Journal: Investigative Ophthalmology & Visual Science

Article Title: Dose-Related Side Effects of Intravitreal Injections of Humanized Anti-Vascular Endothelial Growth Factor in Rats: Glial Cell Reactivity and Retinal Ganglion Cell Loss

doi: 10.1167/iovs.65.4.10

Figure Lengend Snippet: Microglial cell reactivity. Representative magnifications taken from the retinal flat mounts of naïve left retinas ( A ) and right retinas ( B ), and from the left retinas of the different experimental groups that received an intravitreal injection of PBS ( C , I ), ranibizumab 10 µg/µL ( D , J ) Ranibizumab 0.38 µg/µL ( E , K ), aflibercept 40 µg/µL ( F , L ), aflibercept 1.5 µg/µL ( G , M ) and anti-rat VEGF ( H , N ) showing the Iba-1 + immunoreactive cells (green) at 7 (second row) and 30 (third row) days after the intravitreal injection (IVI). Scale bar, 100 µm.

Article Snippet: The following substances were injected in the different experimental groups: (1) PBS (Sigma Aldrich, Madrid, Spain); (2) ranibizumab (humanized anti-VEGF, Lucentis, Novartis) at two different concentrations: the human clinical concentration (higher concentration: 10 μg/μL) and a lower concentration (0.38 μg/μL) (see next paragraph); (3) aflibercept (humanized anti-VEGF, Bayer) at two different concentrations: the human clinical concentration (higher concentration; 40 μg/μL) and a lower concentration (1.5 μg/μL; see next paragraph); and (4) goat polyclonal anti-rat VEGF 164 diluted in PBS (15 μg/mL = 0, 015 μg/μL; MGC70609; Leinco Technologies, Inc., St. Louis, MO, USA).

Techniques: Injection

Journal: Investigative Ophthalmology & Visual Science

Article Title: Dose-Related Side Effects of Intravitreal Injections of Humanized Anti-Vascular Endothelial Growth Factor in Rats: Glial Cell Reactivity and Retinal Ganglion Cell Loss

doi: 10.1167/iovs.65.4.10

Figure Lengend Snippet: Macroglial cell reactivity. Representative magnifications taken from the retinal flat mounts of naïve left retinas ( A ) and right retinas ( B ), and from the left retinas of the different experimental groups that received intravitreal injections of PBS ( C , I ), ranibizumab 10 µg/µL ( D , J ), ranibizumab 0.38 µg/µL ( E , K ), aflibercept 40 µg/µL ( F , L ), aflibercept 1.5 µg/µL ( G , M ), and anti-rat VEGF ( H , N ) showing the GFAP immunoreactivity (red) at 7 days (second row) and 30 days (third row) after the intravitreal injection (IVI). Scale bar, 100 µm.

Article Snippet: The following substances were injected in the different experimental groups: (1) PBS (Sigma Aldrich, Madrid, Spain); (2) ranibizumab (humanized anti-VEGF, Lucentis, Novartis) at two different concentrations: the human clinical concentration (higher concentration: 10 μg/μL) and a lower concentration (0.38 μg/μL) (see next paragraph); (3) aflibercept (humanized anti-VEGF, Bayer) at two different concentrations: the human clinical concentration (higher concentration; 40 μg/μL) and a lower concentration (1.5 μg/μL; see next paragraph); and (4) goat polyclonal anti-rat VEGF 164 diluted in PBS (15 μg/mL = 0, 015 μg/μL; MGC70609; Leinco Technologies, Inc., St. Louis, MO, USA).

Techniques: Injection

Journal: Investigative Ophthalmology & Visual Science

Article Title: Dose-Related Side Effects of Intravitreal Injections of Humanized Anti-Vascular Endothelial Growth Factor in Rats: Glial Cell Reactivity and Retinal Ganglion Cell Loss

doi: 10.1167/iovs.65.4.10

Figure Lengend Snippet: Brn3a ± RGC survival. Representative magnifications taken from the retinal flat mounts of the naïve left retinas ( A ) and right retinas ( B ), and from the left retinas of the different experimental groups that received an intravitreal injection of PBS ( C , I ), ranibizumab 10 µg/µL ( D , J ), ranibizumab 0.38 µg/µL ( E , K ), aflibercept 40 µg/µL ( F , L ), aflibercept 1.5 µg/µL ( G , M ), and anti-rat VEGF ( H , N ), showing Brn3a + cells (red) at 7 days (second row) and 30 days (third row) after the intravitreal injection (IVI). Scale bar, 100 µm. ( O ) Bar graph showing mean numbers ± SD of Brn3a + RGCs per retina in naïve animals (N, grey; n = 3 animals: 6 retinas, 3 left retinas and 3 right retinas), right retinas of the experimental animals (right eyes, red; n = 106 retinas), and left retinas of eyes of experimental animals that received IVI of PBS (purple; n = 6 at each survival time), ranibizumab 10 µg/µL (Rbz, dark blue; n = 10 at each survival time), ranibizumab 0.38 µg/µL (Rbz, light blue; n = 10 at each survival time), aflibercept 40 µg/µL (Afbt, dark green; n = 10 at each survival time), aflibercept 1.5 µg/µL (Afbt, light green; n = 10 at each survival time), and anti-rat VEGF (orange; n = 10 at each survival time). *Statistically significant differences between the left injected retinas and the naïve eyes, the right eyes, and the other experimental groups at the same survival time ( P ≤ 0.0001; one-way- ANOVA, Tukey's test). † Statistically significant difference with the lower dose of the same substance (ranibizumab) at 7 days ( P ≤ 0.0001; t test). ‡Statistically significant difference with the lower dose of the same substance (ranibizumab) at 30 days ( P ≤ 0.0001; t test). § Statistically significant difference with the lower dose of the same substance (aflibercept) at 7 days ( P = 0.0018; t test). # Statistically significant difference with the lower dose of the same substance (aflibercept) at 30 days ( P ≤ 0.0001; t test).

Article Snippet: The following substances were injected in the different experimental groups: (1) PBS (Sigma Aldrich, Madrid, Spain); (2) ranibizumab (humanized anti-VEGF, Lucentis, Novartis) at two different concentrations: the human clinical concentration (higher concentration: 10 μg/μL) and a lower concentration (0.38 μg/μL) (see next paragraph); (3) aflibercept (humanized anti-VEGF, Bayer) at two different concentrations: the human clinical concentration (higher concentration; 40 μg/μL) and a lower concentration (1.5 μg/μL; see next paragraph); and (4) goat polyclonal anti-rat VEGF 164 diluted in PBS (15 μg/mL = 0, 015 μg/μL; MGC70609; Leinco Technologies, Inc., St. Louis, MO, USA).

Techniques: Injection

Journal: Investigative Ophthalmology & Visual Science

Article Title: Dose-Related Side Effects of Intravitreal Injections of Humanized Anti-Vascular Endothelial Growth Factor in Rats: Glial Cell Reactivity and Retinal Ganglion Cell Loss

doi: 10.1167/iovs.65.4.10

Figure Lengend Snippet:

Article Snippet: The following substances were injected in the different experimental groups: (1) PBS (Sigma Aldrich, Madrid, Spain); (2) ranibizumab (humanized anti-VEGF, Lucentis, Novartis) at two different concentrations: the human clinical concentration (higher concentration: 10 μg/μL) and a lower concentration (0.38 μg/μL) (see next paragraph); (3) aflibercept (humanized anti-VEGF, Bayer) at two different concentrations: the human clinical concentration (higher concentration; 40 μg/μL) and a lower concentration (1.5 μg/μL; see next paragraph); and (4) goat polyclonal anti-rat VEGF 164 diluted in PBS (15 μg/mL = 0, 015 μg/μL; MGC70609; Leinco Technologies, Inc., St. Louis, MO, USA).

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: Dose-Related Side Effects of Intravitreal Injections of Humanized Anti-Vascular Endothelial Growth Factor in Rats: Glial Cell Reactivity and Retinal Ganglion Cell Loss

doi: 10.1167/iovs.65.4.10

Figure Lengend Snippet: Topography of Brn3a ± RGC after intravitreal injection (IVI) of different anti-VEGF agents. Representative isodensity maps showing the topography of Brn3a + RGC in naïve left retinas ( A ) and right retinas ( B ), and the left retinas of the experimental groups that received an intravitreal injection of PBS ( C , I ), ranibizumab 10 µg/µL ( D , J ), ranibizumab 0.38 µg/µL ( E , K ), aflibercept 40 µg/µL ( F , L ), aflibercept 1.5 µg/µL ( G , M ), and anti-rat VEGF ( H , N ). The color scale bar at the bottom of ( B ) indicates cell density from 0 (purple) to ‡3500 (red). Scale bar, 100 µm.

Article Snippet: The following substances were injected in the different experimental groups: (1) PBS (Sigma Aldrich, Madrid, Spain); (2) ranibizumab (humanized anti-VEGF, Lucentis, Novartis) at two different concentrations: the human clinical concentration (higher concentration: 10 μg/μL) and a lower concentration (0.38 μg/μL) (see next paragraph); (3) aflibercept (humanized anti-VEGF, Bayer) at two different concentrations: the human clinical concentration (higher concentration; 40 μg/μL) and a lower concentration (1.5 μg/μL; see next paragraph); and (4) goat polyclonal anti-rat VEGF 164 diluted in PBS (15 μg/mL = 0, 015 μg/μL; MGC70609; Leinco Technologies, Inc., St. Louis, MO, USA).

Techniques: Injection

Journal: Investigative Ophthalmology & Visual Science

Article Title: Dose-Related Side Effects of Intravitreal Injections of Humanized Anti-Vascular Endothelial Growth Factor in Rats: Glial Cell Reactivity and Retinal Ganglion Cell Loss

doi: 10.1167/iovs.65.4.10

Figure Lengend Snippet: Intrinsically photosensitive retinal ganglion cell (ipRGC) survival. Representative magnifications taken from retinal flat mounts of naïve: left eyes ( A ) and right eyes ( B ), and the different experimental groups that received an intravitreal injection of PBS ( C , I ), ranibizumab 10 µg/µL ( D , J ), ranibizumab 0.38 µg/µL ( E , K ), aflibercept 40 µg/µL ( F , L ), aflibercept 1.5 µg/µL ( G , M ), and anti-rat VEGF ( H , N ) showing IpRGC + cells (green). Scale bar, 100 µm. ( O ) Bar graphs showing mean ± SD of Brn3a + RGCs in naïve animals (N, grey; n = 3 animals: 6 retinas, 3 left retinas and 3 right retinas). Right retinas of the experimental animals (right eyes, red; n = 106 retinas), and left retinas of experimental animals that received intravitreal injection (IVI) of PBS (purple; n = 6 at each survival time), ranibizumab 10 µg/µL (Rbz, dark blue; n = 10 at each survival time), ranibizumab 0.38 µg/µL (Rbz, light blue; n = 10 at each survival time), aflibercept 40 µg/µL (Afbt, dark green; n = 10 at each survival time), aflibercept 1.5 µg/µL (Afbt, light green; n = 10 at each survival time), and anti-rat VEGF (Orange; n = 10 at each survival time). There were no significant differences between the groups.

Article Snippet: The following substances were injected in the different experimental groups: (1) PBS (Sigma Aldrich, Madrid, Spain); (2) ranibizumab (humanized anti-VEGF, Lucentis, Novartis) at two different concentrations: the human clinical concentration (higher concentration: 10 μg/μL) and a lower concentration (0.38 μg/μL) (see next paragraph); (3) aflibercept (humanized anti-VEGF, Bayer) at two different concentrations: the human clinical concentration (higher concentration; 40 μg/μL) and a lower concentration (1.5 μg/μL; see next paragraph); and (4) goat polyclonal anti-rat VEGF 164 diluted in PBS (15 μg/mL = 0, 015 μg/μL; MGC70609; Leinco Technologies, Inc., St. Louis, MO, USA).

Techniques: Injection