Date of Graduation

5-2013

Document Type

Thesis

Degree Name

Bachelor of Science in Biological Engineering

Degree Level

Undergraduate

Department

Biological and Agricultural Engineering

Advisor/Mentor

Wochok, Jeffrey C.

Committee Member/Reader

Carrier, Danielle J.

Committee Member/Second Reader

Costello, Thomas A.

Abstract

For severe muscle damage, tissue engineering has become a viable option to culture muscle tissues with the ultimate goal of implantation. However, the tissues are typically cultured on permanent scaffolds which would elicit a foreign body response if placed in the body. To avoid this, biological scaffolds are ideal for tissue culture and limiting any negative reactions. A sacrificial scaffold can be used to gather extracellular matrix (ECM), which can then be used as the primary scaffold for culturing tissues for implantation. To reach the ultimate goal of implanting muscle tissue, an understanding of how to produce ECM on a larger scale must also be gained. Supplementing culture media with pro-fibrotic molecules is an option to consider for enhancing ECM formation. Two conditions, molecular crowding and the addition of transforming growth factor beta 1 (TGF-β1), were considered. In addition, the effect of the concentration of TGF-β1 on material yield was also analyzed. In the first experiment, molecular crowding macromolecules, TGF-β1, and a control with no additives were considered. . For the second experiment, concentrations of TGF-β1 at 0.5, 1, and 5 ng/mL were added to culture media and material yield was compared to a control set with no TGF-β1. Cells were cultured for three weeks on sacrificial scaffolds under varied culture conditions, the scaffolds were removed, and the mass of ECM collected was measured. The material yields were then analyzed using an analysis of variance (ANOVA) and linear regression. The results from the first experiment indicated that neither molecular crowding nor TGF-β1 increase the material yield. In the second experiment, it appeared that the addition of TGF-β1 at all concentrations increased material yield. This could indicate even less TGF-β1 could be added to culture media to increase material yield. Additionally, the amount of fetal bovine serum (FBS) added to the culture media was different between the two experiments, which could explain the conflicting results regarding the effect of TGF-β1 on material yield in the two experiments. Based on the results of the data, ECM formation can be enhanced by supplementing the culture media with TGF-β1 at a concentration as low as 0.5 ng/mL.

Keywords

extracellular matrix; tissue engineering; muscle damage

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