025006 (14 pages)
Deep wounds in the gingiva caused bytrauma or surgery require a rapid and robust healing of connective tissues. Wepropose utilizing gas-brushed nanofibers coated with collagen and fibrin for that purpose. Our hypotheses are that protein-coated nanofibers will: (i) attract and mobilize cells in various spatial orientations, and (ii) regulate the expression levels ofspecific extracellular matrix (ECM)-associated proteins, determining the initial conformational nature ofdense and soft connective tissues. Gingival fibroblast monolayers and3D spheroids were cultured onECMsubstrate and covered with gas-blown poly-(DL-lactide-co-glycolide)(PLGA) nanofibers (uncoated/coated with collagen and fibrin). Cell attraction and rearrangement was followed byF-actin staining and confocal microscopy. Thicknesses ofthe cell layers, developed within the nanofibers, were quantified byImageJ software. The expression ofcollagen1α1 chain (Col1α1), fibronectin, and metalloproteinase 2 (MMP2) encoding genes was determined byquantitative reverse transcription analysis. Collagen- and fibrin- coated nanofibers induced cell migration toward fibers and supported cellular growth within the scaffolds. Both proteins affected the spatial rearrangement offibroblasts byfavoring packed cell clusters or intermittent cell spreading. These cell arrangements resembled the structural characteristic ofdense and soft connective tissues, respectively. Within three days ofincubation, fibroblast spheroids interacted with the fibers, and grew robustlybyincreasing their thickness compared to monolayers. While theECMkeycomponents, such as fibronectin andMMP2encoding genes, were expressed in both protein groups, Col1α1 was predominantlyexpressed in bundled fibroblasts grown on collagen fibers. This enhanced expression ofcollagen1 is typical for dense connective tissue. Based on results ofthis study, our gas-blown, collagen- and fibrin-coated PLGA nanofibers are viable candidates for engineering soft and dense connective tissues with the required structural characteristics and functions needed for wound healing applications. Rapid regeneration of these layers should enhance healing ofopen wounds in a harsh oral environment.
Original Publication Citation
Kaufman, G., Whitescarver, R. A., Nunes, L., Palmer, X. L., Skrtic, D., & Tutak, W. (2018). Effects of protein-coated nanofibers on conformation of gingival fibroblast spheroids: Potential utility for connective tissue regeneration. Biomedical Materials, 13(2), 025006. doi:10.1088/1748-605X/aa91d9
Kaufman, Gili; Whitescarver, Ryan A.; Nunes, Laiz; Palmer, Xavier-Lewis; Skrtic, Drago; and Tutak, Wojtek, "Effects of Protein-Coated Nanofibers on Conformation of Gingival Fibroblast Spheroids: Potential Utility for Connective Tissue Regeneration" (2018). Engineering Technology Faculty Publications. 63.