Breast Cancer Stem Cell Culture and Enrichment Using Poly(ε-Caprolactone) Scaffolds.

The cancer stem cell (CSC) population displays self-renewal capabilities, resistance to conventional therapies, and a tendency to post-treatment recurrence. Increasing knowledge about CSCs' phenotype and functions is needed to investigate new therapeutic strategies against the CSC population. Here, poly(ε-caprolactone) (PCL), a biocompatible polymer free of toxic dye, has been used to fabricate scaffolds, solid structures suitable for 3D cancer cell culture. It has been reported that scaffold cell culture enhances the CSCs population. A RepRap BCN3D+ printer and 3 mm PCL wire were used to fabricate circular scaffolds. PCL design and fabrication parameters were first determined and then optimized considering several measurable variables of the resulting scaffolds. MCF7 breast carcinoma cell line was used to assess scaffolds adequacy for 3D cell culture. To evaluate CSC enrichment, the Mammosphere Forming Index (MFI) was performed in 2D and 3D MCF7 cultures. Results showed that the 60° scaffolds were more suitable for 3D culture than the 45° and 90° ones. Moreover, 3D culture experiments, in adherent and non-adherent conditions, showed a significant increase in MFI compared to 2D cultures (control). Thus, 3D cell culture with PCL scaffolds could be useful to improve cancer cell culture and enrich the CSCs population.

A novel approach to low-cost, rapid and simultaneous colorimetric detection of multiple analytes using 3D printed microfluidic channels.

This research paper presents an inventive technique to swiftly create microfluidic channels on distinct membrane papers, enabling colorimetric drug detection. Using a modified DIY RepRap 3D printer with a syringe pump, microfluidic channels (µPADs) are crafted on a flexible nylon-based substrate. This allows simultaneous detection of four common drugs with a single reagent. An optimized blend of polydimethylsiloxane (PDMS) dissolved in hexane is used to create hydrophobic channels on various filter papers. The PDMS-hexane mixture infiltrates the paper's pores, forming hydrophobic barriers that confine liquids within the channels. These barriers are cured on the printer's hot plate, controlling channel width and preventing spreading. Capillary action drives fluid along these paths without spreading. This novel approach provides a versatile solution for rapid microfluidic channel creation on membrane papers. The DIY RepRap 3D printer integration offers precise control and faster curing. The PDMS-hexane solution accurately forms hydrophobic barriers, containing liquids within desired channels. The resulting microfluidic system holds potential for portable, cost-effective drug detection and various sensing applications.

Parametric nasopharyngeal swab for sampling COVID-19 and other respiratory viruses: Open source design, SLA 3-D printing and UV curing system.

Access to nasopharyngeal swabs for sampling remain a bottleneck in some regions for COVID-19 testing. This study develops a distributed manufacturing solution using only an open source manufacturing tool chain consisting of two types of open source 3-D printing and batch UV curing, and provides a parametric fully free design of a nasopharyngeal swab. The swab was designed using parametric OpenSCAD in two components (a head with engineered break point and various handles), which has several advantages: i) minimizing print time on relatively slow SLA printers, ii) enabling the use of smaller print volume open source SLA printers, iii) reducing the amount of relatively expensive UV resin, and iv) enabling production of handle on more accessible material extrusion 3-D printers. A modular open source UV LED box was designed, fabricated for $45 and tested for batch curing. Swabs can be fabricated for $0.06-$0.12/swab. The results of the mechanical validation tests showed that the swabs could withstand greater forces than would be expected in normal clinical use. The swabs were also able to absorb a significant amounts of synthetic mucus materials and passed abrasion and handling tests. The results show the open source swab are promising candidates for clinical trials.

Conversion of self-contained breathing apparatus mask to open source powered air-purifying particulate respirator for fire fighter COVID-19 response.

To assist firefighters and other first responders to use their existing equipment for respiration during the COVID-19 pandemic without using single-use, low-supply, masks, this study outlines an open source kit to convert a 3M-manufactured Scott Safety self-contained breathing apparatus (SCBA) into a powered air-purifying particulate respirator (PAPR). The open source PAPR can be fabricated with a low-cost 3-D printer and widely available components for less than $150, replacing commercial conversion kits saving 85% or full-fledged proprietary PAPRs saving over 90%. The parametric designs allow for adaptation to other core components and can be custom fit specifically to fire-fighter equipment, including their suspenders. The open source PAPR has controllable air flow and its design enables breathing even if the fan is disconnected or if the battery dies. The open source PAPR was tested for air flow as a function of battery life and was found to meet NIOSH air flow requirements for 4 h, which is 300% over expected regular use.

Open source high-temperature RepRap for 3-D printing heat-sterilizable PPE and other applications.

Thermal sterilization is generally avoided for 3-D printed components because of the relatively low deformation temperatures for common thermoplastics used for material extrusion-based additive manufacturing. 3-D printing materials required for high-temperature heat sterilizable components for COVID-19 and other applications demands 3-D printers with heated beds, hot ends that can reach higher temperatures than polytetrafluoroethylene (PTFE) hot ends and heated chambers to avoid part warping and delamination. There are several high temperature printers on the market, but their high costs make them inaccessible for full home-based distributed manufacturing required during pandemic lockdowns. To allow for all these requirements to be met for under $1000, the Cerberus - an open source three-headed self-replicating rapid prototyper (RepRap) was designed and tested with the following capabilities: i) 200 °C-capable heated bed, ii) 500 °C-capable hot end, iii) isolated heated chamber with 1 kW space heater core and iv) mains voltage chamber and bed heating for rapid start. The Cereberus successfully prints polyetherketoneketone (PEKK) and polyetherimide (PEI, ULTEM) with tensile strengths of 77.5 and 80.5 MPa, respectively. As a case study, open source face masks were 3-D printed in PEKK and shown not to warp upon widely home-accessible oven-based sterilization.

Partially RepRapable automated open source bag valve mask-based ventilator.

This study describes the development of a simple and easy-to-build portable automated bag valve mask (BVM) compression system, which, during acute shortages and supply chain disruptions can serve as a temporary emergency ventilator. The resuscitation system is based on the Arduino controller with a real-time operating system installed on a largely RepRap 3-D printable parametric component-based structure. The cost of the materials for the system is under $170, which makes it affordable for replication by makers around the world. The device provides a controlled breathing mode with tidal volumes from 100 to 800 mL, breathing rates from 5 to 40 breaths/minute, and inspiratory-to-expiratory ratio from 1:1 to 1:4. The system is designed for reliability and scalability of measurement circuits through the use of the serial peripheral interface and has the ability to connect additional hardware due to the object-oriented algorithmic approach. Experimental results after testing on an artificial lung for peak inspiratory pressure (PIP), respiratory rate (RR), positive end-expiratory pressure (PEEP), tidal volume, proximal pressure, and lung pressure demonstrate repeatability and accuracy exceeding human capabilities in BVM-based manual ventilation. Future work is necessary to further develop and test the system to make it acceptable for deployment outside of emergencies such as with COVID-19 pandemic in clinical environments, however, the nature of the design is such that desired features are relatively easy to add using protocols and parametric design files provided.

Open source arc analyzer: Multi-sensor monitoring of wire arc additive manufacturing.

Low-cost high-resolution metal 3-D printing remains elusive for the scientific community. Low-cost gas metal arc wire (GMAW)-based 3-D printing enables wire arc additive manufacturing (WAAM) for near net shape applications, but has limited resolution due to the complexities of the arcing process. To begin to monitor and thus control these complexities, the initial designs of the open source GMAW 3-D printer have evolved to include current and voltage monitoring. Building on this prior work, in this study, the design, fabrication and use of the open source arc analyzer is described. The arc analyzer is a multi-sensor monitoring system for quantifying the processing during WAAM, which includes voltage, current, sound, light intensity, radio frequency, and temperature data outputs. The open source arc analyzer is tested here on aluminum WAAM by varying wire feed rate and measuring the resultant changes in the sensor data. Visual inspection and microstructural analysis of the printed samples looking for the presence of porosity are used as the physical indicators of quality. The value of the sensors was assessed and the most impactful sensors were found to be the light and radio frequency sensors, which showed arc extinction events and a characteristic "good weld" peak frequency.

Economic savings for scientific free and open source technology: A review.

Both the free and open source software (FOSS) as well as the distributed digital manufacturing of free and open source hardware (FOSH) has shown particular promise among scientists for developing custom scientific tools. Early research found substantial economic savings for these technologies, but as the open source design paradigm has grown by orders of magnitude it is possible that the savings observed in the early work was isolated to special cases. Today there are examples of open source technology for science in the vast majority of disciplines and several resources dedicated specifically to publishing them. Do the tremendous economic savings observed earlier hold today? To answer that question, this study evaluates free and open source technologies in the two repositories compared to proprietary functionally-equivalent tools as a function of their use of Arduino-based electronics, RepRap-class 3-D printing, as well as the combination of the two. The results of the review find overwhelming evidence for a wide range of scientific tools, that open source technologies provide economic savings of 87% compared to equivalent or lesser proprietary tools. These economic savings increased slightly to 89% for those that used Arduino technology and even more to 92% for those that used RepRap-class 3-D printing. Combining both Arduino and 3-D printing the savings averaged 94% for free and open source tools over commercial equivalents. The results provide strong evidence for financial support of open source hardware and software development for the sciences. Given the overwhelming economic advantages of free and open source technologies, it appears financially responsible to divert funding of proprietary scientific tools and their development in favor of FOSH. Policies were outlined that provide nations with a template for strategically harvesting the opportunities provided by the free and open source paradigm.

Design and implementation of a low cost, modular, adaptable and open-source XYZ positioning system for neurophysiology.

In recent years, open-source 3D printing technologies have become increasingly applied to biological research. We have created a fully open-source, versatile and low cost XYZ positioning system using 3D printer components. As this system is controlled by a Python3 based operating system running on a Raspberry Pi 3 Model B, its behaviour can be adapted to meet multiple needs in neurophysiology. We have developed two main applications of this system. First, we have created an automated microscopy script that links seamlessly with image stitching plugins in ImageJ (Fiji) allowing the user to create high resolution montages. Second, we have created a series of movement scripts allowing the application of graded rates of stretch to muscle spindles. Here we outline the construction and implementation of this system and discuss how we have utilised this tool in our research.

Design of a Scaffold Parameter Selection System with Additive Manufacturing for a Biomedical Cell Culture.

Open-source 3D printers mean objects can be quickly and efficiently produced. However, design and fabrication parameters need to be optimized to set up the correct printing procedure; a procedure in which the characteristics of the printing materials selected for use can also influence the process. This work focuses on optimizing the printing process of the open-source 3D extruder machine RepRap, which is used to manufacture poly(ε-caprolactone) (PCL) scaffolds for cell culture applications. PCL is a biocompatible polymer that is free of toxic dye and has been used to fabricate scaffolds, i.e., solid structures suitable for 3D cancer cell cultures. Scaffold cell culture has been described as enhancing cancer stem cell (CSC) populations related to tumor chemoresistance and/or their recurrence after chemotherapy. A RepRap BCN3D+ printer and 3 mm PCL wire were used to fabricate circular scaffolds. Design and fabrication parameters were first determined with SolidWorks and Slic3r software and subsequently optimized following a novel sequential flowchart. In the flowchart described here, the parameters were gradually optimized step by step, by taking several measurable variables of the resulting scaffolds into consideration to guarantee high-quality printing. Three deposition angles (45°, 60° and 90°) were fabricated and tested. MCF-7 breast carcinoma cells and NIH/3T3 murine fibroblasts were used to assess scaffold adequacy for 3D cell cultures. The 60° scaffolds were found to be suitable for the purpose. Therefore, PCL scaffolds fabricated via the flowchart optimization with a RepRap 3D printer could be used for 3D cell cultures and may boost CSCs to study new therapeutic treatments for this malignant population. Moreover, the flowchart defined here could represent a standard procedure for non-engineers (i.e., mainly physicians) when manufacturing new culture systems is required.