Dynamic changes of ocular biometric parameters:a modified form-deprivation myopia model of young guinea pigs

AIM: To evaluate the dynamic ocular biometric changes of a modified form-deprivation myopia model in young guinea pigs. METHODS: The animals were randomly assigned to two groups: the monocularly deprived facemask group (MDF, with all the right eyes covered, n=24) and the normal control group(free of facemask, n=24). Each group was then equally divided into four subgroups which were followed up for 2, 4, 6 and 8 weeks, respectively. Parameters measured from every eye included refraction, corneal curvature, axial length and the dry weight of sclera at the posterior pole. RESULTS: All the facemasks remained in place during the follow-up. The covered eyes developed myopia with the vitreous chamber lengthening and the dry weight of posterior sclera reduced at each time point compared with the contralateral uncovered (P<0.05 at all time points). The changes had a linear correlation with the deprivation time (P<0.05). There were no significant differences in all the parameters between the uncovered eyes of MDF group and the normal control group (P>0.05 at all time points). CONCLUSION: Monocular form deprivation with the facemask is highly effective and non-invasive in inducing axial myopia in guinea pigs. The axial myopia is mainly caused by the increased vitreous chamber length and the weakened posterior sclera rigidity. The form-deprivation eye didn't interfere with the natural development of the contralateral eye.

Scleral TGF-β1 and Smad3 expression is altered by TCM Bu Jing Yi Shi Tablets in guinea pigs with form-deprivation myopia

Objective:To explore whether the traditional Chinese medicine(TCM)Bu Jing Yi Shi Tablets alters the expression of scleral TGF-b1 and Smad3 in guinea pigs with formdeprivation myopia(FDM).Methods:Sixty-five guinea pigs were randomly divided into control,model,low-,medium-,and high-dose treatment groups.Except for the control group,FDM was induced by covering the right eye of each animal with opaque latex.The treatment groups were gavaged with different suspension concentrations of Bu Jing Yi Shi Tablets.Refraction and axial length were performed before and after myopia induction.At the end of the experiment,all right eyes were extracted,and scleral sections were prepared for staining and TGF-b1 and Smad3 immunohistochemistry.Scleral thickness and area,the scleral fibroblast quantity,and scleral TGFb1 and Smad3 expressions were measured.Results:The 5 FDM groups had the same initial axial length and diopter,the final diopter and axial length of the model group were significantly increased compared with the control group and treatment groups(P<.01).The axial length of each treatment group decreased as the dose decreased compared with the model group(P<.01);the total scleral area(P<.05 e.01)and the number of scleral fibroblasts(P<.01)in the model group were significantly lower than the treatment groups.Both the TGF-b1 and Smad3 integral optical densities in the model group were significantly lower than the control and medium-and high-dose treatment groups(P<.01).TGF-b1 and Smad3 mRNAs in the model group were decreased compared with the control group,but increased in expression after treatment.

Mechanism of the DL-alpha-aminoadipic acid inhibitory effect on form-deprived myopia in guinea pig

AIM:To investigate the effect of intravitreal injection of DL-alpha-aminoadipic acid (DL-α-AAA) on ocular refractive state and retinal dopamine, transforming growth factor-β2 (TGFβ2 ), vasoactive intestinal polypeptide (VIP) in guinea pig form-deprived myopia. METHODS:Four-week-old pigmented guinea pigs were randomly assigned to 4 groups:normal control, deprivation, deprivation plus DL-α-AAA, deprivation plus saline. Form deprivation was induced with the self-made translucent eye shields, and lasted for 14 days. 8μg DL-α-AAA was injected into the vitreous chamber of deprived eyes. The corneal radius of curvature, refraction and axial length were measured. Retinal dopamine content was evaluated by the high-performance liquid chromatography with electrochemical detection, and TGFβ2 and VIP protein were detected by Western blotting. RESULTS:Fourteen days of eye occlusion caused the axial length to elongate and become myopic in the form-deprived eyes, with the decrease of retinal dopamine and the increase of TGFβ 2 and vasoactive intestinal polypeptide (VIP) protein. Intravitreal injection of DL-α-AAA could inhibit the myopic shift from (-3.65±1.06)D to (-1.48 ±0.63)D, P 【0.01 due to goggles occluding and cause the decrease of retinal TGFβ2 protein in the deprived eyes. However, intravitreal injection of DL-α-AAA had no significant effect on retinal dopamine and VIP protein in deprived eyes. Retinal TGFβ2 protein correlated highly with the ocular refraction (y =-3.34 + 0.31/x , F =74.75, P 【0.001) and axial length (y =8.39-0.02/ x , F =48.32, P 【0.001) in different treatment groups. ·CONCLUSION:Intravitreal injection of DL-α-AAA is effectively able to suppress the development of form deprivation myopia, which may be associated with retinalTGFβ2 protein in guinea pigs.

Comparison of form-deprived myopia and lens-induced myopia in guinea pigs

AIM:To study the efficacy difference between formdeprived myopia(FDM)and lens-induced myopia(LIM),the degree of myopia,axial length and pathological changes of the posterior sclera from guinea pigs were evaluated.METHODS:Four-week pigmented guinea pigs were randomly assigned into 3 groups,including normal control(n=6),FDM group with monocular cover(n=11)and LIM group with monocular-7D lens treatment(n=11).FDM group was form-deprived while LIM group was lens-induced for 14d.Refractive error and axial length were measured prior to and post treatment,respectively.Morphological changes of sclera were examined using both light and electronic microscopes.RESULTS:After 14d treatment,refractive errors for FDM group and LIM group were-3.05±0.71D and-2.12±1.29D,respectively,which were significantly more myopic than that of normal controls and fellow control eyes(P【0.01).As for axial length,it was 7.93±0.03 mm for FDM group and 7.89±0.06 mm for LIM group,which were significantly longer than both normal and fellow controls(P【0.01).With respect to both refractory error and axial length,the differences between FDM group and LIM group were not significant(P】0.05).Under light microscope,both FDM group and LIM group showed thinned sclera,disarrangement of fibrosis and enlargeddisassociation between fibers.Consistently,ultrastructural examination showed degenerated fibroblasts and thinned fibers in posterior sclera.CONCLUSION:Following two weeks of myopia induction in guinea pigs,with regard to the degree of myopia,axial length and pathological alterations,there was no significant difference between FDM and LIM models.Therefore,FDM and LIM are equally effective and useful as a model of experimental myopia and guinea pigs are ideal animals for induction of experimental myopia because their high sensitivity to both formdeprivation and lens-induction.

Protective effects of riboflavin-UVA-mediated posterior sclera collagen cross-linking in a guinea pig model of form-deprived myopia

AIM:To evaluate the effect of posterior sclera collagen cross-linking induced by riboflavin-ultraviolet A(UVA)on form-deprived myopia in guinea pigs.METHODES:Twenty-five pigmented guinea pigs of 3-week-old were randomly assigned into 4 groups that included normal control(NOR,n=7),form-deprived(FDM,n=7),normal with riboflavin-UVA cross-linking(NOR+CL,n=5)and form-deprived with cross-linking(FDM+CL,n=6).The NOR+CL group and the FDM+CL group received the riboflavin-UVA induced cross-linking at day 0.FDM was induced by monocularly deprived with facemask in the right eyes.The refraction,axial length and corneal curvature were measured by retinoscopy,A-scan and keratometer respectively in scheduled time points(day 0 and 1,2,3,4 wk after form-deprivation).At the end of 4 weeks’experiment,stress-strain tests of sclera were measured and morphological changes of sclera and retina were examined.RESULTS:After 4 wk,the interocular difference of refractive error were-0.11±0.67,-2.93±0.56,1.10±0.58,and-1.63±0.41 D in the NOR,FDM,NOR+CL,and FDM+CL groups respectively.Mixed-effect linear model revealed significant effect of FDM(P<0.01)and CL(P<0.001).Also,after 4 wk,the interocular difference of axial length were 0.01±0.04,0.29±0.07,-0.13±0.06,and 0.11±0.05 mm in the NOR,FDM,NOR+CL,and FDM+CL group.Mixedeffect linear model revealed significant effect of FDM(P<0.001)and CL(P<0.01).As for corneal curvature,significant interocular difference have not found between any of the two groups.At the end of this experiment,the ultimate stress and elastic modulus were found significantly increased in both CL groups.But no difference was found in the groups without cross-linked.There was no abnormality observed in the retina and RPE cells of the treated eyes.CONCLUSION:The posterior sclera collagen crosslinking induced by riboflavin-UVA can slow down the progress of myopia and increase the sclera biomechanical strength in the guinea pig model of form-deprived myopia.

Expression and role of specificity protein 1 in the sclera remodeling of experimental myopia in guinea pigs

AIM：To study the expression of collagen I and transcription factor specificity protein 1（Sp1）,a transforming growth factor-β1（TGF-β1） downstream target,and reveal the impact of the TGF-β1-Sp1 signaling pathway on collagen remodeling in myopic sclera.METHODS：Seventy-five 1-week-old guinea pigs were randomly divided into normal control,form deprivation myopia（FDM）,and self-control groups.FDM was induced for different times using coverage with translucent latex balloons and FDM recovery was performed for 1wk after 4wk treatment;then,changes in refractive power and axial length were measured.Immunohistochemistry and reverse transcription-polymerase chain reaction were used to evaluate dynamic changes in collagen I and Sp1 expression in the sclera of guinea pigs with emmetropia and experimental myopia,and the relationship between collagen I and Sp1 levels was analyzed.RESULTS：In the FDM group,the refractive power was gradually changed（from 2.09±0.30 D at week 0 to-1.23±0.69 D,-4.17±0.59 D,-7.07±0.56 D,and-4.30±0.58 D at weeks 2,4,6,and 1wk after 4wk,respectively;P〈0.05）,indicating deepening of myopia.The axial length was increased（from 5.92±0.39 mm at week 0 to 6.62±0.36 mm,7.30±0.34 mm,7.99±0.32 mm,and 7.41±0.36 mm at weeks 2,4,6,and 1wk after 4wk;P〈0.05）.The m RNA and protein expression of Sp1 and collagen I in the sclera of the FDM group was lower than that of the control groups（P〈0.05）,and the reduction was eye-coverage time-dependent.Furthermore,correlation between Sp1 and collagen I down-regulation in the myopic sclera was observed.CONCLUSION：Our data indicate that transcription factor Sp1 may be involved in the regulation of type I collagensynthesis/degradation during myopic sclera remodeling,suggesting that TGF-β1 signaling plays a role in the development and progression of myopia.

Vasoactive intestinal peptide,a promising agent for myopia?

AIM： To investigate the role of vasoactive intestinal peptide（VIP） in form-deprivation myopia（FDM）.METHODS： FDM was created in three groups of eight chicks by placing a translucent diffuser on their right eyes.Intravitreal injections of saline and VIP were applied once a day into the occluded eyes of groups 2 and 3,respectively.Retinoscopy and axial length（AL） measurements were performed on the first and 8^th days of diffuser wear.The retina mR NA levels of the VIP receptors and the ZENK protein in right eyes of the three groups and left eyes of the first group on day 8 were determined using real time polymerase chain reaction（PCR）.RESULTS： The median final refraction（D） in right eyes were-13.75（-16.00,-12.00）,-11.50（-12.50,-7.50）,and-1.50（-4.75,-0.75） in groups 1,2,and 3,respectively（P〈0.001）.The median AL（mm） in right eyes were 10.65（10.00,11.10）,9.90（9.70,10.00）,and 9.20（9.15,9.25） in groups 1,2,and 3,respectively（P〈0.001）.The median delta-delta cycle threshold（CT） values for the VIP2 receptors were 1.07（0.82,1.43）,1.22（0.98,1.65）,0.29（0.22,0.45） in right eyes of groups 1,2,and 3,and 1.18（0.90,1.37） in left eyes of group 1,respectively（P=0.001）.The median delta-delta CT values for the ZENK protein were 1.07（0.63,5.03）,3.55（2.20,5.55）,undetectable in right eyes of groups 1,2,and 3 and 1.89（0.21,4.73） in left eyes of group 1,respectively（P=0.001）.CONCLUSION： VIP has potential inhibitory effects in the development of FDM.