In Vivo Scanning Laser Confocal Microscopy of Conjunctival Goblet Cells in Medically-controlled Glaucoma

Staso, Silvio Di; Agnifili, Luca; Ciancaglini, Marco; Murano, Gianluca; Borrelli, Enrico; Mastropasqua, Leonardo

doi:10.21873/invivo.11259

Cited by 12 publications

(8 citation statements)

References 38 publications

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“…Indeed, studying the gel in its native environment, the equilibrium swollen state is the most optimal option and is possible with laser scanning confocal microscopy (LSCM), which allows the visualization of swollen hydrogels in both 2D and 3D. LSCM allows the direct observation of samples in their hydrated state without the necessity of freezing, collecting z-plane images (z-stacks) with subsequent gel volume reconstruction to provide information about the size and distribution of pores in hydrogels [30]. Moreover, LSCM enables the observation of native hydrogels, as well as hydrogels with growing cells in situ [31,32].…”

Section: Introductionmentioning

confidence: 99%

Revealing the True Morphological Structure of Macroporous Soft Hydrogels for Tissue Engineering

Podhorská

Vetrík

Chylíková-Krumbholcová

et al. 2020

Applied Sciences

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(1) Background: Macroporous hydrogel scaffolds based on poly [N-(2-hydroxypropyl) methacrylamide] are one of the widely studied biocompatible materials for tissue reparation and regeneration. This study investigated the morphological changes during hydrogel characterization which can significantly influence their future application. (2) Methods: Three types of macroporous soft hydrogels differing in pore size were prepared. The macroporosity was achieved by the addition of sacrificial template particles of sodium chloride of various sizes (0–30, 30–50, and 50–90 µm) to the polymerizing mixture. The 3D structure of the hydrogels was then investigated by scanning electron microscopy (SEM) and laser scanning confocal microscopy (LSCM). The SEM was performed with specimens rapidly frozen to various temperatures, while non-frozen gels were visualized with LSCM. (3 and 4) Results and Conclusion: In comparison to LSCM, the SEM images revealed a significant alteration in the mean pore size and appearance of newly formed multiple connections between the pores, depending on the freezing conditions. Additionally, after freezing for SEM, the gel matrix between the pores and the fine pores collapsed. LSCM visualization aided the understanding of the dynamics of pore generation using sodium chloride, providing the direct observation of hydrogel scaffolds with the growing cells. Moreover, the reconstructed confocal z-stacks were a promising tool to quantify the swollen hydrogel volume reconstruction which is not possible with SEM.

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Section: Introductionmentioning

confidence: 99%

Revealing the True Morphological Structure of Macroporous Soft Hydrogels for Tissue Engineering

Podhorská

Vetrík

Chylíková-Krumbholcová

et al. 2020

Applied Sciences

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“…Antiglaucoma eye drops have been reported to be associated with corneal epithelial damage and a significant reduction in conjunctival goblet cells [29]. Preservatives in antiglaucoma eye drops have been reported to affect tear turnover and stability, and result in the infiltration of inflammatory cells and fibroblasts in the conjunctiva [30].…”

Section: Discussionmentioning

confidence: 99%

The Effects of 3% Diquafosol Sodium Eye Drops on Tear Function and the Ocular Surface of Cu, Zn-Superoxide Dismutase-1 (Sod1) Knockout Mice Treated with Antiglaucoma Eye Medications

et al. 2020

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Anti-glaucoma eye drop treatment often induces dry eyes and can lead to poor medication adherence. This study aimed to investigate the effects of 3% diquafosol sodium eye drops on tear function and the ocular surface epithelium in Sod1−/− mice after treatment with anti-glaucoma eye drops. The mice were divided into four groups: group 1, control group; group 2, anti-glaucoma eye drop; group 3, anti-glaucoma eye drops followed by a secretagogue eye drop (3% diquafosol); and group 4, simultaneous anti-glaucoma and secretagogue eye drop. Mice underwent assessments of tear quantity, tear film breakup time, and vital staining score. Mice in groups 3 and 4 showed significantly better tear stability and lower corneal staining scores than mice in group 2 after eye drop instillations (p < 0.05). Mice in group 4 showed significantly better tear stability, lower corneal staining scores, and higher goblet cell densities than those in group 1 after eye drop instillations (p < 0.05). The conjunctival epithelium showed stratification and abundance of Muc5AC-positive goblet cells in group 4, whereas thinning with desquamation was observed with a few goblet cells in group 2. Thus, simultaneous administration of 3% diquafosol sodium eye drops with topical anti-glaucoma drops showed favorable effects on tear stability and the corneal epithelium against the ocular surface toxicity inflicted by the anti-glaucoma eye drops.

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“…In 2018, Di Staso et al . reported results from a clinical study in 55 medically controlled glaucomatous patients, 17 DED patients and 17 healthy individuals aimed to evaluate goblet cell density using non-invasive in-vivo laser scanning confocal microscopy ( 135 ). The study revealed a significant reduction in goblet cell density in both glaucoma and DED groups compared to healthy controls (P<0.001) and the authors suggested that goblet cell reduction “may play a role in pathophysiology of the glaucoma-related disease of ocular surface”.…”

Section: Conjunctivamentioning

confidence: 99%

Review of Biomarkers in Ocular Matrices: Challenges and Opportunities

et al. 2019

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Biomarkers provide a powerful and dynamic approach to improve our understanding of the mechanisms underlying ocular diseases with applications in diagnosis, disease modulation or for predicting and monitoring of clinical response to treatment. Defined as measurable indicator of normal or pathological processes, biomarker evaluation has been used extensively in drug development within clinical settings to better comprehend effectiveness of treatment in ocular diseases. Biomarkers in the eye have the advantage of access to multiple ocular matrices via minimally invasive methods. Repeat sampling for biomarker assessment has enabled reproducible objective measures of disease process or biological responses to a drug treatment. This review describes the usage of biomarkers with respect to four commonly sampled ocular matrices in clinic: tears, conjunctiva, aqueous humor and vitreous. Issues that affect the evaluation of biomarkers are discussed along with opportunities to leverage biomarkers such that ultimately, they can be used for customized targeted therapy.

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In Vivo Scanning Laser Confocal Microscopy of Conjunctival Goblet Cells in Medically-controlled Glaucoma

Abstract: Abstract. Aim: The aim of this study was to evaluate the goblet cell density (GCD) of conjunctiva in medically- GCD with OSDI and with BUT (p<0.001; respectively

Cited by 12 publications

References 38 publications

Revealing the True Morphological Structure of Macroporous Soft Hydrogels for Tissue Engineering

Revealing the True Morphological Structure of Macroporous Soft Hydrogels for Tissue Engineering

The Effects of 3% Diquafosol Sodium Eye Drops on Tear Function and the Ocular Surface of Cu, Zn-Superoxide Dismutase-1 (Sod1) Knockout Mice Treated with Antiglaucoma Eye Medications

Review of Biomarkers in Ocular Matrices: Challenges and Opportunities

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