Droplet based microfluidics (digital microfluidics) with Electrowetting on dielectric (EWOD) has gained popularity with the promise of being technology for a true lab-on-chip device with applications spanning across assays/library prep, next-gen sequencing and point-of-care diagnostics. Most electrowetting device architecture are linear electrode arrays with a shared path for droplets, imposing serious limitations -- cross contamination and limited number of parallel operations. Our work is in addressing these issues through large 2D grid arrays with direct addressability providing flexible programmability.

Scaling electrowetting to larger arrays still remains a challenge due to complex and expensive cleanroom fabrication of microfluidic devices. We take the approach of using inexpensive PCB manufacturing, investigate challenges and solutions for scaling electrowetting to large area droplet manipulation. PCB manufactured electrowetting arrays impose many challenges due to the irregularities from process and materials used. These challenges generally relate to preparing the surface that interfaces with droplets -- a dielectric material on the electrodes and the top most hydrophobic coating that interfaces with the droplets. A requirement for robust droplet manipulation with EWOD is thin (<10um) hydrophobic dielectric material which does not break down at droplet actuation voltages (AC/DC, 60V to 200V) and has a no droplet pinning. For this, we engineered materials specifically for large area PCBs.

Traditionally, digital microfluidic devices sandwich droplets between two plates and have focussed on sub-microliter droplet volumes. In our approach, droplets are on an open surface with which we are able to manipulate droplets in microliter and milliliter volumes. With milliliter droplet manipulation ability on our electrowetting device, we demonstrate “digital millifluidics”. Finally, we report the performance of our device and to motivate the need for large open arrays we show an example of running multiple parallel biological experiments.

Human mesenchymal stem cells were reseeded in decellularized human bone subject to a controlled mechanical loading to create a bone-on-chip that was cultured for over 26 months. The cell morphology and their secretome were characterized using immunohistochemistry and in situ immunofluorescence under confocal microscopy. The presence of stem cell derived osteocytes was confirmed at 547 days. Different cell populations were identified. Some cells were connected by long processes and formed a network. Comparison of the MSCs in vitro reorganization and calcium response to in situ mechanical stimulation were compared to MLOY4 cells reseeded on human bone. The bone-on-chip produced an ECM of which the strength was nearly a quarter of native bone after 109 days and that contained calcium minerals at 39 days and type I collagen at 256 days. The cytoplasmic calcium concentration variations seemed to adapt to the expected in vivo mechanical load at the successive stages of cell differentiation in agreement with studies using fluid shear flow stimulation. Some degree of bone-like formation over a long period of time with the formation of a newly formed matrix was observed.

We propose a 3D-printable soft, stretchable, and transparent hydrogel-elastomer device that is able to detect simulated ‘nerve’ signals. The signal is passed to a conductive hydrogel electrode through a non-contact method of capacitive coupling through polydimethylsiloxane (PDMS). We demonstrate that the device is able to detect sinusoidal waveforms passed through a simulated ‘nerve’ made from conductive hydrogel over a range of frequencies (1 kHz – 1 MHz). Analysis of signal detection showed a correlation to the electrode contact area and a Vin/Vout of larger than 10%. This provides the framework for the future development of a soft, 3D-printable, capacitive coupling device that can be used as a cuff electrode for detecting peripheral nerve signals.

Poly (lactic acid) (PLA) bioplastics are recyclable and biodegradable thermoplastics. They are derived from environmentally friendly sources such as potatoes, cornstarch and sugarcane. However, PLA is inherently brittle with low impact strength. The goal of this study is to improve mechanical properties of PLA by the addition of calcium carbonate (CaCO3) fillers. PLA composites were prepared by injection molding conventional limestone (LS) and white chicken eggshell (WES) powders with particle sizes of 63 μm and 32 μm in amounts of 5 wt. %, 10 wt. % and 20 wt. %. Mechanical properties such as, tensile strength, tensile modulus, and Charpy impact strengths were investigated. These three properties were evaluated and the results statistically analyzed using ANOVA F-test. For both particle sizes, the tensile strength decreased as the filler content increased, but was highest for a filler loading of 5 wt. %. In general, the 32 μm powder fillers had better tensile strengths than 63 μm sized fillers. The tensile modulus increased with filler content and was highest at 20 wt. % for both particle sizes. The LS/PLA composites had better toughness than the WES/PLA composites. The particle filler morphology and fractured surfaces were observed by scanning electron microscopy (SEM) and determined to have well dispersed particles with smooth fractured surfaces. Water absorption behavior of PLA/CaCO3 composites were studied by immersion in distilled water at room temperature for 56 days. Virgin PLA absorbed the least amount of water while the water absorption of CaCO3 composites were a function of powder type and content.

Hydrogels have been widely used in many applications from tissue engineering to drug delivery systems. For both tissue engineering and drug delivery, the mechanical properties are important because they would affect cell-materials interactions and injectability of drugs encapsulated in hydrogel carriers. Therefore, it is important to study the mechanical properties of these hydrogels, particularly at physiological temperature (37°C). This study adopted strain sweep and frequency sweep rotational rheological tests to investigate the rheological characteristics of various tissue engineering relevant hydrogels with different concentrations at 37°C. These hydrogels include alginate, RGD-alginate, and copolymerized collagen/alginate/fibrin. It has revealed that the addition of RGD has negligible effect on the elastic modulus and viscosity of alginate. Alginate gels have demonstrated shear thinning behavior which indicates that they are suitable candidates as carriers for cells or drug delivery. The addition of collagen and fibrin would reinforce the mechanical properties of alginate which makes it a strong scaffold material.