αv p3g8  (Developmental Studies Hybridoma Bank)

Journal: Nature materials

Article Title: Hydrogels with precisely controlled integrin activation dictate vascular patterning and permeability

doi: 10.1038/nmat4954

Figure Lengend Snippet: (a) HUVEC sprouting in the presence of integrin αv, α5, β1 or β3 (5μg/ml changed daily) blocking antibodies. Blocking α5, β1 or β3 completely blocks sprouting, while blocking αv does not. α3/α5β1 specific matrices = gel + 2μM Fn9*10, αvβ3 specific matrices = gel + 2μM Fn9(4G)10. Scale bars, 100 μm. (b) comparison of branch cluster occurrence with and without αv blocking (n=51 HUVEC coated beads from three independent gels). (c) Images showing HUVEC sprouts in RGD (1000μM) modified FXIIIa stabilized Fn depleted fibrin matrices. RGD modified matrices promote HUVEC sprouting and branch cluster formation. Scale bar: 50 μm (d,e) Quantification of HUVEC sprouting in RGD modified Fib 3 matrices at different concentrations. (f) Quantification of branch clusters per bead. For quantification n ≥ 15 HUVEC coated beads, from 3 independent gels. Each bead is treated as its own independent sample. Statistical analyses were performed using Prism (GraphPad, San Diego, CA). Brown-Forsythe test was performed to ensure that the variance of the data is similar between the groups that are being statistically compared. Data were analyzed using a one-way analysis of variance (ANOVA) followed by a Tukey post-hoc test and a 95% confidence interval. All plots represent mean ± SD. *,*** and **** indicate P < 0.05, P < 0.001 and P < 0.0001.

Article Snippet: For Fn9(4G)10 gels, HUVEC beads were suspended at a concentration of 500 beads/ml in 2 mg/ml fibrinogen (Fib1 or Fib3), 1 U/ml factor XIII, 0.04 U/ml aprotinin and 2 μM (high dosage) or 0.239 μM (low dosage) Fn9(4G)10 at a pH of 7.4 with or without 5μg/ml of β3 integrin blocking antibody (9H5, Developmental Studies Hybridoma Bank) or αv integrin blocking antibody (P3G8, Developmental Studies Hybridoma Bank).

Techniques: Blocking Assay, Comparison, Modification

Journal: Nature materials

Article Title: Hydrogels with precisely controlled integrin activation dictate vascular patterning and permeability

doi: 10.1038/nmat4954

Figure Lengend Snippet: A middle cerebral artery occlusion ischaemic stroke model was utilized to look at the effects of injected integrin-specific HA matrices on stroke repair 10 days post-injection (injection was performed 5 days post-stroke). a, Fibronectin location in normal and stroke brain 15 days post-stroke. Scale bars, 50 μm. b, Schematic illustration of the hydrogel injected into the mouse brain after stroke. VEGF nanocapsules (nV) were designed to slowly release VEGF. c, Fluorescent microscopy showing brain vasculature in both the infarct and peri-infarct (stained for Glut-1 or Glut-1 plus tomato lectin intravascular perfusion) as well as leaked red blood cells (stained for Ter-119). The asterisk indicates the stroke site while the white dashed curve indicates the boundary between the infarct (stroke site) and peri-infarct (the area adjacent to the stroke site). Scale bars, 100 μm. d,e, Quantification of the total vessel area (perfused and not prefused) in the infarct (inside the stroke) and peri-infarct (around the stroke) areas. f, Quantification of Ter-119-positive red blood cell area. g, The morpho-analysis of growing vessels in the peri-infarct area for vessel ramification. All plots represent mean ± s.d. Each dot in the plots represents an individual mouse. Statistical analyses were performed using Prism (GraphPad). Data in d,e were analysed using a one-way analysis of variance followed by a Tukey post hoc test and a 95% confidence interval. Data in f,g were analysed using a two-tailed unpaired test. *, ** and *** indicate P < 0.05, P < 0.01 and P < 0.001; NS, not significant.

Article Snippet: For Fn9(4G)10 gels, HUVEC beads were suspended at a concentration of 500 beads/ml in 2 mg/ml fibrinogen (Fib1 or Fib3), 1 U/ml factor XIII, 0.04 U/ml aprotinin and 2 μM (high dosage) or 0.239 μM (low dosage) Fn9(4G)10 at a pH of 7.4 with or without 5μg/ml of β3 integrin blocking antibody (9H5, Developmental Studies Hybridoma Bank) or αv integrin blocking antibody (P3G8, Developmental Studies Hybridoma Bank).

Techniques: Injection, Microscopy, Staining, Two Tailed Test

Journal: Nature materials

Article Title: Hydrogels with precisely controlled integrin activation dictate vascular patterning and permeability

doi: 10.1038/nmat4954

Figure Lengend Snippet: α3/α5β1 and αvβ3 integrin-specific materials regulate vascular patterning in vitro and in vivo

Article Snippet: For Fn9(4G)10 gels, HUVEC beads were suspended at a concentration of 500 beads/ml in 2 mg/ml fibrinogen (Fib1 or Fib3), 1 U/ml factor XIII, 0.04 U/ml aprotinin and 2 μM (high dosage) or 0.239 μM (low dosage) Fn9(4G)10 at a pH of 7.4 with or without 5μg/ml of β3 integrin blocking antibody (9H5, Developmental Studies Hybridoma Bank) or αv integrin blocking antibody (P3G8, Developmental Studies Hybridoma Bank).

Techniques: In Vitro, In Vivo