Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Kendra Swain - Alabama State University
Co-Author(s): Vinoy Thomas, Department of Materials Science and Engineering, University of Alabama at Birmingham
Over the past few years, tissue engineering and regenerative applications have yielded many neoteric tissue replacements and implementations. Electrospinning is a valuable technique for the fabrication of 3D fiber scaffolds for advanced wound healing and in this manner reclamation of the skin. Scaffolds are comprised of biocompatible biomaterials, such as collagen, manufactured materials, or in the current study a combination of polycaprolactone (PCL), and polydioxanone (PDO). PCL is a long-degrading polymer and is FDA approved with high porosity and PDO which has been used for biomedical applications, mostly in the form of surgical sutures. These polymers were fabricated into a bioresorbable scaffolds through electrospinning. This newly designed functionally graded composition will be used for skin growth.
Scaffolds were synthesized using PDO and PCL which were dissolved in hexaflouro -2- propanol (HFIP) and stirred 24h. Different polymer solutions of PDO, PCL, PDO+PCL, and PDO+PCL+ Collagen were used for making the scaffolds used. Electrospinning parameters were optimized to get fibers evenly dispersed across metal spiked plate. Scaffolds were then vacuumed dried for 2 days before characterization. Morphology and fiber composition were analyzed with Scanning electron microscopy (SEM). Tensile properties were measured with a dynamic mechanical analyzer (DMA). Scaffolds were sterilized by UV light and afterwards washing with ethanol, sterile distilled water, and finally HBSS. The growth and proliferation of normal human keratinocytes were evaluated in keratinocyte growth serum and our EpiLife medium. Fibroblast was grown in DMEM 10 media until confluent. Scaffolds were then placed into a 48 well plate and seeded with keratinocytes and fibroblast in separate wells for 21 days.
Synthesized scaffolds showed cell growth on scaffolds were 100% viable on days 3 and 5 with a decrease to 90% on days 7 and down to 80% on day 15. Keratinocytes and Fibroblast were viable on all variable scaffolds used.
In conclusion scaffolds were successfully spun for PDO and PCL synthesized polymers. SEM showed ECM mimicking fibrous morphology for each blend that was used. Cell studies confirms cell adhesion and growth on scaffolds used for extended periods. Future studies are to develop procedures for establishing confluent, layers of co-cultured human keratinocytes and fibroblast on the surface of synthesized PDO and PCL scaffolds.
Funder Acknowledgement(s): Dr. Komal Vig (PI) and by National Institues of Health-MBRS-RISE(1R25GM106995-01)grants and Dr. Shree Singh (PI)
Faculty Advisor: Komal Vig, komalvig@alasu.edu
Role: All parts of the research was done by myself