Preparation of endothelial keratoplasty lamellae from donated whole eyes post vitreous humour aspiration at the central eye bank of Iran.

To compare the endothelial parameters and thickness profiles of endothelial keratoplasty (EK) lamellae from donated whole eyes post vitreous humour aspiration (VHA) with those prepared from their mate control eyes (without VHA). Between March 2019 and March 2020, a few steps were added when aspirating the vitreous humour and also before dissecting the corneal tissue with microkeratome. EK lamellae were prepared from whole eyes that underwent VHA and their corresponding endothelial and thickness profiles were compared with those prepared from their respective fellow control eyes. Post-operative data in terms of graft attachment and clarity and the rate of reoperation were also reviewed. 115 eyes that underwent VHA and 115 of their respective fellow eyes were enrolled. No significant difference was noted in the endothelial parameters between the two groups. Mean central thickness of the EK lamellae and increase of thickness towards the periphery were not significantly different between the groups. Both groups did not show a significant difference with respect to the anticipated dissection depth, post-operative graft clarity, graft attachment, and the rate of regraft. This study demonstrates that aspirating vitreous humour has no adverse effect on the endothelial and thickness profiles of the EK lamellae that are prepared from these donated whole eyes, once the specific steps outlined by the eye bank are adhered to when aspirating the vitreous humor before performing the microkeratome dissection. The success rate of the grafted lamellae was comparable between the study groups.

Diagnostic accuracy of confocal scan in detecting acanthamoeba keratitis and fungal keratitis: a systematic review and meta-analysis.

PURPOSE: Acanthamoeba keratitis (AK) and fungal keratitis (FK) are two microbial keratitis that cause serious damage and, without early accurate diagnosis and treatment, may lead to blindness. In vivo corneal confocal scan, as an emerging ocular diagnostic method in comparison with microbiological smears and cultures as the gold standard, may assist in accelerating appropriate diagnosis. OBJECTIVE: To determine the diagnostic accuracy of confocal scan for the diagnosis of AK and FK. METHODS: Data were collected via a comprehensive literature search of PubMed, Web of Science, Cochrane Library, Embase and Scopus using keywords related to diagnostic accuracy of confocal scan in AK and FK up to October 2022. Pooled data underwent meta-analysis in terms of sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and overall diagnostic odds ratio (DOR) of confocal scan for the diagnosis of AK and FK. RESULTS: The final 14 relevant studies were identified, including 1950 eyes. Meta-analysis in AK group revealed 94% sensitivity, 87% specificity, 89% PPV, 92% NPV, and DOR of 143.32, and in FK group disclosed 88% sensitivity, 85% specificity, 85% PPV, 88% NPV, and DOR of 75.98. CONCLUSION: The accuracy of confocal scan for the diagnosis of AK was significantly more than that for detecting FK; despite the limitations such as limited numbers of available retrospective studies for the detection of FK, confocal scan had an acceptable performance in detecting FK eyes. The overall performance of NCS was similar with that of HRT-RCM for the detection of both types of keratitis.

Epibulbar solitary myofibroma in an elderly patient: a case report.

A 93-year-old male patient presented with abrupt expansion of an old epibulbar mass at the temporal area of the left eye. He had a medical history of previously treated laryngeal cancer with surgery and radiotherapy. The tumor, despite being firmly attached to the underlying sclera, was excised completely and histopathological examinations revealed a solitary myofibroma. The patient had a 4-month uneventful follow-up with excellent wound healing. Solitary myofibroma may be a differential diagnosis for epibulbar masses in elderly patients.

Huge heterotopic brain cyst of the orbit: report of a case and its management.

Heterotopic brain tissues are a group of extracerebral neuroglial tissues. Heterotopic brain tissue in the orbit remains a rare clinical entity. This article presents a 7-year-old male child who presented with severe globe displacement, proptosis, and vision loss in the right eye. The orbital imaging showed a huge orbital cystic mass displacing the globe. The cyst was excised entirely from the orbit. The histopathological investigations revealed the presence of a cystic lesion containing brain tissue that was immune reactive for S-100 and glial fibrillary acidic proteins. The diagnosis was confirmed to be heterotopic brain tissue due to the lack of visible bony defect. The relevant literature was also reviewed.

Inhibition of matrix metalloproteinase-9 for the treatment of dry eye syndrome; a review study.

Dry eye syndrome (DES) and tear dysfunction are multifactorial conditions affecting meibomian glands, lacrimal glands, and ocular surface. This ocular disorder can cause eye irritation, irregular cornea, corneal barrier disruption, and blurred vision. Uncontrolled increase in matrix metalloproteinase-9 (MMP-9) level and activity has been detected in the tears and ocular surface in the patients with DES, which has been proved to be related to disruption of tight junctions in apical corneal epithelium associated with severe signs of DES. These uncontrolled activities of MMP-9 lead to desquamation of ocular surface epithelia. Therefore, this review study was conducted to summarize the evidence regarding MMP-9 contribution in DES, and inhibition of MMP-9, as a therapeutic target for treatment of DES. For this purpose, herein, the related studies designed novel pharmaceutical compounds for direct and indirect inhibition of MMP-9 as treatment approaches for DES were reviewed. These compounds were designed to improve corneal barrier function, reduce inflammation on ocular surface, and restore tear production.

Intravitreal connective tissue growth factor neutralizing antibody or bevacizumab alone or in combination for prevention of proliferative vitreoretinopathy in an experimental model.

Connective tissue growth factor (CTGF) is released by retinal pigment epithelial (RPE) cells and detectable in proliferative membranes (PrMs). This experimental study was performed to investigate the mRNA and protein levels of both CTGF and vascular endothelial growth factor A (VEGF-A) in a rabbit model of proliferative vitreoretinopathy (PVR). In addition, the effects of a single intravitreal injection of the safe dose of anti-CTGF or bevacizumab as monotherapy and in combination were evaluated. PVR was induced in the right eye of albino rabbits by intravitreal injection of cultured adult human RPE cells. Quantitative real-time reverse transcription PCR (qRT-PCR) and Western blot analysis of CTGF and VEGF-A were performed on whole eye tissue in the PVR model versus controls at different time points. In the next step, the PVR models were assigned to five groups. The monotherapy groups received a single intravitreal injection of 0.1 ml of anti-CTGF 100 µg/ml (final concentration of 6.6 µg/ml in the vitreous) or 0.03 ml of 25 mg/ml bevacizumab. In the combined group, the abovementioned amounts of anti-CTGF and bevacizumab were injected intravitreally from separate sites in one session. No antibody injection was performed in the control group. Intravitreal injection of 0.1 ml of control IgG (1 mg/ml of isotype matched) antibody was performed in the placebo group. After 2 weeks, histologic evaluation including, trichrome staining for collagen, immunostaining by anti-alpha-smooth muscle actin for myofibroblasts, and anti-collagen type-1 antibody on paraffin embedded anterior-posterior sections was done. In addition, fundus photography was performed for clinically equivalent PVR staging. Twenty-four hours following PVR induction, CTGF mRNA and protein levels increased five- and- three-fold compared to controls, respectively (P < 0.001). VEGF-A mRNA and protein levels decreased significantly after 72 h of PVR induction compared to controls (P < 0.05). Means of PrM thickness and myofibroblast cell counts significantly decreased in the anti-CTGF group (P < 0.001 and P < 0.05, respectively). The mean area of collagen type-1 fibers of PrM in the mono- and combination therapy groups that received intravitreal anti-CTGF was significantly reduced (P < 0.001); in addition, mild PVR (stage-1 and 2) formation occurred in comparison with moderate to severe PVR (stage-4 and higher) in other groups. In conclusion, we found that intravitreal injection of CTGF neutralizing antibody resulted in a reduction in PrM thickness, collagen fibers and myofibroblast density in the PVR model. CTGF inhibition may represent a potential therapeutic target for PVR.

Intraconal Orbital Hidrocystoma in an Infant.

ABSTRACT: Intraconal orbital hidrocystoma is a very rare entity. Herein, a 6-month old boy with an intraconal apocrine hidrocystoma will be reported. The patient presented with a left-sided mild proptosis and significant anisometropic hypermetropia. Clinical examinations revealed choroidal folds and optic disc blurring in the left eye. Orbital MRI disclosed an intraconal well-defined cystic lesion that was hypointense in T1 and hyperintense in T2 weighted images. The lesion was excised completely through a lateral orbitotomy and diagnosed histologically as an apocrine hidrocystoma. Excepting partial improvement of anisometropia, other clinical signs were improved after surgery.

Effects of topical erythropoietin on healing experimentally-induced avascular scleral damage in a rabbit model.

The present study was designed to investigate the effect of topical erythropoietin on the healing process of induced necrotizing scleritis and to evaluate the ocular side effects of this treatment modality in a rabbit model. Necrotizing scleritis was induced in 8 New Zealand albino rabbits. The animals were then randomly divided into one of two groups: a treated group administered a topical erythropoietin-containing cellulose-based gel every 8 h or a control group treated with a cellulose-based gel without erythropoietin every 8 h. The sizes of the lesions measured at different time points were compared between the groups. After three months, the rabbits' eyes were enucleated and histologically and immunohistochemically evaluated for angiogenesis and apoptosis. The lesions were completely vascularized in all eyes of the treated group and 50% of eyes of the control group. The mean interval from the induction of scleral necrosis to a complete improvement was 28 days in the treated group and 62.5 days in the control group (P = 0.04). Histological examination revealed that erythropoietin enhanced the improvement of necrotizing scleritis by stimulating angiogenesis and reducing apoptosis. Neovascularization of the cornea, iris, or retina was not observed in the treated group. We observed a significantly faster recovery to complete improvement of necrotizing scleritis in rabbit eyes treated with erythropoietin compared to those of the control group. Treated eyes had a higher rate of complete healing and had no ocular safety concerns. This therapeutic modality represents a promising treatment for scleral necrosis following various types of ocular surgery.

Trimethyl chitosan-hyaluronic acid nano-polyplexes for intravitreal VEGFR-2 siRNA delivery: Formulation and in vivo efficacy evaluation.

As vascular endothelial growth factor in choroidal neovascularization is a major cause of visual loss of the elderlies and diabetics, gene therapy may offer an alternative treatment. However, siRNA instability and inefficient delivery are the main hindrances. To address this issue, we developed a nano-sized siRNA loaded therapeutic delivery system. The chitosan-hyaluronic acid nano-polyplexes were prepared by the modified ionic gelation method. The obtained nano-polyplex with a narrow size distribution, indicated no significant cytotoxicity in the MTT test and proper cellular uptake in confocal images. The RT-PCR analysis indicated remarkable gene silencing on HUVEC cells. The intravitreally administered nano-polyplexes in rabbits overcame both the vitreous and retina barriers and reached the posterior tissues efficiently. Intravitreal injections of the VEGFR-2 siRNA nano-polyplexes significantly reduced the size of the laser-induced choroidal neovascularization, compared to the control group. Consequently, the developed formulation can be a promising candidate for intravitreal delivery of siRNA.

Proposed risk factors for failure of transplanted eye bank-prepared descemet stripping automated endothelial keratoplasty tissues.

To investigate the possible risk factors for failure of transplanted eye bank-prepared Descemet stripping automated endothelial keratoplasty (DSAEK) tissues. In a retrospective study between March 2011 and March 2019, all the failed DSAEK cases (131 cases) reported to the Central Eye Bank of Iran were compared with a surgeon-matched successful DSAEK group (control, 126 cases) in terms of the donor, DSAEK tissue, and recipient characteristics. Univariate analysis indicated that the DSAEK tissue preparation from excised corneoscleral tissues (OR 2.17; P = 0.026) and donor conjunctival hyperemia (OR 1.62; P = 0.042) were more common in the re-DSAEK group than in the controls. Other donor and recipient characteristics as well as other DSAEK tissue criteria were not significantly different between the re-DSAEK and control groups. DSAEK lenticules in both re-DSAEK and control groups demonstrated symmetric contours. Subgroup analyses in the re-DSAEK group revealed a higher rate of surgeons' low to moderate experience in comparison to those of high experience (P < 0.001). Additionally, failed DSAEK was observed more commonly in donors aged over 50 years than among those under 30 years (P < 0.001). Our study showed that DSAEK tissues prepared from excised corneoscleral tissues and from donors with conjunctival hyperemia were associated with higher rates of re-DSAEK due to failed DSAEK. An increasing trend of re-DSAEK was observed with shorter surgeons' experience and more advanced donor's age.

Effects of intravitreal connective tissue growth factor neutralizing antibody on choroidal neovascular membrane-associated subretinal fibrosis.

Connective tissue growth factor (CTGF) plays an essential role in the regulation of extracellular matrix proteins and pro-fibrotic and angiogenic factors. This experimental research was conducted to evaluate if CTGF is elevated after induction of a choroidal neovascular membrane (CNVM) and whether intravitreal anti-CTGF without and with intravitreal bevacizumab (IVB) may have any effect on the CNVM associated sub-retinal fibrosis. In adherence to ARRIVE guidelines, CNVM was induced by laser spots in the right eye retinas of ninety-four pigmented rats. Quantitative real-time reverse transcription PCR (qRT-PCR) and western-blot analysis were performed on sclerochoroidal tissues of forty-four rats before and at different time intervals after laser application. The remaining fifty rats were randomly divided into five groups after laser application. Group A received intravitreal injection of 2  µl of the 50 µg/ml anti-CTGF. In group B, intravitreal injection of 2  µl of 25 mg/ml bevacizumab was performed. Group C received 1  µl intravitreal anti-CTGF and 1  µl IVB. Group D did not receive any intravitreal injection as the control group. In group E, intravitreal injection of 2  µl of nonspecific purified mouse IgG antibody was performed as the placebo group. After two weeks, double immunohistochemistry was performed by isolectin B4 and anti-collagen type1 on the sclerochoroidal flat-mounts. Masked measurement of the fluorescent images of the CNVM and CNVM associated sub-retinal fibrosis areas was performed using the image J software. Ctgf mRNA and CTGF protein levels increased to the maximum level in 24 h after laser application and remained higher than the control level up to the 14th day for the Ctgf mRNA and up to the 7th day for the CTGF protein level. Means of CNVM associated sub-retinal fibrosis areas in three treatment groups (A, B and C) were significantly less than the control (D) and placebo (E) groups (P < 0.001, <0.05, <0.001 respectively). For groups A and C, mean CNVM associated sub-retinal fibrosis areas were also significantly less than group B (P < 0.05 and < 0.01, respectively). In conclusion, this study showed significant reduction of the CNVM associated sub-retinal fibrosis via inhibition of the CTGF which mediates the final steps of fibrosis in various inflammatory and angiogenic pathways.

Trephine- and dye-free technique for eye bank preparation of pre-stripped Descemet membrane endothelial keratoplasty tissue.

We describe a rapid and easy technique for the preparation of pre-stripped Descemet membrane (DM) endothelial keratoplasty (DMEK) tissue in the eye bank, without the use of dye or trephining by the eye bank technician. After drawing 2 radial marks 90° apart on the scleral rim of the cornea, a circumferential incision was made internal to the trabecular meshwork and the edge of the DM opposite to the marked area was scored and gently peeled towards the marked area. Finally, the peeled DM was gently unrolled and attached to the posterior stromal bed via absorbing excess liquid from the periphery. The prepared DMEK tissue while reattached on the posterior aspect of the cornea was subjected to slit-lamp biomicroscopic and specular microscopic examinations. In case of proper preparation of DMEK tissue and suitability of endothelial cells for DMEK surgery, the tissue was distributed for transplantation. At the time of surgery, trypan blue staining was performed by the cornea surgeon before DM trephination. Duration of DMEK tissue preparation, uneventful preparation, post-preparation endothelial cell loss, surgeon's satisfaction, and intra- and post-operative attachment of the tissue were investigated variables in this study. Over a 2-year period, 28 DMEK tissues were successfully prepared from 30 corneas using our new technique. The median duration of DMEK preparation was 100 s. The area of endothelial cell loss was 1.8% ± 1.2% centrally and 6.2% ± 1.8% in areas close to the forceps contacts. Seven out of the 28 (25%) DMEK tissues were ruined by surgeons during surgery and the overall rate of re-bubbling in 19 transplanted DMEK tissues was 16%. The presented method, which uses no trephining or vital dye, has simplified and expedited the preparation process of pre-stripped DMEK tissue in the eye banks.

Systematic review and meta-analysis investigating autograft versus allograft cultivated limbal epithelial transplantation in limbal stem cell deficiency.

PURPOSE: Currently, regenerative medicine has attracted much attention among researchers investigating new methods to treat ocular surface diseases. Based on this new concept, cultivated limbal epithelial transplantation (CLET), whether in the form of autograft or allograft, has emerged as a promising surgical procedure for treating limbal stem cell deficiency (LSCD). Given that there is no updated comparison between autograft and allograft CLETs, the present review and meta-analysis aims to compare and determine the efficacy of two different CLET techniques, autologous versus allogeneic, based on a literature review of relevant studies. METHODS: A comprehensive search of electronic databases, including PubMed, Web of Science, Cochrane Library, Embase and Scopus, for related articles was performed in March 2018 to obtain relevant articles and to conduct a meta-analysis investigating the success rate of ocular surface regeneration and two-line improvement in best-corrected visual acuity (BCVA) using autograft versus allograft transplantations. RESULTS: A total of 30 studies, including 1306 eyes from 1288 patients with LSCD, with a sample size ranging from 6 to 200 and follow-up period of 0.6-156 months, were reviewed. Of 1306 eyes, 982 (75.2%) underwent autograft and 324 (24.8%) received allografts from living or deceased donors. Meta-analysis revealed that there was no significant difference between autograft and allograft CLETs in terms of success rate and two-line BCVA improvement. The prospective studies showed a zero difference between the two groups; only two retrospective studies included in the analysis pulled the autografts up to 1.82 and 1.2 times more than allografts in terms of success rate and two-line BCVA improvement, respectively [pooled OR 1.82 (95% CI 0.80-4.11); pooled OR 1.2 (95% CI 0.54-2.65)]. There was no statistically significant evidence of bias in the meta-analysis in terms of success rates and two-line BCVA improvement. CONCLUSIONS: The present analysis revealed no significant differences in success rates or visual improvement between autograft and allograft surgical techniques.

Gamma irradiation of ocular melanoma and lymphoma cells in the presence of gold nanoparticles: in vitro study.

The aim of this work was to determine whether conjugation of cultivated choroidal melanoma and Burkitt's lymphoma cells with gold nanoparticles (GNPs) is beneficial for these series of ocular cancer patients. GNPs are radiosensitizers and can sensitize tumors to radiotherapy.This application has been examined in several tumor types, but not in choroidal melanoma. This study shows the results of in vitro study on the choroidal melanoma and also Burkitt's lymphoma cells in the presence of GNPs during continuous gamma irradiation. Cytotoxicity of GNPs were assessed for five different concentrations then cultured melanoma and Burkitt's lymphoma cells were irradiated with a Gamma source in the presence and absence of NPs. Incubation of melanoma cells with GNP concentrations below 100 µg/ml, accompanied by gamma irradiation, increased cell death (P value = 0.016) . In the absence of irradiation, GNPs at these concentrations did not affect cultured melanoma cell metabolism. Reduced cell viability resulted from a significant increase in absorbed energy by the tumor. Moreover, GNP concentrations higher than 200 µg/ml induced cytotoxicity in melanoma cells. Cytotoxicity assay in GNPs-loaded Burkitt's lymphoma cells showed a slight decrease in cell viability at 50 µg/ml and clear cytotoxicity at concentrations higher than 100 µg/ml (P value = 0.035). Concentration and proper injection doses of GNPs in sensitive tissues such as the human eye are important variables yet to be determined.This is the first report of choroidal melanoma dosimetry performed in the presence of GNPs and provides valuable insights into future therapeutic approaches. Further in vitro study with more different sizes and concentrations is needed to determine the optimum size and concentration before any clinical research in this regard.

Single versus double pass technique for preparation of ultrathin Descemet's stripping automated endothelial keratoplasty tissues from donated whole eyes.

This study was conducted to analyze the preoperative thickness profile and endothelial rating of ultrathin Descemet's stripping automated endothelial keratoplasty (UT-DSAEK) tissues prepared with a single versus double microkeratome pass from donated whole eyes and corresponding eye bank postoperative results. Microkeratome-assisted UT-DSAEK tissues were prepared from freshly donated whole eyes with single-pass (SP) and double-pass (DP) technique in the Central Eye Bank of Iran. Preoperative thickness profiles and endothelial cell densities of UT-DSAEK tissues were obtained from optical coherence tomography and specular microscopy, respectively, and compared between groups. Corneal perforation rates during the eye bank preparation and postoperative reports of transplanted UT-DSAEK tissues were also compared. Over a 15-month period, 342 UT-DSAEK tissues were prepared: 248 via SP and 94 with DP technique. Mean donor corneal central thickness was 610 ± 58 µm with SP and 790 ± 100 µm with DP technique. Mean central thickness of UT-DSAEK tissues was not statistically different between the groups (84.8 ± 11.0 µm with SP and 85.1 ± 10.5 µm with DP technique, P = 0.857). Mean increase of UT-DSAEK thickness from central to pericentral and peripheral cornea was not significantly different with both techniques. Mean differences between thicknesses of 2 pericentral locations and between those of 2 peripheral locations were not statistically different in the study groups. Corneal perforation of 1.6 and 1.1% occurred in SP and DP groups, respectively. Failed graft was reported 6 months postoperatively in 4 (1.6%) cases with SP and in 1 (1.1%) case with DP technique. Preoperative thickness profiles of UT-DSAEK tissues prepared from donated whole eyes via SP technique were not significantly different from those prepared with DP, showing a symmetric increase of thickness towards peripheral locations.

Measurements of donor endothelial keratoplasty lenticules prepared from fresh donated whole eyes by using ultrasound and optical coherence tomography.

This study was conducted to analyze the profile and thickness of endothelial keratoplasty lenticules prepared from fresh donated whole eyes with Visante optical coherence tomography (V-OCT) compared to measurements obtained from ultrasound pachymetry (USP) at the Central Eye Bank of Iran. Microkeratome-assisted precut corneas were prepared for Descemet stripping automated endothelial keratoplasty by using standard eye bank protocol. Central posterior lenticule thickness (CPLT) on fresh whole eye, before excising corneoscleral disc and transferring to Optisol-GS, was measured by USP. V-OCT was used to measure central, paracentral, and midperipheral thicknesses of lenticules after transferring the tissue to Optisol-GS. Chi Square and Bonferroni tests were respectively used to uncover the differences between the USP and V-OCT measurements and also the thickness profile of lenticules. Postoperative reports for the entire transplanted lenticules were recorded. Accordingly, on evaluation of 312 enrolled precut corneas, CPLT measurements by V-OCT versus USP were statistically different (mean: 136 µm vs 165 µm, respectively; P = 0.008). Thickness profile of the posterior lenticules revealed increased thickness from the central to the peripheral parts of the cornea (mean increase of 16 µm at the pericentral and 64.2 µm at the peripheral locations, respectively); however, the increase in the thickness was relatively symmetrical. Postoperative reports of transplanted lenticules were unremarkable, since there were no posterior flap detachments. In essence, V-OCT measurements of microkeratome-assisted precut lenticules prepared from fresh donated whole eyes averaged 29 µm thinner than USP measurements and revealed a significant but symmetric increase of thickness towards the peripheral parts of the corneas. However, the variation in the thickness profile did not affect the attachment or the clarity of transplanted precut lenticlues.

Safety and efficacy of autologous serum eye drop for treatment of dry eyes in graft-versus-host disease.

PURPOSE: To evaluate the treatment of autologous serum eye drops (ASED) on dry eyes in patients with graft-versus-host disease (GVHD). METHODS: A retrospective chart review of 35 patients with a history of ocular GVHD following hematopoietic stem cell transplantation that used ASED to alleviate dry eye symptoms was performed. Patients were categorized into three different groups. If patients had available ophthalmic data before and after starting treatment was group 1 (n = 14), had available ophthalmic data after starting treatment in group 2 (n = 10) and had available ophthalmic data before treatment or did not have any data after starting treatment in group 3 (n = 11). Data were collected on patient's age, gender, primary diagnosis, visual acuity and fluorescein corneal staining were collected on individual eyes in order to evaluate the efficacy of the ASED on alleviating dry eye-related signs and symptoms. RESULTS: No adverse ocular effect from the ASED was found in our series (except one fungal keratitis). All patients reported either improvement (55%) or stability (45%) in their ocular symptoms upon the use of ASED. In patients with available data before and after starting treatment, the corneal staining score improved by a median of 1 (p = 0.003) and the LogMAR visual acuity had a non-significant improvement. CONCLUSION: In our study, ASED used by patients with ocular GVHD were both safe and effective. ASED should be considered in patients with GVHD who suffer from dry eyes.

Simultaneous application of bevacizumab and anti-CTGF antibody effectively suppresses proangiogenic and profibrotic factors in human RPE cells.

PURPOSE: Retinal pigment epithelial (RPE) cells play key roles in the development of choroidal neovascularization and subsequent fibrosis. We investigated the impact of bevacizumab, antihuman vascular endothelial growth factor (VEGF) antibody, and anticonnective tissue growth factor (anti-CTGF) neutralizing antibody, individually or in combination, on proangiogenic and profibrotic properties of RPE cells. METHODS: Primary cultures of human RPE cells were incubated with different concentrations of bevacizumab (0.25, 0.5, and 0.8 mg/ml) and/or anti-CTGF (10 µg/ml), and cell proliferation and apoptosis were determined. Expression and activity of proangiogenic and profibrotic genes including matrix metalloproteinases (MMP)-2 and 9, VEGFA, CTGF, vascular endothelial growth factor receptor-1 (VEGFR-1), cathepsin D, tissue inhibitor of metalloproteinases (TIMP) -1 and -2, and alpha smooth muscle actin (α-SMA) were assessed with slot blot, real-time RT-PCR, and zymography. RESULTS: Bevacizumab alone inhibited proliferation of RPE cells while anti-CTGF or bevacizumab and anti-CTGF combined had no inhibitory effect in this regard. Bevacizumab increased MMP-2, MMP-9, and cathepsin D but decreased VEGFA and VEGFR-1 expression. The CTGF level was increased by using 0.25 mg/ml bevacizumab but decreased at the 0.8 mg/ml concentration of bevacizumab. Treatment with anti-CTGF antibody decreased MMP-2 expression whereas combined treatment with bevacizumab and anti-CTGF resulted in decreased expression of MMP-2, TIMP-1, cathepsin D, VEGFA, CTGF, and α-SMA in the treated cultures. CONCLUSIONS: Treatment of RPE cells with the combination of bevacizumab and anti-CTGF could effectively suppress the proangiogenic and profibrotic activity of RPE cells.

Molecular imaging reveals elevated VEGFR-2 expression in retinal capillaries in diabetes: a novel biomarker for early diagnosis.

Diabetic retinopathy (DR) is a microvascular complication of diabetes and a leading cause of vision loss. Biomarkers and methods for early diagnosis of DR are urgently needed. Using a new molecular imaging approach, we show up to 94% higher accumulation of custom designed imaging probes against vascular endothelial growth factor receptor 2 (VEGFR-2) in retinal and choroidal vessels of diabetic animals (P<0.01), compared to normal controls. More than 80% of the VEGFR-2 in the diabetic retina was in the capillaries, compared to 47% in normal controls (P<0.01). Angiography in rabbit retinas revealed microvascular capillaries to be the location for VEGF-A-induced leakage, as expressed by significantly higher rate of fluorophore spreading with VEGF-A injection when compared to vehicle control (26±2 vs. 3±1 µm/s, P<0.05). Immunohistochemistry showed VEGFR-2 expression in capillaries of diabetic animals but not in normal controls. Macular vessels from diabetic patients (n=7) showed significantly more VEGFR-2 compared to nondiabetic controls (n=5) or peripheral retinal regions of the same retinas (P<0.01 in both cases). Here we introduce a new approach for early diagnosis of DR and VEGFR-2 as a molecular marker. VEGFR-2 could become a key diagnostic target, one that might help to prevent retinal vascular leakage and proliferation in diabetic patients.

Comparing quantitative and qualitative indices of the donated corneas maintained in Optisol-GS with those kept in Eusol-C.

This study was conducted to compare the quantitative and qualitative indices of the donated corneas maintained in either Optisol-GS or Eusol-C storage media. In an ante-grade single blind study, two corneas from each donor with a death to preservation time of less than 30 h and with a minimum of "an apparent good cornea rating" were maintained in corneal storage media; randomly one in Optisol-GS and the other in Eusol-C. Slit-lamp biomicroscopic and specular microscopic examinations were performed on days 1 and 7. The results of the qualitative and quantitative indices and the final cornea rating were recorded. Statistical analyses were performed to evaluate any differences between the two media. 180 corneas from 90 donors with an age range of 29.3 ± 22.4 years were allocated in two groups: 90 corneas in Optisol-GS and the other 90 in Eusol-C. Five corneas in Optisol-GS and four corneas in Eusol-C were excluded from the study due to lack of specular images. There was no significant change of endothelial rating from day 1 to day 7 between two media (P = 0.175). As the maintenance time of the donated corneas increased, no significant difference was noted between the two groups in terms of endothelial indices, stromal edema, and Descemet's folding. This study shows no superiority between Eusol-C and Optisol-GS in short term preservation of donated corneas.