Date of Graduation

12-2014

Document Type

Thesis

Degree Name

Master of Science in Biomedical Engineering (MSBME)

Degree Level

Graduate

Department

Biomedical Engineering

Advisor/Mentor

Wolchok, Jeffrey C.

Committee Member

Muldoon, Timothy J.

Second Committee Member

Washington, Tyrone A.

Keywords

Decellularization; Device; Extracellular Matrix; Skeletal Muscle

Abstract

Decellularization of skeletal muscle is a process that removes cellular components of skeletal muscle tissue while leaving behind the intact extracellular matrix (ECM). Skeletal muscle ECM is currently being studied as a biologic scaffold for repairing volumetric muscle loss (VML) because the removal of cells greatly reduces the antigenicity of the donor tissue. Decellularization usually relies on passive diffusion of detergents, surfactants and/or osmotic solutions to strip cells from the ECM. However, passive diffusion alone is usually not sufficient for complete removal of cells from the interior of large pieces of skeletal muscle using detergents, such as sodium dodecyl sulfate (SDS). The goal of this study was to develop a device that not only removes cells by perfusion from the interior of skeletal muscle, but also monitors the progress of decellularization in real-time. The device, based around a Raspberry Pi, is a standalone system that does not require a desktop computer or expensive software packages. Different flow rates (0.1, 1.0 and 10 mL/hr) along with different concentrations of SDS (0.2% and 1.0%) were tested. Decellularization progress was monitored and logged to an online spreadsheet. The device was found to be capable of decellularizing the medial gastrocnemius of a rat in under 10 hours. Complete decellularization was validated using fluorescent imaging. Perfusion decellularized muscle samples were found to have no significant differences in collagen or sulfated glycosaminoglycan (sGAG) content when compared to samples that were decellularized using current passive diffusion protocols. The ECM obtained through the use of this device is currently being used for the repair of VML in a rat model.

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