Comparison of the trabecular meshwork length between open and closed angle with evaluation of the scleral spur location.

This study analysed the trabecular meshwork (TM) length in open and closed angle patients and presented a better method for locating the scleral spur using Schwalbe's line method in anterior segment optical coherence tomography (AS-OCT). Patients who underwent AS-OCT at Yonsei University Gangnam Severance Hospital we enrolled; 58 and 57 open and closed angle eyes, respectively. We measured the distance from the scleral spur to Schwalbe's line as the TM length and compared it between open and closed angle patients, and to previous studies. TM length was applied to locate the scleral spur using Schwalbe's line method. Mean TM lengths were 811 ± 83 µm and 575 ± 96 µm in the open and closed angle groups, respectively (p < 0.001). Comparing the actual TM heights using Schwalbe's line method with an updated reference distance significantly increased accuracy to locate the scleral spur (open: 811 µm, closed: 575 µm) when compared with the 1000 µm reference distance (p < 0.001). TM length was significantly different between open and closed angle patients. Further, the reference distance for Schwalbe's line method should be distinguished according to open and closed angle eyes in order that the scleral spur can be located properly.

Nerve terminals at the human corneoscleral limbus.

AIMS: To demonstrate and characterise distinct subepithelial compact nerve endings (CNE) at the human corneoscleral limbus. METHODS: Ten fresh human donor corneoscleral discs (mean age, 67 years) and 26 organ-cultured corneoscleral rims (mean age, 59 years) were studied. All samples were subjected to enzyme histochemical staining related to endogenous acetylcholinesterase present in nerve tissue and H&E staining. Whole-mount en face imaging with NanoZoomer digital pathology microscope and serial cross-section imaging with light microscope were undertaken. RESULTS: Nerves entering the corneoscleral limbus and peripheral cornea terminate under the epithelium as enlarged multiloculated and multinucleated ovoid structures within a 2 mm zone. They are closely associated with the rete pegs of the limbal palisades and the limbal epithelial crypts, often located within characteristic stromal invaginations of these structures. Their numbers ranged from 70 to 300 per corneoscleral rim. The size ranged from 20 to 100 µm. They had one or more nerve connections and were interconnected to other similar endings and to the limbal nerve plexus. CONCLUSION: Human corneoscleral limbus demonstrates a population of nerve terminals resembling CNE with distinct morphological features. They are closely associated with the limbal stem cell niches, suggesting a potential contribution to the niche environment.

Shear behavior of bovine scleral tissue.

Ocular tissue properties have been widely studied in tension and compression for humans and a variety of animals. However, direct shear testing of the tissues of the sclera appear to be absent from the literature even though modeling, analyses, and anatomical studies have indicated that shear may play a role in the etiology of primary open angle glaucoma (POAG). In this work, the mechanical behavior of bovine scleral tissue in shear has been studied in both out-of-plane and in-plane modes of deformation. Stress-strain and relaxation tests were conducted on tissue specimens at controlled temperature and hydration focusing on trends related to specimen location and orientation. There was generally found to be no significant effect of specimen orientation and angular location in the globe on shear stiffness in both modes. The in-plane response, which is the primary load carrying mode, was found to be substantially stiffer than the out-of-plane mode. Also, within the in-plane studies, tissue further from the optic nerve was stiffer than the near tissue. The viscosity coefficient of the tissue varied insignificantly with distance from the optic nerve, but overall was much higher in-plane than out-of-plane.

Ultrasound biomicroscopy diagnosis of an axenfeld loop.

A 67-year-old man was referred for evaluation of a small mass on the sclera of the right eye. Ultrasound biomicroscopy of the mass suggested the diagnosis of Axenfeld loop.

Examination of the anterior uveoscleral pathway in domestic species.

OBJECTIVE: To investigate the uveoscleral (US) pathway in the normal eyes of four domestic spp.: the cat, pig, cow and horse by examining the comparative anatomical structure of anterior US pathway. ANIMALS STUDIED: Four cats, ten pigs, four cows, eight horses. PROCEDURES: Formalin-preserved specimens from anterior uveas of the cat, pig, cow and horse were embedded and serially sectioned sagittally and tangentially and stained with H&E, Masson's trichrome, smooth muscle actin immunolabel, or elastin stain. RESULTS: Spaces between the endings of the outer anterior ciliary body musculature (CBM) formed avenues for the beginning of the US pathway and varied in the amount of extracellular matrix (ECM) material being most developed in the pig. In the cow, other anterior muscle bundles attached the CBM to the sclera concomitant with the presence of an anterior elastic sheath. In the horse, these muscle bundles were connected to branching connective tissue trabeculae within the US pathway that were attached radially to the sclera. In the cat, muscle bundles were more abundant and formed a fine meshwork of trabecular associations with the posterior ICA. Supraciliary development was most pronounced in the horse and least in the pig. CONCLUSION: All species possessed clearly developed and unique US pathways. The outermost muscle bundles of the CBM appeared to have close interaction with the US pathway and the degree of these muscle associations differed with species. The species specific anatomical variations within the US pathway could play a pivotal role in the variability of aqueous outflow along this pathway.

Diffuse corneoscleral limbal neuromas with prominent corneal nerves in multiple endocrine neoplasia syndrome type IIB.

This report describes a 7-year-old boy with multiple endocrine neoplasia type 2B who presented to the ophthalmology department because of peripheral corneal changes. In addition to prominent corneal nerves and eyelid neuromas, the patient was found to have substantial corneoscleral limbal thickening with corneal neovascularization.

Comprovação anatomopatológica da presença de tecido vascular e nervoso evidenciado à ecografia como eco tangencial ao descolamento de coróide: relato de dois casos / Histopathologic confirmation of vascular and nervous tissue identified on ultrasound as an tangential image to choroidal detachment: report of two cases

São apresentados dois casos de descolamento de coróide em olhos com endoftalmite submetidos a enucleação. O estudo anatomopatológico permitiu identificar a presença de tecido vascular e nervoso interpondo-se entre a esclera e coróide deslocada, como havia sido evidenciado pela ecografia.

Two cases of endophthalmitis with choroidal detachment submitted to enucleation are presented. The histopathologic study allowed the identification of vascular and nervous tissue between the choroid and sclera, as it was seen on ultrasound examination.

[Histopathologic confirmation of vascular and nervous tissue identified on ultrasound as an tangential image to choroidal detachment: report of two cases]. / Comprovação anatomopatológica da presença de tecido vascular e nervoso evidenciado à ecografia como eco tangencial ao descolamento de coróide: relato de dois casos.

Two cases of endophthalmitis with choroidal detachment submitted to enucleation are presented. The histopathologic study allowed the identification of vascular and nervous tissue between the choroid and sclera, as it was seen on ultrasound examination.

Promotion of angiogenesis in tissue engineering: developing multicellular matrices with multiple capacities.

One of the aims of tissue engineering is to be able to develop multi-tissue organs in the future. This requires the optimization of conditions for the differentiation of multiple cell types and maintenance of the differentiated phenotype within complex engineered tissues. The goal of this study was to develop prototype tissue engineered matrices to support the simultaneous growth of different cell types with a particular focus on the angiogenic process. We examined two different matrix compositions for the promotion of blood vessel and tube formation. A fibrin-based matrix with the addition of a combination of growth factors supported vascular growth and the invasion of inflammatory cells. Using this fibrin matrix, in combination with a collagen-based hydrogel, a simple in vitro model of the cornea with adjacent sclera was developed that was complete with innervation and vascular structures. In addition, we showed that collagen-based matrices were effective in delivering mononuclear endothelial progenitor cells to ischemic tissue in vivo, and allowing these cells to incorporate into vascular structures. It is anticipated that with further development, these matrices have potential for use as delivery matrices for cell transplantation and for in vitro study purposes of multiple cell types.

Angioarchitecture and innervation of the primate anterior episclera.

PURPOSE: To investigate the primate episcleral vasculature and its innervation with respect to morphological specializations. METHODS: Serial sections of the anterior episclera of 8 monkey eyes and 20 human eyes were investigated enzyme- and immunohistochemically using antibodies against smooth-muscle alpha-actin (SMA), neurofilament, synaptophysin, substance P (SP), calcitonin gene-related peptide (CGRP), vesicular acetylcholine transporter (VACHT), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), tyrosine hydroxylase (TH), vesicular monoamine transporter II (VMAT II), as well as the NADPH-diaphorase reaction. Arteriovenous anastomoses (AVA) were quantified. RESULTS: All episcleral vessels including veins showed intense staining for SMA. Capillary loops were only seen in the limbal arcades, not in the episclera itself. Instead, AVA connected the episcleral arteries with the veins, which formed an interlacing vascular network. In the monkey episclera, 4-6/mm2 AVA were found; in the human episclera, 0.5-1/mm2. Numerous nerve endings staining for NADPHd (NADPHdiaphorase) and TH surrounded all episcleral vessels including anastomoses and veins. NPY, VIP, and VACHT-immunoreactive (IR) nerve terminals were less numerous. CGRP and SP-IR terminals were seen both at the vessels and in the intervascular connective tissue. CONCLUSIONS: The episcleral vasculature shows a specialized morphology with absence of capillaries, numerous arteriovenous anastomoses, a muscle-rich venous network, and intense innervation by vasodilative and vasoconstrictive nerves. This might allow regulation of blood flow and volume in the episcleral vessels and Voigt's capillaries for thermoregulation and modulation of episcleral venous pressure and thereby outflow facility.

Substance P and opioid peptidergic innervation of the anterior eye segment of the rat: an immunohistochemical study.

Recently discovered endogenous opioid peptides such as nociceptin are known to modulate neurotransmitter release of primary afferent neurons (especially substance P, SP) and they have also been demonstrated in peripheral nerve fibres. The aim of this study was to investigate the opioid peptidergic innervation of the anterior eye segment and to compare it with the innervation pattern of SP in order to shed light on the functional relationship between these peptides. Anterior eye segments of 20 rat eyes were cut in a tangential plane and the sections stained with antibodies against SP, nociceptin, nocistatin, endomorphin 1 and 2, leu-enkephalin and met-enkephalin. Sections of the spinal cord or brain were used as positive controls. Numerous SP-immunoreactive nerve fibres were found in the conjunctiva, cornea, episclera, trabecular meshwork, iris and ciliary body. A weak staining for met-enkephalin and leu-enkephalin could only be found in the iris and anteriormost ciliary body. Nerve fibres immunoreactive for nociceptin, nocistatin, and endomorphin 1 or 2 could not be detected in any part of the anterior eye segment. It is tempting to speculate that the opioid peptidergic innervation of the anterior ciliary body may play a role in the modulation of intraocular inflammation.

Expression of protein gene product 9.5 in lepromatous eyes showing ciliary body nerve damage and a "dying back" phenomenon in the posterior ciliary nerves.

BACKGROUND/AIM: Peripheral nerve destruction is the hallmark of leprosy. Ocular complications form a substantial part of the clinical manifestations but histopathology of nerve destruction within ocular structures has not been shown satisfactorily. The role of protein gene product (PGP) 9.5 in identifying nerve destruction in the ciliary body and posterior ciliary nerves of lepromatous eyes is shown. METHODS: Serial sections from two lepromatous eyes and two non-lepromatous eyes were stained with PGP 9.5. Histopathological comparison was done on the expression of the PGP 9.5 stain in nerves within the ciliary body, posterior ciliary nerves adjacent to the optic nerve, and nerves tracking through the sclera. RESULTS: In non-lepromatous eyes, PGP 9.5 was expressed in nerves within the ciliary body, the nerves within the sclera, and posterior ciliary nerves adjacent to the optic nerve. In lepromatous eyes no PGP 9.5 was expressed, signifying nerve destruction. CONCLUSIONS: Nerve destruction in lepromatous eyes has been confirmed histopathologically by the absence of or patchy staining with PGP 9.5. Nerve destruction in the ciliary body can extend to the posterior ciliary nerves by an ascending axonopathy. This "dying back" phenomenon is akin to the "glove and stocking" anaesthesia found in lepromatous leprosy.

Activation of scleral cold thermoreceptors by temperature and blood flow changes.

PURPOSE: To study the response of scleral cold receptors located in areas of the eye unexposed to temperature and blood flow changes. METHODS: In anesthetized cats, the neural activity was recorded from single, cold-sensory fibers of the ciliary nerves innervating the sclera and limbus. Controlled temperature changes of the receptive field were performed with a contact thermode. Ocular blood flow reductions were obtained by occluding the ipsilateral common carotid artery for 30 to 60 seconds with a compressor placed around the artery. Local blood flow was measured with a laser Doppler flowmeter. Temperature was measured with a microprobe introduced in the subscleral space. Ocular sympathetic stimulation was performed with a pair of silver electrodes placed on the preganglionic cervical sympathetic trunk. To induce local hypoxia, N(2) was applied on the scleral surface with a specially designed chamber. For systemic hypoxia the breathing air was replaced with a gas mixture containing 10% O(2) in N(2). RESULTS: Sensory nerve fibers identified as cold receptors exhibited ongoing nerve activity in bursts at 35 degrees C and responded to cooling pulses applied to their receptive fields with an increase in the impulse discharge that reached a peak and decayed gradually to a lower level. When temperature was reduced from 35 degrees C to 34 degrees C, frequency increased monotonically with decreasing temperature of the sclera. Between 35 degrees C and 30 degrees C, peak and mean frequencies were roughly proportional to temperature of the sclera. The characteristics of burst discharges also depended on scleral temperature. Electrical stimulation of the cervical sympathetic trunk induced a decrease in blood flow and temperature and evoked an increase in the firing frequency of cold-sensory fibers that was proportional to the frequency of stimulating pulses. Carotid occlusion also elicited an increase of the discharge of cold thermoreceptor fibers that occurred in parallel with a decrease in blood flow and temperature in the receptive field area. Local or systemic hypoxia did not modify appreciably the spontaneous firing frequency of scleral cold-sensory fibers. CONCLUSIONS: Scleral and episcleral cold-sensory fibers encoded as a change in their impulse frequency and firing pattern temperature reductions of less than 1 degrees C in scleral tissues. Activation of scleral and episcleral cold-sensory fibers by sympathetic vasoconstriction and acute arterial pressure reductions appear to be secondary to the temperature decrease that accompanies the reduction in ocular blood flow caused by these maneuvers. Scleral thermosensory fibers are located in ocular territories not directly exposed to external temperature changes. Thus, the sensory information on local blood flow variations provided by these receptors may be involved in a reflex regulation of choroidal blood flow that functions to maintain a constant temperature and blood supply to the retina.

Neuroanatomy of the extraocular muscle tendon enthesis in macaque, normal human, and patients with congenital nystagmus.

PURPOSE: To search for the presence of, and qualitatively characterize the microscopic anatomy of, nerve terminals in the tendino-scleral (enthesial) area of the extraocular muscle tendon. METHODS: Extraocular rectus muscle tendino-scleralspecimens from a macaque monkey, a normal human, cadavers, and patients with congenital nystagmus were obtained and studied with transmission electron microscopy. RESULTS: Enthesial neurovascular structures were identified in all specimens. The enthesial area showed structures consisting of myelinated and unmyelinated nerve fibers with associated supporting vascular capillaries. Qualitative analysis showed anomalous neurovascular structures were in the enthesis of patients with congenital nystagmus. CONCLUSIONS: This study confirms the presence of normal and abnormal neural anatomy in the enthesial area of the extraocular muscle tendon. Neurovascular abnormalities in the enthesial part of the extraocular muscle tendon may be a result of, or contribute to, disease pathogenesis. If these structures are related to afferent central nervous system input, then disruption of them during surgery might influence postoperative outcome.

Three-dimensional reconstruction of scleral cold thermoreceptors of the cat eye.

Sensory endings that respond to local cooling were identified electrophysiologically in the cat's sclera. Functionally identified scleral thermal fibers were then used to analyze the structural characteristics of cold receptor endings. Four Adelta units sensitive to controlled cooling of their scleral receptive fields were recorded. The receptive areas were mapped, demarcated with pins and examined electron microscopically using extensive three-dimensional reconstructions. The supporting tissue within the receptive areas of cold units consisted of dense collageneous tissue with a small number of blood vessels that were either veins or capillaries. Adelta nerve fibers were found within these tissue blocks presumably corresponding with cold sensitive fibers. Small nerves and single nerve fibers devoid of a perineurium were found in all parts of the tissue, only occasionally passing a blood vessel. The terminal portions showed axonal swellings all along the unmyelinated segment filled with mitochondria, glycogen particles, and some vesicles. About 30% of the terminal axonal membrane is not covered by Schwann cells. In the unmyelinated distal portion, the mitochondrial content ranged from 0.012 to 0.038 microm(3) mitochondrial volume per microm(2) nerve fiber membrane. In comparison with sensory endings in the cat's knee joint, cold receptors in the cat sclera showed many similarities in their three-dimensional structure with polymodal nociceptor endings of the knee joint but contain less mitochondria. This suggests that cold sensory endings do not require specialized cellular processes for the transduction of cold stimuli, as is the case for multimodal transduction and sensitization in the terminal portion of polymodal nociceptors.

Occurrence of catecholaminergic nerve fibers in the human uveoscleral tissue in conditions of normal and raised intraocular pressure.

The distribution of catecholaminergic nerve fibers (CNF) in human uveoscleral tissue was studied in six human eyes with normal intraocular pressure and in five eyes with increased pressure. The eyes with increased pressure had no visual field alterations and the patients did not have any glaucoma-related optic neuropathies. The amount of norepinephrine in these structures was also analysed. Catecholaminergic nerve fibers were detected by means of fluorescence microscopy and were counted using the quantitative analysis of images. Our results demonstrate that the occurrence of catecholaminergic nerve fibers (expressed in Conventional Units = C.U.) in human uveoscleral tissue is 15.4 +/- 1.6 C.U. in eyes with normal intraocular pressure. In eyes with increased intraocular pressure, these values were 12.2 +/- 1.2 C.U. Moreover, the amount of norepinephrine in tissue homogenates of the same eyes was evaluated and found to be 21.7 +/- 1.3 microg/gr tissue fresh weight of the human uveoscleral tissue in eyes with normal intraocular pressure. This value decreased to 18.8 +/- 1.1 microg/gr tissue fresh weight in the same tissue in conditions of raised intraocular pressure. In these experiments, the small number of eyes examined made it difficult to draw general conclusions. However, the role of human uveoscleral tissue was emphasized by the rich catecholaminergic innervation. A decrease of catecholaminergic nerve fibers and norepinephrine occurs when intraocular pressure is elevated. The modifications of these parameters, involved in the sympathetic control of aqueous humor outflow, may support the hypothesis of a possible relevant role for the human uveoscleral tissue in different pathological conditions.

Efferent and afferent innervation of primate trabecular meshwork and scleral spur.

PURPOSE: To determine the correlation between nerve terminals and cells or extracellular matrix (ECM) components in different portions of the primate trabecular meshwork (TM) and scleral spur (SS). METHODS: Serial sagittal and tangential sections through the anterior segments of 10 cynomolgus monkey eyes and 12 human eyes were investigated immunohistochemically with antibodies against the vesicular acetylcholine transporter (VACHT), vasoactive intestinal polypeptide (VIP), tyrosine-hydroxylase (TH), neuropeptide Y (NPY), substance P (SP), calcitonin gene-related peptide (CGRP), and galanin (GAL) and with a reduced nicotinamide adenine dinucleotide phosphate-diaphorase (NADPHd) reaction. The distribution of the terminals was compared with that of alpha-smooth-muscle actin (SMA) staining in TM and SS. The relationship between terminals and adjacent cells or ECM components was also studied in ultrathin sections through the TM and SS of 11 monkey eyes cut in sagittal, tangential, and frontal planes. RESULTS: NADPHd-positive nerve terminals were present, especially in the outer portion of both human and monkey TM and in the SS. VACHT-immunoreactive (IR) fibers were found in human but not in monkey SS and TM. The fibers were most numerous in the elongated SS and posterior TM where most cells also stained for SMA. SP- and CGRP-IR nerve endings were also more numerous in the outer TM and SS than in the inner TM. Ultrastructurally, staining for SP was seen in nerve endings containing mitochondria and dense core vesicles and was in contact with the cribriform elastic network. In the posterior SS of monkey eyes were large terminals similar to those previously described in human eyes. CONCLUSIONS: The results show for the first time that in the primate TM and SS, there are cholinergic and nitrergic nerve terminals that could induce contraction and relaxation of TM and SS cells. Terminals in contact with the elastic-like network of the TM and containing SP-IR resemble afferent mechanoreceptor-like terminals in other parts of the body. These findings raise the possibility that the TM may have some ability to self-regulate aqueous humor outflow.

Senile scleral plaques: CT.

I studied the CT appearances and the distribution of 109 asymptomatic calcified senile scleral plaques in 49 patients (98 eyes). The vast majority were just anterior to the insertion of the horizontal rectus muscles. The calcified plaques are variable in size, can be single or multiple, involve one or both eyes, and are only seen in elderly patients. Beam-hardening artifact and forward protrusion beyond the confines of the sclera are uncommon but potentially confusing features. Confusion with other pathology is unlikely with awareness of the plaques' asymptomatic nature and their clinical and CT appearances.

Autonomic nerves containing substance P in the aqueous outflow channels and scleral spur of the guinea pig.

PURPOSE: To study the innervation of the aqueous outflow channels and scleral spur by autonomic nerves containing substance P. METHODS: The experiments were conducted on guinea pigs. Immunohistochemical techniques and capsaicin-ablation of the sensory nerves were used to investigate nerves containing substance P at the light and electron microscopic level. RESULTS: Nerves containing substance P were observed in the aqueous outflow channels and scleral spur regions. The fine structures of these nerves had a similar pattern in those regions, and the labeled elements had abundant small vesicles, a few large vesicles, and numerous neurotubuli. Following capsaicin treatment, these nerves remained intact and no degenerated substance P-like immunoreactive nerves were found. CONCLUSIONS: Nerves containing substance P are most likely of autonomic origin in view of their ultrastructural features. These nerves innervate the aqueous outflow channels and scleral spur, and are probably important for neurogenic influences on the intraocular pressure by the autonomic nervous system.