Ability of Nonswelling Polyethylene Glycol-Based Vitreous Hydrogel to Maintain Transparency in the Presence of Vitreous Hemorrhage.

Purpose: Postoperative vitreous hemorrhage is a vision-impacting complication of vitrectomy. This preclinical in vitro study assessed the potential ability of a nonswelling polyethylene glycol-based artificial vitreous hydrogel to maintain transparency in the vitreous cavity in the presence of vitreous hemorrhage. Methods: Samples (1 mL) of diluted blood at concentrations of 0.1%, 0.25%, 0.5%, and 1.25% were added to 1 mL samples of polymerized hydrogel in cuvettes (gel + blood group); 2 mL samples of diluted blood at the same concentrations were prepared as controls (blood only group). Spectral transmission curves for the hydrogel (gel + blood group) and diluted blood (blood only group) were obtained before and on days 1, 2, 5, 7, 14, and 28 of the experiment. Between-group comparisons were made using the Student's t-test. The percentage of transmittance in the visible light spectrum (400-700 nm) was measured at each time point. Results: Mean light transmittance was maintained at >90% until day 7 in the gel + blood group and was significantly greater in the gel + blood than in the blood only groups in samples containing blood diluted to 0.25%, 0.5%, and 1.25% during the 28-day study period (P < 0.05). Conclusions: A nonswelling polyethylene glycol-based artificial vitreous hydrogel maintained high optical transparency in the presence of blood through the study period. Injection of this hydrogel into the vitreous cavity at the end of surgery might help to prevent or mitigate vitreous hemorrhage-associated postoperative visual loss. Translational Relevance: The hydrogel may prevent visual loss due to postoperative vitreous hemorrhage.

Periocular injection of in situ hydrogels containing Leu-Ile, an inducer for neurotrophic factors, promotes retinal ganglion cell survival after optic nerve injury.

Intraocular administration of neurotrophic factors has been shown to delay irreversible degeneration of retinal ganglion cells (RGCs). It would be beneficial for the treatment of optic nerve (ON) injury if such neurotrophic factors could be delivered in a less-invasive manner. The dipeptide leucine-isoleucine (Leu-Ile) appears to induce the production of neurotrophic factors, including brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF), in the brain. We therefore administered Leu-Ile via periocular depot injection in rats and investigated the dipeptide's ability to induce BDNF and GDNF in the retina and to delay RGC loss in an ON injury model. Poloxamer-alginate hydrogels containing Leu-Ile were injected into the subconjunctival space of intact or ON-injured rats. BDNF and GDNF levels in the retina were determined by an enzyme immunoassay. Survival of RGCs was assessed in retinal flatmounts. Activation of extracellular signal-regulated kinases (ERK) and cAMP response element binding protein (CREB) in the retina was examined by Western blotting. At 2 h after injection of fluorescein isothiocyanate-conjugated Leu-Ile, the fluorescence intensities in the retina were 4.3-fold higher than those in the saline control. Treatment with Leu-Ile significantly increased the retinal levels of BDNF at 6 h and GDNF at 6-72 h after injection. Treatment with Leu-Ile significantly increased RGC survival to 14 days after ON injury and enhanced the activation of ERK at 72 h and CREB at 48 h after injection in the ON-injured retina. These results suggest that periocular delivery of Leu-Ile induces BDNF and GDNF production in the retina, which may eventually enhance RGC survival after ON injury.

Neuroprotective effect of PACAP against kainic acid-induced neurotoxicity in rat retina.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is well known to protect delayed neuronal cell death in the brain of rodents. In order to investigate the neuroprotective action of PACAP in the retina, we examined the effects of PACAP on kainic acid (KA)-induced neurotoxicity in the rat retina. Many ganglion cells in the retina died after KA injection in the control group and PACAP treatment significantly promoted cell survival. These findings strongly suggest that PACAP plays very important roles in preventing cell death in the retina.

Pituitary adenylate cyclase-activating peptide (PACAP) stimulates production of interleukin-6 in rat Müller cells.

Pituitary adenylate cyclase-activating peptide (PACAP) is known to regulate not only neurons but also astrocytes. Here, we investigated, both in vitro and in vivo, the effects of PACAP38 on rat Müller cells, which are the predominant glial element in the retina. Müller cells isolated from juvenile Wistar rats were treated with PACAP38 or PACAP6-38, a PACAP selective antagonist. Cell proliferation was determined by measuring the incorporation of bromodeoxyuridine with ELISA. Interleukin-6 (IL-6) levels in the culture medium were determined by a bioassay using B9 cells, IL-6 dependent hybridoma. In adult Wistar rats, the expression of IL-6 in the retina after intravitreal injection of PACAP38 (10 pmol) was assessed by immunohistochemistry. PACAP38 stimulated IL-6 production in Müller cells at a concentration as low as 10(-12) M, which did not induce cell proliferation. This elevation of IL-6 production was inhibited by PACAP6-38. Radial IL-6 expression was observed throughout the retina at 2 and 3 days after PACAP38 injection. These data demonstrate that Müller cells are one of the target cells for PACAP. IL-6, which is released from Müller cells with stimulation by PACAP, may play a significant role in the retina.