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Our project aims to standardize the decellularization of a porcine lung by creating a bioreactor to house the lung, automating the decellularization process and developing a protocol that will increase the precision and the repeatability of the process. Our deliverables include a working prototype, an automated system that will inform the user when the decellularization process is complete, a pressure sensor to control perfusion, and automated pressurized pulses that will increase the rate of decellularization.
Our accomplishments thus far include: a design for a working prototype that will decellularize a porcine lung, determining the proper rate to perfuse the lung, finalizing the list of chemicals and enzymes, and finding a colorimetric cellular assay to determine when decellularization has been completed.
Our research has shown that some amount of degradation of the extracellular matrix (ECM) will occur in the decellularization process. The degradation of the ECM will be minimized by controlling the flow rate to mimic physiological pressure and eliminating any air bubbles trapped within the lung thus allowing a faster perfusion rate of the decellularization chemicals. We can also minimize degradation by modifying existing protocols that already in use and by using a new method, such as N-TIRE, that has yet to be fully investigated.
The first problem we encountered was the identification of an existing automated method to decellularize a porcine lung. To overcome this, we have improved on the functionality by included a method to verify complete decellularization, modifying the protocol to reduce ECM degradation and reducing pressure during perfusion. The second problem that we encountered involved determining which assay could be used to determine if the lung had been fully decellularized by analyzing the fluid expelled from the lung. We chose the Bradford assay due to the visible color change. The third problem was with the lack of communication amongst team members. This was resolved following a meeting and discussion about more effective avenues of communication. The final problems we encountered were with using the N-TIRE method. These include the temporary vasoconstriction induced by the pulses, utilizing the process on an organ the size of the lung, and the possibility of damaging the lung tissue.
biomedical engineering, porcine lung
Biomedical Engineering and Bioengineering | Engineering
VCU Capstone Design Expo Posters
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