Eugene D. Boland

TAILORING TISSUE ENGINEERING SCAFFOLDS USING ELECTROSTATIC PROCESSING TECHNIQUES: A STUDY OF POLY(GLYCOLIC ACID) ELECTROSPINNING

Poly(glycolic acid) (PGA) has long been a popular polymer in the tissue engineering field. PGA possesses many favorable properties such as biocompatibility, bioabsorbability, and tensile strength. The traditional fiber formation techniques of melt extrusion and cold-drawing are generally limited to fibers of 10–12 μm in diameter. Electrostatic spinning, or electrospinning, is an attractive approach for the production of much smaller diameter fibers which are of interest as tissue engineering scaffolds. We demonstrate the ability to control the fiber diameter of PGA as a function of solution concentration and fiber orientation, as well as show a correlation between the fiber orientation, elastic modulu, and strain to failure of PGA in a uniaxial model.

Direct-write Bioprinting Three-Dimensional Biohybrid Systems for Future Regenerative Therapies

Regenerative medicine seeks to repair or replace dysfunctional tissues with engineered biological or biohybrid systems. Current clinical regenerative models utilize simple uniform tissue constructs formed with cells cultured onto biocompatible scaffolds. Future regenerative therapies will require the fabrication of complex three-dimensional constructs containing multiple cell types and extracellular matrices. We believe bioprinting technologies will provide a key role in the design and construction of future engineered tissues for cell-based and regenerative therapies. This review describes the current state-of-the-art bioprinting technologies, focusing on direct-write bioprinting. We describe a number of process and device considerations for successful bioprinting of composite biohybrid constructs. In addition, we have provided baseline direct-write printing parameters for a hydrogel system (Pluronic F127) often used in cardiovascular applications. Direct-write dispensed lines (gels with viscosities ranging from 30 mPa s to greater than 600 × 10⁶ mPa s) were measured following mechanical and pneumatic printing via three commercially available needle sizes (20 ga, 25 ga, and 30 ga). Example patterns containing microvascular cells and isolated microvessel fragments were also bioprinted into composite 3D structures. Cells and vessel fragments remained viable and maintained in vitro behavior after incorporation into biohybrid structures. Direct-write bioprinting of biologicals provides a unique method to design and fabricate complex, multicomponent 3D structures for experimental use. We hope our design insights and baseline parameter descriptions of direct-write bioprinting will provide a useful foundation for colleagues to incorporate this 3D fabrication method into future regenerative therapies.

Electrospinning of Collagen Type II: A Feasibility Study

Collagen is the natural scaffolding found in all tissues and has been explored extensively for use as a tissue engineering scaffold with limited success. In this feasibility study, the electrospinning of collagen type II and subsequent chondrocyte seeding was investigated for potential use in cartilage tissue engineering. The electrospinning process utilized lyophilized, chicken sternal cartilage collagen type II suspended in 1,1,1,3,3,3 hexaflouro-2-propanol and demonstrated that collagen type II could be electrospun to form nonwoven fibrous mats composed of type II fibers that ranged from 110 nm to 1.8μm in diameter. The fiber diameter was dependant on the type II concentration in solution with a higher concentration producing the larger diameters. The preliminary chondrocyte seeding study demonstrated that electrospun collagen type II scaffolds support cell growth and are readily infiltrated. In conclusion, the feasibility of collagen type II electrospinning has been demonstrated and the novel scaffolds produced are composed of nano- to micron-scale fiber diameters that have been shown to be compatible with chondrocytes.

Electrospun nanofibre fibrinogen for urinary tract tissue reconstruction

The purpose of this study was to demonstrate that human bladder smooth muscle cells (HBSM) remodel electrospun fibrinogen mats. Fibrinogen scaffolds were electrospun and disinfected using standard methods. Scaffolds were seeded with 5 x 10(4) HBSM per scaffold. Cultures were supplemented with aprotinin concentrations of 0 KIU ml(-1) (no aprotinin), 100 KIU ml(-1) or 1000 KIU ml(-1) and incubated with twice weekly media changes. Samples were removed for evaluation at 1, 3, 7 and 14 days. Cultured scaffolds were evaluated with a WST-1 cell proliferation assay, scanning electron microscopy and histology. Cell culture demonstrated that HBSM readily migrated into and initiated remodelling of the electrospun fibrinogen scaffolds by deposition of collagen. Proliferation was suppressed during this initial phase with respect to a 2D control due to cell migration. Histology confirmed that proliferation increased during the later stages of remodelling. Remodelling was slower at higher aprotinin concentrations. These results demonstrate that HBSM rapidly remodel an electrospun fibrinogen scaffold and deposit native collagen. The process can be modulated using aprotinin, a protease inhibitor. These initial findings indicate that there is tremendous potential for electrospun fibrinogen as a urologic tissue engineering scaffold with the ultimate goal of producing an implantable acellular product that would promote cellular in-growth and in situ tissue regeneration.

Biomedical Nanoscience: Electrospinning Basic Concepts, Applications, and Classroom Demonstration

Electrospinning is an old polymer processing technique that has recently been rediscovered. It allows for the easy creation of nano- to micro-fibers that can be collected to form a non-woven structure, which can then be used to fabricate novel structures for various applications including tissue engineering scaffolds, clothing, drug delivery vehicles, and filtration media. Current research in our laboratories is focused on the processing of synthetic and biological polymers to create materials with tailored properties and functions for tissue engineering scaffolds and various other medical applications. This technology is revolutionizing the biomaterials and nanotechnology fields and has prompted us to incorporate its history, basic concepts, and applications into diverse courses such as Biomaterials, Tissue Engineering, Polymers in Medicine, and Senior Design in Chemical and Biomedical Engineering. This Innovation of the Curriculum is timely and crucial for multiple reasons. There is a need for a systematic approach to course structure that ties historical concepts to new materials and processes and, ultimately, to practical applications. Combining this lecture organization with active learning in the forms of open discussions and hands-on experiments/demonstrations will enhance learning outcomes (including retention and critical thinking) at all levels of education. At the undergraduate and graduate levels in the courses mentioned, discussions of electrospinning can create a classroom atmosphere of creative thinking, and an actual demonstration of nanomaterial fabrication can serve as a visual aid to the students. More importantly, this curriculum innovation can be used at the high school level to demonstrate nanotechnology and its applications to medicine, which will aid in sparking the interest of future generations of tissue engineers, biomaterial scientists, nanotechnologists, and scientists and engineers in general.