recombinant mouse endostatin  (R&D Systems)

Journal: Nature materials

Article Title: Dual-function injectable angiogenic biomaterial for the repair of brain tissue following stroke

doi: 10.1038/s41563-018-0083-8

Figure Lengend Snippet: (A) Fluorescent images of neuroblasts (Dcx) and the proliferation marker BrdU, and (B) axonal neurofilaments (NF200) in and around the stroke site (*) at 2 weeks and (G) at 16 weeks after gel transplantation. (C) Quantification of neuroblasts (Dcx) and proliferating neuroblasts (Dcx/BrdU) in the ipsilateral ventricle, (D) the number of neuroblasts migrating and their migration distance and number, (E) the axonal area (NF200) in and around the stroke site, and (F) infiltration distance and penetration angle in the stroke site. (H) Quantification of axonal area (NF200) in and (I) around the stroke site, (J) infiltration distance and (K) axonal penetration angle 16 weeks after gel injection. In all plots, the dotted red line and red number indicates the value for the give quantification of the contralateral side. Empty gel = gel = HA hydrogel, Vs = 200 ng of soluble VEGF, lcV = 1μg nH loaded with 200 ng VEGF, hcV = 0.01 μg nH loaded with 200 ng VEGF and 0.99 μg unloaded nH, Endo = a daily i.p injection of endostatin day 5 to 15. Data is presented using a min to max box plot. Each dot in the plots represents one animal and p values were determined by One-way ANOVA with a Tukey’s post-hoc test, with *, ** and **** indicating p < 0.05, p < 0.01 and p < 0.0001, respectively. Scale bar: 100 μm.

Article Snippet: Endostatin (Recombinant mouse, 100 μg/ml; Alpha Diagnostic, San Antonio, TX), a VEGF-independent angiogenesis inhibitor, was injected subcutaneously daily during days 5–15 after stroke to hcV-treated mice. hcV-Endostatin mice were submitted to behavioral tests and their brain used for immunohistology at 16 weeks time point.

Techniques: Marker, Transplantation Assay, Migration, Injection

Journal: Nature materials

Article Title: Dual-function injectable angiogenic biomaterial for the repair of brain tissue following stroke

doi: 10.1038/s41563-018-0083-8

Figure Lengend Snippet: (A) Fluorescent images of vessels (Glut-1, red) and axonal neurofilaments (NF200, green) in and around the stroke site (*) 16 weeks after gel transplantation. (B) Quantitative assessment of the proximity between the 2 networks with the quantification of NF200 positive signal on vessels and (C) positive area a distance of 50 μm from vessels. (D) Fluorescent images of the peri-infarct astrocytic scar (GFAP, green) and BDA-traced neurons (red) in the ipsilateral hemisphere of gel + hcV injected mice 16 weeks after gel transplantation. (E) Fluorescent images of astrocytes (GFAP) co-stained with vessels (Glut-1) and pericytes/smooth muscle cells (PDGFR-β), or (F) with end-feet astrocytes (Aquaporin-4) in the stroke site of hcV-treated mice, 16 weeks after gel transplantation. Empty gel = gel = HA hydrogel, Vs = 200 ng of soluble VEGF, lcV = 1μg nH loaded with 200 ng VEGF, hcV = 0.01 μg nH loaded with 200 ng VEGF and 0.99 μg unloaded nH, Endo = a daily i.p injection of endostatin days 5 to 15. Data is presented using a min to max box plot. Each dot in the plots represents one animal and p values were determined by One-way ANOVA with a Tukey’s post-hoc test, with ** and **** indicating p < 0.01 and p < 0.0001, respectively. Data represent the average. $$$ indicates p < 0.001 vs Gel+hcV. Scale bar: 100 μm.

Article Snippet: Endostatin (Recombinant mouse, 100 μg/ml; Alpha Diagnostic, San Antonio, TX), a VEGF-independent angiogenesis inhibitor, was injected subcutaneously daily during days 5–15 after stroke to hcV-treated mice. hcV-Endostatin mice were submitted to behavioral tests and their brain used for immunohistology at 16 weeks time point.

Techniques: Transplantation Assay, Injection, Staining

Journal: Nature materials

Article Title: Dual-function injectable angiogenic biomaterial for the repair of brain tissue following stroke

doi: 10.1038/s41563-018-0083-8

Figure Lengend Snippet: (A) Experiment timeline for the behavioral tests. Mice were injected 5 days after stroke with one of the following treatments: empty gel, gel + Vs, gel + lcV, and gel + hcV. Mice were subjected to different behavioral tests (illustrated by vertical grey boxes on the timeline) On week 0, 1, 4, 8, 12 and 16 after stroke. (B1) The Cylinder test was used to measure the dexterity of their contralateral forelimb, (B2) the Grid test for the contralateral hindlimb, and (B3, B4) the Pasta test for the contralateral forepaw, normally sensitive to post-stroke lateralized impairments. (C1–C4) In order to determine the role of gel+hcV -induced vascularization on behavioral recovery, a supplemental set of gel+hcV animals was administered with endostatin, a VEGF-independent angiogenic inhibitor for 10 days after the gel injection, and submitted to the same behavioral tests: Cylinder (C1), Grid (C2), and Pasta (C3–C4). (D1–D4) In order to determine the role of gel+hcV –induced axonal growth on recovery, a supplemental set of gel+hcV animals received a brain injection of an AAV5 viral construct expressing hM4 DREADD receptors (designer receptors exclusively activated by a designer drug) directly in the stroke area on week 13. Transfected neurons are silenced after i.p administration of the DREADD ligand, clozapine-N-oxide (CNO) on week 16. Mice are then submitted to the same behavioral tests: Cylinder (D1), Grid (D2), and Pasta (D3–D4). Empty gel = HA hydrogel, Vs = 200 ng of soluble VEGF, lcV = 2μg nH loaded with 200 ng VEGF, hcV = 0.01 μg nH loaded with 200 ng VEGF and 0.99 μg unloaded nH. Data represent the average ± SEM (n = 12 mice) and p values were determined by One-way ANOVA, Tukey’s post-hoc test, * indicating P < 0.05.

Article Snippet: Endostatin (Recombinant mouse, 100 μg/ml; Alpha Diagnostic, San Antonio, TX), a VEGF-independent angiogenesis inhibitor, was injected subcutaneously daily during days 5–15 after stroke to hcV-treated mice. hcV-Endostatin mice were submitted to behavioral tests and their brain used for immunohistology at 16 weeks time point.

Techniques: Injection, Construct, Expressing, Transfection