PURPOSE: To investigate the role of hydrogen sulfide in the survival and collagen gel contraction of cultured human Tenon's capsule fibroblasts (HTCFs). METHODS: Primarily cultured HTCFs were exposed to 0, 100, 200, or 300 µM hydrogen sulfide (sodium hydrogen sulfide, NaHS) for 2 days. Cellular survival was assessed by MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay. Degree of apoptosis was assessed with flow cytometry using annexin-V/propidium iodide double staining. To evaluate the effect of NaHS on cellular transdifferentiation, HTCFs were stimulated with 5 ng/mL TGF-β1 and the level of expression of α-smooth muscle actin (SMA) mRNA was assessed using reverse-transcription polymerase chain reaction. The cells were embedded in collagen gel, and the amount of gel contraction was measured. RESULTS: NaHS at 300 µM reduced HTCF survival (p = 0.013); NaHS at both 200 and 300 µM increased apoptosis in a dose-dependent manner (p = 0.013 and p = 0.016). TGF-β1 increased the expression of α-SMA mRNA (p = 0.041); co-treatment with 100 µM NaHS decreased TGF-β1-induced α-SMA mRNA expression (p = 0.039) and inhibited collagen gel contraction. CONCLUSIONS: NaHS at high concentration reduced cellular survival and increased HTCF apoptosis. NaHS decreased TGF-β 1-induced increases in α-SMA mRNA expression and collagen gel contraction. Thus, hydrogen sulfide may suppress scar formation by inhibiting HTCF transdifferentiation and contraction of collagen gels.
Actins ; Apoptosis ; Cicatrix ; Collagen ; Fibroblasts ; Flow Cytometry ; Gels ; Humans ; Hydrogen Sulfide ; Hydrogen ; Polymerase Chain Reaction ; RNA, Messenger ; Tenon Capsule

PURPOSE: To investigate the effects of valproic acid on the survival of cultured human Tenon's capsule fibroblasts (HTFBs). METHODS: Primary cultured HTFBs were exposed to 0, 0.25, 0.5, and 1.0 mM valproic acid with or without 0, 1.0, 2.5 µg/mL mitomycin C, and incubated for 5 days. Cell survival was assessed using an MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay and the degree of apoptosis was assessed by flow cytometry using annexin-V/propidium iodide double staining. RESULTS: Valproic acid decreased the survival of HTFBs in a dose-dependent manner, and survival was further decreased by adding mitomycin C to valproic acid. Both valproic acid and mitomycin C induced apoptosis of HTFBs. Valproic acid induced less apoptosis than mitomycin C. CONCLUSIONS: Valproic acid decreased the cellular survival of HTFBs and induced apoptosis. The antiproliferative effects of valproic acid were further enhanced by the addition of mitomycin C.
Apoptosis ; Cell Survival ; Fibroblasts ; Flow Cytometry ; Humans ; Mitomycin ; Tenon Capsule ; Valproic Acid

PURPOSE: To evaluate the anti-fibrotic effects of nilotinib on the survival of cultured human Tenon's capsule fibroblasts (HTFs). METHODS: HTF primary cultures were obtained from samples following glaucoma surgery. Primarily cultured HTFs were exposed to 1, 5, 10, and 20 µM nilotinib for 24 hours. The effects of nilotinib on HTF proliferation and cell viability were determined using the 3-(4,5-dimethylthiazone-2-yl)-2,5-diphenyl tetrazolium (MTT) assay, and apoptosis was determined by flow cytometry using annexin-V/propidium iodide (PI) double staining. Apoptosis-related proteins were detected by western blotting. RESULTS: The MTT assay showed that nilotinib induced an inhibition of HTF proliferation at concentrations of 10 and 20 µM (p < 0.001 and p < 0.001, respectively). Annexin V/PI double staining showed significantly increased apoptosis in cells treated with nilotinib. Nilotinib activated caspase-3, -9, and poly adenosine diphosphate ribose polymerase cleavage, and downregulated the expression of B-cell lymphoma-extra large and Bax, which indicated that nilotinib-induced apoptosis was partly mediated through the mitochondrial pathway. In addition, treatment with nilotinib decreased the expression of α-smooth muscle actin and transforming growth factor-β. CONCLUSIONS: Nilotinib decreased cell survival of cultured HTFs and induced mitochondria-mediated apoptosis. The results suggested that nilotinib may exert antiproliferative effects on HTFs, making it a possible agent to control postoperative fibrosis in patients undergoing glaucoma surgery.
Actins ; Apoptosis ; B-Lymphocytes ; Blotting, Western ; Caspase 3 ; Cell Survival ; Fibroblasts ; Fibrosis ; Flow Cytometry ; Glaucoma ; Humans ; In Vitro Techniques ; Poly Adenosine Diphosphate Ribose ; Tenon Capsule

PURPOSE: We report a case of infraduction limitation improvement and orthotropia at primary position in a patient with traumatic isolated inferior rectus muscle rupture after immediate repair surgery of the ruptured muscle. CASE SUMMARY: A 47-year-old man came to our emergency department complaining of right eyelid contusion and conjunctival laceration after trauma due to the metal part of a high pressure hose. Right hypertropia of 20 prism diopters was observed at primary gaze and right hypertropia of 30 prism diopters was observed at down gaze. The right eye showed a -4 infraduction limitation. We sutured the distal part of the ruptured inferior rectus muscle with surrounding tissue, including the Tenon's capsule and the part of the muscle stump, to its original insertion. Orthotropia at primary position and 8-prism-diopters of right hypertropia on down gaze was observed one day after surgery. At 40 days after surgery, the patient had orthotropia at primary position and 4-prism-diopter right hypertropia on down gaze. Infraduction limitation of the right eye improved to -1. CONCLUSIONS: Immediate repair of complete traumatic isolated inferior rectus muscle rupture can correct the limitation of extraocular movement and achieve orthotropia at primary position.
Contusions ; Emergency Service, Hospital ; Eyelids ; Humans ; Lacerations ; Middle Aged ; Rupture* ; Strabismus ; Tenon Capsule

PURPOSE: To evaluate the effects of posterior subtenon triamcinolone acetonide injection on refractory diabetic macular edema (DME) after intravitreal bevacizumab (IVB) injection failure. METHODS: Patients with DME and central subfield thickness (CST) >300 microm who did not respond to IVB injections were retrospectively included. Specifically, we enrolled patients who were diagnosed with refractory DME and who experienced an increase in CST after 1 to 2 IVB injections or no decrease after > or =3 consecutive IVB injections. One clinician injected 20 mg of triamcinolone acetonide into the posterior subtenon space. All patients received ophthalmic examinations at baseline and at 2, 4, and 6 months post-baseline. Examinations included Snellen visual acuity, intraocular pressure, and spectral-domain optical coherence tomography. RESULTS: Forty eyes of 34 patients were included. The average baseline CST was 476 microm. The average CST decreased to 368 microm at 2 months, 374 microm at 4 months, and 427 microm at 6 months (p < 0.001 for all results, Wilcoxon signed-rank test). The average intraocular pressure increased from 15.50 to 16.92 mmHg at 2 months but decreased to 16.30 mmHg at 4 months and 15.65 mmHg at 6 months. Logarithm of the minimum angle of resolution visual acuity improved from 0.56 to 0.50 at 2 months (p = 0.023), 0.50 at 4 months (p = 0.083), and 0.48 at 6 months (p = 0.133, Wilcoxon signed-rank test). No complications were detected. CONCLUSIONS: Posterior subtenon triamcinolone acetonide is an effective and safe treatment for reducing CST in DME refractory to IVB.
Aged ; Angiogenesis Inhibitors/*therapeutic use ; Bevacizumab/*therapeutic use ; Diabetic Retinopathy/diagnostic imaging/*drug therapy/physiopathology ; Female ; Glucocorticoids/*administration & dosage ; Humans ; Injections, Intraocular ; Intraocular Pressure/physiology ; Intravitreal Injections ; Macular Edema/diagnostic imaging/*drug therapy/physiopathology ; Male ; Middle Aged ; Retrospective Studies ; Tenon Capsule/*drug effects ; Tomography, Optical Coherence ; Treatment Failure ; Triamcinolone Acetonide/*administration & dosage ; Vascular Endothelial Growth Factor A/antagonists & inhibitors ; Visual Acuity/physiology

PURPOSE: Resveratrol exerts cytoprotective or cytotoxic effects according to cell type. This study was performed to evaluate the effects of resveratrol on the survival of cultured human Tenon's capsule fibroblasts (HTFBs). METHODS: Primarily cultured HTFBs were exposed to 0, 10, or 100 microM resveratrol for 3 days. Cellular survival was assessed using the MTT assay and degree of apoptosis was analyzed with flow cytometry using annexin-V/propidium iodide double staining. RESULTS: Resveratrol decreased the survival of HTFBs after exposure to 10 microM (p = 0.04). In flow cytometric analysis, 10 microM resveratrol did not affect the degree of apoptosis (p = 0.89), but 100 microM resveratrol increased the degree of apoptosis (p = 0.003). Both 10 and 100 microM resveratrol did not affect the degree of necrosis (p = 0.74, 0.33). CONCLUSIONS: Resveratrol decreased cellular survival of cultured HTFBs and induced apoptosis. Thus, resveratrol may exert antiproliferative effects on HTFBs.
Apoptosis ; Fibroblasts* ; Flow Cytometry ; Humans* ; Necrosis ; Tenon Capsule

Objective: To evaluate and compare the effects of bevacizumab, mitoinycin-C (MMC), 5-fluorouracil (5-FU), and triamcinolone acetonide (TA) on the viability of cultured human Tenon's capsule fibroblasts (cHTF) in vitro.

Methods: Human Tenon's fibroblasts (HTF) were harvested and cultured in a Roswell-Park-Memorial 1-Institute (RPMI) media. MMC, 5-FU, bevaciz. umab, and TA were administered to the cHTF at 3-fold decreasing concentrations starting from 20 ug, 5 mg, 25 mg, and 4 mg respectively. A negative control/untreated group containing RPMI media only was included in the study. Fibroblast cell viability was assessed using resazurin fluorim etric assay. Half¬maximal inhibitory concentration (IC50) was computed for agents which showed significant decrease in cHTF viability compared to the untreated group.

Results: There was no significant difference in cH IF viability between the untreated control group compared to 5-FU (p=0.97), bevacizumab (p=0.10), and TA (p=0.06) groups. Mitomycin-C showed a significant decrease in cHTF viability (p<0.001) which was dose dependent. The IC50 of MMC was computed at 12.16 ug using the prism software.

Conclusion: Mitomycin-C demonstrated dose-dependent decrease in viability of cultured human Tenon's fibroblasts. 5-FU, bevacizumab, and triamcinolone did not show this effect.

Key Words: Mitomycin-C, 5-fluorouracil, Bevaciz. umab, Tria. mcinolone acetonide, Fibroblast, Trabeculectomy

Human ; Male ; Female ; .humans ; Mitomycin ; Triamcinolone Acetonide ; Trabeculectomy ; Resazurin ; Bevacizumab ; Fluorouracil ; Cell Survival ; Control Groups ; Inhibitory Concentration 50 ; Tenon Capsule ; Xanthenes ; Oxazines ; Antibodies, Monoclonal, Humanized ; Fibroblasts ; Software

PURPOSE: To evaluate the effect of adjunctive subtenon injection of triamcinolone acetonide (TA) in gas-filled eyes after vitrectomy for complicated proliferative diabetic retinopathy (PDR). METHODS: This nonrandomized comparative study included 27 patients (27 eyes) who underwent pars plana vitrectomy and gas tamponade for treatment of PDR with tractional or combined tractional-rhegmatogenous retinal detachment and who received subtenon injection of TA (40 mg) at the end of surgery. The study group was compared with the control group (29 eyes), which was matched with the study group for preoperative and intraoperative parameters, but underwent pars plana vitrectomy and gas tamponade without a subtenon injection of TA. RESULTS: Retinal reattachments without reoperation were achieved in 25 eyes (92.6%) and 26 eyes (89.7%) at 6 months (p = 1.000) in the study and control groups, respectively. The study group and the control group did not differ significantly in the frequency of postoperative proliferative vitreoretinopathy, retinal redetachment rate, reoperation rate, macular pucker formation, postoperative vitreous hemorrhage, gain in visual acuity, intraocular pressure, and intraocular inflammation (p > 0.05). CONCLUSIONS: The clinical results of pars plana vitrectomy for complicated PDR are not improved significantly by an adjunctive subtenon TA injection in gas-filled eyes.
Diabetic Retinopathy/complications/*therapy ; Female ; Gases ; Glucocorticoids/administration & dosage ; Humans ; Injections ; Male ; Middle Aged ; Tenon Capsule ; Treatment Outcome ; Triamcinolone Acetonide/*administration & dosage ; Visual Acuity ; Vitrectomy/*methods ; Vitreoretinopathy, Proliferative/etiology/*therapy

PURPOSE: To compare the effect of loteprednol etabonate (LE) with prednisolone acetate (PDA) drops on the proliferation of human Tenon's capsule fibroblasts (HTFBs). METHODS: Primarily cultured HTFBs were treated with serially diluted PDA and LE for 3 days. Cellular survival was determined by a rapid colorimetric assay using MTT. RT-PCR was performed to determine the relative expression of TGF-beta mRNA in response to LE and PDA. RESULTS: PDA inhibited proliferation of HTCF in a dose-dependent manner and LE inhibited significantly the proliferation of HTCF at the higher concentration of 50 microg/ml (p < 0.05). Compared to LE, PDA inhibited proliferation of HTCF significantly at each diluted concentration (p < 0.05). Expressions of TGF-beta were decreased as the concentration of both PDA and LE increased. PDA decreased expression of TGF-beta more significantly compared to LE at each concentration (p < 0.05). CONCLUSIONS: Although LE has offered promising anti-inflammatory efficacy with decreased impact on intraocular pressure, LE may be less effective than PDA in inhibiting fibroblast proliferation and may be not comparable to PDA in preventing excessive scarring after glaucoma filtering surgery.
Androstadienes ; Cicatrix ; Fibroblasts ; Filtering Surgery ; Glaucoma ; Humans ; Intraocular Pressure ; Ophthalmic Solutions ; Prednisolone ; RNA, Messenger ; Tenon Capsule ; Transforming Growth Factor beta ; Loteprednol Etabonate

OBJECTIVETo investigate the effect of tetrandrine (Tet) on the expression of bax, bcl-2, and transforming growth factor-β2 (TGF-β2) mRNA in cultured human fibroblasts of Tenon's capsules (TCFS) and explore its possible mechanism.

METHODSThe third passage of TCFS cultured in vitro were exposed to 1×10(-5) mol/L Tet for 24 h, and real-time fluorescence quantitative PCR was used to detect the changes in the expressions of bax, bcl-2, and TGF-β2 mRNA.

RESULTSThe expression level of bax mRNA was obviously higher, while bcl-2 and TGF-β2 mRNA levels were significantly lower in Tet-treated TCFS than those in the control cells (P<0.05).

CONCLUSIONTet can inhibit the proliferation of TCFS possibly by reducing the expressions of bcl-2 and TGF-β2 mRNA, enhancing the expression of bax mRNA and inducing cell apoptosis, suggesting its potential in preventing fibrous scar formation after glaucoma filtration surgery.

Apoptosis ; Benzylisoquinolines ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix ; metabolism ; prevention & control ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Tenon Capsule ; cytology ; Transforming Growth Factor beta2 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism