Junghyuk Ko

Fabrication of poly (ϵ-caprolactone) microfiber scaffolds with varying topography and mechanical properties for stem cell-based tissue engineering applications

Highly porous poly (ϵ-caprolactone) microfiber scaffolds can be fabricated using electrospinning for tissue engineering applications. Melt electrospinning produces such scaffolds by direct deposition of a polymer melt instead of dissolving the polymer in a solvent as performed during solution electrospinning. The objective of this study was to investigate the significant parameters associated with the melt electrospinning process that influence fiber diameter and scaffold morphology, including processing temperature, collection distance, applied, voltage and nozzle size. The mechanical properties of these microfiber scaffolds varied with microfiber diameter. Additionally, the porosity of scaffolds was determined by combining experimental data with mathematical modeling. To test the cytocompatability of these fibrous scaffolds, we seeded neural progenitors derived from murine R1 embryonic stem cell lines onto these scaffolds, where they could survive, migrate, and differentiate into neurons; demonstrating the potential of these melt electrospun scaffolds for tissue engineering applications.

Electrospun biomaterial scaffolds with varied topographies for neuronal differentiation of human-induced pluripotent stem cells

In this study, we investigated the effect of micro and nanoscale scaffold topography on promoting neuronal differentiation of human induced pluripotent stem cells (iPSCs) and directing the resulting neuronal outgrowth in an organized manner. We used melt electrospinning to fabricate poly (ε-caprolactone) (PCL) scaffolds with loop mesh and biaxial aligned microscale topographies. Biaxial aligned microscale scaffolds were further functionalized with retinoic acid releasing PCL nanofibers using solution electrospinning. These scaffolds were then seeded with neural progenitors derived from human iPSCs. We found that smaller diameter loop mesh scaffolds (43.7 ± 3.9 µm) induced higher expression of the neural markers Nestin and Pax6 compared to thicker diameter loop mesh scaffolds (85 ± 4 µm). The loop mesh and biaxial aligned scaffolds guided the neurite outgrowth of human iPSCs along the topographical features with the maximum neurite length of these cells being longer on the biaxial aligned scaffolds. Finally, our novel bimodal scaffolds also supported the neuronal differentiation of human iPSCs as they presented both physical and chemical cues to these cells, encouraging their differentiation. These results give insight into how physical and chemical cues can be used to engineer neural tissue.

Development of a glial cell-derived neurotrophic factor-releasing artificial dura for neural tissue engineering applications

Encapsulated electrospun nanofibers can serve as an artificial dura mater, the membrane that surrounds the brain and spinal cord, due to their desirable drug delivery properties. Such nanofiber scaffolds can be used to deliver drugs such as glial cell-derived neurotrophic factor (GDNF). GDNF promotes the survival of both dopaminergic and motor neurons, making it an important target for treatment of central nervous system injuries and disorders. This work focuses on designing a novel class of encapsulated poly(ε-caprolactone) (PCL) nanofiber scaffolds with different topographies (random and aligned) that generate controlled release of GDNF to potentially serve as a suitable substitute for the dura mater during neurosurgical procedures. Random and aligned scaffolds fabricated using solution electrospinning were characterized for their physical properties and their ability to release GDNF over one month. GDNF bioactivity was confirmed using a PC12 cell assay with the highest concentrations of released GDNF (∼341 ng mL-1 GDNF) inducing the highest levels of neurite extension (∼556 μm). To test the cytocompatibility of aligned GDNF encapsulated PCL nanofibers, we successfully seeded neural progenitors derived from human induced pluripotent stem cells (hiPSCs) onto the scaffolds where they survived and differentiated into neurons. Overall, this research demonstrates the potential of such substrates to act as artificial dura while delivering bioactive GDNF in a controlled fashion. These scaffolds also support the culture and differentiation of hiPSC-derived neural progenitors, suggesting their biocompatibility with the cells of the central nervous system.

Using mathematical modeling to control topographical properties of poly (ε-caprolactone) melt electrospun scaffolds

Melt electrospinning creates fibrous scaffolds using direct deposition. The main challenge of melt electrospinning is controlling the topography of the scaffolds for tissue engineering applications. Mathematical modeling enables a better understanding of the parameters that determine the topography of scaffolds. The objective of this study is to build two types of mathematical models. First, we modeled the melt electrospinning process by incorporating parameters such as nozzle size, counter electrode distance and applied voltage that influence fiber diameter and scaffold porosity. Our second model describes the accumulation of the extruded microfibers on flat and round surfaces using data from the microfiber modeling. These models were validated through the use of experimentally obtained data. Scanning electron microscopy (SEM) was used to image the scaffolds and the fiber diameters were measured using Quartz-PCI Image Management Systems® in SEM to measure scaffold porosity.

Design and fabrication of auxetic stretchable force sensor for hand rehabilitation

Using a melt electrospinning technique, stretchable force sensors were designed for use in an application of hand rehabilitation. The main purpose of this study was to verify that the use of auxetic sensors improved hand rehabilitation practices when compared to their absence. For this study, novel stretchable poly (-caprolactone) (PCL) force sensors were fabricated into the following formations: auxetic microfiber sheets (AMSs), auxetic solid sheets (ASSs), microfiber sheets (MSs), and solid sheets (SSs). A femtosecond laser device was used to make an auxetic structure in the MSs and SSs. Subsequently, these sensors were coated with gold particles to make them conductive for the electrical current resistance assays. Through the cycles of applied stress and strain, auxetic structures were able to retain their original shape once these forces have been dissipated. This stretchable sensor could potentially measure applied external loads, resistance, and strain and could also be attachable to a desired substrate. In order to verify the workability and practicality of our designed sensors, we have attempted to use the sensors on a human hand. The AMS sensor had the highest sensitivity on measuring force and resistance among the four types of sensors. To our knowledge, this is the first study to form a stretchable force sensor using a melt electrospinning technique.

Biomaterial Strategies for Delivering Stem Cells as a Treatment for Spinal Cord Injury

Ongoing clinical trials are evaluating the use of stem cells as a way to treat traumatic spinal cord injury (SCI). However, the inhibitory environment present in the injured spinal cord makes it challenging to achieve the survival of these cells along with desired differentiation into the appropriate phenotypes necessary to regain function. Transplanting stem cells along with an instructive biomaterial scaffold can increase cell survival and improve differentiation efficiency. This study reviews the literature discussing different types of instructive biomaterial scaffolds developed for transplanting stem cells into the injured spinal cord. We have chosen to focus specifically on biomaterial scaffolds that direct the differentiation of neural stem cells and pluripotent stem cells since they offer the most promise for producing the cell phenotypes that could restore function after SCI. In terms of biomaterial scaffolds, this article reviews the literature associated with using hydrogels made from natural biomaterials and electrospun scaffolds for differentiating stem cells into neural phenotypes. It then presents new data showing how these different types of scaffolds can be combined for neural tissue engineering applications and provides directions for future studies.

Design and fabrication of auxetic PCL nanofiber membranes for biomedical applications

The main objective of this study was to fabricate poly (ε-caprolactone) (PCL)-based auxetic nanofiber membranes and characterize them for their mechanical and physicochemical properties. As a first step, the PCL nanofibers were fabricated by electrospinning with two different thicknesses of 40μm (called PCL thin membrane) and 180μm (called PCL thick membrane). In the second step, they were tailored into auxetic patterns using femtosecond laser cut technique. The physicochemical and mechanical properties of the auxetic nanofiber membranes were studied and compared with the conventional electrospun PCL nanofibers (non-auxetic nanofiber membranes) as a control. The results showed that there were no significant changes observed among them in terms of their chemical functionality and thermal property. However, there was a notable difference observed in the mechanical properties. For instance, the thin auxetic nanofiber membrane showed the magnitude of elongation almost ten times higher than the control, which clearly demonstrates the high flexibility of auxetic nanofiber membranes. This is because that the auxetic nanofiber membranes have lesser rigidity than the control nanofibers under the same load which could be due to the rotational motion of the auxetic structures. The major finding of this study is that the auxetic PCL nanofiber membranes are highly flexible (10-fold higher elongation capacity than the conventional PCL nanofibers) and have tunable mechanical properties. Therefore, the auxetic PCL nanofiber membranes may serve as a potent material in various biomedical applications, in particular, tissue engineering where scaffolds with mechanical cues play a major role.

Mathematical model for predicting topographical properties of poly (ε-caprolactone) melt electrospun scaffolds including the effects of temperature and linear transitional speed

Funding support from Natural Sciences and Engineering Research Council (NSERC) Discovery Grants is acknowledged. The authors would also like to acknowledge the Advanced Microscopy Facility at the University of Victoria.

Fabrication and Characterization of Nonwoven Auxetic Polymer Stent

Synthetic biomaterials have better controlled physical and mechanical properties and can be used to tailor both soft and hard tissues. A tiny, expandable mesh tubes called stents keep blood vessels open and allow blood flow and treat blockage to improve quality of patient’s life. The main focus of this work is to (i) fabricate a polymeric sheet of melt electrospun polycaprolactone microfibers; (ii) tailor auxetic geometry by micromachining on polycaprolactone microfiber and polycaprolactone sheet; (iii) fabricate a cylindrical tube to make auxetic stents. Final results for mechanical characterization and performance analysis of auxetic polymer stents are discussed. GRAPHICAL ABSTRACT

The effect of nozzle-exit-channel shape on resultant fiber diameter in melt-electrospinning

In recent decades, electrospinning using a molten poly (e-caprolactone) resin has gained attention for creating fibrous tissue scaffolds. The topography and diameter control of such electrospun microfibers is an important issue for their different applications in tissue engineering. Charge density, initial nozzle-exit-channel cross-sectional area, nozzle to collector distance, viscosity, and processing temperature are the most important input parameters that affect the final electrospun fiber diameters. In this paper we will show that the effect of nozzle-exit-channel shape is as important as the other effective parameters in a resultant fiber diameter. However, to the best of our knowledge, the effect of nozzle-exit-channel shapes on a resultant fiber diameter have not been studied before. Comparing rectangular and circular nozzles with almost the same exit-channel cross-sectional areas in a similar processing condition showed that using a rectangular nozzle resulted in decreasing final fiber diameter up to 50%. Furthermore, the effect of processing temperature on the final fiber topography was investigated.