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Acta Ophthalmologica Aug 2015To confirm the efficacy of liquid-based cytology (LBC) method in the observation of vitreous cells in various vitreoretinal diseases in human.
PURPOSE
To confirm the efficacy of liquid-based cytology (LBC) method in the observation of vitreous cells in various vitreoretinal diseases in human.
METHODS
Vitreous fluid samples from 30 eyes were obtained by 23-gauge 3-port pars plana vitrectomy. After making three ports, we collected vitreous specimen from the core vitreous cavity without infusion. We divided the samples into a quiescent group and an active group based on clinical signs of inflammation. To confirm availability of LBC preparation slides for immunostaining, we also performed immunocytochemistry (ICC) for CD68, RPE65 and DEC-205 (CD205) using LBC slides of 10 cell-rich cases including retinal detachment and endophthalmitis.
RESULTS
Using LBC method, small amounts of vitreous cells were observed efficiently. Vitreous cells were observed in inflammatory quiescent cases including macular pucker and macular hole. The number of vitreous cells increased significantly in the cases with clinically active inflammation (2297 versus 207 cells/ml, respectively, p < 0.01, Mann-Whitney U-test). The ICC results showed the presence of CD68(+) cells in all 10 cases. Large numbers of DEC-205(+) cells were observed in one case with infectious endophthalmitis. In the cases with retinal detachment, the predominant cell type was RPE65(+) . Neutrophils and lymphocytes were also observed.
CONCLUSIONS
The LBC method makes it possible to examine vitreous specimens easily and efficiently, facilitating the expedient diagnosis of vitreoretinal diseases, and the preparation slides are available for immunocytochemistry. This study also showed that vitreoretinal disease involves the migration of various types of cells including macrophages, neutrophils, lymphocytes, RPE65(+) pigmented cells and DEC-205(+) cells.
Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Biomarkers; Cellular Structures; Cytological Techniques; Eye Diseases; Female; Humans; Immunoenzyme Techniques; Lectins, C-Type; Male; Middle Aged; Minor Histocompatibility Antigens; Receptors, Cell Surface; Retinal Diseases; Retrospective Studies; Vitrectomy; Vitreous Body; cis-trans-Isomerases
PubMed: 25752226
DOI: 10.1111/aos.12623 -
Investigative Ophthalmology & Visual... Oct 2017To assess vitreous levels of inflammatory cytokines and neurotrophins (NTs) in diabetic retinopathy (DR) and elucidate their potential roles.
PURPOSE
To assess vitreous levels of inflammatory cytokines and neurotrophins (NTs) in diabetic retinopathy (DR) and elucidate their potential roles.
METHODS
A prospective study was performed on 50 vitreous samples obtained from patients with DR (n = 22) and the nondiabetic controls (n = 28). All patients were candidates for vitrectomy. Inflammatory cytokine and NT levels were determined with ELISA. Potential source and role of NTs was determined by using human retinal Müller glia and mouse photoreceptor cells and challenging them with TNF-α or IL-1β, followed by detection of NTs and cell death.
RESULTS
Vitreous NT levels of all DR patients were significantly higher than those of nondiabetic controls (nerve growth factor [NGF, P = 0.0001], brain-derived neurotrophic factor [BDNF, P = 0.009], neurotrophin-3 [NT-3, P < 0.0001], neurotrophin-4 [NT-4, P = 0.0001], ciliary neurotrophic factor [CNTF, P = 0.0001], and glial cell-derived neurotrophic factor [GDNF, P = 0.008]). Similarly, the levels of inflammatory mediators IL-1β (P < 0.0001), IL-6 (P = 0.0005), IL-8 (P < 0.0001), and TNF-α (P < 0.0001) were also higher in eyes with DR. Interestingly, inflammatory cytokine and NT levels, particularly TNF-α (P < 0.05), IL-8 (P < 0.004), NT-3 (P = 0.012), NGF (P = 0.04), GDNF (P = 0.005), and CNTF (P = 0.002), were higher in eyes with nonproliferative diabetic retinopathy (NPDR) than in eyes with active proliferative diabetic retinopathy (PDR). Cytokine stimulation of Müller glia resulted in production of NTs, and GDNF treatment reduced photoreceptor cell death in response to inflammation and oxidative stress.
CONCLUSIONS
Together, our study demonstrated that patients with DR have higher levels of both inflammatory cytokines and NTs in their vitreous. Müller glia could be the potential source of NTs under inflammatory conditions to exert neuroprotection.
Topics: Adult; Aged; Aged, 80 and over; Biomarkers; Cells, Cultured; Cytokines; Diabetic Retinopathy; Female; Humans; Male; Middle Aged; Nerve Growth Factors; Prospective Studies; Vitreous Body
PubMed: 29084332
DOI: 10.1167/iovs.17-21973 -
Indian Journal of Ophthalmology May 2015Optical coherence tomography (OCT) has revolutionized imaging of ocular structures and various disease conditions. Though it has been used in the clinic for some... (Review)
Review
Optical coherence tomography (OCT) has revolutionized imaging of ocular structures and various disease conditions. Though it has been used in the clinic for some decades, the OCT has only recently found its way into the operating theater. Early attempts at intraoperative OCT, hand-held and microscope mounted, have already improved our understanding of the surgical pathology and the role it might play in surgical decision-making. The microscope-integrated OCT now allows seamless, high-resolution, real-time imaging of surgical maneuvers from the incision to wound closure. Visualization of instruments and intraoperative tissue manipulation are possible with this in vivo modality and, therefore, help improve the outcome of surgery. In this article, we describe the advantages it offers during various vitreoretinal procedures.
Topics: Humans; Microscopy; Monitoring, Intraoperative; Retinal Diseases; Tomography, Optical Coherence; Vitreoretinal Surgery; Vitreous Body
PubMed: 26139800
DOI: 10.4103/0301-4738.159865 -
Investigative Ophthalmology & Visual... Oct 2013Previous studies that measured liquefaction and oxygen content in human vitreous suggested that exposure of the lens to excess oxygen causes nuclear cataracts. Here, we... (Comparative Study)
Comparative Study
PURPOSE
Previous studies that measured liquefaction and oxygen content in human vitreous suggested that exposure of the lens to excess oxygen causes nuclear cataracts. Here, we developed a computational model that reproduced available experimental oxygen distributions for intact and degraded human vitreous in physiologic and environmentally perturbed conditions. After validation, the model was used to estimate how age-related changes in vitreous physiology and structure alter oxygen levels at the lens.
METHODS
A finite-element model for oxygen transport and consumption in the human vitreous was created. Major inputs included ascorbate-mediated oxygen consumption in the vitreous, consumption at the posterior lens surface, and inflow from the retinal vasculature. Concentration-dependent relations were determined from experimental human data or estimated from animal studies, with the impact of all assumptions explored via parameter studies.
RESULTS
The model reproduced experimental data in humans, including oxygen partial pressure (Po2) gradients (≈15 mm Hg) across the anterior-posterior extent of the vitreous body, higher oxygen levels at the pars plana relative to the vitreous core, increases in Po2 near the lens after cataract surgery, and equilibration in the vitreous chamber following vitrectomy. Loss of the antioxidative capacity of ascorbate increases oxygen levels 3-fold at the lens surface. Homogeneous vitreous degeneration (liquefaction), but not partial posterior vitreous detachment, greatly increases oxygen exposure to the lens.
CONCLUSIONS
Ascorbate content and the structure of the vitreous gel are critical determinants of lens oxygen exposure. Minimally invasive surgery and restoration of vitreous structure warrant further attention as strategies for preventing nuclear cataracts.
Topics: Biological Transport; Humans; Lens, Crystalline; Models, Biological; Oxygen; Oxygen Consumption; Vitreous Body
PubMed: 24008409
DOI: 10.1167/iovs.13-12609 -
Investigative Ophthalmology & Visual... Jul 2013To determine if the accommodative forward movements of the vitreous zonule and lens equator occur in the human eye, as they do in the rhesus monkey eye; to investigate... (Comparative Study)
Comparative Study
PURPOSE
To determine if the accommodative forward movements of the vitreous zonule and lens equator occur in the human eye, as they do in the rhesus monkey eye; to investigate the connection between the vitreous zonule posterior insertion zone and the posterior lens equator; and to determine which components-muscle apex width, lens thickness, lens equator position, vitreous zonule, circumlental space, and/or other intraocular dimensions, including those stated in the objectives above-are most important in predicting accommodative amplitude and presbyopia.
METHODS
Accommodation was induced pharmacologically in 12 visually normal human subjects (ages 19-65 years) and by midbrain electrical stimulation in 11 rhesus monkeys (ages 6-27 years). Ultrasound biomicroscopy imaged the entire ciliary body, anterior and posterior lens surfaces, and the zonule. Relevant distances were measured in the resting and accommodated eyes. Stepwise regression analysis determined which variables were the most important predictors.
RESULTS
The human vitreous zonule and lens equator move forward (anteriorly) during accommodation, and their movements decline with age, as in the monkey. Over all ages studied, age could explain accommodative amplitude, but not as well as accommodative lens thickening and resting muscle apex thickness did together. Accommodative change in distances between the vitreous zonule insertion zone and the posterior lens equator or muscle apex were important for predicting accommodative lens thickening.
CONCLUSIONS
Our findings quantify the movements of the zonule and ciliary muscle during accommodation, and identify their age-related changes that could impact the optical change that occurs during accommodation and IOL function.
Topics: Accommodation, Ocular; Adult; Aged; Aging; Animals; Ciliary Body; Disease Models, Animal; Female; Humans; Lens, Crystalline; Macaca mulatta; Male; Microscopy, Acoustic; Middle Aged; Presbyopia; Vitreous Body; Young Adult
PubMed: 23745002
DOI: 10.1167/iovs.12-10846 -
Transactions of the American... 1968
Topics: Academic Dissertations as Topic; Animals; Biochemical Phenomena; Biochemistry; Cataract; Chondroitin; Colloids; Dogs; Eye Injuries; Fundus Oculi; Gels; Heparin; Hyaluronic Acid; Injections; Ion Exchange; Ion Exchange Resins; Iron; Lens, Crystalline; Models, Biological; Photography; Polysaccharides; Rabbits; Retina; Retinal Detachment; Staining and Labeling; Sulfates; Sulfonic Acids; Tissue Adhesions; Uronic Acids; Vitreous Body
PubMed: 4181020
DOI: No ID Found -
Indian Journal of Ophthalmology Jun 2019To study the clinicopathological findings of Persistent Fetal Vasculature (PFV) in patients with congenital cataract and PFV.
PURPOSE
To study the clinicopathological findings of Persistent Fetal Vasculature (PFV) in patients with congenital cataract and PFV.
METHODS
Six eyes with anterior or combined PFV with cataract underwent phacoaspiration with primary posterior capsulotomy with anterior vitrectomy with intraocular lens implantation followed by histopathological evaluation of the PFV stalk and membrane.
RESULTS
Four and two patients had combined and anterior PFV respectively. There was no postoperative hyphema, vitreous haemorrhage, glaucoma or retinal detachment in six months. Haematoxylin and eosin staining showed inflammatory cells predominantly with extramedullary hematopoeisis and vascularisation.
CONCLUSION
We recommend IOL implantation in PFV, with early and aggressive amblyopia therapy.
Topics: Cataract; Follow-Up Studies; Humans; Infant; Infant, Newborn; Lenses, Intraocular; Male; Microscopy, Acoustic; Persistent Hyperplastic Primary Vitreous; Phacoemulsification; Prognosis; Prospective Studies; Vitrectomy; Vitreous Body
PubMed: 31124487
DOI: 10.4103/ijo.IJO_1375_18 -
Arquivos Brasileiros de Oftalmologia 2012Optical coherence tomography (OCT) is valuable for macula evaluation. However, as this technique relies on light energy it cannot be performed in the presence of opaque... (Comparative Study)
Comparative Study
PURPOSE
Optical coherence tomography (OCT) is valuable for macula evaluation. However, as this technique relies on light energy it cannot be performed in the presence of opaque media. In such cases, the ultrasound (US) may predict some macular features. The aim of this study was to characterize images obtained by ultrasound with 10 and 20-MHz transducers comparing to OCT, as well as to analyze the relationship between the vitreous and retina in eyes with macular hole (MH).
METHODS
29 eyes of 22 patients with biomicroscopic evidence of MH at different stages were included. All patients were evaluated using ultrasonography with 10 and 20-MHz transducers and OCT.
RESULTS
OCT identified signs of MH in 25 of 29 eyes. The remaining 4 cases not identified by US were pseudoholes caused by epiretinal membranes. In MH stages I (2 eyes) and II (1 eye), both transducers were not useful to analyze the macular thickening, but suggestive findings as macular irregularity, operculum or partial posterior vitreous detachment (PVD) were highlighted. In stages III (14 eyes) and IV (5 eyes), both transducers identified the double hump irregularity and thickening. US could measure the macular thickness and other suggestive findings for MH: operculum, vitreomacular traction and partial or complete PVD. In cases of pseudoholes, US identified irregularities macular contour and a discrete depression.
CONCLUSION
10-MHz US was useful for an overall assessment of the vitreous body as well as its relationship to the retina. The 20-MHz transducer allowed valuable information on the vitreomacular interface and macular contour. OCT provides superior quality for fine morphological study of macular area, except in cases of opaque media. In these cases, and even if OCT is not available, the combined US study is able to provide a valid evaluation of the macular area improving therapeutic approach.
Topics: Aged; Child; Female; Humans; Macula Lutea; Male; Microscopy, Acoustic; Middle Aged; Retina; Retinal Perforations; Tomography, Optical Coherence; Transducers; Vitreous Body; Vitreous Detachment
PubMed: 23715145
DOI: 10.1590/s0004-27492012000600009 -
Science Advances Nov 2018The intravitreal delivery of therapeutic agents promises major benefits in the field of ocular medicine. Traditional delivery methods rely on the random, passive...
The intravitreal delivery of therapeutic agents promises major benefits in the field of ocular medicine. Traditional delivery methods rely on the random, passive diffusion of molecules, which do not allow for the rapid delivery of a concentrated cargo to a defined region at the posterior pole of the eye. The use of particles promises targeted delivery but faces the challenge that most tissues including the vitreous have a tight macromolecular matrix that acts as a barrier and prevents its penetration. Here, we demonstrate novel intravitreal delivery microvehicles-slippery micropropellers-that can be actively propelled through the vitreous humor to reach the retina. The propulsion is achieved by helical magnetic micropropellers that have a liquid layer coating to minimize adhesion to the surrounding biopolymeric network. The submicrometer diameter of the propellers enables the penetration of the biopolymeric network and the propulsion through the porcine vitreous body of the eye over centimeter distances. Clinical optical coherence tomography is used to monitor the movement of the propellers and confirm their arrival on the retina near the optic disc. Overcoming the adhesion forces and actively navigating a swarm of micropropellers in the dense vitreous humor promise practical applications in ophthalmology.
Topics: Animals; Biomechanical Phenomena; Fluorocarbons; Intravitreal Injections; Pharmaceutical Preparations; Retina; Swine; Vitreous Body
PubMed: 30406201
DOI: 10.1126/sciadv.aat4388 -
Molecules (Basel, Switzerland) Aug 2021Orexins are two neuropeptides synthesised mainly in the brain lateral hypothalamic area. The orexinergic system provides arousal-dependent cues for a plethora of brain...
Orexins are two neuropeptides synthesised mainly in the brain lateral hypothalamic area. The orexinergic system provides arousal-dependent cues for a plethora of brain centres, playing a vital role in feeding behaviour, regulation of the sleep-wake cycle and circadian rhythms. Recently, orexins were found to be produced in the retina of an eye; however, their content in the vitreous body and possible daily pattern of expression have not yet been explored. In this manuscript, we describe the development and validation of a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method designed for quantitative bioanalysis of orexin in the rat vitreous body. Orexin was extracted from vitreous body samples with a water:acetonitrile:formic acid (80:20:0.1; //) mixture followed by vortexing and centrifuging. Separation was performed on a reverse-phase HPLC column under gradient conditions. Orexin was analysed via multiple-reaction monitoring (MRM) in the positive electrospray mode. The total analysis time for each sample was less than 5.0 min. Once the method was fully optimised, it was then validated, following the 2018 FDA guidance on bioanalytical method validations. The calibration curves for orexin (1-500 ng/mL) were constructed using a linear regression with a 1/x weighting. The lower limit of quantitation for orexin was 1.0 pg/mL for the vitreous body. Intra-day and inter-day estimates of accuracy and precision were within 10% of their nominal values, indicating that the method is reliable for quantitation of orexin in the rat vitreous body. From the physiological perspective, our results are the first to show daily rhythm of orexin synthesis by the retina with possible implications on the circadian regulation of vision.
Topics: Animals; Calibration; Chromatography, Liquid; Circadian Rhythm; Linear Models; Male; Orexins; Rats; Retina; Tandem Mass Spectrometry; Vitreous Body
PubMed: 34443622
DOI: 10.3390/molecules26165036