Long-Term Effects on Retinal Structure and Function in a Mouse Endothelin-1 Model of Retinal Ganglion Cell Degeneration.

Purpose: To study the long-term effects of endothelin-1 (ET-1)-induced retinal pathologies in mouse, using clinically relevant tools. Methods: Adult C57BL/6 mice (7-9 weeks old) were intravitreally injected with PBS (n = 10) or 0.25 (n = 8), 0.5 (n = 8), or 1 nmol ET-1 (n = 9) and examined using electroretinogram, optical coherence tomography (OCT), and Doppler OCT at baseline and postinjection days 10, 28, and 56. Retinal ganglion cell (RGC) survival in retinal whole mount was quantified at days 28 and 56. Results: ET-1 induced immediate retinal arterial constriction. The significantly reduced total blood flow and positive scotopic threshold response in the 0.5- and 1-nmol ET-1 groups at day 10 were recovered at day 28. A-wave magnitude was also significantly reduced at days 10 and 28. While a comparable and significant reduction in retinal nerve fiber layer thickness was detected in all ET-1 groups at day 56, the 1-nmol group was the earliest to develop such change at day 28. All ET-1 groups showed a transient inner retinal layer thinning at days 10 and 28 and a plateaued outer layer thickness at days 10 to 56. The 1-nmol group showed a significant RGC loss over all retinal locations examined at day 28 as compared with PBS control. As for the lower-dosage groups, significant RGC density loss at central and midperipheral retina was detected at day 56 when compared with day 28. Conclusions: ET-1 injection in mice resulted in a transient vascular constriction and reduction in retinal functions, as well as a gradual loss of retinal nerve fiber layer and RGC in a dose-dependent manner.

Potential role of Lycium barbarum polysaccharides in glaucoma management: evidence from preclinical in vivo studies.

In recent years, the pharmacological benefits of herbal extracts have been revisited for their potential neuroprotective effects in glaucoma. The polysaccharides extracted from the fruits of Lycium barbarum L., or Lycium barbarum polysaccharides, exert their anti-aging effect through reducing oxidative stress, modulating the immune response, enhancing neuronal responses, and promoting cytoprotection. The therapeutic efficacy of Lycium barbarum polysaccharides in preserving retinal ganglion cells and their functions was demonstrated in a range of experimental models of optic neuropathies. These include the acute and chronic ocular hypertension models, the partial optic nerve transection model, and the ischemic-reperfusion injuries model. Based on these findings, Lycium barbarum polysaccharides appear to be a good candidate to be developed as a neuroprotective agent for treating multifactorial diseases. This review aims to present a comprehensive review on the latest preclinical evidence on the pre- and post-treatment benefits of Lycium barbarum polysaccharides in retinal ganglion cell neuroprotection. The possible mechanisms of Lycium barbarum polysaccharides mediating retinal ganglion cell neuroprotection will also be described. Moreover, the potential research gaps in the effective translation of Lycium barbarum polysaccharides treatment into clinical glaucoma management will be discussed.

Experimentally induced myopia and myopic astigmatism alter retinal electrophysiology in chickens.

Myopia (or "short-sightedness") and astigmatism are major causes of visual impairment worldwide. Significant amounts of astigmatism are frequently observed in infants and have been associated with myopia development. Although it is well established that both myopia and astigmatism are associated with ocular structural changes from anterior to posterior segments, very little is known on how these refractive errors alter retinal functions. This study investigated the effects of experimentally induced myopia and myopic-astigmatism on retinal electrophysiology by using an image-guided, multifocal global flash stimulation in chickens, a widely used animal model for refractive error development. Myopia and myopic-astigmatism were experimentally induced, respectively, by wearing spherical (- 10 D, n = 12) and sphero-cylindrical lenses (- 6.00 DS/- 8.00 DCx90: Hyperopic With-The Rule, H-WTR, n = 15; - 6.00 DS/- 8.00 DCx180: Hyperopic Against-The-Rule, H-ATR, n = 11) monocularly for a week (post-hatching day 5 to 12). An aged-matched control group without any lens treatment provided normal data (n = 12). Multifocal electrophysiological results revealed significant regional variation in the amplitude of induced component (IC) (central greater than peripheral; both p < 0.05) in the normal and H-ATR groups, but not in the - 10 D and H-WTR groups. Most importantly, for the first time, our results showed that both H-WTR and H-ATR groups exhibited a significantly longer implicit time of the inner retinal response at the central region when compared to the normal and - 10 D groups, highlighting a significant role of astigmatism in retinal physiology.

Selective blue-filtering spectacle lens protected primary porcine RPE cells against light emitting diode-induced cell damage.

This study aimed to investigate whether use of a selective-blue-filtering (S-BF) lens can protect cultured primary porcine RPE cells against photo-irradiation. Transmittance of S-BF and UV-filtering (UVF) lenses was characterised spectrophotometrically. RPE cells were exposed to 1700 lux of white (peak λ at 443 and 533 nm; 0.44 mW/cm2) or blue (peak λ at 448 and 523 nm; 0.85 mW/cm2) LED light for 16 h to evaluate the influence of light source on the culture. The effect of the S-BF and UVF ophthalmic lenses on RPE cell cultures under blue light irradiation was then investigated. Cell viability was compared using trypan blue and MTT assays. Intracellular ROS production was detected by a fluorescein probe CM-H2DCFDA. Expression levels of catalase and Prdx3 were analysed by western blot. Trypan blue staining showed blue light caused more cell death than no light (p = 0.001) or white light (p = 0.005). MTT assay supported the hypothesis that exposure to blue light damaged RPE cells more severely than no light (p = 0.002) or white light (p = 0.014). Under blue light, use of the S-BF lens, which blocked 17% more blue light than the UVF lens, resulted in higher cellular viability (S-BF: 93.4±1.4% vs UVF: 90.6±1.4%; p = 0.022; MTT: 1.2-fold; p = 0.029). Blue and white light both significantly increased ROS production. The S-BF lens protected cells, resulting in lower levels of ROS and higher expression of catalase and Prdx3. To conclude, blue LED light exposure resulted in significant cytotoxicity to RPE cells. Partial blockage of blue light by an S-BF lens led to protective effects against retinal phototoxicity, which were mediated by reduction of ROS and increased levels of antioxidant enzymes.

Longitudinal outcomes of circumlimbal suture model-induced chronic ocular hypertension in Sprague-Dawley albino rats.

PURPOSE: To characterise longitudinal structural and functional changes in albino Sprague-Dawley rats following circumlimbal suture ocular hypertension (OHT) induction. METHODS: Ten-week-old rats (n = 24) underwent suture implantation around the limbal region in both eyes. On the next day, the suture was removed from one eye (control eyes) and left intact in the other eye (OHT eyes) of each animal. Intraocular pressure (IOP) was monitored weekly twice for the next 15 weeks. Optical coherence tomography (OCT) and electroretinogram (ERG) were measured at baseline and weeks 4, 8, 12, and 15, and eyes were then collected for histological assessment. RESULTS: Sutured eyes (n = 12) developed IOP elevation of ~ 50% in the first 2 weeks that was sustained at ~ 25% above the control eye up to week 15 (p = 0.001). Animals with insufficient IOP elevation (n = 6), corneal changes (n = 3), and attrition (n = 3) were excluded from the analysis. OHT eyes developed significant retinal nerve fibre layer (RNFL) thinning (week 4: - 19 ± 14%, p = 0.10; week 8: - 17 ± 12%, p = 0.04; week 12: - 16 ± 10%, p = 0.04, relative to baseline) and reduction in retinal ganglion cell (RGC) density (- 32 ± 26%, p = 0.02). At week 15, both inner (9 ± 7%, p = 0.01) and outer retinal layer thicknesses (6.0 ± 5%, p = 0.001) showed a mild increase in thicknesses. The positive scotopic threshold response (- 28 ± 25%, p = 0.04) and a-wave were significantly reduced at week 12 (- 35 ± 21%; p = 0.04), whereas b-wave was not significantly affected (week 12: - 18 ± 27%, p = 0.24). CONCLUSION: The circumlimbal suture model produced a chronic, moderate IOP elevation in an albino strain that led to RNFL thinning and reduced RGC density along with the reductions in ganglion and photoreceptoral cell functions. There was a small thickening in both outer and inner retinal layers.

Posttreatment Intervention With Lycium Barbarum Polysaccharides is Neuroprotective in a Rat Model of Chronic Ocular Hypertension.

Purpose: To investigate the neuroprotective effects of Lycium barbarum polysaccharides (LBP) against chronic ocular hypertension (OHT) in rats and to consider if effects differed when treatment was applied before (pretreatment) or during (posttreatment) chronic IOP elevation. Methods: Sprague-Dawley rats (10-weeks old) underwent suture implantation around the limbus for 15 weeks (OHT) or 1 day (sham). Four experimental groups were studied, three OHT groups (n = 8 each) treated either with vehicle (PBS), LBP pretreatment or posttreatment, and a sham control (n = 5) received no treatment. LBP (1 mg/kg) pre- and posttreatment were commenced at 1 week before and 4 weeks after OHT induction, respectively. Treatments continued up through week 15. IOP was monitored twice weekly for 15 weeks. Optical coherence tomography and ERG were measured at baseline, week 4, 8, 12, and 15. Eyes were collected for ganglion cell layer (GCL) histologic analysis at week 15. Results: Suture implantation successfully induced approximately 50% IOP elevation and the cumulative IOP was similar between the three OHT groups. When compared with vehicle control (week 4: -23 ± 5%, P = 0.03), LBP pretreatment delayed the onset of retinal nerve fiber layer (RNFL) thinning (week 4, 8: -2 ± 7%, -11 ± 3%, P > 0.05) and arrested further reduction up through week 15 (-10 ± 4%, P > 0.05). LBP posttreatment intervention showed no significant change in rate of loss (week 4, 15: -25 ± 4.1%, -28 ± 3%). However, both LBP treatments preserved the retinal ganglion cells (RGC) and retinal functions up to week 15, which were significantly reduced in vehicle control. Conclusions: LBP posttreatment arrested the subsequent neuronal degeneration after treatment commencement and preserved RGC density and retinal functions in a chronic OHT model, which was comparable with pretreatment outcomes.

Lycium Barbarum Polysaccharides Rescue Neurodegeneration in an Acute Ocular Hypertension Rat Model Under Pre- and Posttreatment Conditions.

Purpose: To investigate the posttreatment neuronal rescue effects of Lycium barbarum polysaccharides (LBP) in an acute ocular hypertensive (AOH) model. Methods: Intraocular pressure (IOP) was elevated manometrically to 80 mm Hg (AOH) or 15 mm Hg (sham) for 120 minutes in adult Sprague-Dawley rats. Five experimental groups were considered: Three AOH groups were pretreated with PBS (vehicle) (n = 9), LBP 1 mg/kg (n = 8), or 10 mg/kg (n = 13), and one AOH group was posttreated with LBP 10 mg/kg (n = 8), once daily. The sham cannulation group (n = 5) received no treatment. Pretreatments commenced 7 days before and posttreatment 6 hours after AOH, and continued up through postcannulation day 28. All the animals underwent optical coherence tomography and electroretinogram measurements at baseline and postcannulation days 10 and 28. The ganglion cell layer (GCL) densities were quantified at day 28. Results: Both inner retinal layer thickness (IRLT) and positive scotopic threshold response (pSTR) underwent significant reduction (≥50% of thickness and amplitude) in the vehicle group (P < 0.05). Pretreatment with LBP 1 and 10 mg/kg retained 77 ± 11% and 89 ± 8% of baseline IRLT, respectively, and preserved pSTR functions. The posttreatment group showed a significant reduction in IRLT (-35 ± 8%, P < 0.001) and pSTR (∼48% of baseline, P < 0.001) on day 10. By day 28, there was an improvement in functional pSTR (∼72% of baseline, P > 0.05) with no significant further thinning (-40 ± 8%, P = 0.15) relative to day 10. GCL density was reduced in vehicle control (P = 0.0001), but did not differ between sham and pre- and posttreated AOH groups. Conclusions: The rescue effect of LBP posttreatment was observed later, which arrested the secondary degeneration and improved the retinal function.

Delay of cone degeneration in retinitis pigmentosa using a 12-month treatment with Lycium barbarum supplement.

ETHNOPHARMACOLOGICAL RELEVANCE: Lycium barbarum L. (also known as "Goji berry"), a traditional Chinese herbal medicine, has been a common herb in the traditional Chinese pharmacopoeia for centuries. The main active component is the Lycium barbarum polysaccharides and its antioxidative effect has been widely shown to provide neuroprotection to the eye, and it would, therefore, be interesting to determine if Lycium barbarum help delay vision deterioration in patients with retinitis pigmentosa. AIM OF THE STUDY: Cone rescue is a potential method for delaying deterioration of visual function in Retinitis pigmentosa (RP). This study aimed to investigate the treatment effect of Lycium barbarum L. (LB) supplement on retinal functions and structure in RP patients after a 12-month intervention trial. METHODS: The investigation was a double-masked and placebo-controlled clinical study. Each of forty-two RP subjects who completed the 12-month intervention (23 and 19 in the treatment and placebo groups respectively) received a daily supply of LB or placebo granules for oral administration. The primary outcome was change of best corrected visual acuity (VA) (90% and 10% contrast) from the baseline to the end of treatment. The secondary outcomes were sensitivity changes of the central visual field, amplitude of full-field electroretinogram (ffERG) (including scotopic maximal response and photopic cone response), and average macular thickness. RESULTS: The compliance rates for both groups exceeded 80%. There were no deteriorations of either 90% or 10% contrast VA in the LB group compared with the placebo group (p = 0.001). A thinning of macular layer was observed in the placebo group, which was not observed in the LB group (p = 0.008). However, no significant differences were found in the sensitivity of visual field or in any parameters of ffERG between the two groups. No significant adverse effects were reported in the treatment group. CONCLUSIONS: LB supplement provides a neuroprotective effect for the retina and could help delay or minimize cone degeneration in RP. CLASSIFICATIONS: Clinical Studies (1.05). TRIAL REGISTRATION: clinicaltrials.gov Identifier NCT02244996.

Subclinical Decrease in Central Inner Retinal Activity Is Associated With Myopia Development in Children.

Purpose: To investigate the characteristics of retinal electrophysiological activity in relation to early myopia development in children. Methods: Fifty-six children aged 6 to 9 years with emmetropic refractive error (defined as ≥ -0.5 diopter [D] and ≤ +0.5 D) were recruited. Cycloplegic refraction, axial length, and global flash multifocal electroretinogram (MOFO mfERG) at 49% and 96% contrast levels were recorded in all children at their first visit. The refraction and axial length measurements were repeated after 1 year. The amplitudes and implicit times of the direct component (DC) and the induced component (IC) of the MOFO mfERG obtained at the initial visit were analyzed. Correlations between the MOFO mfERG parameters and changes in refractive error and axial length were investigated. Results: The mean spherical equivalent refractive error and axial length of the eyes of the children at the first visit were +0.19 ± 0.33 D and 23.14 ± 0.6 mm, respectively. After 1 year, the mean refractive error increased by -0.55 ± 0.53 D, whereas axial length increased by 0.37 ± 0.22 mm. The changes in refractive error and axial length were significantly correlated with the central IC amplitudes at 49% contrast level measured at the initial visit (ρ = 0.46, P < 0.001 and ρ = -0.34, P = 0.01, respectively). Conclusions: The prospective changes we have shown are believed to derive from central inner retina. These changes appear to precede myopia and could be a potential reference for juvenile myopia development.

Childhood exposure to constricted living space: a possible environmental threat for myopia development.

PURPOSE: People in Hong Kong generally live in a densely populated area and their homes are smaller compared with most other cities worldwide. Interestingly, East Asian cities with high population densities seem to have higher myopia prevalence, but the association between them has not been established. This study investigated whether the crowded habitat in Hong Kong is associated with refractive error among children. METHODS: In total, 1075 subjects [Mean age (S.D.): 9.95 years (0.97), 586 boys] were recruited. Information such as demographics, living environment, parental education and ocular status were collected using parental questionnaires. The ocular axial length and refractive status of all subjects were measured by qualified personnel. RESULTS: Ocular axial length was found to be significantly longer among those living in districts with a higher population density (F2,1072  = 6.15, p = 0.002) and those living in a smaller home (F2,1072  = 3.16, p = 0.04). Axial lengths were the same among different types of housing (F3,1071  = 1.24, p = 0.29). Non-cycloplegic autorefraction suggested a more negative refractive error in those living in districts with a higher population density (F2,1072  = 7.88, p < 0.001) and those living in a smaller home (F2,1072  = 4.25, p = 0.02). After adjustment for other confounding covariates, the population density and home size also significantly predicted axial length and non-cycloplegic refractive error in the multiple linear regression model, while axial length and refractive error had no relationship with types of housing. CONCLUSIONS: Axial length in children and childhood refractive error were associated with high population density and small home size. A constricted living space may be an environmental threat for myopia development in children.

Stereoacuity in mild, moderate and severe glaucoma.

PURPOSE: To determine the association between stereoacuity and severity of glaucoma. METHODS: Stereoacuity was measured using the Titmus stereo test for 97 glaucoma subjects with visual acuity better than 0.3 logMAR (6/12, 20/40 Snellen) and reliable Humphrey visual fields (24-2 SITA standard). The severity of visual fields was graded with the Hodapp-Anderson-Parrish (HAP) classification and visual fields index (VFI) score and the stereoacuities for different grades were compared. Presence of a significant field defect (p < 1%) in any one of the paracentral points in 10-2 SITA standard/Macular threshold was considered as a threat to fixation. RESULTS: Mean ± SD age of the subjects was 57.28 ± 14.0 years. The median [inter-quartile range, (IQR)] stereoacuity of the 97 subjects was 50 (40-100) arc sec with 13 subjects having stereoacuity of >3000 arc sec. Median (IQR) stereoacuity of 97 subjects classified with HAP as mild, moderate and severe were 40 (40-40), 50 (40-60) and 60 (40-400) arc sec; similarly median (IQR) stereoacuity of the subjects classified using VFI as mild, moderate and severe were 40 (40-55), 60 (40-100) and 120 (50-3000) s of arc and the difference between groups were significant (p < 0.001). Median (IQR) stereoacuity in subjects with no threat to fixation (45) and threat to fixation in at least one eye (52) were 40 (40-60) and 70 (40-800) arc sec (p < 0.001). CONCLUSIONS: Decreased stereoacuity was associated with greater glaucomatous visual field loss, although it was normal with early visual field defects. Relatively moderate defects can be associated with decreased stereoacuity and it is more pronounced in the severe stages of the disease when there is a threat to fixation.