Fig. 1: Nr-CWS can treat endothelial cell dysfunction caused by Transient hyperglycemia.

A Schematic representation of the experimental model in vitro. (Image produced by Figdraw). B, C The tube formation assay was used to assess the ability of endothelial cells to form three-dimensional capillary-like structures in vitro (B, scale bars = 100 µm) and to quantify the tube formation and cell growth (C) (n = 3 per group). D EDU staining of HUVECs from different treatment groups to detect cell proliferation function. Positive cells were imaged (scale bars = 100 µm, left panel) and quantified (right panel) (n = 3 per group). E Cellular ROS levels in Control, DM, MM and Nr-CWS of HUVECs. The ROS was quantified (right panel) (n = 3 per group). F Schematic representation of the experimental model in vivo. G Representative wound images at day 0 and 14 during the healing process, And KRT14 immunohistochemical staining of 14-day wound tissue. The wound-healing rate was measured (right panel) every d and is shown as the percentage of the initial wound area (n = 8 per group). H, I Histological changes and collagen deposition in wounds were evaluated by H H&E staining and I Masson staining. bars = 100 µm. J CD31 and CDH5 (red for CD31; green for CDH5; blue for nuclei) in skin sections from different experimental groups. bars = 100 µm. K, L A laser Doppler imager show the blood flow perfusion in the wounds that were subjected to different treatments (K) and quantification of wound perfusion in the wounds (L) (n = 8 per group). M Epithelial thickness measured by H&E staining (n = 3 per group). The data are represented as mean ± SD of at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 are considered significant. HUVECs human umbilical vein endothelial cells, Nr-CWS Nocadia rubra cell wall skeleton preparation, DM diabetic memory, MM metabolic memory.