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Objective: To investigate the inhibitive effects of 5-fluorouracil-loaded polylactic acid nanoparticles (5-FU-PLA-NPs) on human Tenon's capsule fibroblasts in vitro., Methods: This paper was experimental study. MTT assay was performed to estimate the effects of 0.1 mg/L, 1.0 mg/L, 10.0 mg/L, 100.0 mg/L, 1000.0 mg/L unloaded polylactic acid nanoparticles (PLA-NPs) on fibroblasts proliferation on 48 h, 72 h. MTT assay was performed to estimate the effects of 0.1 mg/L, 1.0 mg/L, 10.0 mg/L, 100.0 mg/L, 1000.0 mg/L original 5-FU and 5-FU-PLA-NPs on fibroblasts' proliferation on 7 consecutive days respectively. The inhibitory rate of the two drugs against the fibroblasts was calculated. Cells were exposed to 100 mg/L original 5-FU and 5-FU-PLA-NPs respectively for 7 days. Semi-quantitative RT-PCR was used to observe the mRNA expression of type III procollagen at the 1st day, 5th day and 7th day., Results: PLA-NPs had no cytotoxicity on the fibroblasts. Both 5-FU-PLA-NPs and original 5-FU could inhibit the proliferation of the fibroblasts and make the expression of type III procollagen mRNA lower in dose-dependent and time-dependent manner. The inhibitive effect of original 5-FU was more effective than 5-FU-PLA-NPs in the initial period, but the inhibitive effect of 5-FU-PLA-NPs was more effective than original 5-FU in long time., Conclusion: PLA has good biocompatibility and safety, and can be used as a carrier of ophthalmic drugs. 5-FU-PLA-NPs shows slow-releasing function. 5-FU-PLA-NPs can be proposed as a potential controlled and targeted ophthalmic delivery system for the treatment of antifibrosis after glaucoma filtering surgery.

Objective: To investigate the effect of Rosiglitazone on the proliferation and the production of transforming growth factor-beta 1 (TGF-beta 1) in cultured humans Tenon's capsule fibroblasts (HTFs)., Methods: It was a experimental study. HTFs were cultured and subcultured in vitro. After HTFs were treated with different concentrations of Rosiglitazone, the proliferation was detected by MTT assay. The migration ability was detected by a scratch method. The production of TGF-beta 1 was determined by ELISA. TGF-beta 1 mRNA expression was examined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR)., Results: Compared with control group, Rosiglitazone significantly (F = 178.293, P < 0.01) inhibited the proliferation (0.33 +/- 0.01 vs 0.29 +/- 0.02) and the migration of HTFs in a dose-dependent manner with the concentrations ranged from 20 to 500 mg/L. Rosiglitazone also significantly (F = 86.404, P < 0.01) reduced the production of TGF-beta 1 in HTFs (323.48 +/- 13.69 vs 267.48 +/- 20.31). TGF-beta 1 mRNA was significantly (F = 176.102, P < 0.01) down-regulated (0.69 +/- 0.02 vs 0.55 +/- 0.02)., Conclusions: This study suggests that the inhibition of the proliferation of Tenon's capsule and the migration of HTFs by Rosiglitazone is due, at least in part, to decrease the production of TGF-beta 1.

Objective: To investigate the effect of connective tissue growth factor's antisense oligonucleotides (ASODN) on the growth of human tenon' s capsule fibroblasts (HTF) induced by transforming growth factor beta2 (TGF-beta2) in vitro., Methods: It was a experimental study. HTF was collected from glaucoma patients and cultured. The 5-6 passage was used for experiments. The HTF induced by TGF-beta2 was divided into the following groups: N group: normal HTF; T group: HTF induced by TGF-beta2; A group: CTGF ASODN antisense:5'-TACTGGCGGCGGTCAT-3' encapsulated with liposome; S group: sense 5'-ATGACCGCCGCCAGTA-3' encapsulated with liposome; D group: HTF encapsulated with liposome only. The activity of HTF treated by different concentrations of liposome was detected using methylthianolyldiphenyl tetrazolium bromide (MT) colorimetry. The expression of CTGF was detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry assays. The expression of fibronectin (Fn) was examined by Western blot and immunocytochemistry assays., Results: Liposome-ASODN (A group) significantly (F=15.25, 204.88, 19.73, 90.00; P <0.05) inhibit the expression of CTGF and Fn in HTF induced by TGF-beta2 compared with S and D group. However, Liposome alone (T group) has no significant impact in HTF growth compared with T group (t = 0.90, 2.32, 0.75, 2.11; P > 0.05)., Conclusion: CTGF-ASODN inhibits the CTGF and Fn expression of HTF induced by TGF-beta2, which may delay the formation of scar in glaucoma filtering surgery.

Objective: To investigate type I collagen and fibronectin expressions in the cultured human Tenon's capsular fibroblasts (HTFs) transfected with Smad 7 vector and to elucidate the possibility of Smad 7 in blocking tissue fibrosis after filtration surgery., Methods: Nucleofector(TM) was used to transfect Smad 7 vector into HTFs. The expressions of alpha2-type I procollagen (COL1A2) mRNA and fibronectin mRNA were detected by reverse transcription real-time quantitative polymerase chain reaction (real-time RT-PCR). The concentration of carboxyterminal propeptide of type I procollagen (PICP) in culture media was detected by radioimmunoassay. The above mentioned experiments were also detected before and after HTFs stimulated by TGF-beta(2) (10 ng/ml). HTFs and vector-HTFs were used as control groups., Results: Smad 7 was successfully transfected into HTFs evidenced by Smad 7 over expression. The expression of COL1A2 mRNA in Smad 7-HTFs was decreased 40.47% and 37.94%, respectively, when compared with the control HTFs and pCMV5-HTFs group. The PICP concentration of Smad 7-HTFs in the culture medium was decreased 52.10% and 57.35%, respectively, comparing to the two control groups. Furthermore, Smad 7-HTFs attenuated the increase of COL1A2 mRNA expression and PICP concentration of HTFs stimulated by TGF-beta(2). There was no difference in the fibronectin mRNA expressions between Smad 7-HTFs and the two control groups., Conclusions: Smad 7 reduces the expression of COL1A2 mRNA and the synthesis of type I collagen, but has no effect on fibronectin mRNA expression, which may imply its ability to inhibit tissue fibrosis after filtration surgery.

Objective: To test the safety of guidelines for limiting exposure to low power level microwave radiation and to try to explore the mechanisms of early damages induced by such radiation in rabbit lens capsule epithelial cells, using flow cytometry as a quantitative method of evaluation of these damages., Methods: The rabbit eyes were exposed to 5 mW/cm(2) (group I) and 10 mW/cm(2) (group II) power densities of microwave for 3 h. Annexin V-PI two-color flow cytometry was used to detect the changes of rabbits lens epithelial cells, apoptosis cells and secondary necrotic cells after radiation. The other eyes served as self-controls., Results: In group I, the percentage of early apoptosis cells in the experimental eyes was significantly greater than that of the controls (P < 0.01), whereas no significantly difference in the percentage of secondary necrotic cells could be observed (P > 0.05). In group II, there was no significant difference between the percentage of early apoptosis cells in experimental eyes and the controls (P > 0.05), whereas the percentage of secondary necrotic cells in the experimental eyes was significantly greater than that of the controls (P < 0.05). The percentage of secondary necrotic cells (16.2 +/- 5.6)% in group II is greater than that in group I (10.3 +/- 6.8)% (P < 0.05), though there is no significant difference in the percentage of early apoptosis cells between these two groups (P > 0.05)., Conclusion: Low power density of microwave radiation (5 mW/cm(2) and 10 mW/cm(2)) can induce irreversible damage of rabbit lens epithelial cells in a dose-dependent manner. Therefore, the safety dosage criterion of microwave exposure needs further investigation.

Objective: To elucidate the possible mechanism of Somatostatin 14 in the treatment of Graves' ophthalmopathy, thus providing laboratory data with the intent to underpin therapeutic developments., Methods: The human retro-ocular fibroblasts (HROFb) was cultured and MTT colorimetry assay was used to determine the proliferation of HROFb., Results: MTT colorimetry assay could precisely reflect the proliferation of HROFb. The absorbance was directly proportional to the numbers of the cells (r = 0.996, P < 0.001). Somatostatin 14 could enhance the cell proliferation percentage at lower levels (< 25 nmol/L) and inhibit that at higher levels (> 25 nmol/L). The cell proliferation correlated positively to Somatostatin 14 levels significantly (r = -0.9054, P < 0.001)., Conclusion: Somatostatin 14 has a dual effect on HROFb.

This is a study amied at the effects of homoharringtonine on the proliferation of human retro-ocular fibroblasts. The human retroocular fibroblasts (HROFb) were cultured and MTT colorimetry assay was used to determine the proliferation of HROFb. The influence of homoharringtonine on the proliferation of HROFb was observed in vitro. The results showed that MTT colorimetry assay could precisely reflect the proliferation of HROFb. The absorbance was directly proportional to the number of the cells (r = 0.996, P < 0.001). The proliferation of HROFb was inhibited by the homoharringtonine and the cell proliferation percentage was correlated positively to the homoharringtonine level (r = 0.6932, P < 0.001).