AREG is involved in scleral remodeling in form-deprivation myopia via the ERK1/2-MMP-2 pathway.

Previous studies have found that amphiregulin (AREG) may participate in eye elongation during the development of myopia, but the mechanism remains unclear. Here, we tested tear concentrations of AREG in adults and detected the role of AREG in scleral remodeling in form-deprivation myopia (FDM) in guinea pigs. We found the tear concentrations of AREG in myopes were significantly higher than those in emmetropes using enzyme-linked immunosorbent assay (ELISA). Tear concentrations of AREG were negatively correlated with spherical equivalent refraction and positively correlated with axial length (AL) and AL/corneal radii. We then used RNAi, DNA transfection and PD98059 treatments to determine the effects of AREG on extracellular signal-regulated kinase 1/2 (ERK1/2) and matrix metalloprotease-2 (MMP-2) in primary scleral fibroblasts (SFs). The hypothesis was further verified via loss- and gain-of-function experiments by intravitreal application of anti-AREG antibody (anti-AR) or AREG in form-deprivation eyes in guinea pigs. Immunofluorescence assay was used for cell type identification. Western-blot and q-PCR were used for the detection of relative expressions. Transmission electron microscopy was performed for posterior scleral observation. In vitro, we found AREG overexpression increased phospho-ERK1/2 and MMP-2 expression, while depletion of AREG inhibited their expressions. PD98059 (an effective ERK1/2 inhibitor) inhibited AREG-induced MMP-2 upregulation. In vivo, we found anti-AR treatments suppressed FDM by inhibiting scleral remodeling, while AREG treatments promoted FDM. Our results suggest that AREG in tear fluids can serve as a potential biomarker in myopes. AREG is involved in scleral remodeling through the ERK1/2-MMP-2 pathway. AREG is a potential target for myopia control.

Relationship between dry eye and expressions of CXCR3 and CCR5 after ocular acid burn.

OBJECTIVE: To investigate the manifestation of dry eye and its relationship with CXCR3 and CCR5 expression in patients with ocular acid burns. METHODS: This is a case-control study. A total of 27 eyes of 22 cases ocular with acid burns of I-V degrees from Jan.2020 to Feb.2021 in Jinshan Hospital of Fudan University were selected as observation group, and 8 eyes of 8 cases of normal people were selected as control group. The follow-up period was 3 months. The visual acuity, intraocular pressure (IOP), corneal fluorescein staining scores (CFS), breakup time of tear film (BUT), Schirmer I test, corneal thickness and tear meniscus height (TMH) were observed at 1 day, 1 and 3 months after injury. The protein expressions of CXCR3 and CCR5 were examined by ELISA and compared among groups at each time point. RESULTS: BUT and Schirmer I tests value in the observation group were lower than those in the control group 3 months after injury (BUT: Group I ~ IV p = 0.0266, p = 0.0222, p = 0.0003, p = 0.0059, respectively; Schirmer I test: Group I ~ IV p = 0.0027, p = 0.0033, p = 0.0016, p = 0.0032, respectively). CFS scores were higher than those in the control group at 1 day after injury (all p < 0.0001), but decreased gradually at 1 and 3 months after injury (Group I ~ IV p = 0.0042, p = 0.0096, p < 0.0001, p < 0.0001, respectively). The corneal thickness and TMH 1 day after injury were higher than those in the control group (corneal thickness: Group II ~ IV p = 0.0010, p < 0.0001, p < 0.0001, respectively; TMH: Group II ~ IV p = 0.0002, p < 0.0001, p < 0.0001, respectively), and also higher than those at 1 month and 3 months after injury (corneal thickness: Group II ~ IV p = 0.0010, p < 0.0001, p < 0.0001, respectively; TMH: Group II ~ IV p = 0.0345 and p = 0.0045, p = 0.0005 and p < 0.0001, p = 0.0114 and p = 0.0019, respectively). The expression levels of CXCR3 and CCR5 protein were significantly negatively correlated with BUT (all p < 0.0001), and CXCR3 and CCR5 were also significantly negatively correlated with Schirmer I test value (p < 0.0001, p = 0.0004, respectively). CONCLUSION: Ocular acid burns can cause dry eye, and the expression of CXCR3 and CCR5 protein in tears may be related to the occurrence of dry eye after ocular acid burn.

Relationship between age, refractive errors and motor fusion in a normal Chinese adult population: a cross-sectional study.

BACKGROUND: To investigate the relationships between motor fusion and sex, age and spherical equivalent (SE). METHODS: This observational study enrolled 243 healthy, nonstrabismic adults, including 94 men and 149 women aged 20 to 59 years. The subjects were divided into three groups according to SE: myopic, emmetropic and hyperopic groups. The subjects were also divided into four groups according to age: 20-29, 30-39, 40-49 and 50-59 years groups. Motor fusion was measured with a synoptophore, including subjective angle (SA), divergence, convergence and fusional vergence range (FVR). RESULTS: The mean values of divergence, convergence and FVR for the whole sample group were 9.72 ± 0.26°, 19.34 ± 0.54°, and 29.06 ± 0.62°, respectively. A higher value of divergence was found in the myopic group than in the emmetropic group (p < 0.05). SE and divergence were significantly different among age groups (all p < 0.05). In addition, linear regression analysis showed that SE was correlated with divergence (p = 0.003). Age was correlated with SE, divergence and FVR (p < 0.001, p = 0.005, p = 0.002, respectively). In addition, the proportion of SA being in the comfort zone (defined as the value of SA satisfying Percival's criterion) in the age groups was significantly different (χ2 = 8.283, p = 0.041). CONCLUSIONS: Motor fusion is associated with age and SE in the normal Chinese adult population.

Time trend of axial length and associated factors in 4- and 5-year-old children in Shanghai from 2013 to 2019.

PURPOSE: To evaluate the time trend of axial length (AL) and associated factors in 4- and 5-year-old children in Shanghai from 2013 to 2019. METHODS: This was a 7-year observational study of 985 four-year-old and 1059 five-year-old children in Shanghai. AL, horizontal and vertical corneal curvature, spherical equivalent (SE), and body height and weight were measured. Furthermore, a questionnaire was collected, including time outdoors and bad eyesight habits. RESULTS: In 4-year-old children, no significant difference was found in AL (P = 0.526), but significant differences were observed in SE (P = 0.001), horizontal corneal curvature (P = 0.006), vertical corneal curvature (P = 0.004), height (P < 0.001), and weight (P = 0.022) from 2013 to 2019. In 5-year-old children, no significant differences were found in AL (P = 0.304), SE (P = 0.200), or weight (P = 0.292), but significant differences were observed in horizontal corneal curvature (P = 0.040), vertical corneal curvature (P = 0.015), and height (P < 0.001) from 2013 to 2019. Multivariate analyses revealed that AL was mainly significantly associated with boys and time outdoors in the 4- and 5-year-old children. CONCLUSIONS: The AL of 4- and 5-year-old children remained relatively stable in Shanghai from 2013 to 2019. Longitudinal studies are needed to confirm the relationship between AL elongation and environmental risk factors.

Relative peripheral refraction in myopic children wearing orthokeratology lenses using a novel multispectral refraction topographer.

CLINICAL RELEVANCE: Orthokeratology (OK) lens is commonly used to control myopia progression of children. Understanding the relationship between relative peripheral refraction (RPR) and the growth rate of axial length (AL) may assist in explaining myopic progression. BACKGROUND: The aim of this work is to investigate the RPR in myopic children wearing OK lenses, and to evaluate its relationship with the growth rate of AL. METHODS: RPRs of 31 children wearing OK lenses and 31 children wearing single-vision glasses were measured with multispectral refraction topography (MRT). MRT shows the total RPR (TRPR), RPR in the superior area (RPR-S), RPR in the inferior area (RPR-I), RPR in the temporal area (RPR-T) and RPR in the nasal area (RPR-N), respectively. It also shows RPR in the visual field of 15° (RPR-15), 30° (RPR-30) and 45° (RPR-45), respectively. RPRs in the visual field from 15° to 30°, 30° to 45° and 15° to 45° are recorded as RPR-(30-15), RPR-(45-30) and RPR-(45-15), respectively. According to the growth rate of AL, children wearing OK lenses were further divided into slow and fast growth groups. RESULTS: TRPR, RPR-I, RPR-T, RPR-N, RPR-15, RPR-30, RPR-45, RPR-(30-15), RPR-(45-30), and RPR-(45-15) of children in the OK lens group were significantly smaller than in the control group (all P < 0.05). TRPR, RPR-N, RPR-15, RPR-30, RPR-45, RPR-(30-15), and RPR-(45-15) of the slow growth group were significantly smaller than in the fast growth group (all P < 0.05). The growth rate of AL were positively correlated with TRPR (R = 0.383, P = 0.040), RPR-N (R = 0.395, P = 0.034), RPR-30 (R = 0.408, P = 0.028), RPR-45 (R = 0.377, P = 0.044), RPR-(30-15) (R = 0.390, P = 0.036). CONCLUSIONS: RPRs of children show relative myopic defocus after wearing OK lenses. Furthermore, the growth rate of AL is smaller with more negative RPR.

Dynamic Changes of AREG in the Sclera during the Development of Form-Deprivation Myopia in Guinea Pigs.

PURPOSE: To investigate amphiregulin (AREG) expression in the sclera during the development of form-deprivation myopia (FDM) and after the recovery of FDM in guinea pigs. METHODS: Sixty-four 2-week-old guinea pigs were randomly divided into the control and FDM groups. The right eyes of animals in FDM group were covered for 2 weeks (2 W) and 4 weeks (4 W), or were covered for 4 weeks and then uncovered for the subsequent 2 weeks (6 W). The diopters and axial lengths (AL) in the right eyes of guinea pigs were measured. Reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence, and western blotting assays were used to detect the relative mRNA and protein expressions of AREG in the sclera of guinea pigs. RESULTS: Before masking (0 W), the refraction and AL in the right eyes of guinea pigs in the control and FDM groups did not differ significantly (both p > .05). Myopic shift was induced in guinea pigs with the diopters gradually decreased and AL increased in the FDM group. While no significant difference was found in control group at different time points, the relative AREG mRNA and protein expression levels in the FDM group were significantly increased in 2 W and 4 W and then decreased after 4 weeks of covering followed by uncovering for 2 weeks (all p < .05). CONCLUSIONS: AREG was expressed in the sclera of guinea pigs. Moreover, the expression levels of AREG increased during the development of FDM and downregulated after recovery of FDM. Therefore, AREG may be involved in the regulation of scleral remodeling in myopia.

Modulation of the ERK1/2-MMP-2 pathway in the sclera of guinea pigs following induction of myopia by flickering light.

It has been shown that flickering light can affect the development of eyeballs. However, the exact mechanism remains unclear. The ERK1/2-MMP-2 pathway is a classic pathway involved in the modulation of the extracellular matrix (ECM) in cancer tissues. However, to the best of our knowledge, the role of this pathway in modulating the scleral ECM in myopia has not been previously examined. The present study aimed to determine the effects of the ERK1/2-MMP-2 pathway on the formation of flickering light-induced myopia (FLM). Guinea pigs were raised under illumination at a flash rate of 0.5 Hz for 6 weeks to induce FLM. Peribulbar injections of dimethylsulfoxide or PD98059 (an inhibitor of phospho-ERK1/2) were administered starting at the third week of FLM modeling. Refraction was measured prior to and following treatments. The thickness of the posterior sclera (PS) was measured under a light microscope following H&E staining. The mRNA levels of MMP-2 were detected by the reverse transcription-quantitative PCR assay. The expression levels of MMP-2 and ERK1/2 were assayed by western blot and immunohistochemical analyses. Following 6 weeks of treatment, the refraction of the FLM group became more myopic compared with that of the control group, while PD98059 treatment inhibited the changes noted in the refraction. A marked reduction in the thickness of PS was observed in the FLM group, while PD98059 inhibited the remodeling of PS. In addition, the expression levels of MMP-2 and protein levels of phospho-ERK1/2 were increased in the FLM group, while PD98059 significantly inhibited MMP-2 mRNA and protein levels. These results indicated that ERK1/2-MMP-2 may be involved in the formation of FLM in guinea pigs by regulating the remodeling of PS.

Dataset on microstructural characteristics and mechanical performance of homogeneous and functionally graded fibrous scaffolds.

Data in this article are supplementary to the corresponding research article [1]. Morphological features of homogeneous and graded nanofibrous electrospun gelatin scaffolds were observed using scanning electron microscopy. Microstructural properties including fiber diameter and pore size were determined via image analysis, using ImageJ. Uniaxial tensile and fracture tests were performed on both homogeneous and graded scaffolds using a universal testing machine. Stress-strain curves of all scaffolds are presented. Computing software, MATLAB, was used to design fibrous networks with thickness-dependent density and alignment gradients (DAG). Finite element analysis software, Abaqus, was used to determine the effect of the number of layers on the fracture properties of DAG multilayer scaffolds.