Comparison of fibrotic response in the human lens capsular bag after femtosecond laser-assisted cataract surgery and conventional phacoemulsification.

PURPOSE: To compare the effect of different laser pulse energy settings in femtosecond laser-assisted cataract surgery with that of standard phacoemulsification and no energy at all used on posterior capsule opacification (PCO) in vitro. SETTING: Cell and Molecular Biology Research Laboratory, Department of Ophthalmology, Ludwig-Maximilians-University Munich, Real Eyes, Ophthalmology Center, Munich, and Institute for Clinical Pathology, Goethe University Frankfurt, Frankfurt, Germany. DESIGN: Experimental study. METHODS: Fifteen cadaver capsular bags were cultivated from 8 human donors under standard cell culture conditions. For preparation of the capsular bag, 4 groups were established as follows: femtosecond laser-assisted cataract surgery standard energy (n = 3), femtosecond laser-assisted cataract surgery high energy (n = 3), phacoemulsification (n = 6), and hydrodissection without energy (extracapsular cataract extraction) (n = 3). Growth of lens epithelial cells was observed and photodocumented. The days until full cell coverage of the posterior capsule were documented. Capsular bags were stained for fibronectin, α-smooth muscle actin, and collagen type 1. RESULTS: Cell growth patterns in all treatment groups were comparable, with no statistically significant differences detected at any timepoint measured (P = .81, Kruskal-Wallis). The markers for fibrosis were equally distributed in all groups, indicating an equal fibrotic reaction in all groups. CONCLUSION: Femtosecond laser-assisted cataract surgery did not increase different cellular responses in PCO formation comparison with phacoemulsification in vitro, even when higher laser pulse energy levels were used.

Efficacy and safety of recombinant tissue plasminogen activator and gas versus bevacizumab and gas for subretinal haemorrhage.

PURPOSE: To report the 12 months efficacy of initial intravitreal bevacizumab or intravitreal recombinant tissue plasminogen activator (rtPA) combined with expansile gas in patients with subretinal haemorrhage caused by neovascular age-related macular degeneration (AMD). METHODS: Forty-five eyes of 45 patients with subretinal haemorrhage (1-5 disc diameters) involving the fovea secondary to neovascular AMD were evaluated retrospectively consecutively. Thirty-two eyes underwent treatment with rtPA (50 µg/0.05 ml) combined with intravitreal sulphur hexafluoride (SF6). The other 13 eyes were treated with bevacizumab (1.25 mg/0.05 ml) and SF6. Thereafter, all patients received Vascular Endothelial Growth Factor (anti-VEGF) treatment according to modified PrONTO criteria. Main outcome was change of best-corrected visual acuity (VA) at 12 months as determined by Early Treatment Diabetic Retinopathy (ETDRS). RESULTS: There was more improvement in patients initially treated with rtPA and gas (14 letters; bevacizumab and gas eight letters) and not suffering from adverse events. The incidence of vitreous haemorrhages was significantly higher in the rtPA group (nine of 32 versus one of 13, p < 0.01). In both groups, an average of 3.5 anti-VEGF injections were performed per patient during 12 months (no difference between both groups). CONCLUSION: Both initial treatment regimen lead to improved functional results after 1 year. However, patients, not suffering from adverse events, who underwent initial treatment with rtPA and gas showed better results. To maintain VA, controlling neovascular AMD by anti-VEGF treatment regime after initial treatment with rtPA+gas is important for all cases.

Modulation of TNF-alpha expression in bone marrow macrophages: involvement of vitamin D response element.

The calcium-regulating hormone, 1,25(OH)(2)D(3), induces tumor necrosis factor-alpha (TNF-alpha) synthesis and release from bone marrow macrophages (BMMs). To investigate the mechanism of this regulation, we have examined the effects of 1,25(OH)(2)D(3) on the cytokine message. 1,25(OH)(2)D(3) increased TNF-alpha mRNA abundance in a dose- and time-dependent manner. The combined treatment of BMMs with LPS and 1,25(OH)(2)D(3) resulted in a synergistic increase of TNF-alpha. The steroid also increased the expression of CD14 (LPS receptor). Vitamin D receptors (VDRs) mediate 1,25(OH)(2)D(3) genomic effects by forming homodimers or heterodimers with retinoic acid receptors (RARs) or retinoic X receptors (RXRs). The RXR ligand, 9-cis retinoic acid (9cRA), reduced TNF-alpha mRNA abundance in BMMs, but increased CD14 mRNA levels. 1,25(OH)(2)D(3) or LPS did not affect TNF-alpha transcript stability. 9cRA, however, caused TNF-alpha mRNA destabilization. Next, we searched for potential vitamin D response elements (VDREs) in the promoter region (1.2 kb) of the TNF-alpha gene, and identified six such sequences. Using electrophoresis mobility shift assay (EMSA) we identified one of those sequences (-1008 to -994) as a likely candidate to be a VDRE (tnfVDRE). The binding of tnfVDRE to BMM-derived nuclear extract was increased following cell treatment with 1,25(OH)(2)D(3). No induction was observed with 9cRA treatment, but the retinoid enhanced the activity of 1,25(OH)(2)D(3) when added together. Previously characterized VDREs (mouse osteopontin and rat osteocalcin) competed effectively with tnfVDRE, demonstrating the nature of the TNF-alpha-derived sequence as a VDRE. We observed super-shift and block-shift of the complex in the presence of either anti-VDR or anti-RXR antibodies. Our data suggest that 1,25(OH)(2)D(3) increases TNF-alpha transcript abundance in BMMs via a transcriptional mechanism; 9cRA decreases TNF-alpha mRNA by destabilizing the transcript, and possibly also by forming transcriptionally inactive complex with 1,25(OH)(2)D(3) on the tnfVDRE. The receptor complex interacting with tnfVDRE found in the promoter of the cytokine gene is probably composed of VDR-RXR heterodimer.