Efficient retinal ganglion cells transduction by retro-orbital venous sinus injection of AAV-PHP.eB in mature mice.

Gene therapy is one of the strategies that may reduce or reverse progressive neurodegeneration in retinal neurodegenerative diseases. However, efficiently delivering transgenes to retinal ganglion cells (RGCs) remains hard to achieve. In this study, we innovatively investigated transduction efficiency of adeno-associated virus (AAV)-PHP.eB in murine RGCs by retro-orbital venous sinus injection. Five doses of AAV-PHP.eB-EGFP were retro-orbitally injected in venous sinus in adult C57/BL6J mice. Two weeks after administration, RGCs transduction efficiency was quantified by retinal flat-mounts and frozen section co-labeling with RGCs marker Rbpms. In addition, safety of this method was evaluated by RGCs survival rate and retinal morphology. To conform efficacy of this new method, AAV-PHP.eB-CNTF was administrated into mature mice through single retro-orbital venous injection after optic nerve crush injury to evaluate axonal elongation. Results indicated that AAV- PHP.eB readily crossed the blood-retina barrier and was able to transduce more than 90% of RGCs when total dose of virus reached 5 × 1010 vector genomes (vg). Moreover, this technique did not affect RGCs survival rate and retinal morphology. Furthermore, retro-orbital venous delivery of AAV-PHP.eB-CNTF effectively transduced RGCs, robustly promoted axonal regeneration after optic nerve crush injury. Thus, novel AAV-PHP.eB retro-orbital injection provides a minimally invasive and efficient route for transgene delivery in treatment of retinal neurodegenerative diseases.

Autosomal Dominant Weill-Marchesani-Like Syndrome in a Chinese Family due to Novel Haplotypic Mutations in LTBP2.

INTRODUCTION: Weill-Marchesani syndrome (WMS) is a hereditary connective tissue disorder with substantial heterogeneity in clinical features and genetic etiology, so it is essential to define the full mutation spectrum for earlier diagnosis. In this study, we report Weill-Marchesani-like syndrome (WMS-like) change to autosomal dominance inheritance caused by novel haplotypic mutations in latent transforming growth factor beta-binding protein 2 (LTBP2). METHODS: Twenty-five members from a 4-generation Chinese family were recruited from Guangzhou, of whom nine were diagnosed with WMS-like disease, nine were healthy, and seven were of "uncertain" clinical status because of their young age. All members received detailed physical and ocular examinations. Whole-exome sequencing, Sanger sequencing, and real-time PCR were used to identify and verify the causative mutations in family members. RESULTS: Genetic sequencing revealed novel haplotypic mutations on the same LTBP2 chromosome associated with WMS-like, c. 2657C>A/p.T886K in exon 16 and deletion of exons 25-36. Real-time PCR and Sanger sequencing verified both mutations in patients with clinically diagnosed WMS-like, and in one "uncertain" child. In these patients, the haplotypic mutations led to ectopia lentis, short stature, and obesity. CONCLUSION: Our study revealed that WMS-like may be associated with haplotypic LTBP2 mutations with autosomal dominant inheritance.

Ocular trauma from Lawn mower accidents: Clinical insights, visual outcomes and microbial profiles.

AIMS: To characterize the clinical features of ocular trauma resulting from lawn mower, identify determinants of unfavorable final visual acuity (FVA), and assess the spectrum of microbial in posttraumatic endophthalmitis. METHODS: This retrospective study enrolled patients who experienced ocular trauma due to lawn mower at Zhongshan Ophthalmic Center from January 2013 to August 2021. Demographics, clinical features, isolated microorganisms, risk factors influencing reduced visual acuity, treatment regimens, and utilization of eyewear were collected. RESULTS: The study included 140 participants (140 eyes) (49.47 ± 12.03 years, 95% male). The predominant injury manifestations were penetrating globe injuries (75.7%) and intraocular foreign bodies (51.4%). Endophthalmitis occurred in 35 cases (25%) and Bacillus cereus (23.5%) was the primary pathogen, followed by Staphylococcus epidermidis (11.8%) and Streptococcus species (11.8%). Following the initial assessment, where 77.9% of patients had initial visual acuity (IVA) at grade IV (ranging from light perception to 4/200) and only 0.7% attained grade I (better than 20/40), post-treatment results revealed that 5.7% achieved FVA at grade I, with a concurrent decrease in patients with grade IV vision to 64.3%. Multivariate logistic regression revealed that injury protection (p < 0.001, OR = 0.237, 95% CI = 0.126-0.446), IVA (p = 0.001, OR = 4.102, 95% CI = 1.730-9.729), and retinal detachment (p = 0.042, OR = 8.105, 95% CI = 1.075-61.111) were significant independent risk factors impacting FVA. CONCLUSION: Lawn mower often cause severe ocular injuries, with high-velocity metal foreign bodies that can lead to infections, most commonly caused by Bacillus cereus. Correct use of protective gear, initial vision assessment, and detecting retinal detachment are crucial for visual prognosis.

Comparison of retinal parameters between rhesus and cynomolgus macaques.

Nonhuman primates are important research models for basic vision research, preclinical pathogenesis, and treatment studies due to strong similarities in retinal structure and function with humans. We compared retinal parameters between 10 healthy normal rhesus macaques (Macaca mulatta) and 10 cynomolgus macaques (Macaca fascicularis) by optical coherence tomography and electroretinography. The Heidelberg Spectralis® HRA+OCT and Roland multifocal electrophysiometer were used to analyze retinal morphology, multifocal electroretinograms (mfERGs), and full-field electroretinograms (ff-ERGs). Mean retinal thickness was lowest in the central fovea of macaques and did not differ significantly between species, but the retinal thicknesses of the nerve fiber ganglion cell layer and the inner plexiform layer were significantly different. The amplitude density of the N1 wave was lower in rhesus macaques than in cynomolgus macaques in ring and quadrant areas. Dark-adapted 3.0 oscillatory potentials (reflection of amacrine cell activity) and light-adapted 30-hz flicker ERG (a sensitive cone-pathway-driven response) waveforms of the ff-ERG were similar in both species, while the times to peaks in dark-adapted 0.01 ERG (the rod-driven response of bipolar cells) and dark-adapted 3.0 ERG (combined rod and cone system responses) as well as the implicit times of the a- and b-waves in light-adapted 3.0 ERG (the single-flash cone response) were substantially different. This study provides normative retinal parameters for nonhuman primate research on basic and clinical ophthalmology, as well as a reference for researchers in the appropriate selection of rhesus or cynomolgus macaques as models for ophthalmology studies.

Corrigendum: Characteristics of eye-related emergency visits and triage differences by nurses and ophthalmologists: perspective from a single eye center in southern China.

[This corrects the article DOI: 10.3389/fmed.2023.1091128.].

EE-Explorer: A Multimodal Artificial Intelligence System for Eye Emergency Triage and Primary Diagnosis.

PURPOSE: To develop a multimodal artificial intelligence (AI) system, EE-Explorer, to triage eye emergencies and assist in primary diagnosis using metadata and ocular images. DESIGN: A diagnostic, cross-sectional, validity and reliability study. METHODS: EE-Explorer consists of 2 models. The triage model was developed from metadata (events, symptoms, and medical history) and ocular surface images via smartphones from 2038 patients presenting to Zhongshan Ophthalmic Center (ZOC) to output 3 classifications: urgent, semiurgent, and nonurgent. The primary diagnostic model was developed from the paired metadata and slitlamp images of 2405 patients from ZOC. Both models were externally tested on 103 participants from 4 other hospitals. A pilot test was conducted in Guangzhou to evaluate the hierarchical referral service pattern assisted by EE-Explorer for unspecialized health care facilities. RESULTS: A high overall accuracy, as indicated by an area under the receiver operating characteristic curve (AUC) of 0.982 (95% CI, 0.966-0.998), was obtained using the triage model, which outperformed the triage nurses (P < .001). In the primary diagnostic model, the diagnostic classification accuracy (CA) and Hamming loss (HL) in the internal testing were 0.808 (95% CI 0.776-0.840) and 0.016 (95% CI 0.006-0.026), respectively. In the external testing, model performance was robust for both triage (average AUC, 0.988, 95% CI 0.967-1.000) and primary diagnosis (CA, 0.718, 95% CI 0.644-0.792; and HL, 0.023, 95% CI 0.000-0.048). In the pilot test in the hierarchical referral settings, EE-explorer demonstrated consistently robust performance and broad participant acceptance. CONCLUSION: The EE-Explorer system showed robust performance in both triage and primary diagnosis for ophthalmic emergency patients. EE-Explorer can provide patients with acute ophthalmic symptoms access to remote self-triage and assist in primary diagnosis in unspecialized health care facilities to achieve rapid and effective treatment strategies.

Characteristics of eye-related emergency visits and triage differences by nurses and ophthalmologists: Perspective from a single eye center in southern China.

Purpose: To describe characteristics of eye-related emergency department (ED) visits and investigate differences in priorities assigned to patients by triage nurses and ophthalmologists. Methods: A prospective survey was conducted at the ED of Zhongshan Ophthalmic Center from January 1, 2021, to May 31, 2021. Clinical data from patients with acute ophthalmic conditions lasting less than 7 days were collected via a standard questionnaire and the urgency levels assigned by nurses and physicians were also recorded. Binary logistic regression was performed to identify characteristics associated with truly emergency conditions and up- or down-triage. Results: A total of 1907 patients were enrolled, with 582 (30.5%) classified as "non-emergency." Red eye (69.7%), eye pain (53.0%), ocular trauma (44.1%), tearing (43.6%), and blurred vision (43.1%) were the most common complaints. Truly emergency tended to be male (OR 2.019, p < 0.001) and with unilateral eye involvement (OR 2.992, p < 0.001). Nurses prioritized conjunctival, scleral, closed ocular trauma and eyelid diseases over doctors while giving less priority to open ocular trauma, cornea, uveitis, and vitreoretinal diseases (p < 0.05). Overemphasis on mild blurred vision (OR 3.718, p = 0.001) and insufficient understanding of conjunctival diseases without red eye (OR 0.254, p = 0.001) were associated with conjunctival disease "up-triage." Insufficient awareness of moderate and severe blurred vision was associated with "down-triage" for ocular trauma (OR 3.475, p = 0.001 and OR 2.422, p = 0.020, respectively). Conclusion: Ophthalmic EDs are typically flooded with patients suffering from acute ocular problems, with a considerable portion for non-emergency conditions. The identification of characteristics associated with truly emergency cases and nurses' triage preferences is valuable in providing target guidance for future ED practice and facilitating the proper allocation of emergency resources.

High-Mobility Group Box 1 Inhibitor BoxA Alleviates Neuroinflammation-Induced Retinal Ganglion Cell Damage in Traumatic Optic Neuropathy.

Traumatic optic neuropathy (TON) is a significant cause of vision loss and irreversible blindness worldwide. It is defined as retinal ganglion cell death and axon degeneration caused by injury. Optic nerve crush (ONC), a well-validated model of TON, activates retinal microglia and initiates neuroinflammation. High-mobility group box 1 (HMGB1), a non-histone chromosomal binding protein in the nucleus of eukaryotic cells, is an important inducer of microglial activation and pro-inflammatory cytokine release. The purpose of this study was to examine the protective effects and mechanism of the HMGB1 inhibitor BoxA to neuroinflammation-induced retinal ganglion cells (RGCs) damage in traumatic optic neuropathy. For that purpose, an optic nerve crush model was established in C57BL/6J mice at 10-12 weeks. Model mice received an intravitreal injection of PBS and the HMGB1 inhibitor BoxA. Our data demonstrated that HMGB1 expression increased after optic nerve crush. Retinal ganglion cell function and morphology were damaged, and retinal ganglion cell numbers were reduced after optic nerve crush. Intravitreal injection of BoxA after ONC can alleviate damage. Furthermore, BoxA reduced microglial activation and expression levels of nuclear factor κB (NF-kB), nucleotide-binding domain, leucine-rich repeat containing protein 3 (NLRP3), and apoptosis-associated speck-like protein containing a CARD (ASC) in experimental ONC mice. In summary, HMGB1 mediates NLRP3 inflammasome via NF-kB to participate in retinal inflammatory injury after ONC. Thus, intravitreal injection of BoxA has potential therapeutic benefits for the effective treatment of RGC death to prevent TON.

Comparison of vascular parameters between normal cynomolgus macaques and healthy humans by optical coherence tomography angiography.

BACKGROUND: The metabolic activity of retina is higher than other human tissues and is crucial to the vision. Cynomolgus macaques is widely used in ophthalmic disease research. The evaluation and comparison of macular and optic disc vascular circulation parameters between normal adult cynomolgus macaques and healthy adult humans using OCT-A can promote better use of nonhuman primate models in studies of ophthalmic vascular disease. METHODS: Twelve normal adult cynomolgus macaques with a mean age of 4.91 ± 0.43 years were studied for data collection. The macula of 28 adult healthy humans (14 males and 14 females), with a mean age of 25.11 ± 6.21 years and the optic discs of 9 adult healthy humans (4 males and 5 females) with a mean age of 28.56 ± 6.78 years were measured. The vessel density (VD) was measured using an RTVue XR with AngioVue. The scan sizes of the macular and optic discs were 3 × 3 mm and 4.5 × 4.5 mm, respectively. RESULTS: OCT-A can image the superficial and deep capillary plexuses and radial peripapillary capillary network. In RPC layer of the optic disc, the VD in the nasal quadrant was lower than the VD in the inferior temporal quadrant. Similarities and significant differences in VD between healthy humans and cynomolgus macaques were obtained using OCT-A. CONCLUSIONS: This study provides normal vascular parameters for adult cynomolgus macaques using OCT-A to help establish an optical parameter database for cynomolgus macaques and compare VD between healthy humans and cynomolgus macaques to promote choroid-retinopathy research. TRIAL REGISTRATION: Current Controlled Trials NCT03692169 , retrospectively registered on 26 sept 2018.

Unplanned readmission after ocular trauma ambulatory surgery: A retrospective study of the Zhongshan Ophthalmic Center.

PURPOSE: Ambulatory surgery (AS) has become common in recent years. This study explored the causes of unplanned readmission after AS and the trends in the rate of unplanned readmission in the Ocular Trauma Department of Zhongshan Ophthalmic Center. METHODS: We collected information on patients who underwent AS from 1 January 2015 to 31 December 2017, summarized the causes of ambulatory hospitalization, and analysed the type of initial surgical procedure and the reasons for 31-day unplanned readmission. The Ocular Trauma Department started to perform AS in around mid-2015, and we collected information on patients who underwent ambulatory pars plana vitrectomy (APPV) from 1 January 2016 to 31 December 2017 to explore the predictive factors for the 31-day unplanned readmission of APPV patients. RESULTS: In 2015, 2016, and 2017, a total of 3443 patients underwent AS, among whom the three most common causes of ambulatory hospitalization were eye wall and appendages injury, aphakia, and silicone oil-filled eye. Of the 3443 patients, 32 returned for 31-day unplanned readmission, a readmission rate of 0.93%. The most common surgery (34.4%, 11/32) that resulted in unplanned readmission was pars plana vitrectomy (PPV). The major reasons for unplanned readmission were retinal detachment (RD) (34.4%, 11/32) and high intraocular pressure (HIP) (18.8%, 6/32). There were 860 patients who underwent APPV from 1 January 2016 to 31 December 2017, among whom 10 needed 31-day unplanned readmission, and the readmission rate of APPV for ocular trauma in 2016 and 2017 was 1.16%. Length of procedure (P < 0.05) and use of silicone oil filling (P < 0.05) were significantly related to the risk of unplanned readmission for APPV surgery. CONCLUSION: It is generally safe to perform AS on patients with ocular trauma. However, longer operative times and silicone oil filling increase the risk of unplanned readmission for APPV surgery.

Influence of super-hydrophobic silicone rubber substrate on the growth and differentiation of human lens epithelial cells.

Materials with low cell adhesion are advantageous for production of replacement intraocular lens (IOL) to prevent posterior capsular opacification (PCO). We evaluated the feasibility of compression molding for manufacture of silicone rubber with super-hydrophobic surface and low cell infiltrative characteristics compared to ordinary hydrophobic silicone rubber. Silicone specimens with complex surface topology (super-hydrophobic) or smooth surfaces (hydrophobic) were manufactured by vacuum deforming and molding. Contact angle, microscopic surface structure, and transparency were evaluated. Super-hydrophobic and smooth samples were compared for effects on proliferation, adhesion, and morphology of human lens epithelial cells (hLECs). Epithelial-mesenchymal transition (EMT) was examined by immunofluorescence expression of fibronectin (Fn), Alpha-smooth muscle actin (α-SMA), and vimentin. The surface contact angle of super-hydrophobic silicone was greater than that of smooth silicone (153.8° vs. 116°). The super-hydrophobic surface exhibited a micron-scale palisade structure under scanning electron microscopy (unit length, width, and height of 80, 25, and 25 µm, respectively). However, cell number per 50 × microscopic field on super-hydrophobic surfaces was markedly reduced 24 and 72 h post-seeding compared to smooth surfaces (p < 0.01). Cells were cuboidal or spherical after 72h on super-hydrophobic surfaces, and exhibited numerous surface microvilli with fluff-base polarity, while cells on smooth surfaces exhibited morphological characteristics of EMT. Expression levels of the α-SMA and vimentin were reduced on super-hydrophobic surfaces compared to smooth surfaces. Super-hydrophobic silicon inhibits proliferation, adhesion, and EMT of hLECs, properties that may prevent fibrosis following cataract surgery.

Oxygen Saturation in Closed-Globe Blunt Ocular Trauma.

Purpose. To evaluate the oxygen saturation in retinal blood vessels in patients after closed-globe blunt ocular trauma. Design. Retrospective observational case series. Methods. Retinal oximetry was performed in both eyes of 29 patients with unilateral closed-globe blunt ocular trauma. Arterial oxygen saturation (SaO2), venous oxygen saturation (SvO2), arteriovenous difference in oxygen saturation (SO2), arteriolar diameter, venular diameter, and arteriovenous difference in diameter were measured. Association parameters including age, finger pulse oximetry, systolic pressure, diastolic pressure, and heart rate were analyzed. Results. The mean SaO2 in traumatic eyes (98.1% ± 6.8%) was not significantly different from SaO2 in unaffected ones (95.3% ± 7.2%) (p = 0.136). Mean SvO2 in traumatic eyes (57.1% ± 10.6%) was significantly lower than in unaffected ones (62.3% ± 8.4%) (p = 0.044). The arteriovenous difference in SO2 in traumatic eyes (41.0% ± 11.2%) was significantly larger than in unaffected ones (33.0% ± 6.9%) (p = 0.002). No significant difference was observed between traumatic eyes and unaffected ones in arteriolar (p = 0.249) and venular diameter (p = 0.972) as well as arteriovenous difference in diameter (p = 0.275). Conclusions. Oxygen consumption is increased in eyes after cgBOT, associated with lower SvO2 and enlarged arteriovenous difference in SO2 but not with changes in diameter of retinal vessels.

Using surgical microscope for sclera buckling and transscleral cryopexy: an alternative procedure of treatment for rhegmatogenous retinal detachment.

Purpose. To observe the long-term effectiveness of scleral buckling and transscleral cryopexy conducted under a surgical microscope in the treatment of uncomplicated rhegmatogenous retinal detachment. Methods. This was a retrospective analysis in a total of 227 consecutive patients (244 eyes) with uncomplicated rhegmatogenous retinal detachment (proliferative vitreoretinopathy ≤ C2). All patients underwent scleral buckling and transscleral cryopexy under a surgical microscope without using a binocular indirect ophthalmoscope or a contact lens. Results. After initial surgery, complete retinal reattachment was achieved in 226 eyes (92.6%), and retinal redetachment developed in 18 eyes (7.4%). The causes of retinal redetachment included presence of new breaks in eight eyes (44%), failure to completely seal the breaks in five eyes (28%), missed retinal breaks in four eyes (22%), and iatrogenic retinal breaks in one eye (6%). Scleral buckling surgery was performed again in 12 eyes (66%). Four eyes (22%) developed proliferative vitreoretinopathy and then were treated by vitrectomy. The sealing of retinal breaks and complete retinal reattachment were achieved in 241 eyes (98.8%). Conclusion. Probably because of clear visualization of retinal breaks and being controllable under a surgical microscope, the microsurgery of scleral buckling and transscleral cryopexy for uncomplicated retinal detachment exhibits advisable effectiveness.

[Clinical study of the auxiliary management with iris retractor for subluxated lens combined with cataract].

OBJECTIVE: To study the clinical effects and safety of the auxiliary management with iris retractor for subluxated lens combined with cataract. METHODS: Thirty-one eyes lens subluxation with cataract come from 27 patients were reviewed, all of them were 30 - 78 years old and III - IV grade of nuclear. Among these patients, 14 were traumatic cataracts, 12 Marfan syndromes, two over-mature phase of cataracts, one caused suspensory ligament broken during phacoemulsification, and one unclear reason. 1 to 4 iris retractors were used during surgeries, according to the extension of zonular loss and nuclear hardness. Anterior vitrectomy would be taken if vitreous prolapsed. Visual acuities and reactions post-operative were observed, too. And correlation analysis would be used to study the relationship of those objections. RESULTS: There were no serious complications during the surgeries such as tearing of capsular, falling of nuclear, bleeding of intraocular, and so on. Only 9 of 31 eyes (29.0%) had vitreous prolapsed interoperation, which appeared a significant correlated with range of zonular loss (Spearman r = 0.453, P = 0.010). However, none of them happened during the phacoemulsification for lens nucleus and only 6 eyes took the anterior vitrectomy. The best corrected postoperative visual acuity of these patients attended to 0.2 - 1.2 at a half to 1 month, increased by 1 - 12 lines [average of (6.0 ± 2.7) lines] compared with preoperative. Twenty-three eyes visual acuity of them were no less than 0.5. Corneal edema postoperatively appeared a significant correlated with vitrectomy (Spearman r = 0.398, P = 0.026), while seemed no associated with the range of zonular loss, the number of iris retractors, energy and duration of ultrasound burst or type of intraocular lens (IOL) (P > 0.05). CONCLUSIONS: Phacoemulsification assists with iris retractor can simplify the process of cataract surgery of subluxated lens. Individualized surgical planning and flexible and decisive measures intra-operative achieved good results and well safety in these dangerous and complex lens diseases.