Xiaofen Mo

Rescue of Photoreceptors by BDNF Gene Transfer Using In Vivo Electroporation in the RCS Rat of Retinitis Pigmentosa

Purpose: To investigate the feasibility of introducing brain-derived neurotrophic factor (BDNF) gene into retinal pigment epithelial cells in vivo by electroporation and whether this method can rescue photoreceptors of retinitis pigmentosa in Royal College Surgeons (RCS) rats. Methods: The BDNF-GFP fusion eukaryotic-expressing plasmid was constructed and subretinally or intravitreously injected into the eyes of RCS rats followed by in vivo electroporation. The expression of BDNF mRNA and protein was detected by RT-PCR and Western immunoblot analysis. The number of surviving photoreceptors was counted, and the TdT-dUTP terminal nick-end labeling (TUNEL) method was used to detect the apoptotic retinal cells at different timepoints after introduction of BDNF plasmid. Results: Treated eyes showed a significantly higher rescue ratio and a lower number of TUNEL-positive photoreceptors than did the control eyes at various timepoints. Conclusion: These findings provide evidence that electroporation is an effective method for gene transfer into retinal pigment epithelial cells, and the rescue of photoreceptors can be achieved by BDNF gene transfection with electroporation.

Neuroprotective effect of transcorneal electrical stimulation on ischemic damage in the rat retina

Some previous studies have showed that transcorneal electrical stimulation (TES) could protect retinal neurons in certain rodent models. However, it is not yet clear whether TES could also definitely protect retinal neurons against ischemic insults. In the present study, we hypothesized that TES had such a neuroprotective effect and further investigated its underlying mechanism. Adult female Sprague-Dawley (SD) rats received TES treatment every other day after ocular ischemia was induced by elevating the intraocular pressure to 120 mm Hg for 60 min. Retinal ganglion cells (RGCs) were labeled retrogradely 7 days before ischemia and were counted 7 and 14 days later. At the same time points, retinal function was assessed by scotopic electroretinography (ERG), combined with retinal histological analysis. The glutamine synthetase (GS) immunoreactivity was compared between ischemic retinas with TES and those with sham stimulation under identical confocal laser microscope conditions. The immunohistochemical indications were confirmed by Western blot analysis. Higher mean density of RGCs was quantified in TES treated retinas compared to retinas with sham stimulation on days 7 and 14 after ischemia. Similarly, histological analysis showed that TES better preserved the mean thickness of separate retinal layers. ERG studies indicated that by undergoing TES treatment, the b-wave amplitude was also significantly preserved on day 7 after ischemia and recovered robustly on day 14. Immunohistochemical and Western blot analysis both revealed that GS levels remarkably increased after TES and lasted for at least 7 days. Our results indicate that TES can protect retinal neurons against ischemic insults, probably related to increasing levels of GS localized in Müller cells. These findings suggest a new approach for potential clinical application to ocular ischemic diseases.

A new type of Schwann cell graft transplantation to promote optic nerve regeneration in adult rats

Like other parts of the central nervous system, the adult mammalian optic nerve is difficult to regenerate after injury. Transplantation of the peripheral nerve or a Schwann cell (SC) graft can promote injured axonal regrowth. We tried to develop a new type of tissue‐engineered SC graft that consisted of SCs seeded onto a poly(lactic‐co‐glycolic acid)/chitosan conduit. Meanwhile, SCs were transfected along the ciliary neurotrophic factor (CNTF) gene in vitro by electroporation to increase their neurotrophic effect. Four weeks after transplantation, GAP‐43 labelled regenerating axons were found in the SC grafts, and axons in the CNTF–SC graft were longer than those in the SC graft. Tissue‐engineered SC grafts can provide a feasible environment for optic nerve regeneration and may become an alternative for bridging damaged nerves and repairing nerve defects in the future. Copyright

Safety evaluation of poly(lactic-co-glycolic acid)/poly(lactic-acid) microspheres through intravitreal injection in rabbits

Poly(lactic-co-glycolic acid) (PLGA) and/or poly(lactic-acid) (PLA) microspheres are important drug delivery systems. This study investigated eye biocompatibility and safety of PLGA/PLA microspheres through intravitreal injection in rabbits. Normal New Zealand rabbits were randomly selected and received intravitreal administration of different doses (low, medium, or high) of PLGA/PLA microspheres and erythropoietin-loaded PLGA/PLA microspheres. The animals were clinically examined and sacrificed at 1, 2, 4, 8, and 12 weeks postadministration, and retinal tissues were prepared for analysis. Retinal reactions to the microspheres were evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick end staining and glial fibrillary acidic protein immunohistochemistry. Retinal structure changes were assessed by hematoxylin and eosin staining and transmission electron microscopy. Finally, retinal function influences were explored by the electroretinography test. Terminal deoxynucleotidyl transferase-mediated dUTP nick end staining revealed no apoptotic cells in the injected retinas; immunohistochemistry did not detect any increased glial fibrillary acidic protein expression. Hematoxylin and eosin staining and transmission electron microscopy revealed no micro- or ultrastructure changes in the retinas at different time points postintravitreal injection. The electroretinography test showed no significant influence of scotopic or photopic amplitudes. The results demonstrated that PLGA/PLA microspheres did not cause retinal histological changes or functional damage and were biocompatible and safe enough for intravitreal injection in rabbits for controlled drug delivery.

Objective optical quality and intraocular scattering in myopic adults.

PURPOSE To evaluate objective optical quality and intraocular scattering in adults with myopia. METHODS This was a cross-sectional study. Patients between 18 and 40 years of age were recruited from those undergoing routine preoperative examinations prior to myopic refractive surgery. The spherical equivalent refraction (SE) ranged from -14.25 to -0.63 diopters (D). Right eyes of 274 subjects were included for factor analyses: 25 eyes were super-high myopia (SE < -9 D), 88 eyes were high myopia (-6 D > SE ≥ -9 D),133 eyes were moderate myopia (-3 D > SE ≥ -6 D), and 28 eyes were low myopia (SE ≥ -3 D). A double-pass system was used to measure the modulation transfer function cutoff frequency (MTFcutoff) and objective scatter index (OSI). RESULTS Mean MTFcutoff was 32.38 ± 9.73 and 27.61 ± 8.11 cycles per degree (cyc/deg) in the high and super-high myopia groups, significantly lower than 39.92 ± 10.53 and 37.39 ± 8.74 cyc/deg in the low and moderate groups (P < 0.01). Mean OSI was 0.89 ± 0.61 and 1.33 ± 0.65 in the high and super-high myopia groups, significantly higher than in the low and moderate groups, 0.50 ± 0.39 and 0.57 ± 0.44 (P < 0.01). No significant difference was found between the low myopia and the moderate myopia group in any of the parameters (P > 0.05). Other factors such as eye side, sex, and age did not significantly affect optical quality parameters or OSI in this study population (P > 0.05). CONCLUSIONS Optical quality and intraocular scattering varied among individuals with myopia. High myopia has more influence on retinal image quality and scattering than moderate and low myopia. Our study also helps to establish MTFcutoff and OSI standards for Chinese refractive surgery candidates.

Intra- and Intersession Repeatability of an Optical Quality and Intraocular Scattering Measurement System in Children

Purpose To evaluate intra- and intersession repeatability of objective optical quality and intraocular scattering measurements with a double-pass system in children. Methods Forty-two eyes of 42 children were included in the study. An optical quality analysis system (OQAS) was used to measure optical quality parameters, including modulation transfer function cutoff frequency (MTFcutoff), Strehl ratio (SR), OQAS values (OV) at 3 different contrasts and objective scatter index (OSI). Three measurement sessions with 10-min intervals were operated by the same technician, and in each session four consecutive measurements were obtained. Results Mean values for MTFcutoff, SR and OSI were 46.85 ± 7.45cpd, 0.27 ± 0.06 and 0.34 ± 0.22 respectively. 1) The intraclass correlation coefficients were ranged from 0.89 to 0.97 and coefficients of variation from 0.06 to 0.16 for all the parameters in the first session; the relative repeatability were 11.1% (MTFcutoff), 22.5% (SR), 10.9% (OV100%), 16.6% (OV2%), 22.4% (OV9%) and 56.3% (OSI). Similar results were found in the second and third sessions. 2) Bland-Altman analysis showed that narrow 95% confidence intervals (compared between the first and second sessions) ranged from -5.42 to 5.28 (MTFcutoff), -0.05 to 0.07 (SR), -0.18 to 0.18 (OV100%), -0.26 to 0.29 (OV20%), -0.33 to 0.39 (OV9%) and -0.11 to 0.09 (OSI); the comparison between any two of the three sessions showed similar results. Conclusion Measurements of optical quality and intraocular scattering in children by the double-pass system showed good intra- and intersession repeatability. Retinal image quality is high and intraocular scattering is low in children.