Vitreous Samples Research Articles

The Association of Intraocular Efavirenz Concentrations and HIV-1 Viral Load Among Persons With HIV.

Efavirenz (EFV) is commonly used in combination antiretroviral therapy. However, in our previous study, many persons living with HIV exhibited ocular complications despite undergoing effective combination antiretroviral therapy. Here, we aimed to determine the intraocular EFV concentrations in the vitreous and analyze the factors affecting viral load in the vitreous in patients with HIV-associated retinopathies. Observational, retrospective study. Fourteen patients receiving EFV in combination with an antiretroviral therapy who underwent pars plana vitrectomy were enrolled between January 2019 and August 2022. The patients were divided into 2 groups based on presence or absence of retinal detachment (RD). Patient characteristics and HIV-1 RNA levels in plasma and vitreous were recorded during pars plana vitrectomy. Paired blood plasma and vitreous samples were obtained for EFV concentration analysis using ultra-high-performance liquid chromatography/tandem mass spectrometry. The median age of the enrolled patients was 48 years (interquartile range, 32.25-53.25), including 12 men and 2 women. Median vitreous and plasma EFV concentrations were 141.5 (interquartile range, 69.63-323.75) and 2620 ng/mL (1680-4207.5), respectively. Median ratio of vitreous/plasma EFV concentrations in the paired samples among all participants was 0.053 (0.018-0.118). Median vitreous/plasma EFV concentrations significantly differed between the non-RD and RD groups (0.04 vs 0.12, P = 0.042). The vitreous EFV concentrations were insufficient to inhibit viral replication in intraocular tissues, which may be because of poor penetration of the blood-retinal barrier. High vitreous EFV concentrations were associated with RD, indicating a correlation between the EFV concentration and the severity of blood-retinal barrier disruption. It implied that EFV was not a suitable antiviral drug to inhibit HIV-1 replication in ocular tissues.

Surgical therapy and pathogen detection in endogenous endophthalmitis : Analysis of data from five German eye hospitals

Endogenous endophthalmitis results from hematogenous spread of bacterial or fungal infection in severely diseased patients. Specific systemic and intraocular therapy is required. The basis for this treatment is causal pathogen detection in blood culture or vitreous sample. However, functional results are limited. The current article provides practical hints for surgical therapy and pathogen detection in patients with endogenous endophthalmitis. Aretrospective analysis of anonymous data of 68male and female patients from 2018-2023 from five ophthalmology clinics in Germany was performed. Mean age of affected patients was 71.4years (31-96years). Surgical therapy included pars plana vitrectomy (ppV) and intravitreal injection (IVOM). In 44of 68patients (65%), 1-3 surgeries were performed, 4-6surgeries were required in 14/68 (21%) of patients, and 10or more surgeries were required in 4/68patients (6%). Pathogen detection was possible in 34% of vitreous specimens and in 11% of anterior chamber samples. Mean initial visual acuity was logMAR1.5. After treatment and amean follow-up of 2.5months, mean visual acuity was logMAR1.3. Preanalytical methods for specimen collection like the Freiburg endophthalmitis set to optimize pathogen detection are presented. Severe inflammatory intraocular reactions in endogenous endophthalmitis necessitate acombination of ppV and repeated IVOM. In addition to providing avitreous sample, ppV also serves to remove inflammatory fibrin membranes. Early pars plana vitrectomy with specific antibiotic or antifungal therapy should be sought in addition to the focus search and systemic therapy.

Serum and Vitreous Levels of Placenta Growth Factor in Diabetic Retinopathy Patients: Correlation With Disease Severity and Optical Coherence Tomographic Parameters.

Purpose The primary objective of this study was to compare placenta growth factor (PlGF) levels in the serum and vitreous of diabetic retinopathy (DR) patients to non-diabetic controls. Additionally, the study aimed to establish associations between serum and vitreous PlGF concentrations and to examine the correlation between vitreous PlGF in DR patients and morphological parameters. Methods This study included serum and vitreous samples from 38 patients, including 21 patients with DR and 17 non-diabetic controls. The control group included non-diabetic patients with rhegmatogenous retinal detachment with retinal tears secondary to posterior vitreous detachment or trauma. PlGF levels were quantified in vitreous and serum samples using an enzyme-linked immunosorbent assay (ELISA). Optical coherence tomography (OCT) scans from DR patients were evaluated to measure the central retinal thickness (CRT) and macular volume (MV). Results DR patients had significantly higher mean vitreous PlGF levels compared to non-DR patients (70.0±39.2 vs. 46.47±9.7 pg/mL, p-value=0.004). However, no significant increase in mean serum PlGF levels was observed in DR patients (p-value=0.232). Within the DR group, proliferative DR (PDR) patients presented significantly higher vitreous PlGF levels than non-PDR (NPDR) patients (76.5±41.0 vs. 42.5±5.0 pg/mL, p-value=0.009). There was no association between serum and vitreous PlGF levels. The correlation between vitreous PlGF levels and morphological parameters was rsp=0.175, p-value=0.488 for CRT, and rsp=0.288, p-value=0.262 for MV. Conclusion This study emphasizes the important role of PlGF in neovascularization, specifically highlighting its overexpression exclusively in vitreous from PDR patients. The observed increase in PlGF levels may be indicative of disease severity. The lack of correlation between vitreous and serum PlGF levels suggests a potential dissociation between intravitreal and systemic PlGF synthesis. Consequently, targeting PlGF in therapeutic approaches may offer an additional strategy for ocular pathologies with a neovascular component.

Revealing new insights: Two-center evidence of microplastics in human vitreous humor and their implications for ocular health

Microplastics (MPs), an emerging environmental contaminant, have raised growing health apprehension due to their detection in various human biospecimens. Despite extensive research into their prevalence in the environment and the human body, the ramifications of their existence within the enclosed confines of the human eye remain largely unexplored. Herein, we assembled a cohort of 49 patients with four ocular diseases (macular hole, macular epiretinal membrane, retinopathy and rhegmatogenous retinal detachment) from two medical centers. After processing the samples with an optimized method, we utilized Laser Direct Infrared (LD-IR) spectroscopy and Pyrolysis Gas Chromatography/Mass Spectrometry (Py-GC/MS) to analyze 49 vitreous samples, evaluating the characteristics of MPs within the internal environment of the human eye. Our results showed that LD-IR scanned a total of 8543 particles in the composite sample from 49 individual vitreous humor samples, identifying 1745 as plastic particles, predominantly below 50 μm. Concurrently, Py-GC/MS analysis of the 49 individual samples corroborated these findings, with nylon 66 exhibiting the highest content, followed by polyvinyl chloride, and detection of polystyrene. Notably, correlations were observed between MP levels and key ocular health parameters, particularly intraocular pressure and the presence of aqueous humor opacities. Intriguingly, individuals afflicted with retinopathy demonstrated heightened ocular health risks associated with MPs. In summary, this research provides significant insights into infiltration of MP pollutants within the human eye, shedding light on their potential implications for ocular health and advocating for further exploration of this emerging health risk.

TIMP3/Wnt axis regulates gliosis of Müller glia

BackgroundPrevious studies have confirmed the expression of tissue inhibitor of metalloproteinase-3 (TIMP3) in Müller glia (MG). However, the role of TIMP3 in MG remains unknown. MethodsA mouse model of laser-induced retinal damage and gliosis was generated using wild-type C57BL/6 mice. TIMP3 and associated proteins were detected using Western blotting and immunofluorescence microscopy. RNA sequencing (GSE132140) of mouse laser-induced gliosis was utilized for pathway analysis. TIMP3 overexpression was induced in human MG. Human vitreous samples were obtained from patients with proliferative diabetic retinopathy (PDR) and healthy controls for protein analysis. ResultsTIMP3 levels increased in mouse eyes after laser damage. Morphology and spatial location of TIMP3 indicated its presence in MG. TIMP3-overexpressing MG showed increased cellular proliferation, migration, and cell nuclei size, suggesting TIMP3-induced gliosis for retinal repair. Glial fibrillary acidic protein (GFAP) and vimentin levels were elevated in TIMP3-overexpressing MG and laser-damaged mouse retinas. RNA sequencing and Western blotting suggested a role for β-catenin in mediating TIMP3 effects on the retina. Human vitreous samples from patients with PDR showed a positive correlation between TIMP3 and GFAP levels, both of which were elevated in patients with PDR. ConclusionsTIMP3 is associated with MG gliosis to enhance the repair ability of damaged retinas and is mediated by the canonical Wnt/β-catenin. Changes in TIMP3 could potentially be used to control gliosis in a range of retinal diseases However, given the multifaceted nature of TIMP3, care must be taken when developing treatments that aim solely to boost the function of TIMP3. FundingNational Cheng Kung University Hospital, Taiwan (NCKUH-10604009 and NCKUH-11202007); the Ministry of Science and Technology (MOST 110-2314-B-006-086-MY3).

THE CLINICAL VALUE OF β-D-GLUCAN TESTING AND NEXT-GENERATION METAGENOMIC SEQUENCING FOR THE DIAGNOSIS OF FUNGAL ENDOPHTHALMITIS.

To explore the clinical value of β-D-glucan (BDG) testing and metagenomic next-generation sequencing (mNGS) for detecting the pathogens of fungal endophthalmitis (FE). This study included 32 cases (32 eyes) with FE and 20 cases (20 eyes) with intraocular inflammation caused by other etiologies. All patients underwent extraction of aqueous humor or vitreous fluid samples for BDG testing and mNGS. The diagnostic performance and total clinical concordance rate of BDG testing and mNGS for FE were evaluated and calculated based on the results of the clinical diagnosis. Among the clinically diagnosed FE, the positivity rates of BDG testing and mNGS (90.63%) were both significantly higher ( P < 0.001) than that of microbial cultures (53.13%). There was 100% consistency in pathogen identification using mNGS and culture identification for culture-positive cases. The area under the curve was 0.927 for BDG testing and 0.853 for mNGS. When the two tests were combined, sensitivity (93.75%), specificity (100.00%), and total clinical concordance rate (96.15%) were all improved, compared with the single tests. The positive rates of BDG test and mNGS were markedly higher than those of cultures in FE identification. The combination of these two tests showed improved performance when compared with individual tests.

CircRNA expression profiles and regulatory networks in the vitreous humor of people with high myopia

Myopia is a global health and economic issue. Circular RNAs (circRNAs) have been shown to play an important role in the pathogenesis of many ocular diseases. We first evaluated the circRNA profiles and possible roles in vitreous humor samples of individuals with high myopia by a competitive endogenous RNA (ceRNA) array. Vitreous humor samples were collected from 15 high myopic (5 for ceRNA array, and 10 for qPCR) and 15 control eyes (5 for ceRNA array, and 10 for qPCR) with idiopathic epiretinal membrane (ERM) and macular hole (MH). 486 circRNAs (339 upregulated and 147 downregulated) and 264 mRNAs (202 upregulated and 62 downregulated) were differentially expressed between the high myopia and control groups. The expression of hsa_circ_0033079 (hsa-circDicer1), hsa_circ_0029989 (hsa-circNbea), hsa_circ_0019072 (hsa-circPank1) and hsa_circ_0089716 (hsa-circEhmt1) were validated by qPCR. Pearson analysis and multivariate regression analysis showed positive and significant correlations for axial length with hsa-circNbea and hsa-circPank1. KEGG analysis showed that the target genes of circRNAs were enriched in the mTOR, insulin, cAMP, and VEGF signaling pathways. GO analysis indicated that circRNAs mainly targeted transcription, cytoplasm, and protein binding. CircRNA-associated ceRNA network analysis and PPI network analysis identified several critical genes for myopia. The expression of circNbea, circPank1, miR-145-5p, miR-204-5p, Nras, Itpr1 were validated by qPCR in the sclera of form-deprivation myopia (FDM) mice model. CircPank1/miR-145-5p/NRAS and circNbea/miR-204-5p/ITPR1 were identified and may be important in the progression of myopia. Our findings suggest that circRNAs may contribute to the pathogenesis of myopia and may serve as potential biomarkers.

Development and application of targeted multiplex polymerase chain reaction to help diagnose infectious uveitis

Abstract&#x0D; Introduction &amp; Objectives : Clinical pattern recognition is paramount in uveitis diagnosis. Pathogen detection from ocular fluid samples is often necessary to support infectious uveitis diagnosis, particularly in cases presenting with atypical clinical appearance. This study aims to present the process of developing targeted multiplex PCR and its application in infectious uveitis.&#x0D; Methods : This was a cross sectional study to evaluate the diagnostic performance of targeted multiplex PCR in infectious uveitis. We obtained ocular fluid samples and reviewed medical records of uveitis patients who underwent ocular fluid analysis at Cipto Mangunkusumo Hospital from February 2022 to March 2023. PCR detection threshold values (DNA copies/mL) were 10.9 for Mycobacterium tuberculosis (Mtb), 672 for Epstein-Barr virus, 4.77 for Cytomegalovirus, 6.37 for Toxoplasma gondii, and 5.53 for Herpes simplex virus. With every two-fold increase of pathogen selection, this method requires a half volume of extracted DNA template from aqueous/vitreous samples than uniplex PCR. The ophthalmologist selected the pathogen combination to be detected, allowing for a tailored examination.&#x0D; Results : Forty-seven aqueous or vitreous samples were analyzed. The positivity rate was 23.4% (11/47) with Mtb yielded the highest positivity (7/34; 20.6%). With final diagnosis as a reference, targeted multiplex PCR resulted in 32.3% sensitivity, 93.8% specificity, 90.9% positive predictive value and 19.2% negative predictive value.&#x0D; Conclusion : With its high specificity, targeted multiplex PCR is useful as a confirmatory but not screening tool in uveitis diagnosis. Ocular fluid analysis is an important part of stepwise diagnostic approach in uveitis.

Triple Gram-negative bacterial endophthalmitis following intravitreal injection

PurposeTo describe a puzzling case of endophthalmitis caused by three unusual bacteria after intravitreal injection, its outcome, and underlying questions.FindingsA 70-year-old female patient was diagnosed with acute endophthalmitis following intravitreal aflibercept injection for age-related macular degeneration. A standard tap and inject procedure was performed. Microbiological analyses on the anterior chamber and vitreous samples yielded the presence of three non-fermenting Gram-negative rods: Pseudomonas stutzeri, Stenotrophomonas maltophilia, and Ochrobactrum anthropi. The outcome was favorable after intravitreal injections of vancomycin and ceftazidime, with an almost complete recovery of the visual acuity to its baseline level. No potential source of infection was identified.ConclusionEndophthalmitis following intravitreal injection can be caused by a wide variety of bacteria, including some rare Gram-negative species. They can sometimes co-exist in a single patient, but their virulence may vary greatly. Due to the variable antibiotic susceptibility and frequent multiresistance associated with non-fermenting Gram-negative rods, a prompt microbiological approach is required. Favorable outcome can be achieved with standard management.

Long-term follow-up of a healthy man with endogenous Streptococcus anginosus endophthalmitis

We report the long-term follow-up of an immunocompetent patient who presented with slowly progressive endogenous endophthalmitis secondary to Streptococcus anginosus. A 46-year-old healthy man presented with a two-month history of right eye iritis. On examination, visual acuity was 20/60 with intraocular pressure of 6 mm Hg. There was a small layer of hypopyon with non-granulomatous anterior uveitis and vitritis. On funduscopy, fluffy white peripheral retinal and pre-retinal lesions were noted in superonasal periphery. The patient denied any present or past illness. Diagnostic pars plana vitrectomy was performed. Culture and polymerase chain reaction of the vitreous sample were positive for Streptococcus anginosus. Intravitreal vancomycin and ceftazidime and systemic ceftriaxone were administered. Work-up which included blood and urine cultures, chest x-ray, echocardiography and abdominal ultrasound was unyielding. Subsequently and because of persistent post-infectious inflammatory reaction, intravitreal and oral steroids were administered in addition to oral azathioprine later on. After one year of follow-up, visual acuity was 20/20 with near vision of Jaeger 3 + and no signs of active uveitis were seen. Therefore, Streptococcus anginosus should be considered in the differential diagnosis of a slowly progressive endophthalmitis also in immunocompetent individuals.

The value of diagnostic vitrectomy: Histocytopathology techniques for the diagnosis of lymphoma of the retina

Abstract Purpose: Primary large B-cell lymphoma of the retina, vitreous, and central nervous system (CNS), is an intraocular tumor with an unspecific and insidious clinical presentation. Pars plana vitrectomy (PPV) and optical coherence tomography (OCT) are useful diagnostic tools for this malignant process. The aim of this study is to evaluate the diagnostic efficacy of PPV for these intraocular lesions under a modified diagnostic protocol with a clinical pathological correlation with OCT imaging. Materials and Methods: A total of 115 samples were collected after a vitrectomy procedure (aspiration or vitrectomy cassette). The samples were centrifuged, and the precipitates were collected. A cell block was prepared and analyzed with multiple stains and an immunohistochemistry (IHC) panel, including B- and T-cell markers, as well as light chain markers, to establish the monoclonal nature of the tumor. Of the 115 samples, 9 (7.83%) were diagnosed with large B-cell lymphoma of the retina, vitreous, and CNS. Conclusion: Diagnostic vitrectomies for the large B-cell lymphoma of the retina, vitreous, and CNS is an excellent tool for the diagnosis of this entity. A negative diagnostic PPV with a strong suspicious OCT image, where the neoplastic cells are located between the retinal pigmented epithelium and Bruch’s membrane, the latter procedure should be either repeated or a chorioretinal biopsy be performed. In contrast, a positive vitrectomy using the IHC panel for large B-cell lymphoma of the retina, vitreous, and CNS is pathognomonic of this condition. In addition, the OCT is an important tool to help in the diagnosis of this difficult entity. Results: The diagnostic PPV provided suitable vitreous samples to all patients with undetermined uveitis and/or intraocular tumor suspicion. A morphological and immunohistichemical (IHC) analysis enabled a conclusive diagnosis of retina, vitreous and CNS lymphoma in all patients submitted to the procedure.

Endogenous Candida endophthalmitis: diagnostics and treatment options based on case studies

Endogenous Candida endophthalmitis is an emergency that can threaten vision and the eye as awhole organ but also the life of the patient due to its systemic cause. Retrospective case evaluation of the University Hospital for Ophthalmology Halle (Saale) and the Eye Clinic of the Ernst von Bergmann Hospital Potsdam from 2017-2022. (Age, gender, side involvement, underlying diseases, symptoms, preoperative and postoperative visual acuity, diagnostics, treatment and complications). The standardized procedures for endogenous Candida endophthalmitis are explained, the data are compared with the literature and treatment suggestions are presented. 8patients with 14eyes were treated for endogenous Candida endophthalmitis. Of the patients 2 were women and 6 were men. The overall average age was 70.25 years (53-82 years), 6 patients had bilateral eye involvement and 2 patients were affected on one side. All patients had several serious immunocompromising underlying diseases that were the cause of the candidemia. All patients underwent apars plana vitrectomy (ppV) on the affected eyes except for one male patient who did not obtain permission for anesthesia. In addition to systemic treatment with antimycotics, voriconazole was routinely administered intravitreally during ppV. The patients received further intravitreal doses of voriconazole based on the findings. Vitreous body samples were taken from all patients during the ppV and detection of Candida albicans was possible in all cases. As part of the local perioperative treatment 1patient received parabulbar administration of atriamcinolone depot in both eyes, 7patients received adexamethasone depot in 11eyes and all 8patients received findings-oriented local treatment with prednisolone acetate eye drops. visual acuity increased from preoperatively 1.2logMar to 1.0logMar. Postoperative retinal detachment did not occur and there were no serious perioperative complications. 2patients died from one of the underlying diseases after 3 and 25months, respectively. Despite an increasing number of case series, there are still no uniform guidelines for ophthalmologists in Germany. There is agreement regarding systemic treatment and the intravitreal administration of antifungal agents. The role of ppV has not yet been clearly defined and the question of the adjuvant use of steroids (systemic and/or local) has also not been conclusively clarified.

Estimation of Time Since Death by Potassium ion Level in the Vitreous Fluids: A Postmoterm Study in a Tertiary Care Center, South India

Background: It is of great value to find time since death in the medicolegal investigations like serious crimes.Vitreous humor was more often used for the biochemical analysis as it is less contaminated or not affected by the putrefactive changes than other fluids of the body. It was stated by many researchers that potassium levels in the vitreous humour is more accurate in predicting time since death.Aim: The aim of the study is to assess the time since death using potassium level in the vitreous humorMethodology: This study includes 54 cases of victims brought to the tertiary care hospital based on inclusion and exclusion criteria. Cases with known time of death and with clear vitreous humor samples were included. Baseline characteristics like name, age, gender, exact time since death was collected. Vitreoushumor was collected through appropriate standardized technique. Sampling time was also noted. The collected data was entered in MS excel and analysis done in SPSS 23 software. P value &lt;0.05 is considered to be significant.Results: Majority of the study participants belongs to 57-67 years of age group (28%) followed by 27-37 years of age (22%).Male predominance was observed in our study 41(76%). There is a linear correlation found between the vitreous potassium concentration and the postmortem interval (R2=0.907). The rate of the vitreous potassium increase was 0.36 meq/hr (Coefficient of regression =2.76 meq/l/hr)Conclusion: There is a linear relationship found between vitreous potassium concentration and time since death.Thus potassium in the vitreous can be used for assessing the time since death.

Role of heparin‐binding mediators in the resistance to anti‐VEGF therapies in diabetic retinopathy

Aims/Purpose: Anti‐vascular endothelial growth factor (VEGF) drugs represent the standard of care for diabetic retinopathy (DR); however, lack of response occurs in approximately 50% of patients. Aim of our work is the identification of the mediators responsible for anti‐VEGF drug resistance.Methods: Vitreous samples were collected from anti‐VEGF non‐responder (non‐RES) patients; non‐RES heat‐inactivated (HI) vitreous were used as control. To develop a new orthotopic model of DR, non‐RES or HI vitreous were injected intravitreally in mice and after 48 h retinas were explanted and analysed. The composition of non‐RES vitreous was assessed by an angio‐inflammatory protein array. The effect of the inhibition of the bioactive heparin‐binding mediators (HBMs) present in non‐RES vitreous was evaluated in the endothelial spheroid sprouting assay.Results: Mouse orthotopic intravitreal injection of non‐RES vitreous upregulated the expression of pro‐angiogenic/pro‐inflammatory mediators; moreover, it induced alterations of retinal vessels, accumulation of inflammatory infiltrate, and activation of astrocyte gliosis. Notably, HI vitreous was ineffective, pointing to a proteinaceous nature of the non‐RES vitreal mediators acting on mouse retinae. In this context, the blockade of vitreal HBMs mediated by the biotechnological heparin‐like molecule K5‐N, OS (H) or by the pentapeptide Boc2, completely abrogated the activity of non‐RES samples. It is worth noticing that the potency of anti‐HBP compounds was significantly higher than the anti‐VEGF ranibizumab, which was partially effective only at the clinically relevant and saturating concentration of 10 μM, with no further improvement at a double dose.Conclusions: Our data show that VEGF is only part of a complex machinery regulating angiogenesis, inflammation and oedema formation during DR and that drug resistance is due to the activation of compensatory mechanisms mediated by HBMs besides VEGF.

Acute zonal occult outer retinopathy as a luetic ocular manifestation

Aims/Purpose: Based on a clinical case seen in our setting, the purpose of this communication is to describe a case of ocular syphilis with an uncommon initial manifestation as well as its clinical and imaging features, diagnosis, treatment and evolution.Methods: We selected a case seen at the University Hospital Infanta Sofía in 2023. A 54‐year‐old male with Acquired Inmune Deficiency Syndrome (AIDS) under antiretroviral (ARV) treatment, diagnosed with cataract, who 1 month after surgery presented a paradoxical worsening of baseline visual acuity (VA) bilaterally accompanied by nyctalopia.Results: During the exam, we found VA from right eye (RE) of 0.03 and from left eye (LE) of 0.03. Fundus examination showed a bilateral central vitreous organization. Macular OCT from both eyes showed preserved foveal profile with pachychoroid associated with patchy absence of external limiting membrane, evident focal loss of ellipsoid zone and cysts in outer layers of the RE. Campimetry with practically abolished fields, neuroimaging study, complete serology including antirecoverin antibodies, and vitreous sample for microbiology were performed. From this study, it was found that the patient had a positive luetic seroconversion within a maximum period of five years. With these data, we oriented the differential diagnosis of the described retinopathy towards an infectious or toxic aetiology due to ARV drugs as the most probable causes. The patient was treated with intravenous penicillin with great improvement of vision and nyctalopia.Conclusions: Ocular syphilis is the great simulator par excellence and although infrequent, it can debut with atypical forms of uveitis.

Evaluating the long-term biological stability of cytokine biomarkers in ocular fluid samples

PurposeThe quality of biological fluid samples is vital for optimal preanalytical procedures and a requirement for effective translational biomarker research. This study aims to determine the effects of storage duration...

Correlation Between Vitreous Level of Angiogenic Growth Factors and Oxygen Saturation in Retinal Vessels in Diabetic Retinopathy.

The aim of this study was to investigate whether there is any association between the levels of the angiogenic growth factors and the vascular oxygen saturation in eyes with diabetic retinopathy. The study was designed as a prospective trial. The cohort consisted of 29 diabetic patients with scheduled vitreous procedures (intravitreal injection or pars plana vitrectomy). The control group included 30 patients scheduled for macular surgery (macular hole or epiretinal membrane). Nine patients (four from the diabetic maculopathy [DM] group and five from the control group) were excluded from the study because of unsuccessful vitreous samples. Retinal oximetry was performed several hours before the vitreous procedure was performed, and vitreous samples were obtained during the procedure. The concentrations of VEGF, Serpin F1/pigment epithelium-derived factor (PEDF), and placental growth factor (PlGF) were measured by ELISA. A negative correlation between level of VEGF and arteriovenous (AV) saturation difference was determined in the DM group (Pearson correlation coefficient r = -0.607; two-tailed test, P = 0.002). Also a negative correlation between level of PlGF and AV saturation difference was determined in the DM group (Pearson correlation coefficient r = -0.521; two-tailed test, P = 0.011) A positive correlation between PlGF level and the vein saturation was not statistically significant (Pearson correlation coefficient r = 0.325; two-tailed test, P = 0.130). We did not find any correlation between vitreous level of PEDF and vascular saturation within the DM group. Our findings in diabetic patients suggests a correlation between the intravitreal level of proangiogenic factors and the AV difference measured by retinal oximetry.

Involvement of Angiopoietin 2 and vascular endothelial growth factor in uveitis.

Angiopoietin (Ang) 2 is released from vascular endothelial cells by the stimulation of vascular endothelial growth factor (VEGF)A. Ang2 increases the expression of leukocyte adhesion molecules on endothelial cells via nuclear factor κB. The aim of this study was to evaluate the effects of Ang2 and VEGFA on ocular autoimmune inflammation. We measured the concentrations of Ang2 and VEGFA in vitreous samples among patients with uveitis. Vitreous samples were collected from 16 patients with idiopathic uveitis (uveitis group) and 16 patients with non-inflammatory eye disease (control group). Experimental autoimmune uveoretinitis (EAU) was induced in B10.BR mice with a human interphotoreceptor retinoid-binding protein-derived peptide. The retinochoroidal tissues of the EAU mice were removed, and the mRNA levels of Ang2 and VEGFA were examined. EAU mice treated with anti-Ang2, anti-VEGFA, a combination of anti-Ang2 and anti-VEGFA, anti-Ang2/VEGFA bispecific, or IgG control antibodies were clinically and histopathologically evaluated. The protein levels of Ang2 and VEGFA were significantly higher in the vitreous samples of patients with uveitis than in controls (P<0.05). The retinochoroidal mRNA levels of Ang2 and VEGFA were significantly upregulated in EAU mice compared to controls (n = 6, P<0.05). Although there was no significant difference, treatment with anti-VEGFA antibody reduced the clinical and histopathological scores. However, treatment with anti-Ang2 antibody reduced the clinical and histopathological scores (n = 18-20, P<0.05). Furthermore, these scores were further decreased when treated by inhibiting both Ang2 and VEGFA. Based on these results, VEGFA and Ang2 were shown to be upregulated locally in the eye of both uveitis patients and models of uveitis. Dual inhibition of Ang2 and VEGFA is suggested to be a new therapeutic strategy for uveitis.

Vitreous Levels of Vascular Endothelial Growth Factor and Platelet-Derived Growth Factor in Patients with Proliferative Diabetic Retinopathy: A Clinical Correlation.

Background: The global epidemic status of diabetic retinopathy (DR) and its burden presents an ongoing challenge to health-care systems. It is of great interest to investigate potential prognostic biomarkers of DR. Such markers could aid in detecting early stages of DR, predicting DR progression and its response to therapeutics. Herein, we investigate the prognostic value of intravitreal concentrations of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) in a DR cohort. Materials and methods: Vitreous sample acquisition was conducted at King Abdullah University Hospital (KAUH) between December 2020 and June 2022. Samples were obtained from any patient scheduled to undergo a pars plana vitrectomy (PPV) for any indication. Included patients were categorized into a DR group or a corresponding non-diabetic (ND) control group. Demographics, clinicopathological variables, standardized laboratory tests results, and optical coherence tomography (OCT) data were obtained for each included individual. Intravitreal concentrations of VEGF and PDGF were assessed using commercial enzyme-linked immunosorbent assay (ELISA). Results: A total of 80 eyes from 80 patients (DR group: n = 42 and ND control group: n = 38) were included in the analysis. The vitreous VEGF levels were significantly higher in the DR group compared to the ND control group (DR group 5744.06 ± 761.5 pg/mL versus ND control group 817.94 ± 403.1 pg/mL, p = 0.0001). In addition, the vitreous PDGF levels were also significantly higher in the DR group than those in the ND control group (DR group 4031.51 ± 410.2 pg/mL versus ND control group 2691.46 ± 821.0 pg/mL, p = 0.001). Bassline differences between test groups and clinical factors impacting VEGF and PDGF concentrations were investigated as well. Multiple regression analysis indicated PDGF as the sole independent risk factor affecting best-corrected visual acuity (BCVA) at the last follow-up visit: the higher the PDGF vitreous levels, the worst the BCVA. Conclusions: Vitreous concentrations of VEGF and PDGF are correlated with DR severity and may exhibit a possible prognostic potential value in DR. Further clinical and experimental data are warranted to confirm the observed findings and to help incorporate them into daily practice.

Highly Sensitive Interleukin Score for Intra-Ocular Lymphoma Diagnosis: Evidence from Analysis of 5-Year Real-Life of Use

Introduction Primary vitreoretinal lymphoma (PVRL) is a rare and aggressive subset of large B-cell lymphoma of immune-privileged sites. It can occur independently or concomitantly to primary central nervous system lymphoma. Symptoms are not specific and can easily be confused with other uveitis. Diagnosis of intra-ocular lymphoma (IOL), including PVRL and secondary lymphoma localization, is based on malignant lymphoid cells identification in vitrectomy(cytology, flow cytometry, molecular analysis). Nevertheless, the fragility of lymphoma cells and low cellularity of samples make diagnosis difficult with these cellular tools. The quantification of interleukin (IL) in intraocular fluids is a promising soluble alternative approach. Based on IL-10 and IL-6 levels, an Interleukine Score for intra-Ocular Lymphoma Diagnosis (ISOLD) designed for aqueous humor (AH) and vitreous was published to guide the diagnosis of IOL (Costopoulos et al, Ophtalmology, 2016). The aim of this work was to evaluate ISOLD performances in real-life practice after five-year of use in a hospital laboratory. Material and methods IL-10 and IL-6 were measured in intra-ocular sample in a single laboratory using the cytometric bead array® kit (BD Biosciences TM) on a FACS Canto II cytometer, with a limit of quantification of 2.5 pg/ml. ISOLD is calculated at diagnosis in AH (AH-ISOLD) or vitreous (V-ISOLD) samples and is associated with a probability of having a large B-cell IOL. This score classifies patients according to two groups: i) reliable zone: ISOLD &amp;lt; -4.6 (&amp;lt;1% probability to have IOL) and ISOLD &amp;gt; +4.6 (&amp;gt;99% probability to have IOL) ; ii) doubtful zone: ISOLD ranging from -4.6 to +4.6 for which the probability and the clinical context have to be considered. All calculated ISOLD from January 2017 to December 2021 were collected. Centers with more than 30 intraocular samples with a calculated ISOLD were contacted to retrieve patients' definitive diagnosis and potential treatments. Results During these 5 years, IL-10 and IL-6 levels have been quantified for 2796 intraocular samples from 70 centers. ISOLD was calculated at diagnosis for 2013 ocular samples. Clinical data were obtained for 1449 samples (1220 AH and 229 vitreous) corresponding to 1243 patients from 13 out of the 18 centers contacted. 9.7% of patients had an IOL, 82.2% had another diagnosis than IOL and 8.1% of patients had no established diagnosis (Figure 1). Considering the 1341 samples with an established diagnosis, 92.7% were in the reliable zone and 7.3% were in the doubtful zone. For the 1128 AH samples, 1050 AH-ISOLD (93.1%) were in the reliable interpretation zone, with a concordant result for 98.6% of them (n=1035 samples) and a discordant result for 1.4% of them (n=15) (Table 1). The sensitivity and specificity were respectively estimated at 90.6% and 99.3% with a positive predictive value (PPV) of 91.7% and a negative predictive value (NPV) of 99.2%. Seventy-eight AH-ISOLD were in doubtful zone including 23 LIO and 55 others diagnoses than IOL. For the 213 vitreous samples, 193 V-ISOLD (90.6%) were in the reliable interpretation zone, with a concordant result for 98.4% of them (n=190 samples) and a discordant result for 1.6% of them (n=3) (Table 1). The sensitivity and specificity were estimated at 97.6% and 99.1%, respectively with a PPV of 98.8% and a NPV of 98.2%. Twenty V-ISOLD were in doubtful zone including 5 LIO and 15 others diagnoses than IOL. Three patients of the 8 who had a false-negative AH-ISOLD had a positive V-ISOLD. For the 17 AH-ISOLD in the doubtful zone, 4 of them had a V-ISOLD concordant with the diagnosis and 2 had an AH-ISOLD in the other eye concordant with the diagnosis. Independently of the sample type, the main cause of false-negative results was administration of corticosteroids prior to sampling and the main causes of false-positive results were infectious diseases (varicella and herpes virus, toxoplasmosis, syphilis). Conclusion This study confirmed on a 5-year period of real life practice that ISOLD, based on IL-10 and IL-6 levels, is a powerful tool to guide the diagnosis towards IOL. In more than 90% of intra-ocular samples, ISOLD was in reliable interpretation zone with very high sensitivity and specificity levels. In case of positive or doubtful AH-ISOLD, a vitrectomy is needed to make the diagnosis. Easy to use, not expansive, this score appears very useful to optimize the diagnostic approach of this rare and aggressive large B-cell lymphoma.