Date Approved

1-10-2024

Embargo Period

1-10-2024

Document Type

Dissertation

Degree Name

Doctor of Philosophy (Ph.D.)

Department

Chemical Engineering

College

Henry M. Rowan College of Engineering

Advisor

Kirtie Yenkie, Ph.D.

Committee Member 1

Gary Thompson, Ph.D.

Committee Member 2

Erik Brewer, Ph.D.

Committee Member 3

Thomas Chistiani, Ph.D.

Committee Member 4

Caleb Roth, Ph.D.

Keywords

Biocompatibility, Decellularization, Electrokinetics, Electroporation, Pulsed electric fields

Subject(s)

Intervertebral disk; Wounds and injuries--Treatments

Disciplines

Biomedical Engineering and Bioengineering | Chemical Engineering

Abstract

Severe back injuries and chronic pain necessitate surgical replacement of damaged intervertebral disc (IVD) cartilage in advanced disease stages. Bovine IVD tissue has been exposed to an upper threshold pulsed electric field (PEF) dose, causing cell death without thermal damage to the tissue. Subsequent PEF exposures at lower magnitudes have accelerated the removal of immunogenic biomolecules though electrokinetic extraction using optimized aqueous solutions. This approach yields a natural scaffold, ready for biocompatibility and mechanical strength assessment. The effects of microsecond pulsed electric fields (µsPEF) on primary bovine AF fibroblast-like cells have been characterized in vitro. PEFs of 10 and 100 µs durations, with varying numbers of pulses and electric field strengths, were applied to the cells. Furthermore, a low-intensity and minimally heating PEF-induced electrokinetic flow for molecular extraction has been established, involving the determination of the electrophoretic mobility of charged molecules within the AF cartilage tissue. Ultimately, a biocompatible AF scaffold has been generated using PEF and electrolyte solutions in a custom-designed bioreactor. Compared to chemical methods that take days, PEF application achieves decellularization of cartilage tissue within three hours, while preserving the desired biomolecules and ultrastructure of the tissue matrix.

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