Expression Of TGF-β2,FGFR-1, Proliferation, And The Study Of Fibroblasts Biomechanical Properties In Experimental Myopia Scleral Fibroblasts On The Impact Of BFGF

Objective: On the impact of bFGF, to determine the cell proliferation and the expression of TGF-β2,FGFR-1 in cultured guinea pig posterior scleral fibroblasts in lens-induced myopia, and also to study the biomechanical properties changes. Some conclusions were valuable for revealing the pathological mechanism and the treatment of myopia.Methods: 10 two-week-old guinea pigs were monocularly fitted with -10.00D lenses randomly,another eye in each animal was untreated as self-control (SC). Before and after the experiment, diopter was measured using retinoscopy and the ocular axial was determined by A-scan ultrasonography. After 45 days, the scleral fibroblasts of each group were cultured and passaged 2 generations in vitro, The cultured cells was determined to be fibroblasts using immunocytolchemistory methods. The scleral fibroblasts of each eye was devided into A, B, C, D four groups, represent for the group of black, the group of 1ng/ml bFGF, the group of 10ng/ml bFGF, the group of 100ng/ml bFGF. MTT was used to detect the proliferation of each group of scleral fibroblasts, immunocytolchemistry was used to detect the expression of TGF-β2 and FGFR-1 in the scleral fibroblasts. And utilized the micropipette aspiration technique coupled with a viscoelastic solid model to investigate the viscoelastic properties of scleral fibroblasts from each group of normal and myopia guinea pigs. All datas were statistically analyzed by SPSS 13.0 system.Results:1 The experimental myopia of guinea pigs induced results: Diopter had no significant difference in binocular before induced. Eye became myopic on average of -6.70±1.93D, eye elongated on average of 1.53±0.31mm after insertion of the lens for 45 days. There was obvious difference between them (Student's t test, P <0.05).2 The results of cells cultured and determined Cultured cells began to grow out of the sclera after 3~5 days, and grew to confluent after 2~3 weeks. The cultured cells was determined to be fibroblasts using immunocytolchemistory methods.3 MTT resultsSeparated the 2 generation cultured cells into A, B, C, D four groups. MTT results: In the group of LIM, there was on obvious difference between group A and group B(P>0.05), but there was statistic significance between group C and group A, group D and group A(P<0.05). The amount of the cells increased 24.9%, 14.2% respectivly. In the group of SC, there was on obvious difference between group A and group B, but there was statistic significance between group C and group A, group D and group A. The amount of the cells increased 27.4%, 12.5% respectivly. 4 The expression of TGF-β2, FGFR-1 in posterior of scleralThe expression of TGF-β2 was located in cytoplasm and nuclear membrane, and it was higher in experimental eye than in control eye. There was significant difference between them (P<0.05). The expression of TGF-β2 is gradually higher follow the concentration of bFGF.The expression of FGFR-1 was located in cytoplasm and nuclear membrane, and it was lower in experimental eye than in control eye. There was significant difference between them (P<0.05). The expression of FGFR-1 is not have a significant change follow the concentration of bFGF.5 Biomechanical research for scleral fibroblasts5.1 Biomechanical research for scleral fibroblasts of SC and LIMExperimental studies have shown that the viscoelastic properties of the scleral fibroblasts in LIM group exhibited a significantly higher equilibrium modulus, and apparent viscosity (E∝=0.3041±0.4658kPa,μ=5.4077±1.041kPa·s)compared with SC group(E∝=0.2125±0.0526kPa,μ=2.8875±0.5138kPa·s).5.2 Biomechanical research for scleral fibroblasts of SC and LIM at the effect of bFGF5.2.1 Biomechanical research for scleral fibroblasts of SC at the effect of bFGF, the equilibrium modulus of group A, B, C, D was E∝=0.2125±0.0526kPa , E∝=0.2158±0.7753kPa, E∝=0.3340±1.062kPa, E∝=0.2963±0.0505kPa respectivly, compared with group A, the group C and D both have a significant difference(P<0.05). The change of equilibrium modulus was gradually higher follow the concentration of bFGF.5.2.2 Apparent viscosity research for scleral fibroblasts of SC at the effect of bFGF, the equilibrium modulus of group A, B, C, D wasμ=2.8875±0.5138kPa·s,μ=2.1557±0.8327kPa·s,μ=4.9425±1.0011kPa.s,μ=3.7078±1.4479kPa.s respectivly, compared with group A, the group C and D both have a significant difference(P<0.05). The change of equilibrium modulus was gradually higher follow the concentration of bFGF.5.2.3 Neither equilibrium modulus nor apparent viscosity research for scleral fibroblasts of LIM on the effect of bFGF have a significant difference(P<0.05).5.2.4 The equilibrium modulus and apparent viscosity research for scleral fibroblasts of LIM was higher compered with the group A and group B of SC, but not higher compered with the group C and group D.Conclusions:1 lenses can induce axial growth and lead to myopia in young guinea pigs.2 bFGF was an effect factor on the proliferation of scleral fibroblasts cells. The concentrations of 10ng/ml, 100ng/ml was useful for the cells proliferation. The concentration of 10ng/ml was the best.3 The expression of TGF-β2 in experimental eye was higher than normal eye, but the expression of FGFR-1 in experimental eye was lower than normal eye.4 Expression of TGF-β2, FGFR-1 were all detected at different bFGF concentrations in guinea pigs scleral fibroblasts. We can conclude that the bFGF plays an important role in the expression of FGFR-1, but not in the expression of TGF-β2.5 There were significant alteration in equilibrium modulus and apparent viscosity of scleral fibroblasts of experimental eye.6 In my experiment only the group of 10ng/ml, 100ng/ml can improve the equilibrium modulus and apparent viscosity of scleral fibroblasts of normal eye.7 In my experiment the bFGF cannot change the equilibrium modulus and apparent viscosity of scleral fibroblasts of LIM. The improve of equilibrium modulus and apparent viscosity research for scleral fibroblasts of SC at the effect of bFGF was not higher than the LIM.