Drug and chemical development along with safety tests rely on the use of numerous clinical models. This is a lengthy process where animal testing is used as a standard for pre-clinical trials. However, these models often fail to represent human physiopathology. This may lead to poor correlation with results from later human clinical trials. Organ-on-a-Chip (OOAC) systems are engineered microfluidic systems, which recapitulate the physiochemical environment of a specific organ by emulating the perfusion and shear stress cellular tissue undergoes in vivo and could replace current animal models. The success of culturing cells and cell-derived tissues within these systems is dependent on the scaffold chosen; hence, scaffolds are critical for the success of OOACs in research. A literature review was conducted looking at current OOAC systems to assess the advantages and disadvantages of different materials and manufacturing techniques used for scaffold production; and the alternatives that could be tailored from the macro tissue engineering research field.
Purpose: Hydrogels with low viscosities tend to be difficult to use in constructing tissue engineering (TE) scaffolds used to replace or restore damaged tissue, due to the length of time it takes for final gelation to take place resulting in the scaffolds collapsing due to their mechanical instability. However, recent advances in rapid prototyping have allowed for a new technology called bioplotting to be developed, which aims to circumvent these inherent problems. This paper aims to present details of the process.Design/methodology/approach: The paper demonstrates how by using the bioplotting technique complex 3D geometrical scaffolds with accurate feature sizes and good pore definition can be fabriated for use as biological matrices. PEG gels containing the cell‐adhesive RGD peptide sequence were patterned using this method to produce layers of directional microchannels which have a functionalised bioactive surface. Seeding these gels with C2C12 myoblasts showed that the cells responded to the topographical features and aligned themselves along the direction of the channels.Findings: This process allows plotting of various materials into a media bath containing material of similar rheological properties which can be used to both support the structure as it is dispensed and also to initiate cross‐linking of the hydrogel. By controlling concentrations, viscosity and the temperature of both the plotting material and the plotting media, the speed of the hydrogel gelation can be enhanced whilst it is cross‐linking in the media bath. TE scaffolds have been produced using a variety of materials including poly(ethylene glycol) (PEG), gelatin, alginic acid and agarose at various concentrations and viscosities.Originality/value: This paper describes one of the very few examples of accurate construction of 3D biological microporous matrices using hydrogel material fabricated by the bioplotting technique. This demonstrates that this technique can be used to produce 3D scaffolds which promote tissue regeneration.
A point of care device utilising Lab-on-a-Chip technologies that is applicable for biological pathogens was designed, fabricated and tested showing sample in to answer out capabilities. The purpose of the design was to develop a cartridge with the capability to perform nucleic acid extraction and purification from a sample using a chitosan membrane at an acidic pH. Waste was stored within the cartridge with the use of sodium polyacrylate to solidify or gelate the sample in a single chamber. Nucleic acid elution was conducted using the RPA amplification reagents (alkaline pH). Passive valves were used to regulate the fluid flow and a multiplexer was designed to distribute the fluid into six microchambers for amplification reactions. Cartridges were produced using soft lithography of silicone from 3D printed moulds, bonded to glass substrates. The isothermal technique, RPA is employed for amplification. This paper shows the results from two separate experiments: the first using the RPA control nucleic acid, the second showing successful amplification from Chlamydia Trachomatis. Endpoint analysis conducted for the RPA analysis was gel electrophoresis that showed 143 base pair DNA was amplified successfully for positive samples whilst negative samples did not show amplification. End point analysis for Chlamydia Trachomatis samples was fluorescence detection that showed successful detection of 1 copy/μL and 10 copies/μL spiked in a MES buffer.
Advances in microfluidics and the introduction of isothermal nucleic acid amplification assays have resulted in a range of solutions for nucleic acid amplification tests suited for point of care and field use. However, miniaturisation of instrumentation for such assays has not seen such rapid advances and fluorescence based assays still depend on complex, bulky and expensive optics such as fluorescence microscopes, photomultiplier tubes and sensitive lens assemblies. In this work we demonstrate a robust, low cost platform for isothermal nucleic acid amplification on a microfluidic device. Using easily obtainable materials and commercial off-the-shelf components, we show real time fluorescence detection using a low cost photodiode and operational amplifier without need for lenses. Temperature regulation on the device is achieved using a heater fabricated with standard printed circuit board fabrication methods. These facile construction methods allow fabrications at a cost compatible with widespread deployment to resource poor settings.
Surface-enhanced Raman spectroscopy (SERS) substrates manufactured using complex nano-patterning techniques have become the norm. However, their cost of manufacture makes them unaffordable to incorporate into most biosensors. The technique shown in this paper is low-cost, reliable and highly sensitive. Chemical etching of solid Ag metal was used to produce simple, yet robust SERS substrates with broadband characteristics. Etching with ammonium hydroxide (NH4OH) and nitric acid (HNO3) helped obtain roughened Ag SERS substrates. Scanning electron microscopy (SEM) and interferometry were used to visualize and quantify surface roughness. Flattened Ag wires had inherent, but non-uniform roughness having peaks and valleys in the microscale. NH4OH treatment removed dirt and smoothened the surface, while HNO3 treatment produced a flake-like morphology with visibly more surface roughness features on Ag metal. SERS efficacy was tested using 4-methylbenzenethiol (MBT). The best SERS enhancement for 1 mM MBT was observed for Ag metal etched for 30 s in NH4OH followed by 10 s in HNO3. Further, MBT could be quantified with detection limits of 1 pM and 100 µM, respectively, using 514 nm and 1064 nm Raman spectrometers. Thus, a rapid and less energy intensive method for producing solid Ag SERS substrate and its efficacy in analyte sensing was demonstrated.