Toward 3D printed microfluidic artificial lungs for respiratory support.

Microfluidic artificial lungs (µALs) are a new class of membrane oxygenators. Compared to traditional hollow-fiber oxygenators, µALs closely mimic the alveolar microenvironment due to their size-scale and promise improved gas exchange efficiency, hemocompatibility, biomimetic blood flow networks, and physiologically relevant blood vessel pressures and shear stresses. Clinical translation of µALs has been stalled by restrictive microfabrication techniques that limit potential artificial lung geometries, overall device size, and throughput. To address these limitations, a high-resolution Asiga MAX X27 UV digital light processing (DLP) 3D printer and custom photopolymerizable polydimethylsiloxane (PDMS) resin were used to rapidly manufacture small-scale µALs via vat photopolymerization (VPP). Devices were designed in SOLIDWORKS with 500 blood channels and 252 gas channels, where gas and blood flow channels were oriented orthogonally and separated by membranes on the top and bottom, permitting two-sided gas exchange. Successful devices were post-processed to remove uncured resin from microchannels and assembled with external tubing in preparation for gas exchange performance testing with ovine whole blood. 3D printed channel dimensions were 172 µm-tall × 320 µm-wide, with 62 µm-thick membranes and 124 µm-wide support columns. Measured outlet blood oxygen saturation (SO2) agreed with theoretical models and rated flow of the device was 1 mL min-1. Blood side pressure drop was 1.58 mmHg at rated flow. This work presents the highest density of 3D printed microchannels in a single device, one of the highest CO2 transfer efficiencies of any artificial lung to date, and a promising approach to translate µALs one step closer to the clinic.

CanVaxKB: a web-based cancer vaccine knowledgebase.

Cancer vaccines have been increasingly studied and developed to prevent or treat various types of cancers. To systematically survey and analyze different reported cancer vaccines, we developed CanVaxKB (https://violinet.org/canvaxkb), the first web-based cancer vaccine knowledgebase that compiles over 670 therapeutic or preventive cancer vaccines that have been experimentally verified to be effective at various stages. Vaccine construction and host response data are also included. These cancer vaccines are developed against various cancer types such as melanoma, hematological cancer, and prostate cancer. CanVaxKB has stored 263 genes or proteins that serve as cancer vaccine antigen genes, which we have collectively termed 'canvaxgens'. Top three mostly used canvaxgens are PMEL, MLANA and CTAG1B, often targeting multiple cancer types. A total of 193 canvaxgens are also reported in cancer-related ONGene, Network of Cancer Genes and/or Sanger Cancer Gene Consensus databases. Enriched functional annotations and clusters of canvaxgens were identified and analyzed. User-friendly web interfaces are searchable for querying and comparing cancer vaccines. CanVaxKB cancer vaccines are also semantically represented by the community-based Vaccine Ontology to support data exchange. Overall, CanVaxKB is a timely and vital cancer vaccine source that facilitates efficient collection and analysis, further helping researchers and physicians to better understand cancer mechanisms.

Low-Viscosity Polydimethylsiloxane Resin for Facile 3D Printing of Elastomeric Microfluidics.

Microfluidics is a rapidly advancing technology with expansive applications but has been restricted by slow, laborious fabrication techniques for polydimethylsiloxane (PDMS)-based devices. Currently, 3D printing promises to address this challenge with high-resolution commercial systems but is limited by a lack of material advances in generating high-fidelity parts with micron-scale features. To overcome this limitation, a low-viscosity, photopolymerizable PDMS resin was formulated with a methacrylate-PDMS copolymer, methacrylate-PDMS telechelic polymer, photoabsorber, Sudan I, photosensitizer, 2-isopropylthioxanthone, and a photoinitiator, 2,4,6-trimethyl benzoyl diphenylphosphine oxide. The performance of this resin was validated on a digital light processing (DLP) 3D printer, an Asiga MAX X27 UV. Resin resolution, part fidelity, mechanical properties, gas permeability, optical transparency, and biocompatibility were investigated. This resin produced resolved, unobstructed channels as small as 38.4 (±5.0) µm tall and membranes as thin as 30.9 (±0.5) µm. The printed material had an elongation at break of 58.6% ± 18.8%, Young's modulus of 0.30 ± 0.04 MPa, and was highly permeable to O2 (596 Barrers) and CO2 (3071 Barrers). Following the ethanol extraction of the unreacted components, this material demonstrated optical clarity and transparency (>80% transmission) and viability as a substrate for in vitro tissue culture. This paper presents a high-resolution, PDMS 3D-printing resin for the facile fabrication of microfluidic and biomedical devices.

Advancing 3D-Printed Microfluidics: Characterization of a Gas-Permeable, High-Resolution PDMS Resin for Stereolithography.

The rapid expansion of microfluidic applications in the last decade has been curtailed by slow, laborious microfabrication techniques. Recently, microfluidics has been explored with additive manufacturing (AM), as it has gained legitimacy for producing end-use products and 3D printers have improved resolution capabilities. While AM satisfies many shortcomings with current microfabrication techniques, there still lacks a suitable replacement for the most used material in microfluidic devices, poly(dimethylsiloxane) (PDMS). Formulation of a gas-permeable, high-resolution PDMS resin was developed using a methacrylate-PDMS copolymer and the novel combination of a photoabsorber, Sudan I, and photosensitizer, 2-Isopropylthioxanthone. Resin characterization and 3D printing were performed using a commercially available DLP-SLA system. A previously developed math model, mechanical testing, optical transmission, and gas-permeability testing were performed to validate the optimized resin formula. The resulting resin has Young's modulus of 11.5 MPa, a 12% elongation at break, and optical transmission of >75% for wavelengths between 500 and 800 nm after polymerization, and is capable of creating channels as small as 60 µm in height and membranes as thin as 20 µm. The potential of AM is just being realized as a fabrication technique for microfluidics as developments in material science and 3D printing technologies continue to push the resolution capabilities of these systems.

Engineered 3D Model of Cancer Stem Cell Enrichment and Chemoresistance.

Intraperitoneal dissemination of ovarian cancers is preceded by the development of chemoresistant tumors with malignant ascites. Despite the high levels of chemoresistance and relapse observed in ovarian cancers, there are no in vitro models to understand the development of chemoresistance in situ. METHOD: We describe a highly integrated approach to establish an in vitro model of chemoresistance and stemness in ovarian cancer, using the 3D hanging drop spheroid platform. The model was established by serially passaging non-adherent spheroids. At each passage, the effectiveness of the model was evaluated via measures of proliferation, response to treatment with cisplatin and a novel ALDH1A inhibitor. Concomitantly, the expression and tumor initiating capacity of cancer stem-like cells (CSCs) was analyzed. RNA-seq was used to establish gene signatures associated with the evolution of tumorigenicity, and chemoresistance. Lastly, a mathematical model was developed to predict the emergence of CSCs during serial passaging of ovarian cancer spheroids. RESULTS: Our serial passage model demonstrated increased cellular proliferation, enriched CSCs, and emergence of a platinum resistant phenotype. In vivo tumor xenograft assays indicated that later passage spheroids were significantly more tumorigenic with higher CSCs, compared to early passage spheroids. RNA-seq revealed several gene signatures supporting the emergence of CSCs, chemoresistance, and malignant phenotypes, with links to poor clinical prognosis. Our mathematical model predicted the emergence of CSC populations within serially passaged spheroids, concurring with experimentally observed data. CONCLUSION: Our integrated approach illustrates the utility of the serial passage spheroid model for examining the emergence and development of chemoresistance in ovarian cancer in a controllable and reproducible format.

Personalized Medicine-Based Approach to Model Patterns of Chemoresistance and Tumor Recurrence Using Ovarian Cancer Stem Cell Spheroids.

Purpose: Chemoresistant ovarian cancers grow in suspension within the ascites fluid. To screen the effect of chemotherapeutics and biologics on resistant ovarian cancers with a personalized basis, we developed a 3D hanging drop spheroid platform.Experimental Design: We initiated spheroids with primary aldehyde dehydrogenase-positive (ALDH+) CD133+ ovarian cancer stem cells (OvCSC) from different patient samples and demonstrated that stem cell progeny from harvested spheroids was similar to the primary tumor. OvCSC spheroids were utilized to initiate tumors in immunodeficient mice. Drug responses to cisplatin and ALDH-targeting compound or JAK2 inhibitor determined whether the OvCSC population within the spheroids could be targeted. Cells that escaped therapy were isolated and used to initiate new spheroids and model tumor reemergence in a personalized manner.Results: OvCSC spheroids from different patients exhibited varying and personalized responses to chemotherapeutics. Xenografts were established from OvCSC spheroids, even with a single spheroid. Distinct responses to therapy were observed in distinct primary tumor xenografts similar to those observed in spheroids. Spheroids resistant to cisplatin/ALDH inhibitor therapy had persistent, albeit lower ALDH expression and complete loss of CD133 expression, whereas those resistant to cisplatin/JAK2 inhibitor therapy were enriched for ALDH+ cells.Conclusions: Our 3D hanging drop suspension platform can be used to propagate primary OvCSCs that represent individual patient tumors effectively by differentiating in vitro and initiating tumors in mice. Therefore, our platform can be used to study cancer stem cell biology and model tumor reemergence to identify new targeted therapeutics from an effective personalized medicine standpoint. Clin Cancer Res; 23(22); 6934-45. ©2017 AACR.

Printing of small molecular medicines from the vapor phase.

There is growing need to develop efficient methods for early-stage drug discovery, continuous manufacturing of drug delivery vehicles, and ultra-precise dosing of high potency drugs. Here we demonstrate the use of solvent-free organic vapor jet printing to deposit nanostructured films of small molecular pharmaceutical ingredients, including caffeine, paracetamol, ibuprofen, tamoxifen, BAY 11-7082 and fluorescein, with accuracy on the scale of micrograms per square centimeter, onto glass, Tegaderm, Listerine tabs, and stainless steel microneedles. The printed films exhibit similar crystallographic order and chemistry as the original powders; controlled, order-of-magnitude enhancements of dissolution rate are observed relative to powder-form particles. In vitro treatment of breast and ovarian cancer cell cultures in aqueous media by tamoxifen and BAY 11-7082 films shows similar behavior to drugs pre-dissolved in dimethyl sulfoxide. The demonstrated precise printing of medicines as films, without the use of solvents, can accelerate drug screening and enable continuous manufacturing, while enhancing dosage accuracy.Traditional approaches used in the pharmaceutical industry are not precise or versatile enough for customized medicine formulation and manufacture. Here the authors produce a method to form coatings, with accurate dosages, as well as a means of closely controlling dissolution kinetics.