Area Of Corneal Neovascularization Research Articles

Available Therapeutic Options for Corneal Neovascularization: A Review

Corneal neovascularization can impair vision and result in a poor quality of life. The pathogenesis involves a complex interplay of angiogenic factors, notably vascular endothelial growth factor (VEGF). This review provides a comprehensive overview of potential therapies for corneal neovascularization, covering tissue inhibitors of metalloproteinases (TIMPs), transforming growth factor beta (TGF-β) inhibitors, interleukin-1L receptor antagonist (IL-1 Ra), nitric oxide synthase (NOS) isoforms, galectin-3 inhibitors, retinal pigment epithelium-derived factor (PEDF), platelet-derived growth factor (PDGF) receptor inhibitors, and surgical treatments. Conventional treatments include anti-VEGF therapy and laser interventions, while emerging therapies such as immunosuppressive drugs (cyclosporine and rapamycin) have been explored. Losartan and decorin are potential antifibrotic agents that mitigate TGF-β-induced fibrosis. Ocular nanosystems are innovative drug-delivery platforms that facilitate the targeted release of therapeutic agents. Gene therapies, such as small interfering RNA and antisense oligonucleotides, are promising approaches for selectively inhibiting angiogenesis-related gene expression. Aganirsen is efficacious in reducing the corneal neovascularization area without significant adverse effects. These multifaceted approaches underscore the corneal neovascularization management complexity and highlight ideas for enhancing therapeutic outcomes. Furthermore, the importance of combination therapies and the need for further research to develop specific inhibitors while considering their therapeutic efficacy and potential adverse effects are discussed.

Platelet Rich Fibrin as a Potential Treatment for Corneal Neovascularization due to Infective Keratitis: An Experimental Study

Abstract
 Introduction & Objectives : Platelet-rich fibrin (PRF) as a source of growth factor has been widely used in various therapeutic fields so that it could accelerate the wound healing process. This study investigated the effect of platelet-rich fibrin PRF in inhibiting neovascularization, inhibiting the expression of vascular endothelial growth factor (VEGF) and stimulating pigment epithelial-derived factor (PEDF) in the infective keratitis animal model.
 Methods : A total of 12 New Zealand white rabbits (Oryctalogus cuniculus) were subjected to keratitis induction by inoculation of Pseudomonas aeruginosa. The rabbits were divided into two groups, the first group as control was treated antibiotic-only, and the second as treatment group was treated with antibiotic and PRF graft. The corneal neovascularization area was clinically measured before and after treatment. Expression of VEGF and PEDF in the corneal epithelium, stroma, and endothelium were evaluated by immunohistochemical examination on day 10 after keratitis induction.
 Results : There were significant differences in VEGF expression between groups in the epithelial layer (p=0.008), but no significant difference in the stromal (p=0.934) and endothelial (p=0.935) layers. PEDF expression showed a significant difference between groups in the epithelial (p = 0.004) and stroma layers (p = 0.029) but no significant difference in the endothelial layer (p = 0.157). However, there was no significant difference in clinical neovascularization between the two groups (p=0.526).
 Conclusion : PRF could inhibit VEGF expression and stimulate PEDF expression in the cornea of Pseudomonas aeruginosa keratitis animal model that potentially reduces the corneal neovascularization in the longer observation.

Bone morphogenetic protein 4 thermosensitive hydrogel inhibits corneal neovascularization by repairing corneal epithelial apical junctional complexes

Corneal neovascularization (CNV) is a heavy attribute of blinding disease changes. Existing medications need numerous infusions and have a limited absorption. Investigating novel drugs with safety, efficacy, and convenience is crucial. In this study, we developed a bone morphogenetic protein 4 (BMP4)-loaded poloxamer-oxidized sodium alginate (F127-OSA) thermosensitive hydrogel. The 14 % F127-OSA hydrogel transformed from sol to gel at 31–32 °C, which might extend the application period on the ocular surface. The hydrogel's porous structure and uniform dispersion made it possible for drugs to release gradually. We used a suture-induced rat CNV model to investigate the mechanism of CNV inhibition by hydrogel. We discovered that F127-OSA hydrogel loaded with BMP4 could significantly reduce the length and area of CNV, relieve corneal edema, and stop aberrant epithelial cell proliferation. The hydrogel's efficacy was superior to that of the common solvent group. Additionally, BMP4 thermosensitive hydrogel repaired ultrastructure, including microvilli, intercellular junctions, and damaged apical junctional complexes (AJCs), suggesting a potential mechanism by which the hydrogel prevented CNV formation. In conclusion, our investigation demonstrates that F127-OSA thermosensitive hydrogel loaded with BMP4 can repair corneal epithelial AJCs and is a promising novel medication for the treatment of CNV.

Clickable corneal neovascularization therapy with ROS-responsive polydopamine silica nanoparticles loaded with fenofibrate

Corneal neovascularization is a severe eye disease that is often associated with an inflammatory cycle and increased oxidative stress. Excessive reactive oxygen species can cause harmful changes in the corneal epithelium and lead to vicious inflammatory cycles, resulting in an increase in corneal neovascularization. Unfortunately, many drugs used in clinic are water-insoluble and have low bioavailability on the ocular surface, which limits their effectiveness. Therefore, it's crucial to design a new drug delivery system. In this study, we designed an oxide-responsive nanoparticle system called MPA, which was consisted of the large pore sized-mesoporous silica nanoparticles (MSNs) core and the polydopamine (PDA) surface shell via the linkage with disulfide bond. Finally, the nanoparticles are endowed with the enhanced loading efficiency and the controllable release of fenofibrate (Feno) in response to the abundant ROS level in the inflammatory environment. In an in vivo study using mice, treatment with MPA@ Feno resulted in minimal corneal neovascularization area and length and significantly reduced expression of pro-angiogenic factors and oxidative stress factors. Therefore, MPA solves the problem of poor drug solubility, clearing oxygen species in the pathological environment, which will improve its therapeutic effect in corneal neovascularization.

Analysis of long-term outcomes of penetrating keratoplasty for congenital corneal opacity

Objective: To investigate the long-term outcomes of corneal grafts after penetrating keratoplasty(PK) for congenital corneal opacity(CCO) in children aged 0 to 5 years and the related influencing factors. Methods: It was a retrospective series case study. Data of 39 children (55 eyes) who underwent PK surgery due to CCO in the keratology Department of Beijing Tongren Hospital from April 2014 to April 2018 and were followed up for more than 30 months were collected. Among them, there were 17 males (43.6%) and 22 females (56.4%). The age at operation was (16.2±13.3) months, and the follow-up time was (46.4±13.8) months. Clinical data such as basic information, preoperative diagnosis, operation age, operation method and postoperative complications were recorded. The corneal graft transparency was analyzed according to preoperative diagnosis, corneal neovascularization area, age at surgery, monocular or binocular surgery interval, primary surgery type and further surgery, and postoperative complications were observed. Results: At 12 months, 24 months and the last follow-up after PK, 78.2% (43/55), 70.9% (39/55) and 58.2% (32/55) of the affected eyes had clear corneal grafts, respectively.There was no statistical significance between Peters anomaly and sclerocornea (P>0.05), while the extent of neovascularization in the limbus had a significant effect on corneal graft transparency, and graft opacity was more likely to occur in patients with vessel area exceeding 2 quadrants (P<0.05).The highest corneal graft transparency was found in children aged 1 to 3 years 80.8%(21/26) (P<0.05), followed by children younger than 6 months (7/15).The translucency rate of the corneal graft was higher in patients undergoing unilateral surgery than in those undergoing bilateral surgery (P<0.05).Translucency of corneal graft was higher in children with simple surgery than with combined surgery (P<0.05), however, cataract surgery after PK had no significant effect on corneal graft transparency (P>0.05).The postoperative complications mainly included immune rejection in 19 eyes (34.5%), complicated cataract in 13 eyes (23.6%), glaucoma in 7 eyes (13.2%), persistent corneal epithelial defect in 7 eyes (13.2%). Conclusions: After PK in children with CCO, the transparent rate of corneal grafts decreases gradually with time, but the long-term translucency of corneal grafts can still be obtained. The range of corneal neovascularization, age at the time of surgery, whether the surgery was binocular and whether the surgery was combined had an effect on the transparency of corneal graft.

Subconjunctival conbercept for the treatment of corneal neovascularization.

To investigate the effectiveness and safety of subconjunctival injection of conbercept in the treatment of corneal neovascularization (CNV). The data on 10 consecutively recruited patients with CNV who received a subconjunctival conbercept (1 mg) once, and measured the area, length, and diameter of neovascularization before and after (1d, 1, 2wk, and 1mo) treatment as well as the occurrence of systemic and ocular complications after treatment were analyzed. There was a statistically significant reduction in the area of CNV one day after treatment (mean±SD: 38.46±11.36 mm2), compared with before treatment (42.46±12.80 mm2, P<0.01). There was also a statistically significant reduction in the length (3.86±1.80 mm vs 4.64±1.77 mm, P<0.01) and diameter (0.044±0.022 vs 0.060±0.026, P<0.05) of CNV, one week after treatment comparing to before treatment. The reduction in all three parameters was maximized at two weeks after treatment (area: 29.49±8.83 mm2, P<0.001; length: 3.50±1.88 mm, P<0.001; and diameter: 0.038±0.017 mm, P<0.01). No severe systemic or ocular complication was observed during the study. During the observation period of one-month, subconjunctival injection of conbercept is an effective and safe method for the reduction of CNV. It may be effective as a preoperative drug for neovascular corneal transplantation.

Effect of topical motesanib in experimental corneal neovascularization model.

This study aimed to compare the efficacy of topical bevacizumab and motesanib in an experimental corneal neovascularization model, and find the most effective motesanib dose. In experiments, 42 Wistar Albino rats were randomly divided into six groups (n = 7). Corneal cauterization was applied to all groups except the group 1. Group 1did not receive any treatment. Topical dimethylsulfoxide was applied toshamgroupthree times a day(tid). Topical bevacizumab drops (5mg/ml) were applied to Group 3 tid. Topical motesanib drops with a dose of 2.5, 5, and 7.5mg/ml were respectively applied in Groups 4, 5, and 6 tid. On the 8th day, corneal photographs of all rats were taken under general anesthesia, and the percentage of corneal neovascular area was calculated. VEGF-A mRNA, VEGFR-2 mRNA, miRNA-21, miRNA-27a, miRNA-31, miRNA-126, miRNA-184, and miRNA-204 were evaluated by the qRT-PCR method in corneas taken after decapitation. The percentage of corneal neovascularization areas and VEGF-A mRNA expression levels were decreased in all treatment groups compared to group 2 (p < 0.05). VEGFR-2 mRNA levels were found to be statistically significantly decreased in groups 4 and 6 compared to group 2 (p < 0.05).Statistically significant changes were detected in the expression levels of only miRNA-126 among all miRNAs. Motesanib with a dose of 7.5mg/ml statistically significantly suppressed the VEGFR-2 mRNA level compared with other treatment doses and may be more effective than bevacizumab. Further, miRNA-126 can be used as a proangiogenic marker.

Effect of Intra-Stromal Bevacizumab on Corneal Vascularization

Purpose: To study the effect of Intra-stromal Bevacizumab on corneal vascularization. Study Design: Quasi experimental study. Place and Duration of Study: Outpatient department of the Combined Military Hospital, Multan from April 2022 to August 2022. Methods: Thirty patients were recruited through non-probability, convenience sampling technique. Ethical approval was taken from Institutional Review Board. Complete ocular examination was performed by a consultant ophthalmologist and patients with corneal neovascularization were included in the study. Area of corneal vascularization was measured on slit lamp. After taking all the aseptic measures and administering anesthetic eye drops, patients were administered 0.2 ml of (2.5 mg/0.1 mL) Bevacizumab (Avastin, Roche) using an ophthalmic microscope. The patient was then called for a follow up after 4 weeks and then the grade of corneal vascularization was checked by a consultant ophthalmologist on slit lamp examination. Data analysis was doneusing SPSS version 25. Results: Mean age of the patients was 43.4 ± 13.7 years (range 25 to 72 years). Male to female ratio was 2:3. There was significant decrease in the area of corneal neovascularization from 2.78mm ± 0.30 to 2.50mm ± 0.27 (p = 0.0003). The mean change in the area of corneal neovascularization was 10.07%. Two patients had sub-conjunctival hemorrhage which settled over due course of time. Conclusion: Intra-stromal Bevacizumab resulted in significant reduction in vascularized area of cornea with minimal side effects indicating that intra-stromal Bevacizumab is an effective and safe modality in the treatment of corneal neovascularization.

Fenofibrate suppresses corneal neovascularization by regulating lipid metabolism through PPARα signaling pathway.

Purpose: The purpose of this study was to explore the potential underlying mechanism of anti-vascular effects of peroxisome proliferator-activated receptor α (PPARα) agonist fenofibrate against corneal neovascularization (CNV) through the changes of lipid metabolism during CNV. Methods: A suture-induced CNV model was established and the clinical indications were evaluated from day 1 to day 7. Treatments of vehicle and fenofibrate were performed for 5days after suture and the CNV areas were compared among the groups. The eyeballs were collected for histological analysis, malondialdehyde (MDA) measurement, terminal deoxynucleotidyl transferase 2'-deoxyuridine 5'-triphosphate nick end labeling (TUNEL) staining, western blot, quantitative real-time PCR (qRT-PCR) assays and immunohistochemical (IHC) staining to elucidate pathological changes and the underlying mechanism. Results: Lipi-Green staining and MDA measurement showed that lipid deposition and peroxidation were increased in the CNV cornea while the expression of long-chain acyl-coenzyme A synthetase 1 (ACSL1), carnitine palmitoyltransterase 1A(CPT1A) and medium-chain acyl-coenzyme A dehydrogenase (ACADM), which are key enzymes of fatty acid β-oxidation (FAO) and targeted genes of peroxisome proliferator-activated receptor alpha (PPARα) pathway, were decreased in CNV cornea. Fenofibrate suppressed lipid accumulation and peroxidation damage in the CNV cornea. Fenofibrate upregulated the expression levels of PPARα, ACSL1, CPT1A, and ACADM compared with vehicle group. IHC staining indicated that fenofibrate also decreased the expression of VEGFa, VEGFc, TNFα, IL1β and CD68. Conclusion: Disorder of lipid metabolism may be involved in the formation of suture-induced CNV and fenofibrate played anti-neovascularization and anti-inflammatory roles on cornea by regulating the key enzymes of lipid metabolism and ameliorating lipid peroxidation damage of cornea through PPARα signaling pathway.

Impaired Autophagy Causes Severe Corneal Neovascularization.

To investigate the role of macrophage autophagy in the process of corneal neovascularization (CNV). In vivo, mice CNV was induced by alkali injury and compared with rapamycin-treated alkaline burn mice. Western blot was used to determine the autophagic status of the macrophages. We quantified the levels of macrophage polarization markers (CD86, INOS, CD163, CD206) by RT-qPCR and measured inflammatory factors through ELISA (IL-6 and TNF-α) in the early phase after injury. In vitro, the human umbilical vein endothelial cells (HUVECs) were co-cultured with macrophage-conditioned medium (MCM) induced by the THP-1 cell line to simulate the neovascular microenvironment. The vascularization capacity of HUVECs was examined using the CCK-8 assay kit, tube formation assay, and scratch wound-healing assay. In vivo, the mRNA expression of Beclin-1 and ATG5 was increased, together with the upregulation of M1 macrophage markers (CD86 and INOS) in corneas after early alkali injury. The area of CNV is effectively relieved in the rapamycin-treated mice. In vitro, upregulation of autophagy level by pretreatment with 3-methyladenine (3-MA) could increase the mRNA expression of the M1 markers. Macrophage-conditioned medium with impaired autophagy contains more IL-6 and TNF-α compared to the M1 macrophage-conditioned medium, promoting HUVEC proliferation, migration, and tube formation capacity. Enhancing the autophagy level with rapamycin (RAPA) could reverse this phenomenon. Impaired autophagy promoted macrophage polarization toward M1 type and increased the expression of IL-6 and TNF-α, which led to severe CNV. Using the autophagy activator (RAPA) could effectively alleviate CNV by promoting autophagy.

Evaluation of effect of nilotinib in an experimental corneal neovascularization model

Aim: This study aims to investigate the neovascularization-inhibiting effect of topical nilotinib and to determine the effective dose of nilotinib.&#x0D; Material and Method: In this study, 42 healthy Wistar albino rats were randomly divided into six groups. The left corneas of all rats except group 1 were cauterized with silver nitrate. Group 1 was the healthy control, with no corneal vascularization, which did not receive any treatment; Group 2 (sham) did not receive treatment, only topical DMSO; Groups 3, 4, and 5 received topical nilotinib at doses of 10, 20, and 40 μM three times a day, respectively; Group 6 received 5 mg/dL topical bevacizumab three times for a day for seven days. On the 8th day, photographs of the corneas were taken, and the percentage of corneal neovascularization area was calculated. Following all rats being killed via anesthesia, the corneas were removed to determine the levels of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) ELISA and corneal immune staining. &#x0D; Results: Other than Group 3, the percentage of neovascular corneal area was lower in the treatment groups compared to Group 2 (p

Dexamethasone and MicroRNA-204 Inhibit Corneal Neovascularization.

This was an in vivo animal study designed to investigate the interaction between dexamethasone (Dex) and microRNA-204 (miR-204) in a mouse alkali burn-induced corneal neovascularization (CNV) model. The function of miR-204 was then investigated in human mammary epithelial cells (HMECs) in vitro. The CNV model was induced by corneal alkali burn in BLAB/c mice. The mice were randomly divided into five groups: normal control (Ctrl), alkali burn-induced corneal injury (Alkali), alkali burn + Dex (Dex), alkali burn + negative control (NTC), and alkali burn + miR-204 agomir (miR-204). Subconjunctival injection of NTC, Dex, or miR-204 agomir was conducted at 0, 3, and 6 days, respectively, after alkali burn. The corneas were collected at day 7 after injury, and the CNV area was observed using immunofluorescence staining. The expression of miR-204 was analyzed with quantitative real time (qRT)-PCR. In HMECs, exogenous miR-204 agomir or antagomir was used to strengthen or inhibit the expression of miR-204. Migration assays and tube formation studies were conducted to evaluate the function of miR-204 on HMECs. At 7 days post-alkali burn, CNV grew aggressively into the cornea. MicroRNA-204 expression was reduced in the Alkali group in contrast with the Ctrl group (P = .003). However, miR-204 was upregulated in the Dex group (vs. alkali group, P = .008). The CNV areas in the NTC and miR-204 groups were 59.30 ± 8.32% and 25.60 ± 2.30%, respectively (P = .002). In vitro, miR-204 agomir showed obvious inhibition on HMEC migration in contrast with NTC (P = .033) and miR-204 antagomir (P = .017). Compared with NTC, miR-204 agomir attenuated tube formation, while miR-204 antagomir accelerated HMEC tube formation (P < .05). The role of Dex in attenuating CNV may be partly attributed to miR-204. MiR-204 may be a potential therapeutic target in alkali burn-induced CNV.

Therapeutic effect of amniotic membrane-fibrin sealant cement on severe ocular surface alkali burn in rabbits

Objective To prepare a biologically active amniotic membrane powder and explore its preservation conditions, and to evaluate the efficacy of the amniotic membrane (AM)-fibrin sealant (FS) cement made from the amniotic powder on the rabbit severe ocular surface alkali burn model. Methods Experimental research. Fresh AM was air-dried, cooled with liquid nitrogen, ground into amniotic powder and sterilized by radiation. The expression of transformed growth factor, nerve growth factor receptor (NGFR), hepatocyte growth factor (HGF), epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) after preparation and 10, 20 and 30 days after storage at room temperature, 4 ℃ and -20 ℃ was tested and compared with that in the fresh AM. The AM-FS cement containing different concentrations of amniotic powder and no amniotic powder was diluted. Rabbit corneal epithelial cells were cultured for 72 hours. The effects of different concentrations of amniotic powder on epithelial cell growth were observed by light microscopy, and the amniotic powder concentration with the largest absorbance value at 450 nm was selected for subsequent animal experiments. Thirty-two right eyes of 32 rabbits as the severe ocular surface alkali burn model were divided using the random counting method into the AM-FS cement group, fresh AM transplantation group, FS group and antibiotic control group (8 rabbits each group) and given different interventions. After weekly observation of corneal repair, hematoxylin and eosin staining, immunohistochemical staining of monocyte chemotaxis protein 1 (MCP-1)and vascular endothelial growth factor (VEGF) were performed and detected by light microscopy at 28 days. The logFC values of the growth factor or receptor expression difference ratio were corrected by BH; the data were analyzed by t-test and analysis of variance. Results: The expression of TGF in the amniotic membrane powder compared with the fresh amniotic membrane group (logFC=-0.11), and the expression of NGFR (HGF, EGF, bFGF) was higher than that of the fresh amniotic membrane group (logFC=-2.07, 0.72, 0.46, 2.62; P<0.05); the expression of HGF, bFGF and EGF in amniotic membrane powder stored for 10 days and 20 days were no lower than fresh amniotic membrane; at 30 days, the expression of growth factors or receptors except HGF and bFGF were decreased, and HGF, bFGF and EGF were no less than 4 ℃ and -20 ℃.The maximum A value was obtained for 0.25 mg/ml of the amniotic membrane powder after 72 hours of the CEC culture 0.98±0.05. The corneal recovery was better in the AM-FS and fresh amniotic membrane transplant groups, with corneal turbidity scores of 3.75±0.46 and 3.50±0.46, respectively, on 28 days, lower than antibiotics (4.29±0.45) (t=2.480, 3.629; P=0.019, 0.001). The corneal neovascular area in the antibiotic control group was compared with the other three groups (t=4.040, 4.339, 2.820; all P<0.001); the corneal neovascular area in the AM-FS group was (9.88±0.20) and (18.96±0.18) mm2 at 7 and 28 days. The corneal neovascularization area at 7 and 28 days in the fresh AM group [(9.54±0.22) and (18.08±0.96) mm2] was smaller than the AM-FS group (t=3.085, 3.017, P=0.005, 0.005). Despite the tiny statistical difference (0.34, 0.88), there was no clinical difference. Hematoxylin and eosin staining showed corneal structures were intact in the AM-FS and fresh AM groups, the epithelial arrangement became normal, and the corneal healing was superior to the FS and antibiotic control groups. Immunohistochemistry showed that the positive expression of VEGF in the fresh AM group was weaker than that in the remaining three groups. MCP-1 was expressed to a similar extent in the AM-FS and fresh AM groups. Conclusions: The active cytokine had high expression and stable properties at room temperature. The AM-FS cement containing 0.25 mg/ml amniotic powder can promote the repair of corneal epithelium, reduce inflammatory reaction and corneal neovascularization after alkali burning in rabbit eyes.

MCC950 attenuates inflammation-mediated damage in canines with Staphylococcus pseudintermedius keratitis by inhibiting the NLRP3 inflammasome

BackgroundBacterial keratitis is a common eye disease in dogs and can seriously affect vision. This study investigated the anti-inflammatory effect of MCC950 in the cornea of canines infected with Staphylococcus pseudintermedius (S. pseudintermedius). MethodsIn vitro, canine cornea epithelial cells were pretreated with MCC950 and PDTC and then infected with S. pseudintermedius. The key proteins of the NF-κB pathway and NLRP3 inflammasome were detected by Western blotting, the levels of inflammatory factors were detected by qPCR, and the levels of MDA and LDH were detected by assay kit. In vivo, the canine keratitis model was established by injecting S. pseudintermedius into the corneal stroma layer. After treatment with MCC950, slit-lamp examinations were performed. Cornea tissue protein and RNA were extracted, and Western blotting was used to detect key proteins of the NF-κB pathway and NLRP3 inflammasome. qPCR was used to detect the inflammatory factors. Paraffin sections of corneal tissue were prepared for HE staining and immunohistochemical staining. ResultsAfter MCC950 treatment, the expression levels of key proteins in the NF-κB pathway and NLRP3 inflammasome in canine cornea epithelial cells and corneal tissues were decreased, and the expression levels of IL-1β, IL-6, IL-8, IL-18 and TNF-α were reduced. Cellular MDA and LDH levels were decreased. In vivo, the degree of corneal opacity, edema, neovascularization and corneal injury area decreased after MCC950 treatment. Canine corneal sections showed that MCC950 attenuated neutrophil infiltration. ConclusionMCC950 alleviates the inflammatory response to canine keratitis caused by S. pseudintermedius by inhibiting the activation of the NLRP3 inflammasome.

The Effect of Bromfenac Sodium Nanopolymer Used in Anterior Segment of the Eye on Corneal Neovascularization.

This study was to explore the inhibitory effect of bromfenac sodium (BF) / chitosan (CS) nanoparticles (NPs) on corneal neovascularization (CNV). 45 New Zealand white rabbits provided by The First Affiliated Hospital of Jinan University were randomly divided into a control group (group A, n = 15), 0.1% BF aqueous solution treatment group (group B, n = 15), and 0.1% BF/CS-NPs suspension treatment group (group C, n = 15). A rabbit corneal alkali burn model was established. The average particle size of BF/CS-NPs with different BF concentrations was mainly 341.6 ± 12.9 nm - 548.7 ± 15.4 nm; and the Zeta potential distribution was 24.3 ± 2.5 mV - 35.7 ± 4.3 mV. When the initial concentration of BF was 1.5 mg/mL, the maximum drug loading was 57.35 ± 5.26%. The area of CNV in group C was significantly lower than that in groups B and A, and the differences were statistically significant (P < 0.05). At 6, 12, 18, and 24 days after surgery, the mRNA expression levels in cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) gene were compared after standardized by β-actin; group A had the highest expression level, followed by group B, and group C had the lowest expression level, showing statistically significant differences (P < 0.05). The BF/CS-NPs granules prepared in this study had stable physical and chemical properties and had a good sustained-release effect, and the release duration can be as long as 48 hours. BF/CS-NPs can inhibit the formation of CNV at different time points after alkali burn, and reduce the expression of VEGF and COX-2 in corneal tissue after alkali burn.

Inhibition of corneal neovascularization by topical application of nintedanib in rabbit models.

To evaluate the potential efficacy and mechanisms of nintedanib in corneal neovascularization (NV) in rabbit models. Corneal NV was induced using 1 mol/L NaOH. Rabbits (n=21) were randomized to 3 groups: Group 1 were treated with 0.9% NaCl, Group 2 with Avastin (5 mg/mL), and Group 3 with nintedanib (1 mg/mL). All treatments started 1d after alkaline burns and were topically performed 3 times a day for 2wk. Photographs were taken on a slit lamp microscope on day 7 and 14. The NV area, the length of the vascularization and angiogenesis index (AI) were used to evaluate the corneal NV. On day 14, the immunohistochemical (IHC) studies of the cornea were examined. Western blot was performed to test the expression levels of vascular endothelial growth factor (VEGF), Akt, p-Akt, P38, p-P38, MMP-2 and MMP-9. The corneal NV area, vessel length and AI in Group 3 were significantly lower than Group 2, with both being lower than Group 1. IHC staining showed that VEGF was significantly overexpressed in the epithelium and stroma of cornea following alkaline burns. In contrast, the level of VEGF was significantly suppressed in both Group 2 and Group 3. Western blot results further confirmed that, compared with Group 1, Group 3 had significantly reduced expressions of VEGF, Akt, p-Akt, p-P38, MMP-2, and MMP-9 in corneal tissues. Trends of lower levels of MMP-2, AKT, and p-AKT in Group 3 than Group 2 were identified. Nintedanib and Avastin can effectively inhibit corneal NV, with P38 MAPK and AKT signaling pathways being possibly involved. Nintedanib seems more effective than Avastin and has the potential to be a novel therapy for preventing corneal NV.

Inhibiting corneal neovascularization by sustainably releasing anti-VEGF and anti-inflammation drugs from silica-thermogel nanohybrids

Corneal neovascularization (CNV) is one of the main factors that induce blindness worldwide. To effectively inhibit CNV, a novel nanohybrid has been developed by incorporating anti-VEGF bevacizumab (BEV)-loaded mesoporous silica nanoparticles (BEV@MSN) into the thermogel matrix with anti-inflammation cyclosporine A (CsA) (BEV@MSN-CsA@Thermogel). This nanohybrid regulates the in vitro release of both bevacizumab and cyclosporine A in a sustainable way for up to four weeks to enhance CNV inhibition through the synergistic anti-VEGF and anti-inflammation. The carrier materials (i.e. silica and thermogel) in this nanohybrid do not show any cytotoxicity to human Tenon's fibroblasts, corneal epithelial cells and corneal endothelial cells. BEV@MSN-CsA@Thermogel effectively prevents proliferation, migration, and tube-like structure formation of human umbilical vein endothelial cells. Moreover, subconjunctival injection of BEV@MSN-CsA@Thermogel significantly inhibits corneal neovascularization in terms of the CNV area, the new vessel length, the corneal opaque area, the corneal inflammation and abnormal fibrosis in a rabbit model. This nanohybrid is thus a promising alternative for effective CNV treatment.

Intrastromal versus subconjunctival anti-VEGF agents for treatment of corneal neovascularization: a rabbit study.

To determine whether subconjunctival or intrastromal administration of anti-VEGF agents is more effective on suture-induced corneal neovascularization (CoNV) in rabbits. CoNV was induced in 48 eyes of 24 New Zealand white rabbits by using an 8/0 silk suture. On the 7th day after suturing, the rabbits were divided into four treatment groups as follows: six rabbits received subconjunctival bevacizumab (group 1), six rabbits received subconjunctival aflibercept (group 2), six rabbits received intrastromal bevacizumab (group 3) and six rabbits received intrastromal aflibercept (group 4). On the 7th and 14th days after suturing, the CoNV area was calculated by standardised analysis of photographs using the Image-J program. On the 14th day after suturing, all rabbits were sacrificed and then corneal tissue was harvested for the analysis of vascular endothelial growth factor (VEGF)-A, VEGF-B and placental growth factor (PIGF) levels. On the 7th day after suturing, CoNV areas were 17.10 ± 2.98, 18.88 ± 3.78, 17.36 ± 4.52, 18.57 ± 4.16 and 17.31 ± 2.81 mm2 in the groups 1-4 and control group, respectively. On the 7th day after intervention and removal of suture, CoNV areas were 4.85 ± 1.99, 6.66 ± 1.73, 2.83 ± 1.08, 2.63 ± 1.16 and 11.93 ± 2.64 mm2 in the group 1-4 and control group, respectively. CoNV area was reduced by 88.1% and 82.5% in eyes receiving intrastromal aflibercept and bevacizumab, respectively (both p < 0.001), and by 64.5% and 69.9% in eyes receiving subconjunctival aflibercept and bevacizumab, respectively (both p = 0.001). Intrastromal anti-VEGF therapy regressed CoNV more effectively than subconjunctival therapy regardless of the type of anti-VEGF agent.

Descemet Membrane Endothelial Keratoplasty in Vascularized Eyes: Outcome and Effect on Corneal Neovascularization

To report the outcomes after Descemet membrane endothelial keratoplasty (DMEK) in vascularized eyes. Consecutive cases of DMEK in vascularized eyes (involving ≥2 vascularized quadrants) were selected from a prospective database. Best corrected visual acuity, endothelial cell density (ECD), central corneal thickness, corneal transplant rejection episode, graft survival, and area of neovascularization (quantified using image analysis software) were evaluated. In this study, 24 eyes of 24 patients were selected [mean age, 65.0 years; mean follow-up duration, 14.8 months (6-36 months)], which consists of 14 vascularized eyes after failed penetrating keratoplasty and 10 vascularized eyes with bullous keratopathy. Best corrected visual acuity improved from 1.60 ± 1.02 LogMAR preoperatively to 0.47 ± 0.37 LogMAR 12 months postoperatively (P < 0.001). Central corneal thickness decreased from 824 ± 193 μm preoperatively to 544 ± 48 μm 12 months postoperatively (P = 0.001). The donor ECD decreased from 2272 ± 723 cells/mm2 preoperatively to 1570 ± 279 cells/mm2 12 months postoperatively. The total loss of ECD at the last visit was 40.7% ± 13.0%. Eight of 24 eyes (33.3%) required rebubbling, which resulted in final attachment. The corneal neovascularization area significantly regressed from 4.68% ± 3.26% preoperatively to 2.28% ± 1.58% (n = 18, P = 0.021). Corneal transplant rejection episodes occurred in 1 eye of 24 patients (4.2%). There was no primary graft failure. DMEK is a feasible option to treat endothelial dysfunction in vascularized eyes.

DCZ3301, an aryl-guanidino agent, inhibits ocular neovascularization via PI3K/AKT and ERK1/2 signaling pathways

Neovascularization is a critical process in the pathophysiology of neovascular eye diseases. Although anti-VEGF therapy has achieved remarkable curative effects, complications, limited efficacy and drug resistance remain the prominent problems. DCZ3301, an aryl-guanidino compound, was reported to have anti-tumor activity in the previous studies. Here, we demonstrated the effects of DCZ3301 on human umbilical vein endothelial cell (HUVEC) in vitro, and performed choroid microvascular sprouting assay ex vivo and alkali-burn induced corneal neovascularization mouse model in vivo. We found that DCZ3301 inhibited the proliferation, migration, and tube formation of HUVECs, while inducing the spontaneous apoptosis of HUVECs by suppressing the activation of PI3K/AKT and ERK1/2 pathways. Furthermore, DCZ3301 inhibited the choroid microvascular sprouting, diminished the area of corneal neovascularization and attenuated the edema of corneal stroma after alkali burn. Together, these results suggested that DCZ3301 exerted anti-angiogenic properties, and might be regarded as a potential candidate for ocular neovascularization.