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. 2020 Feb 21;10:3205. doi: 10.1038/s41598-020-60196-y

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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In-vitro viability and metabolism evaluation in HUVEC line. (a) cell number evaluation at 2 h, 24 h and 48 h after scaffold direct dissolution in: (i) control cell (first column); (ii) cells after 11% of alginate administration (second column); (iii) cell after 11% of alginate administration and ethanol crosslinking (third column); (iv) cell after 11% of alginate administration, ethanol crosslinking and NPs administration (forth column); (v) cell after NPs administration (fifth column). (b) same as (a) for scaffold indirect dissolution. (c) cell viability evaluation at 2 h, 24 h and 48 h after scaffold direct dissolution in: i) control cell (first column); (ii) cells after 11% of alginate administration (second column); iii) cell after 11% of alginate administration and ethanol crosslinking (third column); (iv) cell after 11% of alginate administration, ethanol crosslinking and NPs administration (forth column); (v) cell after NPs administration (fifth column). (d) same as (c) for scaffold indirect dissolution. (e) ATP evaluation at 2 h, 24 h and 48 h after scaffold direct dissolution in: (i) control cell (first column); (ii) cells after 11% of alginate administration (second column); iii) cell after 11% of alginate administration and ethanol crosslinking (third column); (iv) cell after 11% of alginate administration, ethanol crosslinking and NPs administration (forth column); (v) cell after NPs administration (fifth column). (f) same as (e) for scaffold indirect dissolution. White columns: control cells; grey columns: cells with 11% of alginate scaffold; pink columns: cells with 11% of alginate scaffold gelled with EtOH; blue columns: cells with 11% of alginate scaffold gelled with EtOH containing NPs: yellow columns: cells with NPs. (g) Cytofluorimetric analysis of HUVEC control cell after 24 h (black trace), HUVECs and alginate after 24 h (red trace), HUVECs and alginate plus NPs after 2 h (green trace) and after 24 h (blue trace). Kruskal-Wallis (post hoc analyses: Dunn’s multiple comparison) was performed and statistical significance was set at p < 0.05. Data are represented as mean ± SEM.