A miniaturized wash-free microfluidic assay for electrical impedance-based assessment of red blood cell-mediated microvascular occlusion.

The production of HbS - an abnormal hemoglobin (Hb) - in sickle cell disease (SCD) results in poorly deformable red blood cells (RBCs) that are prone to microcapillary occlusion, causing tissue ischemia and organ damage. Novel treatments, including gene therapy, may reduce SCD morbidity, but methods to functionally evaluate RBCs remain limited. Previously, we presented the microfluidic impedance red cell assay (MIRCA) for rapid assessment of RBC deformability, employing electrical impedance-based readout to measure RBC occlusion of progressively narrowing micropillar openings. We describe herein the design, development, validation, and clinical utility of the next-generation MIRCA assay, featuring enhanced portability, rapidity, and usability. It incorporates a miniaturized impedance analyzer and features a simplified wash-free operation that yields an occlusion index (OI) within 15 min as a new metric for RBC occlusion. We show a correlation between OI and percent fetal hemoglobin (%HbF), other laboratory biomarkers of RBC hemolysis, and SCD severity. To demonstrate the assay's versatility, we tested RBC samples from treatment-naïve SCD patients in Uganda that yielded OI levels similar to those from hydroxyurea (HU)-treated patients in the U.S., highlighting the role of %HbF in protecting against microcapillary occlusion independent of other pharmacological effects. The MIRCA assay could also identify a subset of HU-treated patients with high occlusion risks, suggesting that they may require treatment adjustments including a second-line therapy to improve their outcomes. This work demonstrates the potential of the MIRCA assay for accelerated evaluation of RBC health, function, and therapeutic effect in an ex vivo model of the microcapillary networks.

A Multichannel Portable Platform With Embedded Thermal Management for Miniaturized Dielectric Blood Coagulometry.

This article presents a standalone, multichannel, miniaturized impedance analyzer (MIA) system for dielectric blood coagulometry measurements with a microfluidic sensor termed ClotChip. The system incorporates a front-end interface board for 4-channel impedance measurements at an excitation frequency of 1 MHz, an integrated resistive heater formed by a pair of printed-circuit board (PCB) traces to keep the blood sample near a physiologic temperature of 37 °C, a software-defined instrument module for signal generation and data acquisition, and a Raspberry Pi-based embedded computer with 7-inch touchscreen display for signal processing and user interface. When measuring fixed test impedances across all four channels, the MIA system exhibits an excellent agreement with a benchtop impedance analyzer, with rms errors of ≤0.30% over a capacitance range of 47-330 pF and ≤0.35% over a conductance range of 2.13-10 mS. Using in vitro-modified human whole blood samples, the two ClotChip output parameters, namely, the time to reach a permittivity peak (Tpeak) and maximum change in permittivity after the peak (Δϵr,max) are assessed by the MIA system and benchmarked against the corresponding parameters of a rotational thromboelastometry (ROTEM) assay. Tpeak exhibits a very strong positive correlation (r = 0.98, p < 10-6, n = 20) with the ROTEM clotting time (CT) parameter, while Δϵr,max exhibits a very strong positive correlation (r = 0.92, p < 10-6, n = 20) with the ROTEM maximum clot firmness (MCF) parameter. This work shows the potential of the MIA system as a standalone, multichannel, portable platform for comprehensive assessment of hemostasis at the point-of-care/point-of-injury (POC/POI).

Implanted High Density Cuff Electrodes Functionally Activate Human Tibial and Peroneal Motor Units Without Chronic Detriment to Peripheral Nerve Health.

OBJECTIVE: Peripheral nerve stimulation via multi-contact nerve cuff electrodes (NCEs) has proved effective in restoring function to individuals with lower-extremity paralysis. This study investigates clinical measures of nerve health over one year post-implantation of a composite flat-interface nerve electrode (C-FINE) on the tibial and peroneal nerves above the knee in a human volunteer. This represents the first deployment of a novel NCE on new neural targets in a uniquely challenging location prone to prolonged externally applied forces, making acute and chronic postoperative observation critical. MATERIALS AND METHODS: A 27-year-old man with an incomplete spinal cord injury (AIS C) at the C3 to C4 level received eight-contact C-FINEs bilaterally on the tibial and peroneal nerves, proximal to the knee. Access to four contacts per cuff exhibiting the most desirable responses was externalized via temporary percutaneous leads. Percutaneous leads were later removed, with contacts generating the best dorsiflexion (two of four) and plantar flexion (one of four) reconnected to a permanently implanted pulse generator. For 13 months post-implantation, nerve health and cuff performance were assessed through motor nerve conduction velocity (MNCV) studies, clinical needle electromyography, compound motor action potential (CMAP), sensory nerve action potential (SNAP), stimulation-evoked tetanic moment collection, and lower-limb circumference measurements. RESULTS: Tibial and peroneal MNCVs remained stable bilaterally above 40 m/sec, with CMAPs increased or stable after six months. SNAPs remained stable across all measurements. CMAP initial charge thresholds remained below 50 nC, with minimal changes to muscle recruitment order in three of four externalized contacts per cuff. Peak tetanic moments remained stable, with bilateral increases in thigh and calf circumferences of 5% and 14% over one year. CONCLUSIONS: Above-knee tibial and peroneal NCEs can restore stimulated ankle-joint function without chronic nerve health detriments. Alongside previous femoral nerve data, this study demonstrates the ability of NCEs to enhance lower-extremity function with limited neuromuscular impact.