Design Optimization of a 3D Microfluidic Channel System for Biomedical Applications

Peshin S., Madou M., Kulinsky L.

Centrifugal microfluidic platforms (CDs) have opened new possibilities for inexpensive point-of-care (POC) diagnostics. They are now widely used in applications requiring polymerase chain reaction steps, blood plasma separation, serial dilutions, and many other diagnostic processes. CD microfluidic devices allow a variety of complex processes to transfer onto the small disc platform that previously were carried out by individual expensive laboratory equipment requiring trained personnel. The portability, ease of operation, integration, and robustness of the CD fluidic platforms requires simple, reliable, and scalable designs to control the flow of fluids. Valves play a vital role in opening/closing of microfluidic channels to enable a precise control of the flow of fluids on a centrifugal platform. Valving systems are also critical in isolating chambers from the rest of a fluidic network at required times, in effectively directing the reagents to the target location, in serial dilutions, and in integration of multiple other processes on a single CD. In this paper, we review the various available fluidic valving systems, discuss their working principles, and evaluate their compatibility with CD fluidic platforms. We categorize the presented valving systems into either “active”, “passive”, or “hybrid”—based on their actuation mechanism that can be mechanical, thermal, hydrophobic/hydrophilic, solubility-based, phase-change, and others. Important topics such as their actuation mechanism, governing physics, variability of performance, necessary disc spin rate for valve actuation, valve response time, and other parameters are discussed. The applicability of some types of valves for specialized functions such as reagent storage, flow control, and other applications is summarized.

Valle N.M., Nucci M.P., Alves A.H., Rodrigues L.D., Mamani J.B., Oliveira F.A., Lopes C.S., Lopes A.T., Carreño M.N., Gamarra L.F.

This systematic review aimed to analyze the development and functionality of microfluidic concentration gradient generators (CGGs) for toxicological evaluation of different biological organisms. We searched articles using the keywords: concentration gradient generator, toxicity, and microfluidic device. Only 33 of the 352 articles found were included and examined regarding the fabrication of the microdevices, the characteristics of the CGG, the biological model, and the desired results. The main fabrication method was soft lithography, using polydimethylsiloxane (PDMS) material (91%) and SU-8 as the mold (58.3%). New technologies were applied to minimize shear and bubble problems, reduce costs, and accelerate prototyping. The Christmas tree CGG design and its variations were the most reported in the studies, as well as the convective method of generation (61%). Biological models included bacteria and nematodes for antibiotic screening, microalgae for pollutant toxicity, tumor and normal cells for, primarily, chemotherapy screening, and Zebrafish embryos for drug and metal developmental toxicity. The toxic effects of each concentration generated were evaluated mostly with imaging and microscopy techniques. This study showed an advantage of CGGs over other techniques and their applicability for several biological models. Even with soft lithography, PDMS, and Christmas tree being more popular in their respective categories, current studies aim to apply new technologies and intricate architectures to improve testing effectiveness and reduce common microfluidics problems, allowing for high applicability of toxicity tests in different medical and environmental models.

Fattahi Z., Hasanzadeh M.

Microfluidics technology holds a special place in the field of biomedical and analytical chemistry thanks to its properties, such as low consumption of reagent, quick analysis time, and ease of integration. Nanotechnology makes significant progress towards high sensitive and selective detection of analytes. Compared to bulk materials, nanomaterials have distinctive properties which motivate the combination of nanoparticles with biosensors. Nanoparticle-based biosensors can bring a series of advantages in advancement high-throughput and ultrasensitive detection. In recent years, numerous studies have been done on the integration of nanomaterials with microfluidic systems. Given the scope of the nanoparticle-integrated biosensors, the purpose of this review is to provide an overview of various types of nanoparticle-integrated microfluidic biosensors. It mainly focuses on developed microfluidics-based on gold nanoparticles, graphene oxide , quantum dots, magnetic nanoparticles, carbon dots, silica nanoparticles, zinc oxide, and nanocomposites. This review summarizes the latest advancement of nanotechnology-assisted microfluidic biosensing on a smartphone for biochemical analysis. Also, the role of nanomaterials on the detection methods such as electrochemical, photoelectrochemical, electrochemiluminescent, optical piezoelectric, and so on was surveyed. In our opinion, the cooperation of microfluidic biodevices and nanoparticles is very beneficial and is expected to be effective in addressing the challenges related to microfluidic systems. • Recent advances and limitations in nanotechnology-assisted microfluidic biosensing were discussed. • The analytical figures of merit of the developed methods are also evaluated. • Current improvements on the monitoring of biomarkers using microfluidic strategies were investigated.

Oh J.M., Begum H.M., Liu Y.L., Ren Y., Shen K.

In tumors, the metabolic demand of cancer cells often outpaces oxygen supply, resulting in a gradient of tumor hypoxia accompanied with heterogeneous resistance to cancer therapeutics. Models recapitulating tumor hypoxia are therefore essential for developing more effective cancer therapeutics. Existing in vitro models often fail to capture the spatial heterogeneity of tumor hypoxia or involve high-cost, complex fabrication/handling techniques. Here, we designed a highly tunable microfluidic device that induces hypoxia through natural cell metabolism and oxygen diffusion barriers. We adopted a cleanroom-free, micromilling-replica-molding strategy and a microfluidic liquid-pinning approach to streamline the fabrication and tumor model establishment. We also implemented a thin-film oxygen diffusion barrier design, which was optimized through COMSOL simulation, to support both two-dimensional (2-D) and three-dimensional (3-D) hypoxic models. We demonstrated that liquid-pinning enables an easy, injection-based micropatterning of cancer cells of a wide range of parameters, showing the high tunability of our design. Human breast cancer and prostate cancer cells were seeded and stained after 24 h of 2-D and 3-D culture to validate the natural induction of hypoxia. We further demonstrated the feasibility of the parallel microfluidic channel design to evaluate dual therapeutic conditions in the same device. Overall, our new microfluidic tumor model serves as a user-friendly, cost-effective, and highly scalable platform that provides spatiotemporal analysis of the hypoxic tumor microenvironments suitable for high-content biological studies and therapeutic discoveries.

Banik S., Uchil A., Kalsang T., Chakrabarty S., Ali M.A., Srisungsitthisunti P., Mahato K.K., Surdo S., Mazumder N.

Microfluidics is revolutionizing the way research on cellular biology has been traditionally conducted. The ability to control the cell physicochemical environment by adjusting flow conditions, while performing cellular analysis at single-cell resolution and high-throughput, has made microfluidics the ideal choice to replace traditional in vitro models. However, such a revolution only truly started with the advent of polydimethylsiloxane (PDMS) as a microfluidic structural material and soft-lithography as a rapid manufacturing technology. Indeed, before the "PDMS age," microfluidic technologies were: costly, time-consuming and, more importantly, accessible only to specialized laboratories and users. The simplicity of molding PDMS in various shapes along with its inherent properties (transparency, biocompatibility, and gas permeability) has spread the applications of innovative microfluidic devices to diverse and important biological fields and clinical studies. This review highlights how PDMS-based microfluidic systems are innovating pre-clinical biological research on cells and organs. These devices were able to cultivate different cell lines, enhance the sensitivity and diagnostic effectiveness of numerous cell-based assays by maintaining consistent chemical gradients, utilizing and detecting the smallest number of analytes while being high-throughput. This review will also assist in identifying the pitfalls in current PDMS-based microfluidic systems to facilitate breakthroughs and advancements in healthcare research.

Kashaninejad N., Moradi E., Moghadas H.

Micro/nanofluidic drug delivery systems have attracted significant attention as they offer unique advantages in targeted and controlled drug delivery. Based on the desired application, these systems can be categorized into three different groups: in vitro, in situ and in vivo microfluidic drug delivery platforms. In vitro microfluidic drug delivery platforms are closely linked with the emerging concept of lab-on-a-chip for cell culture studies. These systems can be used to administer drugs or therapeutic agents, mostly at the cellular or tissue level, to find the therapeutic index and can potentially be used for personalized medicine. In situ and in vivo microfluidic drug delivery platforms are still at the developmental stage and can be used for drug delivery at tissue or organ levels. A famous example of these systems are microneedles that can be used for painless and controllable delivery of drugs or vaccines through human skin. This chapter presents the cutting edge advances in the design and fabrication of in vitro microfluidic drug delivery systems that can be used for both cellular and tissue drug delivery. It also briefly discusses the in situ drug delivery platforms using microneedles.

Liu Y., Sun L., Zhang H., Shang L., Zhao Y.

Drug development is a long process whose main content includes drug synthesis, drug delivery, and drug evaluation. Compared with conventional drug development procedures, microfluidics has emerged as a revolutionary technology in that it offers a miniaturized and highly controllable environment for bio(chemical) reactions to take place. It is also compatible with analytical strategies to implement integrated and high-throughput screening and evaluations. In this review, we provide a comprehensive summary of the entire microfluidics-based drug development system, from drug synthesis to drug evaluation. The challenges in the current status and the prospects for future development are also discussed. We believe that this review will promote communications throughout diversified scientific and engineering communities that will continue contributing to this burgeoning field.

Vitorino R., Guedes S., da Costa J.P., Kašička V.

Microfluidics is the advanced microtechnology of fluid manipulation in channels with at least one dimension in the range of 1–100 microns. Microfluidic technology offers a growing number of tools for manipulating small volumes of fluid to control chemical, biological, and physical processes relevant to separation, analysis, and detection. Currently, microfluidic devices play an important role in many biological, chemical, physical, biotechnological and engineering applications. There are numerous ways to fabricate the necessary microchannels and integrate them into microfluidic platforms. In peptidomics and proteomics, microfluidics is often used in combination with mass spectrometric (MS) analysis. This review provides an overview of using microfluidic systems for peptidomics, proteomics and cell analysis. The application of microfluidics in combination with MS detection and other novel techniques to answer clinical questions is also discussed in the context of disease diagnosis and therapy. Recent developments and applications of capillary and microchip (electro)separation methods in proteomic and peptidomic analysis are summarized. The state of the art of microchip platforms for cell sorting and single-cell analysis is also discussed. Advances in detection methods are reported, and new applications in proteomics and peptidomics, quality control of peptide and protein pharmaceuticals, analysis of proteins and peptides in biomatrices and determination of their physicochemical parameters are highlighted.

Tayyab M., Sami M.A., Raji H., Mushnoori S., Javanmard M.

COVID-19 has been declared a global pandemic which has brought the world economy and the society to a standstill. The current emphasis of testing is on detection of genetic material of SARS-CoV-2. Such tests are useful for assessing the current state of a subject: Infected or not infected. In addition to such tests, antibody testing is necessary to stratify the population into three groups: never exposed, infected, and immune. Such a stratification is necessary for safely reopening the society and remobilizing the economy. The aim of this review article is to inform the audience of the current diagnostic and surveillance technologies that are being employed for the detection of SARS-CoV-2 antibodies along with their shortcomings, and to highlight microfluidic sensors and devices that show promise of being commercialized for detection and quantification of SARS-CoV-2 antibodies in low-resource and Point-of-Care (POC) settings.

Berkenbrock J.A., Grecco-Machado R., Achenbach S.

Extensive testing of populations against COVID-19 has been suggested as a game-changer quest to control the spread of this contagious disease and to avoid further disruption in our social, healthcare and economical systems. Nonetheless, testing millions of people for a new virus brings about quite a few challenges. The development of effective tests for the new coronavirus has become a worldwide task that relies on recent discoveries and lessons learned from past outbreaks. In this work, we review the most recent publications on microfluidics devices for the detection of viruses. The topics of discussion include different detection approaches, methods of signalling and fabrication techniques. Besides the miniaturization of traditional benchtop detection assays, approaches such as electrochemical analyses, field-effect transistors and resistive pulse sensors are considered. For emergency fabrication of quick test kits, the local capabilities must be evaluated, and the joint work of universities, industries, and governments seems to be an unequivocal necessity.

Sweet E., Yang B., Chen J., Vickerman R., Lin Y., Long A., Jacobs E., Wu T., Mercier C., Jew R., Attal Y., Liu S., Chang A., Lin L.

Microfluidic concentration gradient generators (µ-CGGs) have been utilized to identify optimal drug compositions through antimicrobial susceptibility testing (AST) for the treatment of antimicrobial-resistant (AMR) infections. Conventional µ-CGGs fabricated via photolithography-based micromachining processes, however, are fundamentally limited to two-dimensional fluidic routing, such that only two distinct antimicrobial drugs can be tested at once. This work addresses this limitation by employing Multijet-3D-printed microchannel networks capable of fluidic routing in three dimensions to generate symmetric multidrug concentration gradients. The three-fluid gradient generation characteristics of the fabricated 3D µ-CGG prototype were quantified through both theoretical simulations and experimental validations. Furthermore, the antimicrobial effects of three highly clinically relevant antibiotic drugs, tetracycline, ciprofloxacin, and amikacin, were evaluated via experimental single-antibiotic minimum inhibitory concentration (MIC) and pairwise and three-way antibiotic combination drug screening (CDS) studies against model antibiotic-resistant Escherichia coli bacteria. As such, this 3D µ-CGG platform has great potential to enable expedited combination AST screening for various biomedical and diagnostic applications. Microfluidic concentration gradient generators (μ-CGGs) have been used to identify optimal drug compositions through antimicrobial susceptibility testing (AST) to treat antimicrobial-resistant infections, and a new method has been developed to overcome previous 2-D limitations. Conventional μ-CGGs fabricated through photolithography-based micromachining processes are limited to 2-D fluidic routing, which allows only two distinct antimicrobial drugs to be tested simultaneously. However, a team headed by Eric Sweet at the University of California, Berkeley, United States employed 3-D-printed microchannel networks that are capable of 3-D fluidic routing to generate symmetrical multi-drug concentration gradients. The authors were able to confirm their 3-D μ-CGG prototype by means of both theoretical simulations and experimental validations. The team believes that its 3-D μ-CGG platform offers considerable potential for conducting multi-drug AST evaluations for a variety of biomedical and diagnostic applications.

Huo C., Bai C., Zhang P.

Abstract The development of micropumps are reviewed and their applications are summarized. The micropumps are categorized into indirectly-driven and directly-driven micropumps according to the ways of driven origination from working principles. The actuation principles are introduced in detail including electrostatic, piezoelectric, thermopneumatic actuation, etc. Moreover, the performance influencing parameters on the property and the applications of various micropumps for medical therapy are described such as reciprocating, peristaltic, rotary, electrohydrodynamic, micropumps, etc. The challenge of micropumps is also discussed.

Nielsen A.V., Beauchamp M.J., Nordin G.P., Woolley A.T.

Traditional microfabrication techniques suffer from several disadvantages, including the inability to create truly three-dimensional (3D) architectures, expensive and time-consuming processes when changing device designs, and difficulty in transitioning from prototyping fabrication to bulk manufacturing. 3D printing is an emerging technique that could overcome these disadvantages. While most 3D printed fluidic devices and features to date have been on the millifluidic size scale, some truly microfluidic devices have been shown. Currently, stereolithography is the most promising approach for routine creation of microfluidic structures, but several approaches under development also have potential. Microfluidic 3D printing is still in an early stage, similar to where polydimethylsiloxane was two decades ago. With additional work to advance printer hardware and software control, expand and improve resin and printing material selections, and realize additional applications for 3D printed devices, we foresee 3D printing becoming the dominant microfluidic fabrication method.

Huang W.E., Lim B., Hsu C., Xiong D., Wu W., Yu Y., Jia H., Wang Y., Zeng Y., Ji M., Chang H., Zhang X., Wang H., Cui Z.

The pandemic coronavirus SARS-CoV-2 in the world has caused a large infected population suffering from COVID-19. To curb the spreading of the virus, WHO urgently demanded an extension of screening and testing; thus, a rapid and simple diagnostic method is needed. We applied a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) to achieve the detection of SARS-CoV-2 in 30 min. We designed four sets of LAMP primers (6 primers in each set), targeting the viral RNA of SARS-CoV-2 in the regions of orf1ab, S gene and N gene. A colorimetric change was used to report the results, which enables the outcome of viral RNA amplification to be read by the naked eye without the need of expensive or dedicated instrument. The sensitivity can be 80 copies of viral RNA per ml in a sample. We validated the RT-LAMP method in a hospital in China, employing 16 clinic samples with 8 positives and 8 negatives. The testing results are consistent with the conventional RT-qPCR. In addition, we also show that one-step process without RNA extraction is feasible to achieve RNA amplification directly from a sample. This rapid, simple and sensitive RT-LAMP method paves a way for a large screening at public domain and hospitals, particularly regional hospitals and medical centres in rural areas.

Wan L., Neumann C.A., LeDuc P.R.

Tumor progression is significantly influenced by factors such as mechanical force, shear stress, chemotaxis, and hypoxia. Here, we reviewed recent achievements and presented potential directions for tumor-on-a-chip systems in the future.