivery systems face significant limitations that impede their efficacy and therapeutic outcome. Biohybrid microrobots have shown considerable promise for active in vivo drug delivery, especially for pulmonary applications via intratracheal routes. However, the invasive nature of intratracheal administration poses barriers to its clinical translation. Herein, we report on an efficient non-invasive inhalation-based method of delivering microrobots to the lungs. A nebulizer is employed to encapsulate picoeukaryote algae microrobots within small aerosol particles, enabling them to reach the lower respiratory tract. Post nebulization, the microrobots retain their motility (~55 μm s^-1) to help achieve a homogeneous lung distribution and long-term retention exceeding five days in the lungs. Therapeutic efficacy is demonstrated in a mouse model of acute methicillin-resistant Staphylococcus aureus pneumonia using this pulmonary inhalation approach to deliver microrobots functionalized with platelet membrane-coated polymeric nanoparticles loaded with vancomycin. These promising findings underscore the benefits of inhalable biohybrid microrobots in a setting that does not require anesthesia, highlighting the substantial translational potential of this delivery system for routine clinical applications.