Discipline: Biological Sciences
Jose A. Cordero - University of Puerto Rico
Co-Author(s): Rajnish Sahu, Center for NanoBiotechnology & Life Sciences Research, Alabama State University, Montgomery, AL; Richa Verma, Center for NanoBiotechnology & Life Sciences Research, Alabama State University, Montgomery, AL; Shree R. Singh, Center for NanoBiotechnology & Life Sciences Research, Alabama State University, Montgomery, Alabama; Vida A. Dennis, Center for NanoBiotechnology & Life Sciences Research, Alabama State University, Montgomery, AL
Skin is the largest, complex and fast growing organ in our body, which acts as a barrier against pathogens, toxins and several impacts. Frequent damages to skin are very common and repaired as natural process of body. However, severe damages sustained by deep burns, injury, or illnesses may lead to permanent loss or impairment of natural healing process. Although the surgical methods are advancing but may not be suitable in certain situation. The supplementation of some biomaterials has been proven to aid the repair mechanism. Treating keratinocytes using PLGA nanoparticles in combination and alone with reduced glutathione, N-acetyl cysteine and naringenin may accelerate the wound healing process. Mouse keratinocytes cell line was grown to form confluent monolayer and a scratch was applied with pipette tip to reflect manual wounding in a method known as a scratch assay. Cells were grown for 48 hours post treatment, supernatants were collected and cells were fixed. Fluorescent microscope images were taken to observe the closer of scratch in comparison to untreated cells. In another similar experiment, total RNA was collected after 24 hours post treatment to check the expression of differentiation marker genes involucrin and filaggrin. Our results show that naringenin treatment was effective in closing the scratch followed by the combination of PLGA/naringenin. Quantitative RT PCR showed high upregulation of early differentiation gene involucrin. Thus indicating that naringenin has significant role in accelerating the wound healing by controlling the inflammation.Not Submitted
Funder Acknowledgement(s): NSF-REU (DBI-1659166) to Dr. Komal Vig (PI) NSFCREST (HRD-1241701) to Dr. Shree S. Singh (PI)
Faculty Advisor: Vida Dennis, email@example.com
Role: During this research, I was responsible for all parts. The COCA cell line was used for cell culture. The EMEM media was prepared with certain specifications to initiate differentiation in the COCA cells. Cells were washed with PBS, incubated with DAPI solution, and mounted using 70% glycerol to then by photographed under Nikon fluorescent microscope using specific wavelength filter for DAPI. Total RNA was isolated using RNA isolation kit (Qiagen) and cDNA was synthesized using manufacturer?s protocol (Applied bioscience). cDNA was amplified using Taq-man qRT-PCR to check the mRNA transcripts for gene expression related to keratinocyte differentiation genes involucrin and filaggrin. Results were calculated as fold change over unstimulated cells.