In recent decades, platinum compounds have been many contributions in medicine. Development of new drugs from the active platinum compounds as well as nanocarriers for targeted delivery and reducing side effects of the drugs has paid much attention. In the study, nanocomplexes were prepared from aquated species of cisplatin and pluronic-conjugated heparin which distributed in the range of 80-100 nm by Transmission Electron Microscopy and 134 nm by Dynamic light scattering (DLS). Formation of the complex was confirmed by FTIR and DLS. The nanocomplexes exhibited high drug-loading capacity (approximately 42.5% wt/wt at 37 °C and 37.5% wt/wt at 25 °C). In vitro, drug-loaded nanogels showed much slower release profiles of cisplatin CDDP in pH 7.4 (physiological pH) compared with pH 5.5 condition at 37 °C. Moreover, the cytotoxicity assay results also indicated that Hep-F127 was cytocompatible; meanwhile, CDDP-loaded nanocomplex was able to reduce the cytotoxic ability of free CDDP (IC50 = 5.68 ± 0.73 μg/ml), which still maintain a significantly antiproliferative activity on NCI-H460 lung cancer cell. The in vitro preliminarily obtained results indicate that the nanocomplex is a candidate for CDDP delivery which can be studied further in cancer therapy.
Three-dimensional (3D) bioprinting has become mainstream for precise and repeatable high-throughput fabrication of complex cell cultures and tissue constructs in drug testing and regenerative medicine, food products, dental and medical implants, biosensors, and so forth. Due to this tremendous growth in demand, an overwhelming amount of hardware manufacturers have recently flooded the market with different types of low-cost bioprinter models—a price segment that is most affordable to typical-sized laboratories. These machines range in sophistication, type of the underlying printing technology, and possible add-ons/features, which makes the selection process rather daunting (especially for a nonexpert customer). Yet, the review articles available in the literature mostly focus on the technical aspects of the printer technologies under development, as opposed to explaining the differences in what is already on the market. In contrast, this paper provides a snapshot of the fast-evolving low-cost bioprinter niche, as well as reputation profiles (relevant to delivery time, part quality, adherence to specifications, warranty, maintenance, etc.) of the companies selling these machines. Specifically, models spanning three dominant technologies—microextrusion, droplet-based/inkjet, and light-based/crosslinking—are reviewed. Additionally, representative examples of high-end competitors (including up-and-coming microfluidics-based bioprinters) are discussed to highlight their major differences and advantages relative to the low-cost models. Finally, forecasts are made based on the trends observed during this survey, as to the anticipated trickling down of the high-end technologies to the low-cost printers. Overall, this paper provides insight for guiding buyers on a limited budget toward making informed purchasing decisions in this fast-paced market.
According to the U.S. Department of Health & Human Services, nearly 115,000 people in the U.S needed a lifesaving organ transplant in 2018, while only ~10% of them have received it. Yet, almost no artificial FDA-approved products are commercially available today -three decades after the inception of tissue engineering. It is hypothesized here that the major bottlenecks restricting its progress stem from lack of access to the inner pore space of the scaffolds. Specifically, the inability to deliver nutrients to, and clear waste from, the center of the scaffolds limits the size of the products that can be cultured. Likewise, the inability to monitor, and control, the cells after seeding them into the scaffold results in nonviable tissue, with an unacceptable product variability. To resolve these bottlenecks, we present a prototype addressable microfluidics device capable of minimally disruptive fluid and cell manipulations within living cultures. As proof-of-concept, we demonstrate its ability to perform additive manufacturing by seeding cells in spatial patterns (including co-culturing multiple cell types); and subtractive manufacturing by removing surface adherent cells via focused flow of trypsin. Additionally, we show that the device can sample fluids and perform cell "biopsies" (which can be subsequently sent for ex-situ analysis), from any location within its Culture Chamber. Finally, the on-chip plumbing is completely automated using external electronics. This opens the possibility to perform long-term computer-driven tissue engineering experiments, where the cell behavior is modulated in response to the minimally disruptive observations (e.g. fluid sampling and cell biopsies) throughout the entire duration of the cultures. It is expected that the proof-of-concept technology will eventually be scaled up to 3D addressable microfluidic scaffolds, capable of overcoming the limitations bottlenecking the transition of tissue engineering technologies to the clinical setting.
Microfluidic devices are constructed from polydimethylsiloxane (PDMS) due to their biocompatibility, fabrication ease, well-established protocols, and simplicity. PDMS-based microfluidic devices are constructed by (i) applying liquid PDMS to a negative mold (usually a silicon or 3D-printed mold) and (ii) curing the PDMS with heat exposure over a set time period. Unreacted resin monomers in 3D-printed molds prevent PDMS from fully curing, resulting in improper channel formation in PDMS and reducing the PDMS device’s efficacy. An in-house protocol that uses SU-8 as a “non-stick” coating on 3D-printed molds facilitates the successful casting of PDMS. Contact angle, surface profile, optical profile, and force testing prove that PDMS cast from SU-8-treated molds resembles pristine PDMS, unlike PDMS cast from untreated molds. Further, this method is generalized to commercial 3D prints using different 3D printing resins. To demonstrate this technique’s viability in microfluidic devices, a microfluidic tree using PDMS from treated 3D prints shows vibrant colors and clear lines. This is absent from an untreated PDMS.
A custom-built mask aligner (CBMA), which fundamentally covers all the key features of a commercial mask aligner, while being low cost and light weight and having low power consumption and high accuracy, is constructed. The CBMA is composed of a custom high fidelity light emitting diode light source, a vacuum chuck, a mask holder, high-precision translation and rotation stages, and high resolution digital microscopes. The total cost of the system is under $7500, which is over ten times cheaper than a comparable commercial system. It produces a collimated ultraviolet illumination of 1.8-2.0 mW cm over an area of a standard 4-in. wafer, at the plane of photoresist exposure, and the alignment accuracy is characterized to be <3 m, which is sufficient for most microfluidic applications. Moreover, this manuscript provides detailed descriptions of the procedures needed to fabricate multilayered master molds using our CBMA. Finally, the capabilities of the CBMA are demonstrated by fabricating two- and three-layer masters for micro-scale devices, commonly encountered in biomicrofluidic applications. The former is a flow-free chemical gradient generator, and the latter is an addressable microfluidic stencil. Scanning electron microscopy is used to confirm that the master molds contain the intended features of different heights.
Directed cell migration in complex micro-environments, such as in vivo pores, is important for predicting locations of artificial tissue growth and optimizing scaffold architectures. Yet, the directional decisions of cells facing multiple physiochemical cues have not been characterized. Hence, we aim to provide a ranking of the relative importance of the following cues to the decision-making of individual fibroblast cells: chemoattractant concentration gradient, channel width, mitosis, and contact-guidance. In this study, bifurcated micro-channels with branches of different widths were created. Fibroblasts were then allowed to travel across these geometries by following a gradient of platelet-derived growth factor-BB (PDGF-BB) established inside the channels. Subsequently, a combination of statistical analysis and image-based diffusion modeling was used to report how the presence of multiple complex migration cues, including cell-cell influences, affect the fibroblast decision-making. It was found that the cells prefer wider channels over a higher chemoattractant gradient when choosing between asymmetric bifurcated branches. Only when the branches were symmetric in width did the gradient become predominant in directing which path the cell will take. Furthermore, when both the gradient and the channels were symmetric, contact guidance became important for guiding the cells in making directional choices. Based on these results we were able to rank these directional cues from most influential to the least as follows: mitosis > channel width asymmetry > chemoattractant gradient difference > and contact-guidance. It is expected that these results will benefit the fields of regenerative medicine, wound healing and developmental biology.
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