Regional Summed Corneal Refractive Power Changes in Myopic Orthokeratology and Their Relationships With Axial Elongation.

PURPOSE: To determine the relationship between regional summed corneal refractive power changes (CRPCs) in myopic orthokeratology and axial elongation. DESIGN: This retrospective study included 70 eyes of 70 patients aged 8 years to 13 years who underwent orthokeratology lens (OK lens) treatment, and all patients underwent regular follow-ups at 1 week, 1 month, 6 months, and 12 months at Zhongshan Ophthalmic Center between January 2019 and May 2021. METHODS: Axial length (AL) was measured at baseline and 12 months by an IOLmaster 5.0. Refractive error power was measured using cycloplegia. Regional summed CRPCs were calculated by MATLAB software using difference tangential power maps at the sixth month acquired by corneal topography measurements (Medmont E300 Corneal Topographer; Medmont Pty, Victoria, Australia) and defined as changes in corneal refractive power at the sixth month from baseline. The regional summed CRPCs were then subdivided into 4-mm diameter circles, 4- to 5-mm diameter chords, and 5- to 6-mm diameter chords according to the distance from the central of the pupil and into negative, positive, and total according to the values. Pearson correlation, multiple linear regression analysis, and stepwise multiple linear regression analysis were performed to analyze the relationships among these parameters. RESULTS: Axial elongation had a negative relationship with positive regional summed corneal refractive power in the central 4-mm diameter circle and age (r=-0.282, P=0.018; r=-0.473, P<0.001, respectively) and a positive relationship with negative regional summed corneal refractive power in the 5- to 6-mm diameter chord (r=0.361, P=0.002). Stepwise multiple linear regression analysis identified age (standardized ß=-0.434, P<0.001) and negative regional summed corneal refractive power in the 5- to 6-mm diameter chord (standardized ß=0.305, P=0.004) as factors influencing AL elongation. CONCLUSION: Negative regional summed corneal refractive power in a 5- to 6-mm diameter chord after OK lens treatment may be an important index for evaluating the control effects of axial elongation.

Long-Term Axial Length Shortening in Myopic Orthokeratology: Incident Probability, Time Course, and Influencing Factors.

Purpose: Long-term axial length (AL) shortening in myopia is uncommon but noteworthy. Current understanding on the condition is limited due to difficulties in case collection. The study reported percentage, probability, and time course of long-term AL shortening in myopic orthokeratology based on a large database. Methods: This study reviewed 142,091 medical records from 29,825 subjects in a single-hospital orthokeratology database that were collected over 10 years. Long-term AL shortening was defined as a change in AL of -0.1 mm or less at any follow-up beyond 1 year. Incident probability was calculated based on multivariate logistic regression. Time course was estimated using mixed-effect regression model. Results: A total of 10,093 subjects (mean initial age, 11.70 ± 2.52 years; 58.8% female) with 80,778 visits were included. The number of subjects experienced long-term AL shortening was 1,662 (16.47%; 95% confidence interval, 15.75%-17.21%). Initial age showed significant impact on the incident occurrence (OR, 1.37; 95% confidence interval, 1.34-1.40; P < 0.001). The estimated probability of AL shortening was approximately 2% for subjects with initial age of 6 years and 50% for those aged 18. Among the 1662 AL shortening cases, the median magnitude of the maximum AL reduction was 0.19 mm. The shortening process mostly occurred within the initial 2 years. Subject characteristics had limited associations with the shortening rate. Conclusions: Long-term AL shortening is possible in subjects receiving myopic orthokeratology. Although age notably affect the incident probability, the time course seems to not vary significantly.

Spontaneous fundus lesions in elderly monkeys: An ideal model for age-related macular degeneration and high myopia clinical research.

AIMS: Age-related macular degeneration (AMD) and high myopia are frequent causes of progressive visual impairment, so it is critical to identify animal models with resembling human retinal physiology, AMD and high myopia pathological features for therapeutic studies. MAIN METHODS: We screened elderly cynomolgus monkeys for fundus lesions by slit-lamp biomicroscope combined with fundus pre-set lens and further examined positive cases by color fundus photography (CFP), optical coherence tomography (OCT), fundus fluorescein angiography (FFA), streak retinoscopy, and A-scan ultrasonography. KEY FINDINGS: Among the 156 animals examined, 10 males and 5 females (30 eyes) exhibited fundus abnormalities (9.6% prevalence). Multi-modal imaging revealed drusen in 20 eyes of 11 animals (prevalence rate of 7.1%), tessellated fundus in 22 eyes of 11 animals, and myopia choroidal neovascularization (CNV) in 4 eyes of 3 animals. SIGNIFICANCE: Aged cynomolgus monkeys exhibit spontaneous fundus lesions resembling human AMD and high myopia, which could be an ideal model for clinical research.

Migration of pre-induced human peripheral blood mononuclear cells from the transplanted to contralateral eye in mice.

BACKGROUND: Retina diseases may lead to blindness as they often afflict both eyes. Stem cell transplantation into the affected eye(s) is a promising therapeutic strategy for certain retinal diseases. Human peripheral blood mononuclear cells (hPBMCs) are a good source of stem cells, but it is unclear whether pre-induced hPBMCs can migrate from the injected eye to the contralateral eye for bilateral treatment. We examine the possibility of bilateral cell transplantation from unilateral cell injection. METHODS: One hundred and sixty-one 3-month-old retinal degeneration 1 (rd1) mice were divided randomly into 3 groups: an untreated group (n = 45), a control group receiving serum-free Dulbecco's modified Eagle's medium (DMEM) injection into the right subretina (n = 45), and a treatment group receiving injection of pre-induced hPBMCs into the right subretina (n = 71). Both eyes were examined by full-field electroretinogram (ERG), immunofluorescence, flow cytometry, and quantitative real-time polymerase chain reaction (qRT-PCR) at 1 and 3 months post-injection. RESULTS: At both 1 and 3 months post-injection, labeled pre-induced hPBMCs were observed in the retinal inner nuclear layer of the contralateral (left untreated) eye as well as the treated eye as evidenced by immunofluorescence staining for a human antigen. Flow cytometry of fluorescently label cells and qRT-PCR of hPBMCs genes confirmed that transplanted hPBMCs migrated from the treated to the contralateral untreated eye and remained viable for up to 3 months. Further, full-field ERG showed clear light-evoked a and b waves in both treated and untreated eyes at 3 months post-transplantation. Labeled pre-induced hPBMCs were also observed in the contralateral optic nerve but not in the blood circulation, suggesting migration via the optic chiasm. CONCLUSION: It may be possible to treat binocular eye diseases by unilateral stem cell injection.

Corneal Recovery Following Rabbit Peripheral Blood Mononuclear Cell-Amniotic Membrane Transplantation with Antivascular Endothelial Growth Factor in Limbal Stem Cell Deficiency Rabbits.

Background: Limbal stem cell deficiency (LSCD) is a refractory ocular surface disorder characterized by progressive corneal epithelial degeneration, conjunctivalization, and neovascularization, potentially leading to blindness. There are currently no effective therapeutic options for patients experiencing routine symptomatic treatment failure. Transplantation of amniotic membrane (AM) with adherent stem cells (but not bare AM transplantation alone) has shown promise in preclinical studies for ocular surface restoration. A major limitation, however, is finding a reliable stem cell source. Stem cells can be isolated from the peripheral blood mononuclear cell (PBMC) population, and these PBMC-derived stem cells have numerous advantages over allogeneic and other autologous stem cell types for therapeutic application, including relative ease of acquisition, nonimmunogenicity, and the absence of ethical issues associated with embryonic stem cells. Experiment: We examined the efficacy of autologous PBMC-AM sheet cultures combined with postoperative antiangiogenesis treatment for corneal restoration in LSCD model rabbits. Rabbit PBMCs (rPBMCs) were isolated, labeled with EdU for in vivo tracing, and then cultured on AMs in conditioned medium before transplantation. Rabbits were transplanted with bare AMs (group 1), rPBMC-AM sheets (group 2), or rPBMC-AM sheets plus postoperative treatment with the vascular endothelial growth factor antagonist bevacizumab (group 3). Corneal opacity and neovascularization were monitored by slit-lamp imaging for 8 weeks and corneas were examined histologically at 1 and 2 months. Results: Corneal opacity decreased in all three groups over 8 weeks, but was significantly lower in group 2 and even lower in group 3. Corneal neovascularization was significantly higher in group 1 throughout the observation period, and significantly lower in group 3 than group 1 and 2 by 8 weeks post-transplant. At 4 weeks, the corneal surface completed epithelialization (although thinner than normal) in group 3 but still patchy in groups 1 and 2. By 8 weeks, the epithelium in group 3 was complete and smooth, resembling a normal epithelium. Integrin ß1 as a progenitor marker was also generally higher in groups 2 and 3. Conclusions: Autologous rPBMC-AM sheets with post-transplant topical bevacizumab can effectively facilitate corneal epithelium recovery in a LSCD model, suggesting clinical utility for LSCD-related ocular surface diseases. Impact statement Limbal stem cell deficiency (LSCD) increases corneal opacity and vascularization, resulting in severe visual impairment or even blindness. Traditional surgical limbal transplant is currently the main treatment option for LSCD, but carries the risks of rejection and immunosuppressant side effects. Autologous stem cell-based therapy is a promising alternative approach, but a reliable stem cell source is a major limitation. We report that transplantation of autologous rabbit peripheral blood mononuclear cell-amniotic membrane sheets plus antivascular endothelial growth factor restored avascular transparent cornea in a rabbit LSCD model. These results demonstrate a potentially effective approach for ocular surface reconstruction in bilateral LSCD.

Safety Assessment and Comparison of Sodium Selenite and Bioselenium Obtained from Yeast in Mice.

Detailed safety assessment of sodium selenite and bioselenium (bio-Se) was conducted and the results were compared and discussed for the purpose of assessing safety of bio-Se for use in food applications. In this work, acute toxicity studies, micronucleus test, and sperm aberration study in mice, 30-day feeding test of mice, were conducted to evaluate the toxicity of bio-Se obtained from yeast with different fermentation time (transformative time: one month, three months, and six months), and the results were compared with that of inorganic Se (sodium selenite). LD50 of sodium selenite was calculated to be 21.17 mg/kg. LD50 of bio-Se obtained from yeast with different fermentation time was calculated to be 740.2 mg/kg, 915.3 mg/kg, and 1179.0 mg/kg, respectively. In the genotoxicity test, bio-Se did not show cytotoxicity and genotoxicity of mice while sodium selenite at all dose groups was significantly different from the negative group. In the 30-day subchronic oral toxicity study, sodium selenite may slow down the growth of the mice and lead to organic damage to some extent. Bio-Se had facilitated effect towards the body weight of the mice and had no significant effect on the shape and function of the important organs of the mice.

Comparison of Two Rabbit Models with Deficiency of Corneal Epithelium and Limbal Stem Cells Established by Different Methods.

Limbal stem cell defect model is an important animal model that provides a basis for the study of ocular surface diseases. The rabbit cornea is of moderate size and is widely used in such studies as an experimental animal model. At present, the main modeling methods are alkali burns, and corneal limbus girdling and corneal epithelium doctoring. Each method has its own characteristics. In this study, we observed rabbit models with severe ocular surface defect established by the two methods and changes after amniotic membrane transplantation. In the first, second, third, and fourth week after operation, the clinical manifestations, corneal transparency, and new vessels were observed according to the standard rating scale of ocular surface, compared between the two methods, and then statistically analyzed. In the fourth week after operation, the rabbits were sacrificed and their corneas and corneal limbus were extracted from sclera, embedded by optimum cutting temperature compound, frozen, and sliced for hematoxylin and eosin staining and pathological examination. There were two groups in this study. Group 1 (alkali burns) had more severe complications, such as, conjunctiva, nubecula, new vessel hyperplasia, and so on, compared to group 2 (corneal limbus girdling and corneal epithelium doctoring). In addition, there were striking differences in corneal transparency and new vessels between the two groups (p < 0.05). Corneal transparency in group 1 was lower than in group 2. New vessels in group 1 were less in the first 2 weeks, but obviously increased compared to group 2 in the subsequent weeks. Alkaline burn could be used to study new vessel hyperplasia, while corneal limbus girdling and corneal epithelium doctoring are more suitable for studying stem cell transdifferentiation, interactive roles of stem cells and microenvironment, and so on.

Adult Human Peripheral Blood Mononuclear Cells Are Capable of Producing Neurocyte or Photoreceptor-Like Cells That Survive in Mouse Eyes After Preinduction With Neonatal Retina.

: Adult human peripheral blood mononuclear cells (hPBMCs) exhibit pluripotency in vitro and so may be a valuable cell source for regenerative therapies. The efficacy of such therapies depends on the survival, differentiation, migration, and integration capacity of hPBMCs in specific tissues. In this study, we examined these capacities of transplanted hPBMCs in mouse retina as well functional improvement after transplant. We isolated hPBMCs and preinduced them for 4 days in media preconditioned with postnatal day 1 rat retina explants. Preinduction increased the proportions of hPBMCs expressing neural stem cell, neural progenitor cell, or photoreceptor markers as revealed by immunofluorescent staining, flow cytometry, and quantitative real-time polymerase chain reaction. Preinduced hPBMCs were transplanted into the subretinal space of retinal degenerative slow (RDS) and retinal degeneration 1 (RD1) mice. At 1, 3, and 6 months after transplantation, treated eyes of RDS mice were collected and cell phenotype was studied by immunofluorescent staining. Preinduced hPBMCs survived in the subretinal space; migrated away from the injection site and into multiple retinal layers; and expressed neural stem cell, neuronal, and photoreceptor markers. Finally, we assessed RD1 retinal function after subretinal transplantation and found significant improvement at 3 months after transplantation. The ease of harvesting, viability in vivo, capacity to express neuronal and photoreceptor proteins, and capacity for functional enhancement suggest that hPBMCs are potential candidates for cell replacement therapy to treat retinal degenerative diseases. SIGNIFICANCE: This study provides support for the use of peripheral blood mononuclear cells (PBMCs) as a potential source of pluripotent stem cells for treating retinal degeneration. First, this study demonstrated that PBMCs can differentiate into retinal neuron-like cells in vitro and in vivo. Second, some transplanted cells expressed markers for neural progenitors, mature neurons, or photoreceptors at 1, 3, and 6 months after subretinal injection. Finally, this study showed that PBMC transplantation can improve the function of a degenerated retina.

[The flexure of on-eye rigid gas permeable contact lenses and its relationship with anterior corneal surface].

OBJECTIVE: To investigate the effect of corneal irregularity on on-eye rigid gas permeable (RGP) contact lenses and the corneal shape changed by RGP contact lenses. METHODS: Corneal topography was taken for 42 bare eyes of 22 subjects before and 1 month after wearing of the RGP contact lens. Off-eye and on-eye wavefront aberrations of the anterior surface of the RGP contact lens were also measured by corneal topography for the same 42 eyes. Zernike aberrations of the RGP contact lens were examined and compared to those of baseline anterior corneal surface. RESULTS: For bare eyes, C4, C5, C7 and C8 were significantly different from the baseline 30 minutes after the RGP contact lens was removed. For the on-eye RGP contact lens, three mean Zernike aberrations were significantly different from those off-eye. These were defocus C4 [OD: (1.003 ± 0.131) µm off-eye and (1.065 ± 0.160) µm on-eye, P = 0.015; OS: (1.003 ± 0.130) µm off-eye and 1.069 (0.851, 1.594) µm on-eye, P = 0.017], main axis astigmatism C5 [OD: (0.072 ± 0.083) µm off-eye and (-0.312 ± 0.232) µm on-eye, P < 0.001; OS: (0.066 ± 0.056) µm off-eye and (-0.349 ± 0.242) µm on-eye, P < 0.001], and spherical aberration C12 [OD: (0.264 ± 0.039) µm off-eye and (0.295 ± 0.048) µm on-eye, P < 0.001; OS: (0.266 ± 0.035) µm off-eye and 0.290 (0.215, 0.471) µm on-eye, P < 0.001]. All these three Zernike aberrations of on-eye RGP contact lenses were significantly correlated to those of the baseline corneas (OD: rc4=0.557, rc5=0.596, rc12=0.581, P < 0.01; OS: rc4=0.684, rc5=0.497, rc12=0.543, P < 0.01). CONCLUSIONS: The results suggest that the surface shape of on-eye RGP contact lenses is not rigid, and its change depends on irregularity of the anterior corneal surface, even if the cornea itself is changed by the RGP contact lens. It is better not to fit for RGP so flatter than the average curvature of the cornea.

Aberration changes of the corneal anterior surface following discontinued use of rigid gas permeable contact lenses.

AIM: To record aberrations with a corneal topographic device on the anterior surface of the cornea at different time-points prior to wearing and following discontinued use of rigid gas permeable (RGP) contact lenses. The effect of wearing RGP on the anterior surface of the cornea was discussed to provide guidance for clinical refractive error correction. METHODS: The study objects were 24 eyes from 24 patients. All patients underwent identical examination procedures prior to lens use, as well as afterwards, including slit-lamp examination, non-contact tonometer measurement, computer optometry and corneal curvature measurement, subjective refraction test, and corneal topography analysis. The patients wore contact lenses everyday for 1 month and then discontinued. Corneal topographies were recorded at certain time points of 30 minutes, 1 day, 3, 7 and 14 days following use. RESULTS: Total corneal aberration at each time point following discontinued use of RGP contact lenses was less than the time point prior to use. Detailed results were as follows: root mean square (RMS) (pre)=(1.438±0.328)µm, RMS (30 minutes)=(1.076±0.355)µm, RMS (1 day)=(1.362±0.402)µm, RMS (3 days)=(1.373±0.398)µm, RMS (7 days)=(1.387±0.415)µm, and RMS (14 days)=(1.430±0.423)µm. Results showed that at 30 minutes after discontinued use of RGP contact lenses, almost all 2(nd)- and 3(rd)-order aberrations change. Quadrafoil Z10 and spherical Z12 of the 4(th)-order were also changed. Alterations to Z5, Z6, and Z12 at 1 day after discontinued use were significant differences compared with the time period prior to RGP use: Z5 and Z6 decreased, and Z12 increased slightly. Z5 and Z6 remained decreased at 3 days after discontinued use, but Z9 and Z10 continued to increase and Z12 returned to levels prior to RGP use. At 14 days after discontinued use, all aberrations were not significantly different from the values prior to use. CONCLUSION: The use RGP contact lenses greatly reduced total aberration of the anterior surface of the cornea. Changes to 2(nd)- and 3(rd)-order aberrations (including Z3, Z4, Z5, Z6, Z7, and Z8) were more significant. Following discontinued use of RGP contact lenses, the majority of lower order aberrations returned to original levels in a short period of time. During this process, a transient higher order aberration appeared, but all changes disappeared within 14 days after discontinued use of RGP contact lenses.

[Analysis of aberration changes of the corneal anterior surface following discontinued use of rigid gas permeable contact lenses].

OBJECTIVE: The present study used a corneal topographic device to record aberrations on the anterior surface of the cornea at different time-points prior to wearing and following discontinued use of rigid gas permeable (RGP) contact lenses. The effect of wearing RGPCL on the anterior surface of the cornea was discussed to provide guidance for clinical refractive error correction. METHODS: The study objects were 60 eyes from 30 patients. All patients underwent identical examination procedures prior to lens use, as well as afterwards, including slit-lamp examination, non-contact tonometer measurement, computer optometry & corneal curvature measurement, subjective refraction test, and corneal topography analysis. The patients wore contact lenses every day for 1 month and then discontinued. Corneal topographies were recorded at certain time points of 30 min, 1 day, 3 days, 1 week, and 2 weeks following use. RESULTS: Total corneal aberration at each time point following discontinued use of RGP contact lenses was less than the time point prior to use. Detailed results are as follows; root mean square (RMS) (pre) = (1.438 ± 0.328), RMS (30 min) = (1.076 ± 0.355), RMS (1 day) = (1.362 ± 0.402), RMS (3 day) = (1.373 ± 0.398), RMS (7 day) = (1.387 ± 0.415), and RMS (14 day) = (1.448 ± 0.423). Results showed that at 30 minutes after discontinued use of RGP contact lenses, almost all 2(nd)- and 3(rd)-order aberrations were altered. Quadrafoil Z10 and spherical Z12 of the 4(th)-order were also changed. Alterations to Z5, Z6, and Z12 at 1 day after discontinued use were significant compared with the time period prior to RGP use: Z5 and Z6 decreased, and Z12 increased slightly (F = 2.869 ∼ 5.549, P = 0.001 ∼ 0.042). Z5 and Z6 remained decreased at 3 days after discontinued use, but Z9 and Z10 continued to increase and Z12 returned to levels prior to RGP use (P > 0.05). At 2 weeks after discontinued use, all aberrations were not significantly different from the values prior to use (P > 0.05). CONCLUSIONS: The use RGP contact lenses greatly reduced total aberration of the anterior surface of the cornea. Changes to 2(nd)- and 3(rd)-order aberrations (including Z3, Z4, Z5, Z6, Z7, and Z8) were more significant. Following discontinued use of RGP contact lenses, the majority of lower order aberrations returned to original levels in a short period of time. During this process, a transient higher order aberration appeared, but all changes disappeared within 2 weeks after discontinued use of RGP contact lenses.