Journal: Biomedicines

Article Title: Mechanistic Illustration: How Newly-Formed Blood Vessels Stopped by the Mineral Blocks of Bone Substitutes Can Be Avoided by Using Innovative Combined Therapeutics

doi: 10.3390/biomedicines9080952

Figure Lengend Snippet: Vasculoneogenesis in bone substitutes subcutaneously implanted in nude mice. ( A , B ) H&E staining of bone substitutes (green lines) with either empty (NF-) ( A ) or HEP/VEGF- ( B ) SNCs, at 12 dpi. Yellow arrowheads indicate blood vessels. Scale bars: 200 µm. The images at the top right show enlargements of blood vessels. Scale bars: 20 µm. ( C ) Quantitative analysis of the vessels found in the implanted bone substitutes. The average number and diameter of the vessels found, together with the average surface covered are given at 4 and 12 dpi. Values are expressed as mean ± SD of at least five images/section and four sections/sample. **: p ≤ 0.05. ( D , E ) Ultrastructural view of a blood vessel found in the HEP/VEGF-SNCs bone substitute, shown as a transverse section. Mural cells enveloping the blood vessels (e.g., the pericyte indicated with a red asterisk) were found in the nano-active bone substitute. ( E ) This is the enlargement of the area delimited by the yellow frame in ( D ). No blood cells are in the lumen of the vessel, as a contrast agent was perfused (residues indicated with a blue asterisk). E: endothelial cell; eN: nucleus of the endothelial cell; P: pericyte; pN: nucleus of the pericyte. Scale bars: 5 µm in d, 2 µm in e. ( F ) Endothelial cells of human origin in the bone substitute, as revealed with anti-human (top panels) or anti-human/mouse PECAM1 antibody (mid panel). Mouse control bone was negative to the anti-human PECAM1 antibody (lower panel). Scale bars: 200 µm.

Article Snippet: Rabbit anti-human PECAM1/CD31 (Microm, Brignais, France) and non-species-specific anti-PECAM1/CD31 (Abcam, Paris, France), then secondary antibody raised against rabbit antibodies and coupled with Alexa 594 fluorochrome (Molecular Probes, Life Technologies, Thermo Fisher Scientific, Illkirch-Graffenstaden, France), diluted 1:200 in PBS 1% BSA, were used.

Techniques: Staining

Journal: bioRxiv

Article Title: 3-nitrotyrosine shortens axons of a non-dopaminergic neuron by inhibiting mitochondrial motility

doi: 10.1101/2024.05.09.593299

Figure Lengend Snippet: 3-NT reduced the axonal length in CGNs. (A) CGNs at 4 days in vitro (DIV) were exposed to varying concentrations of 3-NT for 1 day, followed by immunocytochemical analysis utilizing Tau-1 (axonal marker; magenta) and MAP2 (dendritic marker; green) antibodies. Nuclear staining using Hoechst dye (blue) is revealed. (B) CGNs at 5 DIV, transduced with a vector expressing EGFP, were subjected to 3-NT treatment for 1 day. (C, D) Quantification of the length of the longest (axonal) process (C) and the rest of the processes (D) in each neuron was performed. Whiskers denote the range extending 1.5 times the interquartile range above and below the box limits. Data points are depicted as dots. 3-NT led to a reduction in the length of the longest process compared to the control condition. The Wilcoxon rank-sum test with the Benjamini-Hochberg correction was employed for comparison against 0 μM (white boxes) of 3-NT (n > 90 neurons from at least 3 experiments per condition). * p < 0.05, ** p < 0.01, *** p < 0.001. Neurons with processes less than 20 μm were excluded from analysis. Scale bars represent 50 μm.

Article Snippet: Primary antibodies used included mouse monoclonal antibodies against Tau-1 (1:1000; AB5622, Merck) or α-tubulin (1:1000; 12G10, AB_1157911, Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), and rabbit polyclonal antibodies against MAP2 (1:1000; AB5622, Merck) or 3-NT (1:1000; 06-284, Merck).

Techniques: In Vitro, Marker, Staining, Transduction, Plasmid Preparation, Expressing, Control, Comparison