Wide-Field In Vivo Confocal Microscopy of Meibomian Gland Acini and Rete Ridges in the Eyelid Margin.
ABSTRACT: Purpose:In vivo confocal microscopy (IVCM) has been widely used to evaluate changes in the meibomian glands (MGs) in response to age and disease. This study examined the structures described as MGs using wide-field IVCM and laser scanning confocal microscopy (LSCM) in situ and characterized their spatial distribution and localization relevant to the eyelid margin. Methods:IVCM was performed on 30 subjects aged 18 to 38 to visualize structures in the eyelid margin. Size, shape, and distribution characteristics were measured, and individual frames were montaged into wide-field images. Structures observed on IVCM were then visualized using LSCM of whole-mount and cryosectioned cadaver eyelids stained with Nile red, mucin-1 (MUC1), laminin-5, and 4',6-diamidine-2'-phenylindole dihydrochloride. Results:The size, distribution, and staining patterns of the reflective structures seen on IVCM did not correspond to the MGs in cadaver eyelids. Instead, staining profiles indicated that these structures corresponded to the rete ridges present at the dermal-epidermal junction. Wide-field imaging showed a densely populated field of rete ridges with distinct size and shape characteristics depending on their location relative to the meibomian orifices. A distal shift of the mucocutaneous junction (MCJ) was evident in some eyelids. Conclusions:IVCM is unable to visualize MGs in the human eyelid margin due to light attenuation at that tissue depth. LSCM confirms that these structures are rete ridges located at the dermal-epidermal junction. Alterations in the structure of the dermal-epidermal junction within the eyelid margin indicate a shifting of the MCJ and may impact tear film dynamics.
Project description:<h4>Importance</h4>The clinical diagnosis of conjunctival and eyelid margin tumors is challenging, and new noninvasive imaging techniques could be valuable in this field.<h4>Objective</h4>To assess the diagnostic accuracy of handheld in vivo reflectance confocal microscopy (IVCM) for the diagnosis of eyelid margin and conjunctival tumors.<h4>Design</h4>A prospective observational study was conducted at University Hospital of Saint-Etienne from January 2, 2011, to December 31, 2016 (inclusion of patients until December 31, 2015, and follow-up until December 31, 2016). A total of 278 consecutive patients with eyelid margin or conjunctival lesions were included. Conjunctival lesions were diagnosed with a conventional clinical examination using a slitlamp and by handheld IVCM. Final diagnoses were established by histopathologic examination for 155 neoformations suspicious for being malignant through clinical and/or IVCM examination that were excised and on follow-up of 12 months or longer for the remaining 140 lesions.<h4>Main outcomes and measures</h4>Sensitivity, specificity, and positive and negative predictive values for malignant tumors of the conjunctiva and eyelid margin were calculated using clinical examination with slitlamp and handheld IVCM.<h4>Results</h4>In the 278 patients (136 [48.9%] females; mean [SD] age, 59  years), a total of 166 eyelid margin and 129 conjunctival lesions were included in the analysis. Of the 155 excised neoformations with a histopathologic diagnosis, IVCM showed higher sensitivity compared with clinical examination conducted with the slitlamp for malignant tumors of the eyelid margin (98% vs 92%) and conjunctiva (100% vs 88%). The specificity for malignant eyelid margin tumors was higher for IVCM than for slitlamp examination (74% vs 46%), but slightly less for malignant conjunctival tumors (78% vs 88%). Analysis of all neoformations (155 excised and 140 in follow-up) confirmed these differences in the diagnostic accuracy of the clinical examination and IVCM. The presence of hyperreflective Langerhans cells mimicking malignant melanocytes was the main cause for misdiagnosis of malignant conjunctival tumors with IVCM.<h4>Conclusions and relevance</h4>Handheld IVCM could be a useful tool for the identification of malignant conjunctival tumors. Further studies are required to confirm the usefulness of this device and identify possible features that can differentiate Langerhans cells from malignant melanocytes to prevent the misdiagnosis of melanoma using IVCM.
Project description:<h4>Purpose</h4>The Meibomian gland (MG) produces the lipid layer of the tear film, and changes to the MG that lead to a decrease or alteration in lipid quality/content may lead to MG dysfunction, a major cause of evaporative dry eye disease with prevalence ranging from 39% to 50%. Little is known about the developmental cues that regulate MG morphogenesis and homeostasis. Our study investigates the role of hyaluronan (HA), a major extracellular matrix component, in eyelid formation and MG development and function.<h4>Methods</h4>Hyaluronan synthase (Has) knockout mice were used to determine the role of HA in the eyelid and MG. Eyelids were obtained during different developmental stages and MG morphology was analyzed. Tet-off H2B-GFP/K5tTA mice and 5-ethynyl-2'-deoxyurdine (EdU) incorporation were used to determine the role of HA in maintaining slow-cycling and proliferating cells within the MG, respectively. Data were confirmed using an in vitro proliferation assay, differentiation assay and spheroid cultures.<h4>Results</h4>Has knockout mice present precocious MG development, and adult mice present MG hyperplasia and dysmorphic MGs and eyelids, with hyperplastic growths arising from the palpebral conjunctiva. Our data show that a highly organized HA network encompasses the MG, and basal cells are embedded within this HA matrix, which supports the proliferating cells. Spheroid cultures showed that HA promotes acini formation.<h4>Conclusions</h4>HA plays an important role in MG and eyelid development. Our findings suggest that Has knockout mice have abnormal HA synthesis, which in turn leads to precocious and exacerbated MG morphogenesis culminating in dysmorphic eyelids and MGs.
Project description:<h4>Purpose</h4>To investigate the indirect effects of intense pulsed light (IPL) on morphological and pathological changes of the meibomian glands (MGs) in apolipoprotein E knockout (<i>ApoE<sup>-/-</sup></i> ) mice and explore the underlying mechanisms.<h4>Methods</h4><i>ApoE<sup>-/-</sup></i> mice were treated with or without IPL three times below the lower eyelids and MGs were not directly exposed to irradiation. The eyelids and ocular surface were observed under a stereoscope. The morphology of MGs was examined by photographing and hematoxylin and eosin staining. Lipid droplets in MGs were examined by Oil Red O staining. The ultrastructure of meibocytes and mitochondria was observed under transmission electron microscopy. The relative gene and protein expression in MGs of upper eyelids was determined by immunostaining, Western blot, and qRT-PCR.<h4>Results</h4>Three IPL treatments decreased the toothpaste-like plugging of orifices and thickening and irregularity of the upper and lower eyelid margins in <i>ApoE<sup>-/-</sup></i> mice. The morphology of some MGs improved after IPL treatments, accompanied by increased proliferation of acinar basal cells and decreased ductal keratinization. Furthermore, the accumulation of hyperchromatic lipid droplets in the acini increased, and the lipid droplets distributed in the cells around the acini were round and small. Compared with untreated <i>ApoE<sup>-/-</sup></i> mice, oxidative stress and apoptosis were downregulated by IPL treatment, accompanied by the improvements in mitochondrial structure. Further research showed that IPL treatments reduced the levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-17A, IL-6 in MGs and inactivated nuclear factor kappa B (NF-κ B).<h4>Conclusion</h4>Collectively, the results demonstrate that indirect effects of IPL can improve the structure and function of MGs and mitigate the progression of MGD, which may be related to the indirect effects of photobiomodulation.
Project description:To describe a standardized technique for acquiring and viewing photographic images of eyelids, assess the reproducibility and validity of a grading protocol for signs of anterior blepharitis, and to explore whether the signs depend on the eyelid or the area of the eyelid assessed.Subjects with anterior blepharitis ranging from none to severe were examined by ophthalmologists at clinical sites. Digital images of the eyelids of subjects were acquired using a protocol that allowed for the calibration of color and luminance. Three ophthalmologists at a centralized reading center applied a novel protocol for grading features of anterior blepharitis from the digital images viewed on color-calibrated monitors. The agreement among graders was assessed using percent agreement and weighted kappa statistics (Kw), and the correlation of photographic and clinical gradings was assessed using Spearman correlation coefficients.Agreement among graders was excellent (Kw > 0.80) on the number of eyelid margin vessels and was substantial (Kw between 0.61 and 0.80) for erythema, collarettes, number of engorged vessels, and number of lashes. Grading of the photographic images and the clinical assessments of erythema and lid debris were moderately correlated (r = 0.27-0.45). The grades for different features depended on whether the upper or lower eyelid, eyelid skin or lid margin, and central or lateral lid were assessed.The application of a protocol to obtain and display calibrated digital images of eyelids supports the standardized assessment of anterior blepharitis in clinical care and research studies.
Project description:Eye is one of the most crucial organs in need of protection. The most important function of eyelids is designed to protect the eyeball. Despite their small and thin surface area, they are among the most sunlight-exposed area of skins, hence, it is prone to develop eyelid tumors. As the malignant tumors have the tendency of growing aggressively and have a high mortality, we have to meticulously remove the lesion along with its surrounding tissue, if needed, to ensure the clearance of tumor margin. To strike a suitable outcome between the cosmetic and function after tumor removal is a quite challenging task. Compromise on aesthetics and eyelids' indispensable function of protecting the eyes during management, may lead to unfavorable cosmetic deformity and loss of vision. Therefore, it is imperative that wise selection of appropriate techniques be discussed and managed accordingly. In this article, the authors reported two case scenarios of eyelid reconstruction which was performed to show the management of full-thickness defect after removal of the malignant neoplasms. Skin flaps and graft technique used to reconstruct the eyelid in each patient was different to emphasize that eyelid malignancies require an individualized management.
Project description:Purpose:To compare the effects of eyelid treatment with the iLUX MGD Treatment System and the LipiFlow Thermal Pulsation System on objective and subjective parameters of meibomian gland function and symptoms. Patients and Methods:In this randomized, open-label, controlled, multicenter clinical trial, both eyes of 142 patients aged ?18 years with Ocular Surface Disease Index (OSDI) scores ?23, total meibomian gland scores (MGS) ?12 in the lower eyelid of each eye, and tear break-up time (TBUT) <10 s were randomized 1:1 to iLUX or LipiFlow treatment, with stratification by test center. The primary effectiveness endpoints were changes in total MGS (masked) and TBUT from baseline to 4 weeks. The secondary effectiveness endpoint was changed in OSDI score from baseline to 4 weeks. Results:Both devices significantly improved effectiveness outcomes, with no differences between the two devices. At the 4-week visit, mean MGS, TBUT, and OSDI scores improved at least 16.9 ± 11.5, 2.6 ± 3.2 s, and 28.0 ± 22.8, respectively, across treatment groups and treated eyes. Four device/procedure-related events occurred in the iLUX group, compared with none in the LipiFlow group, but there were no device-related adverse events that involved changes in lid margins, eyelids, or lash integrity. Corneal staining, intraocular pressure, and visual acuity did not differ in the two groups. Conclusion:Both treatments produced significant improvements in meibomian gland function and symptoms. For all effectiveness measures, there were no statistically significant differences between the two treatments.
Project description:Meibomian glands (MGs) are sebaceous glands of the eyelid margin that secrete lipids needed to avert tear evaporation and to help maintain ocular surface homeostasis. Obstruction of MGs or other forms of MG dysfunction can promote chronic diseases of the ocular surface. Although chronic eyelid inflammation, such as allergic eye disease, is an associated risk factor for obstructive MG dysfunction, it is not clear whether inflammatory processes contribute to the pathophysiology of MG obstruction. We show that polymorphonuclear neutrophils (PMNs) promoted MG obstruction in a chronic inflammatory model of allergic eye disease in mice. Analysis of leukocytes in tears of patients with MG dysfunction showed an increase in PMN numbers compared to healthy subjects. Moreover, PMN numbers in tears positively correlated with clinical severity of MG dysfunction. Our findings point to a role for PMNs in the pathogenesis and progression of MG dysfunction.
Project description:Congenital eyelid imbrication syndrome (CEIS) is an extremely rare, benign, transient, self-limiting eyelid malposition disorder. The classic triad of signs in patients with a CEIS consists of bilateral upper eyelids overriding the lower eyelids when child was in sleep, bilateral medial and lateral canthal tendon laxity and tarsal conjunctival hyperemia. We report a third case of congenital combined eyelid imbrication and floppy eyelid syndrome in healthy neonate that was resolved within a week with conservative treatment.
Project description:<b>Purpose:</b> The present study was designed to observe the curative effect of early intralesional corticosteroid injection to treat unsatisfactory double eyelid. <b>Methods:</b> This retrospective, observational study included 30 female patients (age 22-35 years) receiving intralesional corticosteroid injection after complaining about the unsatisfactory double eyelid post-transcutaneous upper eyelid blepharoplasty. The postoperative double eyelid anomalies included multiple folds, sunken eyelids, and severe postoperative eyelid edema. The evaluation of the clinical effect was based on the bilateral symmetry of the eyelid, the placement of the lid fold, the radian of the double eyelid, and the extent of the swelling and depression. <b>Results:</b> Multiple folds were observed in 11 (36.67%) patients, sunken eyelids in 9 (30.00%) patients, and severe postoperative eyelid edema in 10 (33.33%) patients. Among them, 17 (56.67%) patients received one intralesional injection and experienced significant improvement in the eyelid, while 8 (26.67%) accepted another injection at 1 month after the first injection and achieved marked improvement. The remaining 5 (16.67%) patients were dissatisfied with the results and might need further repair surgery: 2 patients had multiple folds, 2 showed postoperative edema, and 1 presented sunken eyelids. None of the patients reported any adverse reactions. <b>Conclusion:</b> Intralesional corticosteroid injection for the correction of the postoperative unsatisfactory double eyelid is safe and convenient, and provides a novel method for early intervention.
Project description:<h4>Introduction</h4>The World Health Organization has identified management of postoperative trichiasis (PTT) as one of the key remaining areas of focus needed to eliminate blinding trachoma as a public health problem. We developed the Bevel-Rotation Advancement Procedure (B-RAP) to treat individuals who need repeat trichiasis surgery.<h4>Methods</h4>Scarring caused by trichiasis surgery can cause the eyelid to become thick and distorted, making repeat surgery more difficult. To minimize eyelid thickness following B-RAP, a beveled incision of the tarsus is made allowing a marginal rotation of the eyelash fragment. Dissection between the anterior and posterior lamellae above the beveled incision and removal of scar tissue allows the marginal rotation to be combined with a posterior lamellar advancement to treat severely scarred eyelids with PTT and eyelid contour abnormalities (ECAs).<h4>Results</h4>Two surgeons performed B-RAP on 44 eyelids of 30 patients with PTT. The number of prior trachomatous trichiasis (TT) surgeries ranged from 2 to more than 4. At the 3-6 months postoperative visit, 37 eyelids (84%) had no recurrence of PTT. Three eyelids had central lashes touching; the remaining eyelids with recurrent PTT had nasal and temporal lashes touching. Fifteen eyelids (34%) had ECAs, but only 1 was severe.<h4>Conclusions</h4>B-RAP was developed considering the altered eyelid anatomy found in the postsurgical eyelid with TT. Thinning of the eyelash fragment and removal of postoperative scar tissue improves the ability to advance and stabilize the eyelash fragment after external rotation. B-RAP shows promise as a procedure for improving outcomes of repeat trichiasis surgery.