Electrospun nanofibers for 3-D cancer models, diagnostics, and therapy.

As one of the leading causes of global mortality, cancer has prompted extensive research and development to advance efficacious drug discovery, sustained drug delivery and improved sensitivity in diagnosis. Towards these applications, nanofibers synthesized by electrospinning have exhibited great clinical potential as a biomimetic tumor microenvironment model for drug screening, a controllable platform for localized, prolonged drug release for cancer therapy, and a highly sensitive cancer diagnostic tool for capture and isolation of circulating tumor cells in the bloodstream and for detection of cancer-associated biomarkers. This review provides an overview of applied nanofiber design with focus on versatile electrospinning fabrication techniques. The influence of topographical, physical, and biochemical properties on the function of nanofiber assemblies is discussed, as well as current and foreseeable barriers to the clinical translation of applied nanofibers in the field of oncology.

Crosslinked Chitosan-PEG Hydrogel for Culture of Human Glioblastoma Cell Spheroids and Drug Screening.

Two-dimensional monolayer cell cultures are routinely utilized for preclinical cancer drug screening, but the results often do not translate well when drugs are tested in vivo. To address this limitation, a biocompatible chitosan-PEG hydrogel (CSPG gel) was synthesized to create a gel that can be easily dispensed into 96-well plates at room temperature and neutral pH. The stiffness of this gel was tailored to be within the stiffness range of human glioblastoma tissue to promote the formation of tumor spheroids. Differences in cell morphology, proliferation rate, and dose-dependent drug cytotoxicity were compared among cell spheroids grown on CSPG gels, cells in monolayer culture on tissue culture polystyrene and cells cultured on Matrigel. Tumor spheroids on CSPG gels displayed statistically significantly greater resistance to chemotherapeutics than in the conditions where cells did not form spheroids. Gene expression analysis suggests that resistance of cells on CSPG gels to the therapy may be partially attributed to upregulation of ATP-binding cassette transporters and downregulation of DNA mismatch repair genes, which was stimulated by spheroid formation. These findings suggest CSPG gel generates tumor spheroids that better reflect the malignant behavior of GBM and provides a cost-effective substrate for preclinical, high-throughput screening of potential cancer therapeutics.

Chitosan-Poly(caprolactone) Nanofibers for Skin Repair.

Dermal wounds, both acute and chronic, represent a significant clinical challenge and therefore the development of novel biomaterial-based skin substitutes to promote skin repair is essential. Nanofibers have garnered attention as materials to promote skin regeneration due to the similarities in morphology and dimensionality between nanofibers and native extracellular matrix proteins, which are critical in guiding cutaneous wound healing. Electrospun chitosan-poly(caprolactone) (CPCL) nanofiber scaffolds, which combine the important intrinsic biological properties of chitosan and the mechanical integrity and stability of PCL, were evaluated as skin tissue engineering scaffolds using a mouse cutaneous excisional skin defect model. Gross assessment of wound size and measurement of defect recovery over time as well as histological evaluation of wound healing showed that CPCL nanofiber scaffolds increased wound healing rate and promoted more complete wound closure as compared with Tegaderm, a commercially available occlusive dressing. CPCL nanofiber scaffolds represent a biomimetic approach to skin repair by serving as an immediately available provisional matrix to promote wound closure. These nanofiber scaffolds may have significant potential as a skin substitute or as the basis for more complex skin tissue engineering constructs involving integration with biologics.

PEG-chitosan hydrogel with tunable stiffness for study of drug response of breast cancer cells.

Mechanical properties of the extracellular matrix have a profound effect on the behavior of anchorage-dependent cells. However, the mechanisms that define the effects of matrix stiffness on cell behavior remains unclear. Therefore, the development and fabrication of synthetic matrices with well-defined stiffness is invaluable for studying the interactions of cells with their biophysical microenvironment in vitro. We demonstrate a methoxypolyethylene glycol (mPEG)-modified chitosan hydrogel network where hydrogel stiffness can be easily modulated under physiological conditions by adjusting the degree of mPEG grafting onto chitosan (PEGylation). We show that the storage modulus of the hydrogel increases as PEGylation decreases and the gels exhibit instant self-recovery after deformation. Breast cancer cells cultured on the stiffest hydrogels adopt a more malignant phenotype with increased resistance to doxorubicin as compared with cells cultured on tissue culture polystyrene or Matrigel. This work demonstrates the utility of mPEG-modified chitosan hydrogel, with tunable mechanical properties, as an improved replacement of conventional culture system for in vitro characterization of breast cancer cell phenotype and evaluation of cancer therapies.

High-throughput and high-yield fabrication of uniaxially-aligned chitosan-based nanofibers by centrifugal electrospinning.

The inability to produce large quantities of nanofibers has been a primary obstacle in advancement and commercialization of electrospinning technologies, especially when aligned nanofibers are desired. Here, we present a high-throughput centrifugal electrospinning (HTP-CES) system capable of producing a large number of highly-aligned nanofiber samples with high-yield and tunable diameters. The versatility of the design was revealed when bead-less nanofibers were produced from copolymer chitosan/polycaprolactone (C-PCL) solutions despite variations in polymer blend composition or spinneret needle gauge. Compared to conventional electrospinning techniques, fibers spun with the HTP-CES not only exhibited superior alignment, but also better diameter uniformity. Nanofiber alignment was quantified using Fast Fourier Transform (FFT) analysis. In addition, a concave correlation between the needle diameter and resultant fiber diameter was identified. This system can be easily scaled up for industrial production of highly-aligned nanofibers with tunable diameters that can potentially meet the requirements for various engineering and biomedical applications.

Chitosan-PEG hydrogel with sol-gel transition triggerable by multiple external stimuli.

Smart hydrogels play an increasingly important role in biomedical applications, since materials that are both biocompatible and multi-stimuli-responsive are highly desirable. A simple, organic solvent-free method is presented to synthesize a biocompatible hydrogel that undergoes a sol-gel transition in response to multiple stimuli. Methoxy-poly(ethylene glycol) (mPEG) is modified into carboxylic-acid-terminated-methoxy-poly(ethylene glycol) (mPEG-acid), which is then grafted onto chitosan via amide linkages yielding mPEG-g-chitosan. Grafting of mPEG onto hydrophobic chitosan imparts hydrophilic properties to the resultant polymer. The mPEG-g-chitosan gel exhibits a controllable multi-stimuli-responsive property. The balance between hydrophilicity and hydrophobicity is believed to confer mPEG-g-chitosan with stimuli-responsive behavior. The effect of salt concentration, solute concentration, temperature, and pH on the sol-gel transition of mPEG-g-chitosan is evaluated and the underlying mechanisms of mPEG-g-chitosan polymer packing and gelation property is discussed.

Chitosan-based scaffolds for bone tissue engineering.

Bone defects requiring grafts to promote healing are frequently occurring and costly problems in health care. Chitosan, a biodegradable, naturally occurring polymer, has drawn considerable attention in recent years as scaffolding material in tissue engineering and regenerative medicine. Chitosan is especially attractive as a bone scaffold material because it supports the attachment and proliferation of osteoblast cells as well as formation of mineralized bone matrix. In this review, we discuss the fundamentals of bone tissue engineering and the unique properties of chitosan as a scaffolding material to treat bone defects for hard tissue regeneration. We present the common methods for fabrication and characterization of chitosan scaffolds, and discuss the influence of material preparation and addition of polymeric or ceramic components or biomolecules on chitosan scaffold properties such as mechanical strength, structural integrity, and functional bone regeneration. Finally, we highlight recent advances in development of chitosan-based scaffolds with enhanced bone regeneration capability.

Biomaterials for high-throughput stem cell culture.

A cell's microenvironment plays a primary role in defining cell fate during tissue development, physiological function, and pathological dysfunction. Understanding the key components and interactions within these microenvironments is critical for effective use of stem cells for disease modeling and therapeutic applications. Yet cell microenvironments are difficult to study, as there are tens or hundreds of parameters that can influence cell behavior simultaneously. Additionally, parameters such as cell-cell interactions, cell-ECM interactions, cell shape, soluble signals, and mechanical forces vary dynamically in 3-dimensional space and time. The number of relevant experimental conditions in these cell-based biological systems quickly becomes intractable using standard experimental platforms and techniques. A new set of strategies involving high-throughput experimental formats and 3-dimensional culture is required to achieve significant progress in understanding and exploiting stem cell biology. This mini-review describes bioengineering approaches that are enabling for high-throughput stem cell culture, screening and analysis.