Polydimethylsiloxane (PDMS) is nearly ubiquitous in microfluidic devices, being easy to work with, economical, and transparent. A detailed protocol is provided here for using PDMS in the fabrication of microfluidic devices to aid those interested in using the material in their work, with information on the many potential ways the material may be used for novel devices.

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Fabrication of microfluidic devices using polydimethylsiloxane

Microfabricated systems provide an excellent platform for the culture of cells, and are an extremely useful tool for the investigation of cellular responses to various stimuli. Advantages offered over traditional methods include cost-effectiveness, controllability, low volume, high resolution, and sensitivity. Both biocompatible and bio- incompatible materials have been developed for use in these applications. Biocompatible materials such as PMMA or PLGA can be used directly for cell culture. However, for bio- incompatible materials such as silicon or PDMS, additional steps need to be taken to render these materials more suitable for cell adhesion and maintenance. This review describes multiple surface modification strategies to improve the biocompatibility of MEMS materials. Basic concepts of cell-biomaterial interactions, such as protein adsorption and cell adhesion are covered. Finally, the applications of these MEMS materials in Tissue Engineering are presented.

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Cell Culture on MEMS Platforms: A Review

Low oxygen tensions experienced in various pathological and physiological conditions are a major stimulus for angiogenesis. Hypoxic conditions play a critical role in regulating cellular behaviour including migration, proliferation and differentiation. This study introduces the use of a microfluidic device that allows for the control of oxygen tension for the study of different three-dimensional (3D) cell cultures for various applications. The device has a central 3D gel region acting as an external cellular matrix, flanked by media channels. On each side, there is a peripheral gas channel through which suitable gas mixtures are supplied to establish a uniform oxygen tension or gradient within the device. The effects of various parameters, such as gas and media flow rates, device thickness, and diffusion coefficients of oxygen were examined using numerical simulations to determine the characteristics of the microfluidic device. A polycarbonate (PC) film with a low oxygen diffusion coefficient was embedded in the device in proximity above the channels to prevent oxygen diffusion from the incubator environment into the polydimethylsiloxane (PDMS) device. The oxygen tension in the device was then validated experimentally using a ruthenium-coated (Ru-coated) oxygen-sensing glass cover slip which confirmed the establishment of low uniform oxygen tensions (<3%) or an oxygen gradient across the gel region. To demonstrate the utility of the microfluidic device for cellular experiments under hypoxic conditions, migratory studies of MDA-MB-231 human breast cancer cells were performed. The microfluidic device allowed for imaging cellular migration with high-resolution, exhibiting an enhanced migration in hypoxia in comparison to normoxia. This microfluidic device presents itself as a promising platform for the investigation of cellular behaviour in a 3D gel scaffold under varying hypoxic conditions.

/pdf/a-novel-microfluidic-platform-for-high-resolution-imaging-of-3p42l4gq0w.pdf

A novel microfluidic platform for high-resolution imaging of a three-dimensional cell culture under a controlled hypoxic environment.

Low oxygen tensions experienced in various pathological and physiological conditions are a major stimulus for angiogenesis Hypoxic conditions play a critical role in regulating cellular behaviour including migration, proliferation and differentiation This study introduces the use of a microfluidic device that allows for the control of oxygen tension for the study of different three-dimensional (3D) cell cultures for various applications The device has a central 3D gel region acting as an external cellular matrix, flanked by media channels On each side, there is a peripheral gas channel through which suitable gas mixtures are supplied to establish a uniform oxygen tension or gradient within the device The effects of various parameters, such as gas and media flow rates, device thickness, and diffusion coefficients of oxygen were examined using numerical simulations to determine the characteristics of the microfluidic device A polycarbonate (PC) film with a low oxygen diffusion coefficient was embedded in the device in proximity above the channels to prevent oxygen diffusion from the incubator environment into the polydimethylsiloxane (PDMS) device The oxygen tension in the device was then validated experimentally using a ruthenium-coated (Ru-coated) oxygen-sensing glass cover slip which confirmed the establishment of low uniform oxygen tensions (<3%) or an oxygen gradient across the gel region To demonstrate the utility of the microfluidic device for cellular experiments under hypoxic conditions, migratory studies of MDA-MB-231 human breast cancer cells were performed The microfluidic device allowed for imaging cellular migration with high-resolution, exhibiting an enhanced migration in hypoxia in comparison to normoxia This microfluidic device presents itself as a promising platform for the investigation of cellular behaviour in a 3D gel scaffold under varying hypoxic conditions

A novel microfluidic platform for high-resolution imaging of a three-dimensional cell culture under a controlled hypoxic environment

Intracortical microelectrodes (IME) are neural devices that initially were designed to function as neuroscience tools to enable researchers to understand the nervous system. Over the years, technology that aids interfacing with the nervous system has allowed the ability to treat patients with a wide range of neurological injuries and diseases. Despite the substantial success that has been demonstrated using intracortical microelectrodes in neural interface applications, these implants eventually fail due to loss of quality recording signals. Recent strategies to improve interfacing with the nervous system have been inspired by methods that mimic the native tissue. This review focusses on one strategy in particular, nano-architecture, a term we introduce that encompasses the approach of roughening the surface of the implant. Various nano-architecture approaches have been hypothesized to improve the biocompatibility of IMEs, enhance the recording quality and increase the longevity of the implant. This review will begin by introducing IME technology and discuss the challenges facing the clinical deployment of IME technology. The biological inspiration of nano-architecture approaches will be explained as well as leading fabrication methods used to create nano-architecture and their limitations. A review of the effects of nano-architecture surfaces on neural cells will be examined, depicting the various cellular responses to these modified surfaces in both in vitro and pre-clinical models. The proposed mechanism elucidating the ability of nano-architectures to influence cellular phenotype will be considered. Finally, the frontiers of next generation nano-architecture IMEs will be identified, with perspective given on the future impact of this interfacing approach.

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Nano-Architectural Approaches for Improved Intracortical Interface Technologies.

Studies on the effects of variations in temperature and mild temperature gradients on cells, gels, and scaffolds are important from the viewpoint of biological function. Small differences in temperature are known to elicit significant variations in cell behavior and individual protein reactivity. For the study of thermal effects and gradients in vitro, it is important to develop microfluidic platforms which are capable of controlling temperature gradients in an environment which mimics the range of physiological conditions. In the present paper, such a microfluidic thermal gradient system (muTGS) system is proposed which can create and maintain a thermal gradient throughout a cell-seeded gel matrix using the hot and cold water supply integrated in the system in the form of a countercurrent heat exchanger. It is found that a uniform temperature gradient can be created and maintained in the device even inside a high temperature and high humidity environment of an incubator. With the help of a hot and cold circuit controlled from outside the incubator the temperature gradient can be regulated. A numerical simulation of the device demonstrates the thermal feature of the chip. Cell viability and activity under a thermal gradient are examined by placing human breast cancer cells in the device.

Sarit K. Das

Papers

A microfluidic platform for studying the effects of small temperature gradients in an incubator environment.