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Design Summary
Design decisions throughout this project were guided by biocompatibility, antimicrobial effectiveness, and scalability. Bacterial cellulose was selected as the primary scaffold due to its biodegradability, flexibility, and ability to support integration of functional biomaterials. Engineered CsgA curli fibers were chosen because they provide a stable nanofiber structure that can be genetically modified to display antimicrobial peptides.
Heliomicin was selected as the antifungal peptide due to its strong antifungal properties, low cytotoxicity, and stability compared to other antimicrobial peptides considered during the design process. Unlike some peptide candidates that showed limited lifespan or increased toxicity, heliomicin provided a balance between antifungal efficacy and biocompatibility.
Vacuum filtration was selected as the integration method because it enabled efficient incorporation of the engineered nanofibers throughout the bacterial cellulose matrix while preserving the structural integrity of the pellicle. This method also provided a simple, low-cost, and scalable approach for the creation of the final wound patch.
Table 1. Decision matrix to analyze the fitnes of the potential anti-microbial peptides. A 1-5 scale is used to evaluate each potential solution. A total score of 500 would indicate perfect fitness.
Table 2. Decision matrix to analyze potential production methods. A 1-5 scale is usd to evalute each potential solution. A total score of 500 would indicate perfect fitness.
Major Components
Peptide Vizualization and Integration Process
Figure 1. Pymol rendering of the heliomicin anti-microbial peptide
Figure 2. Schematic of the peptide vacuum filtration process
K. rhaeticus Cellulose Pellicle Growth
Figure 3. Pellicle growing in Falcon tube
Figure 4. Final pellicle shape and sizes after 2 week culturing period
Genetic Engineering of CsgA-Heliomicin Fibers and Fiber Integration
Figure 5. Gibson Assembly schematic; depicting the genetic engineering process of creating recombinant DNA for our peptide expression
Figure 6. Concentrated CsgA-Heliomicin fibers
Figure 7. Membrane Filtration setup for filtering nanofibers into cellulose pellicles
Figure 8. Fiber solution surrounding pellicle
Figure 9. Post-integration pellicle
Validation Testing
Figure 10. Pelliciles prepared using Nile blue staining to vizualize heliomicin peptide integration
Figure 11. Nile blue-stained CsgA-Heliomicin fibers visualized through SoRa microscopy
Figure 12. Graph of CFU survival assay results
Economic Evaluation
Figure 13. SuperPro Simulation of an Automated Pilot-Scale Manufacturing Process
Figure 14. Executive Summary of SuperPro Economic Simulation
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