Abstract
For salivary gland (SG) tissue engineering, we cultured acinar NS-SV-AC cell line or primary SG fibroblasts for 14 days in avian egg yolk plasma (EYP). Media or egg white (EW) supplemented the cultures as they grew in 3D-Cryo histology well inserts. In the second half of this manuscript, we measured EYP’s freeze-thaw gelation and freeze-thaw induced gelled EYP (GEYP), and designed and tested further GEYP tissue engineering applications. With a 3D-Cryo well insert, we tested GEYP as a structural support for 3D cell culture or as a bio-ink for 3D-Bioprinting fluorescent cells. In non-printed EYP + EW or GEYP + EW cultures, sagittal sections of the cultures showed cells remaining above the well’s base. Ki-67 expression was lacking for fibroblasts, contrasting NS-SV-AC’s constant expression. Rheological viscoelastic measurements of GEYP at 37 °C on seven different freezing periods showed constant increase from 0 in mean storage and loss moduli, to 320 Pa and 120 Pa, respectively, after 30 days. We successfully 3D-printed GEYP with controlled geometries. We manually extruded GEYP bio-ink with fluorescence cells into a 3D-Cryo well insert and showed cell positioning. The 3D-Cryo well inserts reveal information on cells in EYP and we demonstrated GEYP cell culture and 3D-printing applications.
Highlights
Salivary hypofunction can be induced by side-effects of medicines, Sjogren’s autoimmune disorder, or head and neck radiation therapy
Materials 2019, 12, 3480 cost-effective, multi-compositional soft biomaterial, we recently discovered that a translucent fraction from the egg yolk (EY)—the egg yolk plasma (EYP)—can under certain circumstances permit media-free human cell survival
In addition to the Egg Yolk Plasma (EYP) + Egg White (EW) gel-like biomaterial, we hypothesized that a freeze-thawed EYP ( Gelled EYP (G EYP)) could still possess gel-like properties, if brought to 37 ◦ C
Summary
Salivary hypofunction can be induced by side-effects of medicines, Sjogren’s autoimmune disorder, or head and neck radiation therapy. Materials 2019, 12, 3480 cost-effective, multi-compositional soft biomaterial, we recently discovered that a translucent fraction from the egg yolk (EY)—the egg yolk plasma (EYP)—can under certain circumstances permit media-free human cell survival. This current study builds on our previous data as we continue developing and understanding the EYP biomaterial for soft tissue engineering. In addition to the EYP + EW gel-like biomaterial, we hypothesized that a freeze-thawed EYP ( G EYP) could still possess gel-like properties, if brought to 37 ◦ C (relevant for cell culture applications) To test this hypothesis, we used a rheometer to examine G EYP’s viscoelastic properties at 37 ◦ C after freeze thaw treatment. The two different bio-inks simulate the epithelial parenchymal to mesenchymal stromal interface, both initiators of tubules and acini during development [15]
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