The application of microfluidic devices for viral diagnosis in developing countries.

Whilst diseases such as diabetes and cardiovascular disorders are increasing in the developed world, the main threat to global health remains viral-based infectious disease. Such diseases are notably prevalent in developing countries, where they represent a major cause of mortality; however, their detection and prevention is typically hampered by poor infrastructure and a lack of resources to support the sophisticated diagnostic tools commonly found in modern laboratories. Microfluidic-based diagnostics has the potential to close the gap between developed and developing world medical needs due to the robustness and reduced operating costs such technology offers. The most recent developments in microfluidic diagnostics for viral infections have explored the separation and enumeration of immune cells, the capture and identification of viral particles, and antiviral drug evaluation within microchannels and chambers. Advances in solid-phase separation, isothermal amplification, real-time detection of nucleotide products, and improved efficiency of detection systems in microfluidic platforms have also opened up opportunities for diagnostic innovation. This chapter reviews the potential capability microfluidic technology can offer in addressing the practical challenges of providing diagnostic technology for developing countries, illustrated by research on key viral diseases.

A microfluidic system for testing the responses of head and neck squamous cell carcinoma tissue biopsies to treatment with chemotherapy drugs.

Tumors are heterogeneous masses of cells characterized pathologically by their size and spread. Their chaotic biology makes treatment of malignancies hard to generalize. We present a robust and reproducible glass microfluidic system, for the maintenance and "interrogation" of head and neck squamous cell carcinoma (HNSCC) tumor biopsies, which enables continuous media perfusion and waste removal, recreating in vivo laminar flow and diffusion-driven conditions. Primary HNSCC or metastatic lymph samples were subsequently treated with 5-fluorouracil and cisplatin, alone and in combination, and were monitored for viability and apoptotic biomarker release 'off-chip' over 7 days. The concentration of lactate dehydrogenase was initially high but rapidly dropped to minimally detectable levels in all tumor samples; conversely, effluent concentration of WST-1 (cell proliferation) increased over 7 days: both factors demonstrating cell viability. Addition of cell lysis reagent resulted in increased cell death and reduction in cell proliferation. An apoptotic biomarker, cytochrome c, was analyzed and all the treated samples showed higher levels than the control, with the combination therapy showing the greatest effect. Hematoxylin- and Eosin-stained sections from the biopsy, before and after maintenance, demonstrated the preservation of tissue architecture. This device offers a novel method of studying the tumor environment, and offers a pre-clinical model for creating personalized treatment regimens.

Study of ethanol induced toxicity in liver explants using microfluidic devices.

Current in vitro methodologies for the culture and analysis of liver specific responses lack the sophistication of in vivo dynamics. In this work, a microfluidic based experimental methodology has been utilized to reproduce a biomimetic microenvironment in which pseudo in vivo liver tissue studies can be carried out under in vitro conditions. This innovative technique, which exploits the inherent advantages of microfluidic technology, has been utilised to study the viability and functionality of explant liver tissue over four days in the presence of varying concentrations of ethanol. Concentrations of ethanol as low as 20 mM have produced a decrease in WST-1 metabolism, a marker of mitochondrial activity, and an increase lactose dehydrogenase release, reflecting cell death, in the explant samples; these effects increase with higher ethanol concentrations. A concomitant decrease in albumin and urea synthesis was also observed. We believe the proposed methodology is widely applicable and is clearly of relevance to biological and clinical research including drug development and toxicity, as well as enabling better fundamental understanding of tissue/cell processes.

Development of a microfluidic device for the maintenance and interrogation of viable tissue biopsies.

A microfluidic based experimental methodology has been developed that offers a biomimetic microenvironment in which pseudo in vivo tissue studies can be carried out under in vitro conditions. Using this innovative technique, which utilizes the inherent advantages of microfluidic technology, liver tissue has been kept in a viable and functional state for over 70 h during which time on-chip cell lysis has been repeatedly performed. Tissue samples were also disaggregated in situ on-chip into individual primary cells, using a collagenase digestion procedure, enabling further cell analysis to be carried out off-line. It is anticipated that this methodology will have a wide impact on biological and clinical research in fields such as cancer prognosis and treatment, drug development and toxicity, as well as enabling better fundamental research into tissue/cell processes.