Direct cell imprint lithography in superconductive carbon black polymer composites: process optimization, characterization and in vitro toxicity analysis

Cell imprint lithography (CIL) or cell replication plays a vital role in fields like biomimetic smart culture substrates, bone tissue engineering, cell guiding, cell adhesion, tissue engineering, cell microenvironments, tissue microenvironments, cell research, drug delivery, diagnostics, therapeutics and many other applications. Herein we report a new formulation of superconductive carbon black photopolymer composite and its characterization towards a CIL process technique. In this article, we demonstrated an approach of using a carbon nanoparticle-polymer composite (CPC) for patterning cells. It is observed that a 0.3 wt % load of carbon nanoparticles (CNPs) in a carbon polymer mixture (CPM) was optimal for cell-imprint replica fabrication. The electrical resistance of the 3-CPC (0.3 wt %) was reduced by 68% when compared to N-CPC (0 wt %). This method successfully replicated the single cell with sub-organelle scale. The shape of microvesicles, grooves, pores, blebs or microvilli on the cellular surface was patterned clearly. This technique delivers a free-standing cell feature substrate. In vitro evaluation of the polymer demonstrated it as an ideal candidate for biomimetic biomaterial applications. This approach also finds its application in study based on morphology, especially for drug delivery applications and for investigations based on molecular pathways

Tumor necrosis factor-related apoptosis inducing ligand stimulates migration and proliferation of vascular smooth muscle cells

Introduction: Tumor Necrosis Factor-Related Apoptosis (TRAIL) has the ability to inhibit angiogenesis through programmed cell death, in the same time able to promote pro-angiogenic activity. It seems that TRAIL has an opposite effects make its role in ischemic disease and its function in a normal cell is still unclear. This study determines the effect of TRAIL in Vascular Smooth Muscle Cells (VSMCs). Materials and Methods: In this study, 6 wells in vitro Scratch Assay were conducted for migration and Cellltiter 96® Aqueous One Solution Cell Proliferation Assay (MTS) was performed for proliferation. In both assays, cells were serum arrested for 24, 48 and 72 hours, following stimulation with different concentration (1ng/ml, 20ng/ml, 50ng/ml and 100ng/ml) and different volume (20µL and 100µL) of TRAIL. PDGF-BB was used as positive control. Results of PDGFBB and TRAIL were compared with untreated VSMC. Results: It Is demonstrated that 100µL of 50ng/ml TRAIL-induced VSMC shows significant (p < 0.05) higher migration rate percentage compared to untreated cells, included 100µL of 20ng/ml PDGF-BB after 48 hours. However, in proliferation assay, the lowest concentration of TRAIL (1ng/ml, 20µL) shows significantly higher proliferation rate compared to untreated cells after 24 hours (p < 0.05) meanwhile PDGF-BB (1ng/ml, 20µL) indicates significant higher proliferation after 48 hours. Conclusion: This study demonstrates that TRAIL promotes the VSMC migration and proliferation in vitro. This indicates that migration and proliferation of VSMC upon vascular injury from media to intima not only involves PDGF-BB, but also TRAIL. Therefore, TRAIL contributes to intimal thickening that can lead to the obstruction of the blood vessels and it is a hallmark of atherosclerosis

Pipette petri dish single-cell trapping (PP-SCT) in microfluidic platforms: A passive hydrodynamic technique

Microfluidics-based biochips play a vital role in single-cell research applications. Handling and positioning of single cells at the microscale level are an essential need for various applications, including genomics, proteomics, secretomics, and lysis-analysis. In this article, the pipette Petri dish single-cell trapping (PP-SCT) technique is demonstrated. PP-SCT is a simple and cost-effective technique with ease of implementation for single cell analysis applications. In this paper a wide operation at different fluid flow rates of the novel PP-SCT technique is demonstrated. The effects of the microfluidic channel shape (straight, branched, and serpent) on the efficiency of single-cell trapping are studied. This article exhibited passive microfluidic-based biochips capable of vertical cell trapping with the hexagonally-positioned array of microwells. Microwells were 35 μm in diameter, a size sufficient to allow the attachment of captured cells for short-term study. Single-cell capture (SCC) capabilities of the microfluidic-biochips were found to be improving from the straight channel, branched channel, and serpent channel, accordingly. Multiple cell capture (MCC) was on the order of decreasing from the straight channel, branch channel, and serpent channel. Among the three designs investigated, the serpent channel biochip offers high SCC percentage with reduced MCC and NC (no capture) percentage. SCC was around 52%, 42%, and 35% for the serpent, branched, and straight channel biochips, respectively, for the tilt angle, θ values were between 10–15°. Human lung cancer cells (A549) were used for characterization. Using the PP-SCT technique, flow rate variations can be precisely achieved with a flow velocity range of 0.25–4 m/s (fluid channel of 2 mm width and 100 µm height). The upper dish (UD) can be used for low flow rate applications and the lower dish (LD) for high flow rate applications. Passive single-cell analysis applications will be facilitated using this method

Direct cell imprint lithography in superconductive carbon black polymer composites: Process optimization, characterization and in vitro toxicity analysis

Cell imprint lithography (CIL) or cell replication plays a vital role in fields like biomimetic smart culture substrates, bone tissue engineering, cell guiding, cell adhesion, tissue engineering, cell microenvironments, tissue microenvironments, cell research, drug delivery, diagnostics, therapeutics and many other applications. Herein we report a new formulation of superconductive carbon black photopolymer composite and its characterization towards a CIL process technique. In this article, we demonstrated an approach of using a carbon nanoparticle-polymer composite (CPC) for patterning cells. It is observed that a 0.3 wt % load of carbon nanoparticles (CNPs) in a carbon polymer mixture (CPM) was optimal for cell-imprint replica fabrication. The electrical resistance of the 3-CPC (0.3 wt %) was reduced by 68% when compared to N-CPC (0 wt %). This method successfully replicated the single cell with sub-organelle scale. The shape of microvesicles, grooves, pores, blebs or microvilli on the cellular surface was patterned clearly. This technique delivers a free-standing cell feature substrate. In vitro evaluation of the polymer demonstrated it as an ideal candidate for biomimetic biomaterial applications. This approach also finds its application in study based on morphology, especially for drug delivery applications and for investigations based on molecular pathways

Pipette Petri Dish Single-Cell Trapping (PP-SCT) in Microfluidic Platforms: A Passive Hydrodynamic Technique

Microfluidics-based biochips play a vital role in single-cell research applications. Handling and positioning of single cells at the microscale level are an essential need for various applications, including genomics, proteomics, secretomics, and lysis-analysis. In this article, the pipette Petri dish single-cell trapping (PP-SCT) technique is demonstrated. PP-SCT is a simple and cost-effective technique with ease of implementation for single cell analysis applications. In this paper a wide operation at different fluid flow rates of the novel PP-SCT technique is demonstrated. The effects of the microfluidic channel shape (straight, branched, and serpent) on the efficiency of single-cell trapping are studied. This article exhibited passive microfluidic-based biochips capable of vertical cell trapping with the hexagonally-positioned array of microwells. Microwells were 35 &mu;m in diameter, a size sufficient to allow the attachment of captured cells for short-term study. Single-cell capture (SCC) capabilities of the microfluidic-biochips were found to be improving from the straight channel, branched channel, and serpent channel, accordingly. Multiple cell capture (MCC) was on the order of decreasing from the straight channel, branch channel, and serpent channel. Among the three designs investigated, the serpent channel biochip offers high SCC percentage with reduced MCC and NC (no capture) percentage. SCC was around 52%, 42%, and 35% for the serpent, branched, and straight channel biochips, respectively, for the tilt angle, &theta; values were between 10&ndash;15&deg;. Human lung cancer cells (A549) were used for characterization. Using the PP-SCT technique, flow rate variations can be precisely achieved with a flow velocity range of 0.25&ndash;4 m/s (fluid channel of 2 mm width and 100 &micro;m height). The upper dish (UD) can be used for low flow rate applications and the lower dish (LD) for high flow rate applications. Passive single-cell analysis applications will be facilitated using this method