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Bacterial cellulose (BC) is a versatile biopolymer with significant potential across biomedical, food, and industrial applications. To remove bacterial contaminants, such as protein and DNA, BC pellicles undergo purification, which traditionally relies on harsh alkali treatments, such as sodium hydroxide or strong surfactants, which present environmental concerns. In response, this study evaluates the efficacy of various non-conventional surfactants—both non-biodegradable and biodegradable—as alternatives for BC purification. Among the surfactants tested, sodium cocoyl isethionate (SCI), a mild anionic and biodegradable surfactant, emerged as particularly effective, achieving an 80.7% reduction in protein content and a 65.19% reduction in double-stranded DNA (dsDNA) content relative to untreated samples. However, these advantages were not without additional challenges, such as the appearance of residual surfactants. Given SCI’s promising performance and biodegradability, it was further examined in two-step treatment protocols; additionally, sodium dodecyl sulfate (SDS) was also examined as a more traditional anionic surfactant as well as NaOH. For the two-step treatment protocol, BC pellicles were treated with one reagent for 3 h, followed by a second reagent for an additional 3 h. Notably, by using NaOH as the final step in the two-step treatment protocol, residual surfactant was not detected in the FTIR analysis. Overall, this work demonstrates that SCI, in addition to subsequent NaOH treatment, can be used as a surfactant-based approach for BC purification, representing a potential environmentally friendly alternative to traditional surfactant-based approaches for BC purification.more » « less
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Despite chronic fibrosis occurring in many pathological conditions, few in vitro studies examine how fibrosis impacts lymphatic endothelial cell (LEC) behavior. This study examined stiffening profiles of PhotoCol®—commercially available methacrylated type I collagen—photo-crosslinked with the photoinitiators: Lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), Irgacure 2959 (IRG), and Ruthenium/Sodium Persulfate (Ru/SPS) prior to evaluating PhotoCol® permeability and LEC response to PhotoCol® at stiffnesses representing normal and fibrotic tissues. Ru/SPS produced the highest stiffness (~6 kilopascal (kPa)) for photo-crosslinked PhotoCol®, but stiffness did not change with burst light exposures (30 and 90 s). The collagen fibril area fraction increased, and dextran permeability (40 kilodalton (kDa)) decreased with photo-crosslinking, showing the impact of photo-crosslinking on microstructure and molecular transport. Human dermal LECs on softer, uncrosslinked PhotoCol® (~0.5 kPa) appeared smaller with less prominent vascular endothelial (VE)-cadherin (cell–cell junction) expression compared to LECs on stiffer PhotoCol® (~6 kPa), which had increased cell size, border irregularity, and VE-cadherin thickness (junction zippering) that is consistent with LEC morphology in fibrotic tissues. Our quantitative morphological analysis demonstrates our ability to produce LECs with a fibrotic phenotype, and the overall study shows that PhotoCol® with Ru/SPS provides the necessary physical properties to systematically study LEC responses related to capillary growth and function under fibrotic conditions.more » « less
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