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ABSTRACT Purpose: As superotemporal implantation of the Ahmed glaucoma valve is not always feasible in cases of refractory glaucoma, this study examined the characteristics and surgical outcomes of cases in which the valve was implanted in a nonsuperotemporal quadrant using a modified long scleral tunnel technique. Methods: This retrospective case-control study included 37 eyes with nonsuperotemporal quadrant--Ahmed glaucoma valve implantation in Group 1 and 69 eyes with superotemporal Ahmed glaucoma valve implantation in Group 2. The demographic characteristics of these groups, surgical outcomes, including complications, further surgical interventions, and surgical success rates were compared. Surgical success was defined as an intraocular pressure not exceeding 21 mmHg, accompanied by a minimum reduction of 20% in intraocular pressure from the baseline without any additional intraocular pressure-lowering procedures, and the absence of light perception loss or phthisis bulbi. Results: Group 1 had significantly higher numbers of eyes with secondary glaucoma and preoperative surgical procedures than Group 2 (p<0.05). Both groups had mean preoperative intraocular pressure values, and mean intraocular pressure values at the last visit of 34.2 and 27.9 months, 35.5 ± 1.5 and 35.8 ± 1.2 mmHg, and 14.5 ± 5 and 14.9 mmHg, respectively. Although both groups had 70.2% and 75.8% as their five-year cumulative probability of success, respectively, the rates of complications, revisional surgery, and additional surgical procedures did not differ significantly (p>0.05). Conclusion: The modified long scleral tunnel technique for Ahmed glaucoma valve implantation in nonsuperotemporal quadrants achieves intraocular pressure control and complication rates comparable to superotemporal implantation.
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Background: In acute chemical injury, damage can range from ocular surface epithelial defects to limbal and scleral ischemia. This may subsequently progress to corneal or scleral melting and perforation and finally result in phthisis bulbi. Thus, acute chemical injury is a potentially blinding condition and warrants attention. The accurate technique to assess the damage incurred should be practiced to avoid undertreatment and subsequent complications. Surgical intervention wherever needed should be appropriately timed and should be performed. The primary aim of medical or surgical intervention in acute chemical injury is to attain a stable and epithelized ocular surface. Even a conjunctival phenotype over the cornea is a desirable outcome. Purpose: This video discusses the nuances involved in the assessment and planning of Tenon advancement with amniotic membrane grafting for treating limbal ischemia in acute chemical injury. Synopsis: The video demonstrates the technique of restoration of limbal vascularization by performing Tenon advancement with amniotic membrane grafting and its outcome. Highlights: Ocular surface painting with fluorescein dye is essential to assess the areas of surface involvement. Merely instilling the fluorescein dye in the cul?de?sac will underestimate the extent of the damage. Tenon advancement should ideally be planned between 7 and 10 days following an injury when actual limbal blanching is obvious. A stable and epithelized ocular surface is the desirable outcome irrespective of the epithelial phenotype.
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Purpose: To evaluate the efficacy and safety of posterior sub?Tenon triamcinolone (PSTA) in chronic postoperative cystoid macular edema (PCME) after pars plana vitrectomy (PPV). Methods: Consecutive 22 patients who developed chronic PCME after PPV and underwent PSTA treatment were included in this retrospective study. Best?corrected visual acuity (BCVA) and central macular thickness (CMT) were measured pre injection and post injection at one month, three months, six months, and at last visit. The patients were divided into three groups according to the injection response status: complete, partial, and resistant. Results: The mean follow?up period was 26.4 ± 16.2 months after PSTA. According to pre?injection values, there was a significant improvement in the values of BCVA and CMT at the first, third, and sixth months and at the last examination (P < 0.05). In the final examination, PCME recovered completely in 12 patients, partially in 8 patients, and resistance was observed in 2 patients. Conclusion: Posterior sub?Tenon triamcinolone seems to be effective in chronic PCME following PPV.
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AIM: To investigate the effect of ALK5 inhibitor EW-7197 on the proliferation and migration of human Tenon fibroblasts(HTFs)induced by transforming growth factor-β1(TGF-β1)and its mechanism.METHODS: The cell proliferation rate was detected by MTS assay, and the optimal concentration and time of EW-7197 were explored. Then HTFs were divided into three groups: normal control group, TGF-β1 induced group and TGF-β1+EW-7197 group. Cell migration was observed by Transwell assay. The protein expression levels of Fibronectin, α-SMA, as well as the phosphorylated Smad2, Smad3(p-Smad2, p-Smad3)were measured by Western blot.RESULTS: MTS assay showed that the proliferation rate of cells treated with 6.0 μmol/L EW-7197 for 24h was the lowest(all P<0.01). Transwell assay showed that the migrated number of HTFs in TGF-β1 induced group was 228.0±17.0/field, which was significantly more than that in normal control group(149.0±15.0/field)and TGF-β1+EW-7197 group(46.0±8.0/field; all P<0.01). Western blot showed that the protein relative expression levels of Fibronectin, α-SMA and p-Smad2, p-Smad3 of HTFs in TGF-β1 induced group were significantly higher than that in normal control group and TGF-β1+EW-7197 group(all P<0.001).CONCLUSION:EW-7197 can suppress the proliferation and migration of TGF-β1-induced HTFs through TGF-β/Smad signaling pathways.
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Objective:To study the effect and mechanism of angiotensin type 1 receptor (AGTR1) blocker olmesartan (OMS) on the apoptosis of human Tenon capsule fibroblasts (HTF).Methods:Tenon capsule tissues were obtained from patients during strabismus surgery in the Second Affiliated Hospital of Xi'an Jiaotong University.Primary HTF were cultured by explant culture.Primary cells were identified by vimentin immunofluorescence staining and flow cytometry.The fibrosis model of HTF was established using 10 ng/ml transforming growth factor-β2 (TGF-β2). The cells were divided into normal control group cultured in culture medium, TGF-β2 group in culture medium containing TGF-β2, TGF-β2+ OMS group in culture medium containing TGF-β2 and OMS, and OMS group in culture medium containing OMS, and were cultured for 48 hours.Cell apoptosis was detected by flow cytometry with annexin V/PI staining.The early apoptosis, late apoptosis, and total apoptosis rates were analyzed.The protein expression of procaspase-9, cleaved caspase-9, bax and bcl-2 in the mitochondrial apoptosis pathway was detected by Western blot.The activity of lactate dehydrogenase (LDH) and superoxide dismutase (SOD) was detected by colorimetry.The study protocol was approved by the Ethics Committee of the Second Affiliated Hospital of Xi'an Jiaotong University (No.2019-014).Results:Primary HTF were successfully isolated and cultured.The cultured cells were long spindle-shaped and positive for vimentin.The expression rate of vimentin in the primary cells was greater than 99%.A statistically statistical difference was found in the early apoptosis rate, late apoptosis rate, and total apoptosis rate among the four groups ( F=24.92, 3.96, 41.82; all at P<0.05). The early and total apoptosis rates were significantly higher in TGF-β2+ OMS group than normal control group and TGF-β2 group, and the late apoptosis rate in TGF-β2+ OMS group was significantly higher than that of normal control group (all at P<0.05). There were statistically significant differences in cleaved caspase-9/procaspase-9, bax, and bax/bcl-2 among the four groups ( F=4.40, 7.98, 4.61; all at P<0.05). The bax/bcl-2 expression was significantly increased in TGF-β2+ OMS group in comparison with normal control group, and the expressions of cleaved caspase-9/procaspase-9, bax, and bax/bcl-2 were significantly elevated in TGF-β2+ OMS group compared with TGF-β2 group (all at P<0.05). LDH activity in the normal control group, TGF-β2 group, TGF-β2+ OMS group and OMS group was (783.99±79.97), (913.16±196.86), (2 529.06±240.21), and (2 134.29±138.96) μmol/(min·L), respectively, showing a statistically significant difference ( F=24.95, P<0.05). Compared with normal control group and TGF-β2 group, LDH activity in TGF-β2+ OMS group was increased, and the differences were statistically significant (both at P<0.05). SOD activity in the normal control group, TGF-β2 group, TGF-β2+ OMS group and OMS group was (50.35±0.97), (41.61±4.56), (28.88±3.26), and (37.61±4.83) μmol/(min·L), respectively, showing a statistically significant difference ( F=5.71, P<0.05). SOD activity was reduced in TGF-β2+ OMS group compared with normal control group and TGF-β2 group, reduced in OMS group compared with normal control group, and the differences were statistically significant (all at P<0.05). Conclusions:AGTR1 blocker OMS can promote the apoptosis of HTF effectively.Mitochondrial apoptosis pathway mediated by bax/bcl-2/caspase-9 and oxidative stress pathway are the potential mechanisms that OMS regulates the apoptosis of HTF.
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Purpose: To assess the efficacy and clinical outcome of Tenon’s patch graft (TPG) in corneal perforation and descemetocele. Methods: In this retrospective study, medical records of 83 patients (85 eyes) who underwent TPG for corneal perforation (58, 68%) or descemetocele (27, 32%) between July 2018 and October 2021 were reviewed. Clinical examination and anterior segment optical coherence tomography (AS?OCT) were performed on every follow?up visit. Anatomical success was considered as the restoration of the structural integrity with the formation of scar and anterior chamber (AC). Results: The mean size of the corneal lesions (corneal perforation or descemetocele) was 4.20 ± 1.01 mm. The mean follow?up period was 9.2 ± 5.48 months. The common underlying etiologies were infectious keratitis in 48% and autoimmune disorders in 35% of cases. TPG successfully restored the globe integrity in 74 (87%) eyes (83% in perforation and 96% in descemetocele). Anatomical failure occurred in 11 eyes (13%). The failures were due to graft dehiscence (8 eyes), graft ectasia (1 eye), and scarring with flat AC (2 eyes). The median time to epithelialization and scar formation were 3 and 15 weeks, respectively. Logistic regression analysis showed few predictors for a successful outcome: descemetoceles, noninfective causes, viral keratitis in infectious etiology, and paracentral or peripheral lesions. Conclusion: TPG can be considered an effective and inexpensive treatment for restoring the structural integrity in the eyes with perforations and descemetoceles, particularly when the donor tissue is unavailable. AS?OCT is a valuable noninvasive tool for monitoring the graft status
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Objective:To investigate the effect of epigallocatechin gallate (EGCG) on the activation of human Tenon fibroblasts (HTFs) and its mechanism.Methods:Tenon capsule tissues from nine eyes of nine advanced primary open angle glaucoma patients during trabeculectomy were obtained for primary cell culture.HTFs harvested were identified by immunofluorescence staining for vimentin and keratin.Cells at passage 4-6 were used for experiment.Viability of HTFs treated with EGCG at 0, 10, 20, 30, 40, 50, 60, 70 and 80 μmol/L was detected by cell counting kit-8 (CCK-8) assay.The cells were divided into blank control group, transforming growth factor (TGF)-β1-induced group, and EGCG-treated group, which were cultured in normal medium, medium containing 10 ng/ml TGF-β, medium containing 10 ng/ml TGF-β+ 50 μmol/L EGCG, respectively.The proliferation rate of HTFs was detected by BrdU labeling assay.Cell migration was observed by scratch wound healing assay.The expression of α-smooth muscle actin (α-SMA) was measured by immunofluorescence staining.The protein relative expression levels of Smad2/3, phosphoinositide-3-kinase (PI3K), protein kinase B (Akt) as well as the phosphorylated Smad2/3 (p-Smad2/3) and phosphorylated Akt (p-Akt) were measured by western blot.This study was approved by the Ethics Committee of Guangdong Provincial People's Hospital (NO.GDREC2019331H[R1]).Results:The HTFs harvested had spindle shape, grew regularly and were vimentin-positive.CCK-8 assay showed that there was no significant difference in the variability of HTFs treated with EGCG at 10, 20, 30, 40 and 50 μmol/L in comparison with 0 μmol/L EGCG treatment (all at P<0.05). BrdU labeling assay showed that cell proliferation in the TGF-β1-induced group was (66.37±12.65)%, which was significantly higher than (14.75±12.33)% in EGCG-treated group ( P<0.05). Three days after scratch, the relative scratch area in the TGF-β1-induced group was (47.33±12.22)%, which was significantly lower than (92.67±4.04)% in the EGCG-treated group ( P<0.05). Immunofluorescence assay showed that α-SMA fluorescence was significantly enhanced in the TGF-β1-induced group in comparison with the blank control group, which was reduced to blank control group level in EGCG-treated group.Western blot analysis showed that there were significant differences in the relative expression levels of p-Smad2/3, PI3K and p-Akt protein among the various groups ( F=58.820, 121.153, 69.289; all at P<0.001). The relative expressions of p-Smad2/3, PI3K and p-Akt in the TGF-β1-induced group were significantly higher than those in the blank control group, 10 μmol/L and 50 μmol/L EGCG-treated groups (all at P<0.05). Conclusions:EGCG can suppress TGF-β1-induced HTFs activation through Smad and PI3K/Akt signaling pathways.
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AIM:To investigate the influence of K-115 on the proliferation and migration of human Tenon's fibroblasts(HTFs)and to access the possible mechanism. Furthermore, to provide new ideas for anti-scar treatment after glaucoma surgery.METHODS: The Tenon capsule tissues were collected from patients who underwent glaucoma surgery in Hebei General Hospital from September 2018 to September 2019. Primary culture of HTFs was performed by tissue block method. The transforming growth factor-β1(TGF-β1)was used to induce HTFs activation that can mimic glaucoma filtration surgery. The cells were treated with K-115 and divided into 4 groups: the control group was treated with dimethyl sulfoxide(DMSO); TGF-β1 group was treated with 10μg/L TGF-β1 for 24h; TGF-β1 +5 K-115 group was pretreated with 5μmol/L K-115 for 2h and then treated with 10μg/L TGF-β1 for 24h; TGF-β1+10 K-115 group was pretreated with 10μmol/L K-115 for 2h and then 10μg/L TGF-β1 was added for 24h. Cell proliferation was observed by cell proliferation experiment. The migration ability of cells was detected by scratch test. The formation of autophagosomes was observed by transmission electron microscopy. Apoptosis was visualized by Hoechst 33342/PI staining.RESULTS: Cell proliferation experiment revealed that K-115 could inhibit the proliferation of HTFs induced by TGF-β1. Scratch test suggested that K-115 could inhibit the migration of HTFs induced by TGF-β1. Transmission electron microscope results showed that K-115 could enhance autophagy of HTFs induced by TGF-β1. Hoechst 33342/PI staining suggested that K-115 did not induce apoptosis.CONCLUSIONS: K-115 may regulate the proliferation and migration of HTFs induced by TGF-β1 by increasing autophagy rather than inducing apoptosis.
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Objective:To evaluate the efficacy of combination therapy of intravitreal conbercept (IVC) with posterior sub-Tenon injection of triamcinolone acetonide (PSTA) in treating macular edema secondary to non-ischemic branch retinal vein occlusion (BRVO).Methods:A nonrandomized controlled study was conducted.Fifty-nine patients (59 eyes) diagnosed with macular edema secondary to non-ischemic BRVO were enrolled in Heping Hospital Affiliated to Changzhi Medical College from October 2016 to November 2019.The subjects were divided into IVC group (28 eyes) and combination therapy of IVC with PSTA group (IVC+ PSTA group for short) (31 eyes). IVC group received IVC 0.5 mg and IVC+ PSTA group received IVC 0.5 mg combined with PSTA 40 mg as the initial therapy, then a pro re nata (PRN) IVC administration was adopted for the two groups.The mean best corrected visual acuity (BCVA) converted to the logarithm of the minimum angle of resolution unit, central macular thickness (CMT) and intraocular pressure (IOP) before and 1, 3, 6 months after injection were measured and compared.The number of repeated IVC injections and relevant complications were recorded.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Heping Hospital Affiliated to Changzhi Medical College (No.20160R9). Written informed consent was obtained from each subject prior to any medical examination.Results:There was a statistically significant difference in BCVA between the two groups among different time points ( Fgroup=0.464, P=0.498; Ftime=25.454, P<0.001). Compared with before injection, the BCVA of both groups was significantly improved at each time point after injection (all at P<0.001). There was a statistically significant difference in CMT between the two groups among different time points ( Fgroup=6.208, P=0.016; Ftime=155.505, P<0.001). The CMT of both groups at each time point after injection was significantly smaller than that before injection, and the CMT in IVC+ PSTA group at 1 and 3 months after injection was smaller than that in IVC group (all at P<0.05). There was a statistically significant overall difference in IOP between the two groups ( Fgroup=9.994, P=0.006; Ftime=2.679, P=0.056). At 1 and 3 months after injection, the IOP in IVC+ PSTA group was higher than that in IVC group, showing statistically significant differences (both at P<0.01). There were 4 eyes with an IOP higher than 21 mmHg (1 mmHg=0.133 kPa) in IVC+ PSTA group.Within the 6-month follow-up, the mean number of repeated IVC injections of IVC group was 1.25±0.93, which was higher than 0.61±0.72 of IVC+ PSTA group, and the difference was statistically significant ( P=0.039). No other ocular complication was observed in both groups. Conclusions:Combination therapy of IVC with PSTA is effective in improving the BCVA and macular edema of patients with macular edema secondary to non-ischemic BRVO.A single PSTA injection can enhance the effect of conbercept in reliving macular edema in early months and reduce the number of repeated conbercept injections in the short term.
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Objective:To investigate the effect of lithium chloride (LiCl) on the gap junctional intercellular communication (GJIC) in human Tenon capsule fibroblasts (HTFs) and its underlying mechanism.Methods:The Tenon capsule tissue of a patient who underwent strabismus surgery in Dezhou People's Hospital in April 2019 was collected and cut into tissue blocks of dimensions 1 mm×1 mm×1 mm.Primary culture and subculture were carried out, and the 4th-generation HTFs were taken for experiment.HTFs were divided into the control group and LiCl treatment group and were cultured with cell medium without or with 80 mmol/L LiCl for another 48 hours according to grouping.The cell scratch and dye labeling technique were used to label the coupling index and evaluate the GJIC function.The expression and localization of Cx43 in HTFs were detected by immunofluorescence staining.The expression levels of Cx43 mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot, respectively.The study protocol was approved by an Ethics Committee of Dezhou People's Hospital (No.2019-023). Written informed consent was obtained from the subject.Results:The cultured spindle-shaped HTFs grew adhering to the wall showing radial monolayer or vortexlike, and the cytoplasm was vimentin positive.Results of dye tracer experiment of cell scratch showed that the cell coupling index of LiCl treatment group was 9.04±0.53, which was significantly higher than 4.94±0.39 of the control group ( t=-18.79, P<0.01). Immunofluorescence staining showed that the Cx43 fluorescence was dotted in the cell membrane between adjacent cells in the control group, and Cx43 staining was obviously enhanced in the LiCl treatment group.The results of real-time fluorescence quantitative PCR showed that with relative expression level of Cx43 mRNA in the control group set to 1, the relative expression level of Cx43 in the LiCl treatment group was significantly increased to 1.97±0.23, showing a statistical significance between them ( t=-14.426, P<0.01). Western blot showed that the relative expression level of Cx43 protein was 0.871±0.057 in the LiCl treatment group, which was significantly higher than 0.446±0.028 in the control group ( t=-11.682, P<0.01). Conclusions:LiCl can enhance the GJIC function between HTFs by upregulating the expression levels of Cx43 mRNA and protein, suggesting that the enhanced GJIC function by LiCl may be one of the mechanisms of its inhibition on HTFs proliferation.
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At present, the mechanism of scar formation in filtering area after glaucoma filtering surgery has not been fully understood.It is believed that stimulation of cytokines and growth factors during postoperative wound healing causes excessive activation of fibroblasts in the Tenon capsule, resulting in the deposition of extracellular matrix and scar formation.Long non-coding RNA (lncRNA) is a kind of RNA sequence that does not encode proteins.It can regulate cell proliferation, apoptosis and become a precursor of microRNA, and its specific expression has been proved and plays an important regulatory role in various fibrotic tissue.With the deepening of research, lncRNA has also been found to be involved in the formation and development of filtering bleb scarring after glaucoma filtering surgery.Previous studies have shown that lncRNA is specifically expressed in Tenon capsule tissues after glaucoma filtering surgery, and different lncRNAs can affect the proliferation of fibroblasts in Tenon capsule or the synthesis of extracellular matrix through different ways, which can affect the formation of fibroblasts scarring.In this article, the role of different lncRNAs in the proliferation of Tenon fibroblasts and its potential in treating bleb scarring were summarized so as to provide a new direction for the treatment of postsurgical scarring.
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@#AIM: To explore the effects of heat shock protein 47(HSP47)siRNA on biological behaviors of human Tenon capsule fibroblasts(HTCF)cells cultured <i>in vitro</i> and the expression level of transforming growth factor-β1(TGF-β1).<p>METHODS: HTCF were cultured <i>in vitro</i> and divided into blank control group, empty vector group and transfection group. In transfection group, interfering siRNA sequences were designed and synthesized based on the HSP47 gene sequences, vectors were constructed and introduced into HTCF. The empty vector group was introduced with empty vectors. The expressions of HSP47 mRNA and protein in cells were detected by RT-PCR and Western blot. The proliferation, apoptosis, invasion and migration of cells were detected by clone formation assay, flow cytometry, Transwell method and scratch test. The expressions of proliferation, apoptosis, invasion and migration proteins, and TGF-β1 were detected by Western blot.<p>RESULTS: Compared with empty vector group, expression of HSP47 mRNA and protein, clone formation rate, cell healing rate, number of invasive cells, relative expression levels of Ki67, N-cadherin and TGF-β1 were significantly decreased in transfection group(<i>P</i><0.05), relative expression level of E-cadherin protein was significantly increased(<i>P</i><0.05), but there was no difference in apoptosis rate, and relative expression levels of Bcl-2 and Bax(<i>P</i>>0.05).<p>CONCLUSION: HSP47 siRNA can reduce proliferation, invasion and migration abilities of HTCF cells by inhibiting the expression of TGF-β1 protein, without significant effects on the apoptosis of HTCF cells.
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Background: Blindness due to cataract presents an enormous problem in terms of human morbidity, economical loss, andsocial burden. Retrobulbar anesthesia was commonly used for cataract surgery. Rare but serious complications led manyophthalmologists to replace retrobulbar with peribulbar anesthesia. However, even peribulbar anesthesia does not eliminatethe serious complications totally. These concerns have led to increased use of blunt needle sub-Tenon’s block over the sharpneedle blocks.Materials and Methods: 200 cases were selected, of which 100 were in the sub-Tenon’s group and the remaining 100 werein the peribulbar group. The efficacy of anesthesia between the two groups was compared in terms of analgesia at variousintervals, akinesia of the globe and eyelids attained after the block. They were graded on a subjective scale and recorded.Minor complications such as chemosis, sub-conjunctival hemorrhage, and rise in increased intraocular pressure (IOP) werealso compared and analyzed.Results: Sub-Tenon’s anesthesia provided better analgesia than peribulbar anesthesia although the akinesia was poorer thanthe latter. Minor complications such as sub-conjunctival hemorrhage were more in sub-Tenon’s group while instantaneous risein IOP was more in peribulbar group. The incidence of chemosis was almost comparable in both the groups.Interpretation and Conclusion: Sub-Tenon’s anesthesia is recommended as a safe and effective alternative to peribulbaranesthesia for small-incision cataract surgery as it provides good analgesia, adequate akinesia, and rare minor complications.
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@#AIM: To investigate the effect of connective tissue growth factor(CTGF)on the proliferation and migration and transformation in Tenon's capsule fibroblasts(Tfb)of primary open angle glaucoma(POAG)patients.<p>METHODS: Tfb were cultured from Tenon's tissues in POAG patients <i>in vitro</i>. The free-serum DMEM-F12 containing 1.0, 10.0, 100.0ng/mL of CTGF was added into medium for 24h and 48h in different experimental group respectively, and only equal volume of free-serum DMEM-F12 was added in the negative control group. After 24h, the cell proliferation was analyzed through MTT assay, and migration was evaluated by crutch method. After 48h, Semi-quantitative RT-PCR was used to observed the mRNA expression of α-smooth muscl actin(α-SMA), and expression of α-SMA protein was examined by immunochemistry.<p>RESULTS: The proliferation values <i>A</i> of the cells in 1.0, 10.0, 100.0ng/mL of CTGF group respectively were 0.436±0.009, 0.643±0.009, 0.679±0.006, and 0.423 ±0.156 in the negative control group. The migrated cell number was 34.600±3.507, 70.400±2.074, 80.000±2.915 in different concentrations of CTGF group respectively, and 31.000±3.536 in the negative control group. And also in different experimental groups respectively, the absorbance ratio of α-SMA/β-actin was 0.873±0.161, 1.213±0.312, 1.352±0.376, and 0.851±0.158 in the negative control group, the expressing levels <i>A</i> of α-SMA protein in Tfb were 0.110±0.026, 0.141±0.017, 0.175±0.027, and 0.108±0.020 in the negative control group. The statistics of the above experimental data showed that, comparing with the negative control group, the 10.0 and 100.0ng/mL CTGF groups was statistically significant different(<i>P</i><0.05), but there was no statistical different between the 1.0ng/mL CTGF group and the negative control group(<i>P</i>>0.05). <p>CONCLUSION: The proliferation, migration, and phenotypic transformation of Tfb can be promoted in CTGF group in POAG patients. These findings suggest that CTGF may play a role in the development of filtering bleb scarring.
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@#AIM: To compare the clinical effects of retinal detachment with choroidal detachment(RD/CD)treatment by posterior subcapsular Tenon injection triamcinolone acetonide and intravenous drip of dexamethasone combined vitrectomy.<p>METHODS: Totally 52 cases(52 eyes)of RD/CD patients in our hospital from March 2014 to October 2017 were retrospectively reviewed. According to the preoperative intervention methods, the patients were divided into two groups: A group and B group. Group A(27 cases, 27 eyes)received intravenous drip of dexamethasone once a day 3-5d before operation. Group B(25 cases, 25 eyes)received posterior subcapsular Tenon injection triamcinolone acetonide 5d before operation. The intraocular pressure, CD, retinal reattachment, visual acuity and complications were measured before and after the intervention.<p>RESULTS: After intervention, the intraocular pressure of group B was 8.09+3.56mmHg, which was significantly higher than 5.65+2.19mmHg before intervention in group B and 6.25+2.53mmHg after intervention in group A. The difference was statistically significant(<i>P</i><0.05). After intervention, the CD height of group A and B was 3.98(1.01, 5.34)mm and 0.92(0.03, 3.88)mm, significantly lower than that in group A and B before intervention, which was 5.22(3.14, 6.64)mm and 5.16(3.34, 7.71)mm. CD loci 6.0(3.0, 10.0)and 3.0(0.0, 6.0)were significantly lower than those of 11.0(9.0, 12.0)and 10.0(8.0, 12.0)before intervention. The CD height and the number of CD loci in group B were lower than those in group A(<i>P</i><0.05). From the last follow-up, the success rate of retinal reattachment in groups A and B were 78% and 96%, respectively(<i>P</i>>0.05). At 1, 3mo and the last follow-up, the visual acuity of group A was 1.69±0.79, 1.39±0.72 and 1.38±0.61 better than that of group A before intervention 2.06±0.28. The visual acuity of group B was 1.42±0.66, 1.29±0.56 and 0.97±0.51 better than that of group A before intervention 2.02±0.58. The visual acuity of group B was better than that of group A at the last follow-up, with statistical difference(<i>P</i><0.05). At 1 and 3mo after operation, 4 eyes in group A had high intraocular pressure, which was significantly lower than that of 11 eyes and 12 eyes in group B(<i>P</i><0.05). At the last follow-up, there was still 1 eye with high intraocular pressure in group A and 2 eyes in group B(<i>P</i>>0.05).<p>CONCLUSION: In the treatment of RD/CD, the effect of posterior subcapsular Tenon injection triamcinolone acetonide with vitrectomy is better than that of intravenous drip of dexamethasone combined vitrectomy, the intraocular pressure should be monitored after operation. If high intraocular pressure occurs, appropriate drug control or removal of triamcinolone acetonide from the posterior Tenon capsule is required.
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Objective To investigate the effects of matrine on proliferation and apoptosis of human Tenon capsule fibroblasts (HTFs) in.vitro.Methods After treated with 0,0.3,0.6 and 0.9 g/L matrine in vitro,cell counting kit-8 (CCK-8) method was used to assay the proliferation of HTFs at 24,48 and 72 hours,Western blot and PCR were performed to evaluate the expression of apoptosis-associated factor caspase-3 on both protein and RNA levels.Results The activity of human Tenon capsule fibroblast at 48 hours and 72 hours after treated with 0.3,0.6,0.9 g/L matrine was significantly inhibited when compared with the 0 g/L matrine group,and the inhibitory effect was dose-dependent and time-dependent (F ion =1 019.51,P =0.00;Ftime =5 848.66,P =0.00;Fi ion =147.45,P=0.00).After treated with 0,0.3,0.6 and 0.9 g/L matrine,the early apoptosis rate of HTFs was (2.68±0.30)％,(5.08±0.47)％,(6.97±0.69)％ and (10.30±1.20)％,the grey value ofcaspase-3 protein was 1.00±0.13,1.90±0.19,2.50±0.30 and 2.67±0.30,the relative expression of caspase-3 mRNA was 0.98 ±0.12,2.01 ±0.34,6.15 ± 0.60 and 11.40 ± 1.12,respectively,with significant differences among them (F =55.74,66.01,154.50;all at P＜0.01),the early apoptosis rate of HTFs,the grey value of caspase-3 protein and the relative expression of caspase-3 mRNA were all increased significantly as the concentration of matrine increased,with significant differences between any two groups (all at P＜0.05).Conclusions Matrine can inhibit the proliferation of HTFs and induce the apoptosis of HTFs in a time-and dose-depended manner.
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Objective:To investigate the effects of vascular endothelial growth factor (VEGF) on the expression of secreted protein acidic and rich in cysteine (SPARC) and the fibrosis in cultured human Tenon's fibroblast (HTF) in vitro. Methods:HTF cells were obtained from Tenon's capsule tissues of patients undergoing strabismus surgery. Immunofluorescence was used to identify the HTF cells. HTF cells were cultured with different concentrations of VEGF, and which were divided into four groups, i.e., 0 ng/mL group, 25 ng/mL group, 50 ng/mL group and 100 ng/mL group. The expression of SPARC, collagen- , and matrix metalloprotein 9 (MMP-9) and the activity of extracellular signal-regulated kinase (ERK) pathway were analyzed by Western blotting and real-time quantitative PCR (qPCR). The abilities of proliferation and migration of HTF cells were detected by MTS assay and scratch test, respectively. Results:HTF cells were observed and identified by inverted phase contrast microscope and immunofluorescence. Under the stimulation of VEGF, the expression of protein and mRNA of SPARC, collagen-I and MMP-9 of HTF cells in other three groups were increased compared with 0 ng/mL group; the phosphorylation activities of ERK pathway were up-regulated, and the proliferation and migration abilities of HTF cells were up-regulated. And the effect was the most obvious in the 50 ng/mL group. Conclusion:VEGF is involved in promoting the fibrosis of HTF cells accompanied by the up-regulation of the SPARC, which suggests SPARC may become a potential regulatory site.
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Objective To investigate the effects of hepatocyte growth factor (HGF) on proliferation and transdifferentiation of human Tenon capsule fibroblasts induced by transforming growth factor-β1(TGF-β1) in vitro.Methods Human Tenon capsule fibroblasts were cultured and divided into blank control group,TGF-β1 treated group and different concentrations HGF+TGF-β1 groups.The TGF-β1 at 10 μg/L was added into culture medium of the TGF-β1 treated group,and different concentrations of HGF (25,50,100,200 μg/L) and 10 μg/L TGF-β1 were added into culture medium of the HGF25 μg/L +TGF-β1,HGF50 μg/L +TGF-β1,HGF100 μg/L +TGF-β1,HGF200 μg/L +TGF-β1 group respectively,Methyl thiazolyl tetrazolium (MTT) was employed to measure the cell proliferation (absorbance at 560 nm).Immunofluorescence staining was used to evaluate and locate the expression of α-smooth muscle action (α-SMA) in the cells.The expression of α-SMA protein in the cells was detected by Western blot assay.Results Cultured cells showed fusiform in shape with the positive response for vimentin.The proliferation value of the cells was 0.203±0.025,0.497 ± 0.101,0.426 ± 0.062,0.354 ± 0.040,0.272 ± 0.084,0.241 ± 0.011 in the blank control group,TGF-β1 treated group,HGF25 μg/L +TGF-β1 group,HGF50 μg/L +TGF-β1 group,HGF100 μg/L +TGF-β group and HGF200 μg/L + TGF-β1 group,respectively,showing a significant difference among the groups (F =9.21 0,P =0.003).Compared with the TGF-β1 treated group,the proliferation values of the cells were significantly reduced in the blank control group and HGF+TGF-β1 group (all at P＜0.05).Immunofiuorescence staining showed that α-SMA protein mainly located in cytoplasm with the strong red fluorescence in the cells of the TGF-β1 treated group and weak red fluorescence in HGF+TGF-β1 group,and the expression of α-SMA was absent in the blank control group.The percentage of α-SMA-positive cells was (60.0±4.7)％ in the TGF-β1 treated group and (14.3±3.1)％ in the HGF+TGF-β1 group,with significant difference between the two groups (t =19.856,P＜0.001).The relative expression levels of the α-SMA protein in the cells were 0.642 ±0.032,1.330± 0.069 and 0.884 ±0.040 in the blank control group,TGF-β1 group and HGF100μg/L +TGF-β1 group,respectively,showing a significant difference among the groups (F =13.370,P＜ 0.001),and relative expression levels of the α-SMA protein in the cells were significantly lower in the blank controlgroup and HGF100 μg/L+TGF-β1 group than that in the TGF-β1 treated group (all at P＜0.05).Conclusions HGF can inhibit the proliferation of human Tenon capsule fibroblasts,down-regulate the expression of α-SMA protein induced by TGF-β1 and arrest the phenotype transformation of fibroblasts in vitro.
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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.
Sujet(s)
Humains , Actines , Apoptose , Cicatrice , Collagène , Fibroblastes , Cytométrie en flux , Gels , Sulfure d'hydrogène , Hydrogène , Réaction de polymérisation en chaîne , ARN messager , Capsule de TenonRÉSUMÉ
BACKGROUND: Conjunctival filtering bleb scar formation is the main reason for the failure of glaucoma filtration surgery. Cytoglobin (Cygb) has been reported to play an important role in extracellular matrix (ECM) remodeling, fibrosis and tissue damage repairing. This study aimed to investigate the role of Cygb in anti-scarring during excessive conjunctival wound healing after glaucoma filtration surgery. METHODS: Cygb was overexpressed in human tenon fibroblasts (hTFs) by transfecting hTFs with lentiviral particles encoding pLenti6.2-FLAG-Cygb. Changes in the mRNA and protein levels of fibronectin, collagen I, collagen III, TGF-ß1, and HIF1α were determined by RT-PCR and western blotting respectively. RESULTS: After Cygb overexpression, hTFs displayed no significant changes in visual appearance and cell counts compared to controls. Whereas, Cygb overexpression significantly decreased the mRNA and protein expression levels of collagen I, collagen III and fibronectin compared with control (p < 0.01). There was also a statistically significant decrease in the mRNA and protein levels of TGF-ß1 and HIF-1α in hTFs with overexpressed Cygb compared with control group (p < 0.05). CONCLUSION: Our study provided evidence that overexpression of Cygb decreased the expression levels of fibronectin, collagen I, collagen III, TGF-ß1 and HIF-1α in hTFs. Therefore, therapies targeting Cygb expression in hTFs may pave a new way for clinicians to solve the problem of post-glaucoma surgery scarring.