Fabrication of Microtube-Embedded Chip to Mimic Blood-Brain Barrier Capillary Vessels.

Capillary vessels of the blood-brain barrier (BBB) regulate the transportation of solutes into the brain and provide defense against the disease-causing pathogens and neurotoxins present in the blood. Paradoxically, this regulation also prevents drug transportation into the brain. These unique characteristics of the BBB cause impediment in the treatment of neurological diseases. The development of preclinical models that mimic the BBB capillary vessel is crucial to investigate the complex transport mechanism. Microfluidics-based in vitro models are now extensively investigated for therapeutic applications due to the ability to create a tunable dynamic extracellular microenvironment. One of the main challenges of creating a BBB-on-a-chip is to recapitulate the tubular capillary structure. This chapter presents two novel fabrication methods for microfluidic devices embedded with tubular micro-channels that resemble the diameter and morphology of capillary vessels. These microfluidic devices can be seeded with cells for physiological and pathological studies to support future drug development.

Fabrication of Nanopores Polylactic Acid Microtubes by Core-Sheath Electrospinning for Capillary Vascularization.

There has been substantial progress in tissue engineering of biological substitutes for medical applications. One of the major challenges in development of complex tissues is the difficulty of creating vascular networks for engineered constructs. The diameter of current artificial vascular channels is usually at millimeter or submillimeter level, while human capillaries are about 5 to 10 µm in diameter. In this paper, a novel core-sheath electrospinning process was adopted to fabricate nanoporous microtubes to mimic the structure of fenestrated capillary vessels. A mixture of polylactic acid (PLA) and polyethylene glycol (PEO) was used as the sheath solution and PEO was used as the core solution. The microtubes were observed under a scanning electron microscope and the images were analyzed by ImageJ. The diameter of the microtubes ranged from 1-8 microns. The diameter of the nanopores ranged from 100 to 800 nm. The statistical analysis showed that the microtube diameter was significantly influenced by the PEO ratio in the sheath solution, pump rate, and the viscosity gradient between the sheath and the core solution. The electrospun microtubes with nanoscale pores highly resemble human fenestrated capillaries. Therefore, the nanoporous microtubes have great potential to support vascularization in engineered tissues.

Musculoskeletal Tissue Engineering Using Fibrous Biomaterials.

In tissue engineering, scaffolds should provide the topological and physical cues as native tissues to guide cell adhesion, growth, migration, and differentiation. Fibrous structure is commonly present in human musculoskeletal tissues, including muscles, tendons, ligaments, and cartilage. Biomimetic fibrous scaffolds are thus critical for musculoskeletal tissue engineering. Electrospinning is a versatile technique for fabricating nanofibers from a variety of biomaterials. However, conventional electrospinning can only generate 2D nanofiber mats. Postprocessing methods are often needed to create 3D electrospun nanofiber scaffolds. In this chapter, we present two novel electrospinning-based scaffold fabrication techniques, which can generate 3D nanofiber scaffolds in one-station process: divergence electrospinning and hybrid 3D printing with parallel electrospinning. These techniques can be applied for engineering tissues with aligned fiber structures.

ZnO Nanowire-Anchored Microfluidic Device With Herringbone Structure Fabricated by Maskless Photolithography.

The integration of nanomaterials in microfluidic devices has emerged as a new research paradigm. Microfluidic devices composed of ZnO nanowires have been developed for the collection of urine extracellular vesicles (EVs) at high efficiency and in situ extraction of various microRNAs (miRNAs). The devices can be used for diagnosing various diseases, including kidney diseases and cancers. A major research need for developing micro total analysis systems is to enhance extraction efficiency. This article presents a novel fabrication method for a herringbone-patterned microfluidic device anchored with ZnO nanowire arrays. The substrates with herringbone patterns were created by maskless photolithography. The ZnO nanowire arrays were grown on the substrates by chemical bathing. The patterned design was to introduce turbulent flows as opposed to laminar flow in traditional devices to increase the mixing and contact of the urine sample with ZnO nanowires. The device showed reduced flow rates compared with conventional planar microfluidic channels and successfully extracted urine EV-encapsulated miRNAs.

Laser engineered net shaping of antimicrobial and biocompatible titanium-silver alloys.

Post-surgery infection is one of the main causes of orthopedic implant failure. This paper presents a powder-feed 3D printing strategy for fabrication of silver (Ag) incorporated titanium (Ti) alloys as an antimicrobial solution for orthopedic implants. Alloys with various Ag concentration, ranging from 0.5% to 2% by weight, were fabricated through laser engineered net shaping (LENS) process. The composition and surface of the fabricated alloys were characterized through X-ray diffraction, energy-dispersive X-ray spectroscopy, and 3D surface profiling. The mechanical properties, antimicrobial performance, and biocompatibility of the alloys were also investigated. Results showed that LENS fabricated TiAg alloys had a marginally higher microhardness and a lower ductility compared to pure Ti. Within only 3 h, TiAg alloys significantly reduced the bacterial attachment of both gram-positive and gram-negative strains by one to four orders of magnitudes. These alloys also demonstrated excellent in-vitro biocompatibility to human osteosarcoma cells. For the first time, laser engineered net shaping (LENS) of TiAg alloy has been explored as an antimicrobial solution for orthopedic applications and showed great potential for biomedical instrumentation.

Hybrid Additive Microfabrication Scaffold Incorporated with Highly Aligned Nanofibers for Musculoskeletal Tissues.

BACKGROUND: Latest tissue engineering strategies for musculoskeletal tissues regeneration focus on creating a biomimetic microenvironment closely resembling the natural topology of extracellular matrix. This paper presents a novel musculoskeletal tissue scaffold fabricated by hybrid additive manufacturing method. METHODS: The skeleton of the scaffold was 3D printed by fused deposition modeling, and a layer of random or aligned polycaprolactone nanofibers were embedded between two frames. A parametric study was performed to investigate the effects of process parameters on nanofiber morphology. A compression test was performed to study the mechanical properties of the scaffold. Human fibroblast cells were cultured in the scaffold for 7 days to evaluate the effect of scaffold microstructure on cell growth. RESULTS: The tip-to-collector distance showed a positive correlation with the fiber alignment, and the electrospinning time showed a negative correlation with the fiber density. With reinforced nanofibers, the hybrid scaffold demonstrated superior compression strength compared to conventional 3D-printed scaffold. The hybrid scaffold with aligned nanofibers led to higher cell attachment and proliferation rates, and a directional cell organization. In addition, there was a nonlinear relationship between the fiber diameter/density and the cell actinfilament density. CONCLUSION: This hybrid biofabrication process can be established as a highly efficient and scalable platform to fabricate biomimetic scaffolds with patterned fibrous microstructure, and will facilitate future development of clinical solutions for musculoskeletal tissue regeneration.