Short- and long-term corneal vascular effects of tafluprost eye drops

Background

Prostaglandin analogs are first line therapy in the treatment of glaucoma, but also display side effects during ocular inflammation. In this context, the potential side effects of prostaglandin analogs on the normally avascular cornea, the main application route for eye drops, are so far not fully defined. Therefore, the aim of this study was to evaluate the vascular effects of the prostaglandin analog tafluprost on the healthy and inflamed cornea.

Methods

For in vitro studies, blood and lymphatic endothelial cells were treated with tafluprost; cell proliferation was assessed after 48 h. For long-term in vivo studies under healthy conditions, naïve corneas of BALB/c mice were treated with tafluprost eye drops for 4 weeks. For short-term in vivo studies under inflammatory conditions, corneal inflammation was induced by suture placement; mice then received tafluprost eye drops for 1 week. Afterwards, corneas were stained with CD31 as panendothelial and LYVE-1 as lymphendothelial (and macrophage) marker.

Results

In vitro, tafluprost did not alter blood or lymphatic endothelial cell proliferation. In vivo, there was no change in limbal blood or lymphatic vessel anatomy after long-term treatment with tafluprost. Short-term treatment with tafluprost under inflammatory conditions did not influence the recruitment of LYVE-1 positive macrophages into the cornea. Moreover, treatment of inflamed corneas with tafluprost did not significantly influence corneal hem- and lymphangiogenesis.

Conclusions

Tafluprost does not affect blood and lymphatic vessel growth, neither under resting nor under inflammatory conditions. These findings suggest a safe vascular profile of tafluprost eye drops at the inflammatory neovascularized cornea.     

Inhibition of hemangiogenesis and lymphangiogenesis after normal-risk corneal transplantation by neutralizing VEGF promotes graft survival

PURPOSE: To evaluate the occurrence and time course of hem- and lymphangiogenesis after normal-risk corneal transplantation in the mouse model and to test whether pharmacologic strategies inhibiting both processes improve long-term graft survival. METHODS: Normal-risk allogeneic (C57BL/6 to BALB/c) and syngeneic (BALB/c to BALB/c) corneal transplantations were performed and occurrence and time course of hem- and lymphangiogenesis after keratoplasty was observed, by using double immunofluorescence of corneal flatmounts (with CD31 as a panendothelial and LYVE-1 as a lymphatic vascular endothelium-specific marker). A molecular trap designed to eliminate VEGF-A (VEGF Trap(R1R2); 12.5 mg/kg) was tested for its ability to inhibit both processes after keratoplasty and to promote long-term graft survival (intraperitoneal injections on the day of surgery and 3, 7, and 14 days later). RESULTS: No blood or lymph vessels were detectable immediately after normal-risk transplantation in either donor or host cornea, but hem- and lymphangiogenesis were clearly visible at day 3 after transplantation. Both vessel types reached donor tissue at 1 week after allografting and similarly after syngeneic grafting. Early postoperative trapping of VEGF-A significantly reduced both hem- and lymphangiogenesis and significantly improved long-term graft survival (78% vs. 40%; P < 0.05). CONCLUSIONS: There is concurrent, VEGF-A-dependent hem- and lymphangiogenesis after normal-risk keratoplasty within the preoperatively avascular recipient bed. Inhibition of hem- and lymphangiogenesis (afferent and efferent arm of an immune response) after normal-risk corneal transplantation improves long-term graft survival, establishing early postoperative hem- and lymphangiogenesis as novel risk factors for graft rejection even in low-risk eyes.     

Comparison of deep anterior lamellar keratoplasty and penetrating keratoplasty with respect to postoperative corneal sensitivity and tear film function

Purpose

To investigate tear film function, central and peripheral corneal sensitivity and corneal subbasal nerve morphology in the cornea after deep anterior lamellar keratoplasty (DALK) compared with penetrating keratoplasty (PK).

Methods

This prospective study compared the changes in 16 eyes of 16 patients who underwent DALK (DALK group) with those in 28 eyes of 28 patients who underwent PK (PK group). Thirty healthy volunteers were also included as controls. Tear functions were evaluated using tear break-up time (TBUT), tear meniscus height (TMH) and corneal fluorescein staining. Corneal sensation was measured with a Cochet-Bonnet esthesiometer. Corneal subbasal nerve morphology was evaluated using in vivo confocal microscopy (IVCM). The patients were examined 1, 3, 6, 9 and 12 months after keratoplasty.

Results

Postoperatively, TMH recovered significantly faster in the DALK group than in the PK group (p?p?p?p?Conclusions Tear film function was restored more rapidly after DALK compared with PK, but there was no significant difference in corneal sensitivity between PK and DALK.     

Soluble vascular endothelial growth factor receptor-3 suppresses allosensitization and promotes corneal allograft survival

Purpose

To investigate the effect of VEGF-C and VEGF-D blockade via soluble VEGFR-3 (sVEGFR-3) on T cell allosensitization, corneal neovascularization, and transplant survival.

Methods

Corneal intrastromal suture placement and allogeneic transplantation were performed on BALB/c mice to evaluate the effect of sVEGFR-3 on corneal neovascularization. Soluble VEGFR-3 trap was injected intraperitoneally to block VEGF-C/D (every other day starting the day of surgery). Immunohistochemical staining of corneal whole mounts was performed using anti-CD31 (PECAM-1) and anti-LYVE-1 antibodies to quantify the levels of hem- and lymphangiogenesis, respectively. Mixed lymphocyte reaction (MLR) was performed to assess indirect and direct host T cell allosensitization and the frequencies of IFN-γ-producing T cells in the draining lymph nodes were assessed using flow cytometry. Graft opacity and survival was evaluated by slit-lamp biomicroscopy.

Results

Treatment with sVEGFR-3 resulted in a significant blockade of lymphangiogenesis 2 weeks post-transplantation and significantly prolonged corneal allograft survival compared to the control group at 8 weeks post-transplantation (87.5 % vs. 50 %), and this was associated with significant reduction in the frequencies of allosensitized T cells and decreased frequencies of IFN-γ–producing CD4 T cells.

Conclusions

Soluble VEGFR-3 suppresses corneal lymphangiogenesis and allograft rejection and may offer a viable therapeutic modality for corneal neovascularization and corneal transplantation.     

Inhibitory effects of topical cyclosporine A 0.05 % on immune-mediated corneal neovascularization in rabbits

Background

We aimed to study the inhibitory effects of topical cyclosporine A (CsA) 0.05 % on immune-mediated corneal neovascularization, and to compare its efficacy with those of dexamethasone 0.1 % and bevacizumab 0.5 %.

Methods

Immune-mediated corneal neovascularization was created in 36 right eyes of 36 rabbits. The rabbits were then randomized into four groups. Group I received CsA 0.05 %, Group II received dexamethasone 0.1 %, Group III received bevacizumab 0.5 %, and Group IV received isotonic saline twice a day for 14 days. The corneal surface covered with neovascular vessels was measured on the photographs. The rabbits were then sacrificed and the corneas excised. Paraffin-embedded sections were stained with hematoxylin-eosin and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay.

Results

The means of percent area of corneal neovascularization in Group I, II, III, and IV were 24.4 %, 5.9 %, 37.1 %, and 44.1 %, respectively. The inhibitory effect of CsA 0.05 % was found to be better than the effect found in the bevacizumab 0.5 % and control groups (p?=?0.03 and p?=?0.02, respectively). CsA 0.05 % was found to have significantly lesser inhibitory effects on corneal neovascularization than dexamethasone 0.1 % (p?<?0.001). Apoptotic cell density was higher in Group III and Group IV than in Group I and Group II. There was no difference between Group I and Group II in terms of apoptotic cell density (p?=?0.7).

Conclusions

Topical CsA 0.05 % was shown to have an inhibitory effect on immune-mediated corneal neovascularization in rabbits.     

Anti-neovascular effect of chondrocyte-derived extracellular matrix on corneal alkaline burns in rabbits

Purpose

We investigated the effect of a chondrocyte-derived extracellular matrix (CDECM) on experimental corneal alkaline burns in rabbits.

Methods

Corneal neovascularization (NV) was induced by applying an 8-mm filter paper soaked in 1 N NaOH to the right central corneas of rabbits for 1 minute. Ten days later, the rabbits were randomly divided into three groups: the alkaline burn group, the CDECM transplantation group, and the human amniotic membrane (HAM) transplantation group. The left eyes were used as controls. CDECM and HAM were transplanted onto the corneal surface to completely cover the resected area and were subsequently sutured. On the 10th day after transplantation, the structural changes of the cornea were analyzed histologically. We examined the effects of CDECM on clinical NV features and on the expression of corneal NV markers.

Results

The alkaline burn produced significant NV and increased the corneal thickness. On day 10 after transplantation, the thickness, NV and opacity of the cornea were markedly decreased in the CDECM group (p?<?0.001). However, the HAM transplantation group did not exhibit improvements in these clinical parameters, and there were no significant differences relative to the burn group. In addition, the use of CDECM improved the healing of the cornea following the alkaline burn by disrupting the corneal epithelial proliferation and reducing the fibrotic changes of the stroma. The hallmarks of NV were significantly induced in the subepithelium by the alkaline burn, and these levels were also suppressed by CDECM. The CDECM suppressed corneal NV by inhibiting nuclear factor-kappa B (NF-κB) activation by blocking the PKC and Akt signaling pathways.

Conclusions

CDECM transplantation was markedly effective in healing alkali-burned corneas by modulating the translocation of NF-κB to the nucleus, thereby representing a promising material for the noninvasive treatment of ocular surface disease.     

Fluorescein angiography, optical coherence tomography, and histopathologic findings in a VEGF165 animal model of retinal angiogenesis

Background

To establish an animal model of retinal neovascularization using vascular endothelial growth factor (VEGF165) and analyze the model using optical coherence tomography (OCT), fluorescein angiography (FA), and histopathologic evaluation.

Methods

Twelve rabbits were divided into groups as follows: group 1 (n?=?3), sham intravitreous injections of 0.1?ml of balanced saline; group 2 (n?=?6), one 10-μg intravitreal injection of VEGF165 on day 0; and group 3 (n?=?3), two 10-μg intravitreal injections of VEGF165, one on day 0 and one on day 7. Follow-up evaluations (days 0, 3, 7, 14, 21, 28) included obtaining fundus color photographs and FA, OCT, and histopathologic examinations. Eyes were enucleated and stained with hematoxylin and eosin (H&;E).

Results

One injection of VEGF (group 2) was associated with dilatation and tortuosity of the retinal blood vessels that developed within 72?h. Retinal neovascularization was present by day 7 and regressed by day 14. However, even on day 28, the capillaries were still tortuous. Two VEGF injections (group 3) caused increased leakage and neovascularization up to day 14; severe capillary nonperfusion was seen during week 4. At the end of the follow-up period, OCT and histopathologic examination of group 3 showed peripapillary tractional retinal detachments. By day 7, the differences between the retinal thickness seen on OCT in groups 2 and 3 and the group 1 control group were significant (p?Conclusions FA, OCT, and histopathologic findings showed that this retinal neovascularization model is efficient, sustainable, and reliable. One injection of VEGF165 created neovascularization that peaked after 1?week; two injections created more intense neovascularization that evolved to retinal detachments after 4?weeks.     

Topische antiangiogene Therapie an der Hornhaut

Background

The efficacy and safety of novel topical inhibitors of corneal neovascularisation will be discussed.

Methods

A literature review after a PUBMED search and own clinical and experimental results are presented.

Results

The off-label use of Avastin® eye drops and GS101 eye drops against insulin receptor substrate (IRS)-1, which have been tested in phase II trial, both seem to be relatively efficient and safe ways to inhibit progressive corneal neovascularisation. Other VEGF antagonists, such as ranibizumab and pegaptanib eye drops also inhibit corneal neovascularisation.

Conclusions

Avastin® and GS101 eye drops are the first specific angiogenesis inhibitors for topical inhibition of corneal angiogenesis available for clinical use.     

Corneal lymphangiogenesis facilitates ocular surface inflammation and cell trafficking in dry eye disease

Purpose

While the normal cornea has limited innervation by the lymphatic system, chronic immune-inflammatory disorders such as dry eye (DE) can induce lymphangiogenesis in the ocular surface. Using a conditional knock-down murine model, Lyve-1^Cre;VEGFR2^flox mice, this study investigated the role of lymphangiogenesis in the pathophysiology of DE.

Pharmacokinetics of bevacizumab after topical and intravitreal administration in human eyes

Background

Topical bevacizumab is a potential treatment modality for corneal neovascularization, and several recent studies have demonstrated its efficacy. No previous study of the pharmacokinetics of topical bevacizumab has been performed in human eyes. The purpose of this study is to investigate the pharmacokinetics of topical administration of bevacizumab in human eyes, and also to compare the pharmacokinetics of intravitreal bevacizumab injections with previously reported data.

Methods

Twenty-two (22 eyes) were included in this study, and divided into four groups: eight patients received topical bevacizumab and aqueous samples were obtained 1 hour later during cataract extraction surgery (group 1), eight patients received topical bevacizumab and vitreous samples were obtained 1 day later during pars-plana vitrectomy (PPV) (group 2), three patients received intravitreal bevacizumab and vitreous samples were obtained during PPV (group 3). Vitreous samples from three patients who received no bevacizumab served as controls (group 4). All samples underwent enzyme-linked immunosorbent assay to detect bevacizumab.

Results

No bevacizumab was detected in the aqueous or vitreous of any topically treated eyes. The mean vitreal half-life for intravitreally injected bevacizumab was 4.9 days in four non-vitrectomized eyes and 0.66 days in one previously vitrectomized eye.

Conclusions

Topically administered bevacizumab does not penetrate the cornea into the anterior chamber and vitreous cavity, indicating that topical use for treating corneal neovascularization has minimal risk of intraocular penetration and adverse events related to intraocular vascular endothelial growth factor inhibition. The half-life following intravitreal bevacizumab injection measured in this study is comparable to that of previous reports, and includes the first demonstration of a significantly reduced half-life following intravitreal injection in a previously vitrectomized eye.     

Automated Keratoconus Detection Using Height Data of Anterior and Posterior Corneal Surfaces

Purpose

To develop a keratoconus detection algorithm using the corneal topographic data of the anterior and posterior corneal surfaces.

Methods

Topographic measurements of the cornea were made with a slit-scanning corneal topographer. We examined 120 subjects (165 eyes); keratoconus patients and keratoconus suspect patients comprised the keratoconus group, and post-photorefractive keratectomy patients, with-the-rule astigmatism patients, and controls without disease comprised the nonkeratoconus group. Two variables of the anterior corneal surface, two variables of the posterior corneal surface, and one corneal thickness variable were obtained by applying the Fourier harmonic decomposition formula. By performing a logistic regression analysis with a training set to differentiate the keratoconus group from the nonkeratoconus group, the Fourier-incorporated keratoconus detection Index (FKI) was created. The validity of the FKI was determined by using independent validation sets.

Results

The FKI distinguished the keratoconus group from the nonkeratoconus group with 96.9% sensitivity and 95.4% specificity in the validation set.

Conclusions

A newly developed automated keratoconus classifier can be used to screen keratoconic patients. The index is based on information obtained by Fourier analysis from not only the anterior corneal surface but also from the posterior corneal surface and corneal thickness.?Jpn J Ophthalmol 2006;50:409–416 © Japanese Ophthalmological Society 2006     

In vitro effect of corneal collagen cross-linking on corneal hydration properties and stiffness

Background

The purpose of this study is to evaluate in-vitro the immediate effect of corneal collagen cross-linking (CXL) on corneal hydration and stiffness.

Methods

Forty-two corneal buttons from freshly enucleated porcine eyes were immersed in riboflavin 0.1% in dextran 20% dilution for 3 h in order for their hydration to reach equilibrium. Corneal buttons where divided into two groups; the first group was stored in dark conditions while the other group was irradiated with UV radiation (370 nm) for 30 min to simulate CXL according to the clinically applied protocol. After irradiation, all corneas were immersed in dextran 20% solution for 3 additional hours. Subsequently, each button underwent weighing, thickness measurement, and was mounted in a special device for the measurement of force versus deformation by compression. Finally, all corneal buttons were dehydrated for 48 h in a desiccating oven set at 62 °C and weighed again to obtain their dry mass. Hydration (%) of each button was calculated.

Results

Mean corneal hydration in the irradiated and the non-irradiated group of corneas was 69.8 and 72.2%, respectively (p?<?0.001). Differences in thickness and compressibility were not statistically significant. Thickness and hydration were positively correlated (Pearson’s r?=?0.714, p?<?0.001).

Conclusions

CXL causes corneal dehydration that can be detected immediately after the procedure. This phenomenon may contribute to increased mechanical stiffness of the cornea. A change in stiffness by means of compressibility could not be detected in porcine corneas.     

The effect of posterior corneal flat meridian and astigmatism amount on the total corneal astigmatism estimated from anterior corneal measurements

Background

To evaluate the effects of posterior corneal astigmatism and the absolute flat meridian difference between anterior and posterior corneal surfaces (AMD_Ant-Post) on the estimation of total corneal astigmatism using anterior corneal measurements (simulated keratometry [K]).

Methods

Ninety-nine eyes of 99 healthy participants were enrolled. Anterior, posterior, and total mean corneal power, cylinder power, flat meridian, and vector components J₀, and J₄₅ measured by a dual Scheimpflug camera were analyzed. The correlation between the posterior corneal cylinder power, AMD_Ant-Post, and the difference in the cylinder power between simulated K and total cornea (cylinder power difference_SimK-Tot) were evaluated.

Results

The cylinder power difference_SimK-Tot was positively correlated with the posterior corneal cylinder power (rho?=?0.704 and P?Ant-Post (rho?=??0.717 and P?0 was strongly associated with the posterior corneal cylinder power and the AMD_Ant-Post. When corneal J₀ had a positive value, the cylinder power of simulated K tended to be larger than the total corneal cylinder power. In comparison, the opposite trend was presented in eyes with negative anterior corneal J₀. When anterior corneal J₀ was larger than 1.0 or smaller than ?0.9, the errors from estimating the total corneal cylinder power using anterior corneal measurements tended to be larger than 0.25 D.

Conclusion

Posterior corneal astigmatism should be considered for more accurate corneal astigmatism predictions, especially in eyes with anterior corneal astigmatism greater than 2.0 D of with-the-rule astigmatism or greater than 1.8 D of against-the-rule astigmatism.     

Einfluss der kornealen Biomechanik auf die Myopieregression nach Laser-in-situ-Keratomileusis

Background

Laser in situ keratomileusis is a safe and accepted method for correcting myopia. The operational results in terms of accuracy as well as the subjective acceptance of patients for corrections to – 8 D are now considered to be promising (Seiler, Refraktive Chirurgie der Hornhaut, 2000); however, postoperative results show individual patient problems in long-term stability. It is believed that the preoperative condition of the cornea (e.g. thickness, biomechanical properties) could have an influence on postoperative problems such as myopic regression.

Method

This study included a total of 46 eyes from 25 patients. At 3 months postoperatively, 15 patients (19 eyes) showed a SEQ of ??0.50 D or more. Within this group, 11 patients (15 eyes) developed a regression (regression group) within the first 3 postoperative months. The remainder of the total group did not show any regression (stability group). The subjects of this study were on average 33?±?8 years (stability group) and 31?±?7 years old (regression group). The corneal thickness was tested and refractive error, visual acuity (BCVA/UCVA) and intraocular pressure was measured. In addition, the corneal hysteresis (CH) and corneal resistance factor (CRF) were determined.

Results

The mean preoperative spherical equivalent refraction was ??3.14 D?±?1.41 D (SE) in the stability group and ? 6.47 D?±?1.40 D (p?=?0.001)in the regression group. Also, the postoperative spherical equivalents were statistically significant different (p?<?0.05). In contrast, the mean preoperative corneal thickness showed no differences in both groups (p?=?0.96) (stability group 563?±?36 µm and regression group 563?±?28 µm).

Conclusions

The aim of the study to detect a possible causal relationship between myopia regression after LASIK and the biomechanical properties of the cornea and corneal thickness could not be clearly identified.     

Evaluation of corneal epithelial and stromal thickness in keratoconus using spectral-domain optical coherence tomography

Purpose

We sought to assess the corneal thickness of the epithelium and stroma in keratoconic and normal eyes by spectral-domain optical coherence tomography (SD-OCT).

Methods

Fifty-seven keratoconic and 20 normal eyes were studied. The eyes were examined by SD-OCT, and the keratoconic eyes were subdivided into 2 groups: those showing only smooth corneal thinning and corneal protrusion on the image (KC1 group) and those showing abnormalities in the Bowman layer or in the stroma, or in both (KC2 group). The thicknesses at the corneal vertex and at the superior, inferior, nasal, and temporal cornea 1.5 mm from the corneal vertex in the KC1 group were compared with those in the normal group. The OCT findings in the KC2 group were described.

Results

The epithelial thickness at the corneal vertex and at the inferior and temporal cornea, and the stromal thickness at all points were significantly thinner in the KC1 group than in the normal group (p < 0.05). The epithelial and stromal thicknesses at the corneal vertex were significantly correlated in the KC1 group and the normal group (r ²  = 0.427, p < 0.0001).The epithelial thickness in the KC2 group was not uniform owing to Bowman layer scarring, stromal scars, and secondary corneal amyloidosis.

Conclusions

Although epithelial thinning is associated with stromal thinning, when the cornea remains clear, the epithelial thickness may vary because of the irregularity of the stroma beneath the epithelium in patients with keratoconus.     

Corneal biochemical features of patients with vernal keratoconjunctivitis

Background

Vernal keratoconjunctivitis (VKC) is a chronic, bilateral, seasonally exacerbated, allergic inflammation of the ocular surface, involving bulbar and ? or tarsal conjunctiva and cornea. The ocular response analyzer (ORA) measures corneal biomechanical properties in vivo by monitoring and analyzing the corneal behavior when its structure is submitted to a force induced by an air jet. This study was designed to examine corneal biomechanical properties and intraocular pressure in patients with VKC, and to compare with control eyes.

Methods

ORA measurements were performed on the both eyes of 26 patients with VKC (group I) and 14 healthy children who served as the control group (group II). Corneal hysteresis (CH), corneal resistance factor (CRF) and intraocular pressure [Goldmann correlated (IOPg) and corneal compensated (IOPcc)] were recorded with ORA.

Results

Mean age of patients with VKC and control groups were 11.3?±?5.8 and 10.6?±?1.9 years for groups I and II respectively. Mean (± SD) of the CH and CRF readings were 10.1?±?1.6 versus 10.5 ±1.6 (p?>?0.05) and 9.5?±?1.7 versus 10.8?±?1.7 mmHg (p?<?0.05), in groups I and II respectively. Mean (± SD) of the IOPg and IOPcc recordings were 13.3 ±3.4 versus 16.6 ±3.6 mmHg (p?<?0.05) and 14.3?±?3.4 versus 16.9?±?3.7 mmHg (p?>?0.05) in groups I and II respectively. Statistical analysis revealed significant differences for CRF and IOPg between the study groups.

Conclusion

The mean CRF and IOPg values of patients with VKC were lower than those of controls. According to the results of our study, one can conclude that corneal biomechanical property, CRF, could be different in VKC patients compared to normals.     

Peptidoglycan and muramyl dipeptide from Staphylococcus aureus induce the expression of VEGF-A in human limbal fibroblasts with the participation of TLR2-NFκB and NOD2-EGFR

Background

Keratitis caused by Staphylococcus aureus often leads to Vascular Endothelial Growth Factor (VEGF)-dependent neovascularization, but contribution of peptidoglycan (PGN), muramyl dipeptide (MDP) and lipoteichoic acid (LTA) from S. aureus to VEGF-dependent neovascularization has not been well-studied. This work was focused on the analysis of S. aureus cell wall components in the production of VEGF family members (VEGF-A, VEGF-B, VEGF-C and VEGF-D) in ocular limbal fibroblasts.

Methods

Primary culture of human limbal fibroblasts (PCHLFs) were stimulated with PGN, MDP, and LTA, and VEGF family; toll-like receptor 2 (TLR2), nucleotide-binding oligomerization domain 1 (NOD1), and NOD2 expression were determined by RT-PCR. Anti-TLR2 antibody, epidermal growth factor receptor (EGFR) signaling inhibitors (AG1478 and PD98059), and NFκB activation were used to analyze VEGF-A by ELISA. TLR2 and NOD1 expression were analyzed by flow cytometry.

Results

The stimulation of PCHLFs with PGN and MDP increased the levels of VEGF-A expression (mRNA and protein) in a time-dependent and dose-dependent manner. VEGF-B, VEGF-C and VEGF-D were expressed constitutively, and no further induction was observed in stimulated PCHLFs. LTA did not increase the expression levels of the VEGF family. TLR2 mRNA and protein were increased only when PCHLFs were stimulated with PGN. Treatment with an anti-TLR2 antibody blocked the interaction of PGN with the receptor, inhibiting VEGF-A over-expression; the presence of anti-TLR2 antibodies did not affect the over-production of VEGF-A after MDP treatment. PCHLFs stimulated with PGN and MDP, but not with LTA, activated NFκB. MDP stimulated the production of NOD1 and NOD2 mRNAs in a time-dependent and dose-dependent manner, and NOD2 protein was only increased by MDP. Treatment of PCHLFs with AG1478 and PD98059 inhibitors prior to stimulation with MDP resulted in the inhibition of VEGF-A over-production, compared with PCHLFs stimulated with MDP alone.

Conclusions

Taken together, these results suggest that limbal fibroblasts produce VEGF-A through PGN-TLR2-NFκB and MDP-NOD2-EGFR.     

The 2009 performance report of the German cornea banks

Background

In Germany, human tissue for corneal and amniotic transplantation is supplied by 27 cornea banks.

Methods

The Section for Tissue Transplantation and Biotechnology of the German Ophthalmological Society records the cornea banks?? activities by means of an annual questionnaire.

Results

In 2009, a total of 4,818 corneal grafts were processed by 21 responding cornea banks, and 57% were deemed suitable for transplantation. This ratio is slightly higher than the European average. In addition, German cornea banks released 1,257 amniotic grafts in 2009.

Discussion

German cornea banks are currently facing new regulatory issues due to updated legislation regarding tissue transplantation. Recent updates in European law have limited the cutoff time for postmortem blood sampling to 24 h, and this regulation may lead to a significant reduction in potential donors.     

Anti-VEGF monoclonal antibody-induced regression of corneal neovascularization and inflammation in a rabbit model of herpetic stromal keratitis

Background

To determine the efficacy of bevacizumab (Avastin), an anti-VEGF monoclonal antibody, administrated via subconjunctival injection as a corneal anti-angiogenic treatment.

Methods

Right corneas of rabbits were infected with herpes simplex virus type 1, KOS strain. On day 13 post-infection (p.i.), animals were treated subconjunctivally (sc) with a single 10-μl dose (25 μg/μl) of bevacizumab (group A) or with the same volume of an isotype monoclonal antibody, as negative control (group B). All animals were observed clinically on days 2, 5, 7, 14, 21, and 28 p.i., and two corneas each day were obtained for histological assessment and viral titration.

Results

Viral replication was observed no longer than 5 days after infection. By day 7 a dense neutrophil invasion of the cornea was detected, which significantly increased while herpetic stromal keratitis progressed in severity. Positive outcomes observed following the treatment with bevacizumab, compared to control, included: (1) Total involution of neovascularization, (2) reduction in disease severity, (3) improved corneal translucency, (4) absence of scarring, (5) preservation of corneal thickness, (6) no neutrophil infiltration of the cornea.

Conclusions

Subconjunctival administration of bevacizumab induced involution of new vessels, abolished inflammatory response, and resulted in return of corneal function. Furthermore, bevacizumab is a novel approach for the treatment of herpetic stromal keratitis.     

Comparison of corneal haze and visual outcome in primary DSAEK versus DSAEK following failed DMEK

Background

Descemet membrane endothelial keratoplasty (DMEK) is being proposed as the procedure of choice in corneal endothelial disease as it achieves better visual and refractive outcomes than Descemet stripping automated endothelial keratoplasty (DSAEK). Nevertheless, primary graft failure is frequent, especially during the learning curve, and secondary back-up procedure consists on DSAEK. We aim to compare corneal haze and visual acuity of patients undergoing primary DSAEK vs. patients undergoing DSAEK as a back-up procedure after primary DMEK failure.

Methods

This study is a comparative case series that included 19 eyes from 16 patients with early stages of corneal failure and limitation of daily activities after primary DSAEK or secondary DSAEK. A control group of non-operated corneas included 10 aged-matched normal eyes. The study was conducted at University Hospital Ramón y Cajal and Vissum Hospital, Madrid, Spain. Corneal densitometry readings and postoperative best-corrected visual acuity in subjects with primary and secondary DSAEK were recorded 6 months after the surgery using the Pentacam Scheimpflug system (Oculus, inc.,Wetzlar, Germany).

Results

In primary DSAEK median densitometry values (range) were statistically significantly higher (p?<?0.05) than normal subjects for the full thickness, posterior and anterior part of the paracentral cornea; and the anterior part of the central cornea. In secondary DSAEK, median densitometry values were statistically significantly higher than normal subjects at all levels of the central and paracentral cornea. In secondary DSAEK, median densitometry values (range) were statistically significantly higher than in primary DSAEK in the full-thickness, anterior part and interface of the central cornea and in the full-thickness and posterior part of the paracentral cornea. Median visual acuity between groups (p?=?0.47) was statistically better for the primary DSAEK group, which also had a higher percentage of patients achieving BCVA of ≥ 20/40 and ≥20/25 than the secondary DSAEK group (100 % vs. 62 % and 60 % vs. 0 % respectively).

Conclusions

There is an increase in central corneal light scattering after secondary DSAEK performed after a failed DMEK as compared to primary DSAEK. This has a negative impact on final visual acuity that needs to be considered in each patient when starting DMEK surgery.