Treatment Of Ocular Surface Disorders Research Articles

Role of Eye Platelet-Rich Plasma in the Treatment of Ocular Surface Disorders

The use of blood derivatives represents an alternative therapeutic approach that is gaining interest in regenerative medicine dueto its potential to stimulate and accelerate tissue healing. Platelet-rich plasma is the highly concentrated form of autologoushuman platelets in a small amount of plasma which contains important growth factors and plasma proteins that play a significantrole in the wound healing process by epithelial differentiation and collagen bundle organization.In this article, we aim to provide an update on the current literature regarding the eye platelet-rich plasma, its methods ofpreparation, physiological and biochemical properties, its clinical applications, safety and efficacy as compared to other bloodderived products, etc. In ophthalmology, this product is being used in the management of symptomatic dry eyes, corneal ulcers,periocular chemical and thermal burns, idiopathic macular hole, skin rejuvenation post blepharoplasties and more recentlyactinic elastosis in the lower eyelid regions. The role of eye platelet-rich plasma in ocular surface disorders has been sparselystudied in literature with more studies and reports on the application of autologous and allogeneic serum eye drops therefore, itbecomes very important to update ourselves with more studies in this topic to prove the efficiency of this blood-derived product.

Beneficial Effects of Plasma Rich in Growth Factors (PRGF) Versus Autologous Serum and Topical Insulin in Ocular Surface Cells

Purpose In the last few decades, several blood derived products such as platelet-rich plasma (PRP), plasma rich in growth factors (PRGF) and autologous serum (AS) have been used for the treatment of ocular surface disorders. Recently, insulin has been proposed to be used as an alternative for the treatment of ocular surface diseases. The aim of this study was to evaluate the biological potential of PRGF eye drops in comparison with AS and insulin on ocular surface cells. Methods Blood from three healthy young donors was collected to obtain autologous serum (AS) eye drops and plasma rich in growth factors (PRGF) eye drops. Insulin (Actrapid®) was diluted at 1 and 0.2 IU/mL. The biological potential of PRGF, AS and insulin was assessed by proliferation in HCE, HK and HConF cells. Wound healing assay was performed in HCE cells after incubation with the different treatments. HConF and HK cells were differentiated to myofibroblast after treatment with 2.5 ng/mL of TGF-β1 and then incubated with all treatments. Results PRGF eye drops induced significantly higher proliferation rate compared to AS or insulin in HConF and HK cells, but not in HCE cells. In addition, the percentage of wound healing area was significantly reduced after PRGF treatment in comparison with AS or insulin treatment. Furthermore, PRGF significantly reduced the number of myodifferentiated cells compared to AS and insulin at both concentrations analyzed. Conclusion The results obtained in the present study show that PRGF increases the biological activity of the ocular surface cells and reduces the expression of fibrosis marker compared to insulin or AS. Translational relevance The present study suggests that plasma rich in growth factors eye drops are a more effective therapy than insulin and autologous serum eye drops for the treatment of ocular surface diseases.

Therapeutic Profile of Human Umbilical Cord Blood Serum and Autologous Serum Therapies in Treatment of Ocular Surface Disorders: A Pilot Study

Purpose: Umbilical cord blood serum (UCBS) is an effective adjunctive treatment along with conventional therapy in ocular surface disorders (OSDs). It aids in rapid ocular surface restoration thereby achieving epithelial integrity, in addition to improvement in subjective and objective parameters. The study aims to compare the efficacy of human umbilical cord blood serum and autologous serum (AS) in treatment of OSD. Methods: A prospective randomized study was conducted on 101 eyes diagnosed with OSD resulting from dry eye disease (DED; n = 40), acute chemical burn (ACB; n = 21), and ocular allergy (OA; n = 40). Randomization was done in Group I, administered with AS, and Group II with UCBS. Outcomes evaluated were visual acuity (VA), eye sensation score (ESS), ocular surface disease index (OSDI), tear break-up time (TBUT), Schirmer's value, Corneal Fluorescein Score, epithelial defect, limbal ischemia, corneal clarity (CC), and improvement in grade of severity. Statistical analysis was done using Wilcoxon signed-rank, Wilcoxon rank sum, Chi-square, and Z-test with a significance level (P ≤ 0.05). Results: In DED, Group II showed significant improvement in VA, ESS, and OSDI by the 7th day, whereas the mean Schirmer, TBUT, and corneal fluorescein staining score improved by 3 months. In ACB, Group II showed improvement in VA, reepithelialization, reduction in limbal ischemia, and CC by 3 months. In OA, Group II showed improvement in ESS by day 7. Conclusion: Human umbilical cord blood serum is more effective than AS in restoring ocular surface.

Erratum to Limbal-Rigid Contact Lens Wear for the Treatment of Ocular Surface Disorders: A Review.

Erratum to Limbal-Rigid Contact Lens Wear for the Treatment of Ocular Surface Disorders: A Review.

Limbal-Rigid Contact Lens Wear for the Treatment of Ocular Surface Disorders: A Review.

:Recently, the prescription of large-diameter rigid gas-permeable contact lenses (CLs), also known as “scleral lenses,” “corneoscleral lenses,” and “limbal-rigid CLs,” is on the rise for the treatment of both moderate and severe ocular surface disorders (OSDs). Compared with scleral lenses, the diameter of limbal-rigid CLs is generally smaller, that is, a diameter ranging from 13.0 to 14.0 mm, and they are designed so that the peripheral edge bears on the limbus. The Suncon Kyoto-CS (Sun Contact Lens Co., Ltd.) is a novel limbal-rigid CL design with multistep curves on the peripheral edge for easy tear exchange during blinking that removes debris and prevents lens clouding or fogging, thus allowing patients to enjoy a longer daily duration of CL wear. In severe OSD cases, limbal-rigid CL wear after surgery is a noninvasive therapeutic approach that can neutralize corneal irregularities, decrease dry eye symptoms, prevent the progression or recurrence of symblepharon, and improve the patient's visual acuity and overall quality of life. Thus, surgeries such as amniotic membrane transplantation and cultivated oral mucosal epithelial transplantation, as well as limbal-rigid CL wear, which is noninvasive, are valuable and effective treatment strategies that can now be applied for the management of patients afflicted with severe OSDs.

Proteomic Characterization of Plasma Rich in Growth Factors and Undiluted Autologous Serum.

Over the last three decades, there has been special interest in developing drugs that mimic the characteristics of natural tears for use it in the treatment of several ocular surface disorders. Interestingly, the composition of blood plasma is very similar to tears. Therefore, different blood-derived products like autologous serum (AS) and plasma rich in growth factors (PRGF) have been developed for the treatment of diverse ocular pathologies. However, scarce studies have been carried out to analyze the differences between both types of blood-derived products. In the present study, blood from three healthy donors was drawn and processed to obtain AS and PRGF eye drops. Then, human corneal stromal keratocytes (HK) were treated with PRGF or undiluted AS. Proteomic analysis was carried out to analyze and characterize the differential protein profiles between PRGF and AS, and the differentially expressed proteins in HK cells after PRGF and AS treatment. The results obtained in the present study show that undiluted AS induces the activation of different pathways related to an inflammatory, angiogenic, oxidative stress and scarring response in HK cells regarding PRGF. These results suggest that PRGF could be a better alternative than AS for the treatment of ocular surface disorders.

Ocular Delivery of Predatory Bacteria with Cryomicroneedles Against Eye Infection.

The development of potent antibiotic alternatives with rapid bactericidal properties is of great importance in addressing the current antibiotic crisis. One representative example is the topical delivery of predatory bacteria to treat ocular bacterial infections. However, there is a lack of suitable methods for the delivery of predatory bacteria into ocular tissue. This work introduces cryomicroneedles (cryoMN) for the ocular delivery of predatory Bdellovibrio bacteriovorus (B. bacteriovorus) bacteria. The cryoMN patches are prepared by freezing B. bacteriovorus containing a cryoprotectant medium in a microneedle template. The viability of B. bacteriovorus in cryoMNs remains above 80% as found in long‐term storage studies, and they successfully impede the growth of gram‐negative bacteria in vitro or in a rodent eye infection model. The infection is significantly relieved by nearly six times through 2.5 days of treatment without substantial effects on the cornea thickness and morphology. This approach represents the safe and efficient delivery of new class of antimicrobial armamentarium to otherwise impermeable ocular surface and opens up new avenues for the treatment of ocular surface disorders.

Scleral Contact Lenses for Treatment of Ocular Surface Disorders: A Narrative Review

Scleral lenses have been in the origin of contact lenses back in 1880, when the pioneers did the first glass contact lenses. With the advent of highly oxygen permeable materials and the increasing need for advanced solutions for severe ocular surface disorders has produced the driving pressures for a great amount of different contact lens designs to be available.

Surgical Application of Human Amniotic Membrane and Amnion-Chorion Membrane in the Oral Cavity and Efficacy Evaluation: Corollary With Ophthalmological and Wound Healing Experiences.

Due to its intrinsic properties, there has been growing interest in human amniotic membrane (hAM) in recent years particularly for the treatment of ocular surface disorders and for wound healing. Herein, we investigate the potential use of hAM and amnion-chorion membrane (ACM) in oral surgery. Based on our analysis of the literature, it appears that their applications are very poorly defined. There are two options: implantation or use as a cover material graft. The oral cavity is submitted to various mechanical and biological stimulations that impair membrane stability and maintenance. Thus, some devices have been combined with the graft to secure its positioning and protect it in this location. This current opinion paper addresses in detail suitable procedures for hAM and ACM utilization in soft and hard tissue reconstruction in the oral cavity. We address their implantation and/or use as a covering, storage format, application side, size and number, multilayer use or folding, suture or use of additional protective covers, re-application and resorption/fate. We gathered evidence on pre- and post-surgical care and evaluation tools. Finally, we integrated ophthalmological and wound healing practices into the collected information. This review aims to help practitioners and researchers better understand the application of hAM and ACM in the oral cavity, a place less easily accessible than ocular or cutaneous surfaces. Additionally, it could be a useful reference in the generation of new ideas for the development of innovative protective covering, suturing or handling devices in this specific indication. Finally, this overview could be considered as a position paper to guide investigators to fulfill all the identified criteria in the future.

A single cell atlas of human cornea that defines its development, limbal progenitor cells and their interactions with the immune cells

PurposeSingle cell (sc) analyses of key embryonic, fetal and adult stages were performed to generate a comprehensive single cell atlas of all the corneal and adjacent conjunctival cell types from development to adulthood. MethodsFour human adult and seventeen embryonic and fetal corneas from 10 to 21 post conception week (PCW) specimens were dissociated to single cells and subjected to scRNA- and/or ATAC-Seq using the 10x Genomics platform. These were embedded using Uniform Manifold Approximation and Projection (UMAP) and clustered using Seurat graph-based clustering. Cluster identification was performed based on marker gene expression, bioinformatic data mining and immunofluorescence (IF) analysis. RNA interference, IF, colony forming efficiency and clonal assays were performed on cultured limbal epithelial cells (LECs). ResultsscRNA-Seq analysis of 21,343 cells from four adult human corneas and adjacent conjunctivas revealed the presence of 21 cell clusters, representing the progenitor and differentiated cells in all layers of cornea and conjunctiva as well as immune cells, melanocytes, fibroblasts, and blood/lymphatic vessels. A small cell cluster with high expression of limbal progenitor cell (LPC) markers was identified and shown via pseudotime analysis to give rise to five other cell types representing all the subtypes of differentiated limbal and corneal epithelial cells. A novel putative LPCs surface marker, GPHA2, expressed on the surface of 0.41% ± 0.21 of the cultured LECs, was identified, based on predominant expression in the limbal crypts of adult and developing cornea and RNAi validation in cultured LECs. Combining scRNA- and ATAC-Seq analyses, we identified multiple upstream regulators for LPCs and demonstrated a close interaction between the immune cells and limbal progenitor cells. RNA-Seq analysis indicated the loss of GPHA2 expression and acquisition of proliferative limbal basal epithelial cell markers during ex vivo LEC expansion, independently of the culture method used. Extending the single cell analyses to keratoconus, we were able to reveal activation of collagenase in the corneal stroma and a reduced pool of limbal suprabasal cells as two key changes underlying the disease phenotype. Single cell RNA-Seq of 89,897 cells obtained from embryonic and fetal cornea indicated that during development, the conjunctival epithelium is the first to be specified from the ocular surface epithelium, followed by the corneal epithelium and the establishment of LPCs, which predate the formation of limbal niche by a few weeks. ConclusionsOur scRNA-and ATAC-Seq data of developing and adult cornea in steady state and disease conditions provide a unique resource for defining genes/pathways that can lead to improvement in ex vivo LPCs expansion, stem cell differentiation methods and better understanding and treatment of ocular surface disorders.

Update on the role of Eye platelet-rich plasma (E-PRP) in the treatment of ocular surface disorders

Platelet rich plasma is the highly concentrated form of autologous human platelets in a small amount of plasma which contains important growth factors and plasma proteins that plays a significant role in wound healing process by epithelial differentiation and collagen bundle organization. In this article, we aim to provide an update on the current literature regarding the eye platelet rich plasma, its methods of preparation, physiological and biochemical properties, its clinical applications, safety and efficacy as compared to other blood derived products etc. In ophthalmology, this product is being used in the management of symptomatic dry eyes, corneal ulcers, periocular chemical and thermal burns, Idiopathic macular hole, Skin rejuvenation post Blepharoplasties and more recently actinic elastosis in the lower eyelid regions. The role of eye platelet rich plasma in ocular surface disorders has been sparsely studied in literature with more studies and reports on the application of autologous and allogeneic serum eye drops and therefore, it becomes very important to update ourselves with more studies in this topic to prove the efficiency of this blood derived product. Moreover, the role of other platelet derived products like platelet rich growth factors, platelet lysate in ocular surface disorders have also been discussed.

Case Series Describing the Use of Low-Temperature Vacuum-Dehydrated Amnion (Omnigen) for the Treatment of Corneal Ulcers in Cats and Dogs: 46 Cases (2016-2017)

Amniotic membrane is widely used in the treatment of ocular surface disorders in human and veterinary patients. Preservation and storage of amnion has proven challenging, prompting the development of new preservation techniques. Omnigen, a novel low-temperature vacuum-dehydrated amnion, is reported to possess enhanced structural properties and biochemical stability in vitro, but its clinical use in veterinary patients is not well described. This study aims to document and describe the varied use of Omnigen for the surgical treatment of corneal ulceration in cats and dogs. A total of 45 patients (46 eyes) were recruited from the clinical record system of the Royal Veterinary College (London) between January 2016 and December 2017. Brachycephalic breeds were over-represented (37/45; 82.2%). Omnigen was used as a standalone graft in 5/46 (10.9%) eyes, as a supplementary graft in 29/46 (63.0%) eyes and as a patch in 12/46 (26.1%) eyes. Graft failure occurred in 10/46 eyes (21.7%). At final examination 43/46 eyes (93.5%) had healed and 31/33 eyes (93.9%) were visual. This study demonstrates the successful use of Omnigen for the surgical treatment of corneal ulceration in cats and dogs. Further studies are needed to clarify its properties and benefits in the clinical field.

Quantification of Growth Factors and Fibronectin in Diverse Preparations of Platelet-Rich Plasma for the Treatment of Ocular Surface Disorders (E-PRP).

PurposeThe purpose of this study was to quantify the presence of growth factors (GFs) and fibronectin in autologous platelet-rich plasma for topical ocular use (E-PRP) comparing their concentration when different preparation and preservation procedures were applied.MethodsE-PRP was prepared with blood from healthy volunteers. The count of platelets, leukocytes, and red blood cells in the whole blood and E-PRP were performed. The concentration of the GFs platelet-derived growth factor BB (PDGF-BB), transforming growth factor β1 (TGF-β1), epidermal growth factor (EGF), vascular endothelial growth factor A (VEGF-A), and fibronectin was determined in each of the four procedures applied including fresh, frozen at −20°C for 3 months, fresh-spin, and frozen-spin at −20°C E-PRP samples. Posterior statistical analysis was performed to establish significant differences between groups and between GFs in relation to the amounts of platelets.ResultsPlatelets in the E-PRP doubled in the number of basal values of whole blood (P ≤ 0.01). The blood cells in the E-PRP decreased drastically in red cells (99%) and also in leukocytes (82%). The concentration of PDGF-BB and EGF was significantly higher (P < 0.01) when the E-PRP samples were frozen at −20°C. However, no significant differences were observed for TGF-β1, VEGF-A, and fibronectin (P > 0.05). The concentration of GFs in the E-PRP did not necessarily correlate with the number of platelets.ConclusionsFreezing the E-PRP for 3 months at −20°C increased the concentration of important proteins, such as PDGF-BB and EGF, and maintained the levels of others. These findings are essential because treatments, such as E-PRP, used by patients with ocular surface dysfunctions tend to prolong it in time. In addition, subsequent centrifugation of the E-PRP decreased the values of TFG-β1, but not the other GFs, which would allow adjusting the concentration of TFG-β1, as necessary. This procedure guarantees their correct conservation and viability.Translational RelevanceThis work demonstrates how clinical application can be improved by starting from basic research. The quantification of GFs and fibronectin in platelet-rich plasma (PRP) helps to clarify which is the best mode of preparation and preservation of PRP for clinical applications. This allows to optimize the product that is delivered to the patients as a treatment for the dysfunctions of the ocular surface, guaranteeing that the conservation does not affect at all the quality of the PRP that it is going to be used.

Ocular surface disease in dogs part 2: diagnosis and treatment

Dry eye disease (DED) is a commonly diagnosed disorder in small animals. It has been historically described as a quantitative or qualitative deficiency in tear production; however, the pathogenesis of the disease is much more complex than previously recognised. Ocular inflammation is the hallmark of DED, reflected in early nonspecific conjunctivitis and later in dramatic changes of the ocular surface. Prompt recognition of the disease, accurate diagnostics and appropriate treatment are paramount in preserving the ocular surface integrity and the patient's comfort. Part 1 discussed the aetiopathogenesis of the condition and described the clinical signs. This second article looks at diagnostics and treatment of ocular surface disorders in dogs.

Ocular surface disease in dogs part 1: aetiopathogenesis and clinical signs

Dry eye disease (DED) is a commonly diagnosed disorder in small animals. It has been historically described as a quantitative or qualitative deficiency in tear production; however, the pathogenesis of the disease is much more complex than previously recognised. Ocular surface disorders in people are currently investigated as a dysfunction of what is termed the ‘lacrimal functional unit’, consisting of the ocular surface, including Meibomian glands and conjunctiva, in addition to lacrimal glands, eyelids and sensory and motor innervation. A loss of homeostasis of the tear film leads to the vicious cycle of increased osmolarity and inflammation, with dreadful consequences for the ocular surface in both dogs and humans. Ocular inflammation is the hallmark of DED, reflected in early nonspecific conjunctivitis and later in dramatic changes of the ocular surface. Prompt recognition of the disease, accurate diagnostics and appropriate treatment are paramount in preserving the ocular surface integrity and the patient's comfort. This article discusses the aetiopathogenesis of the condition, and describes the clinical signs, while the second article will look at diagnostics and treatment of ocular surface disorders in dogs.

Utilization of metabolic energy in treatment of ocular surface disorders: polyphosphate as an energy source for corneal epithelial cell proliferation.

Impaired regeneration of the corneal epithelium, as found in many ocular surface diseases, is a major clinical problem in ophthalmology. We hypothesized that corneal epithelial regeneration can be promoted by the physiological, energy-delivering as well as “morphogenetically active” polymer, inorganic polyphosphate (polyP). Corneal limbal explants (diameter, 4 mm) were cultivated on collagen-coated well plates in the absence or presence of polyP (chain length, ∼40 Pi units; 50 μg ml−1) or human platelet lysate (hp-lysate; 5% v/v). Cell outgrowth and differentiation were analyzed after staining with DRAQ5 (nuclei) and rhodamine phalloidin (cytoskeleton), as well as by environmental scanning electron microscopy (ESEM). Cell growth/viability of hCECs was assessed by XTT assay. The expression of SDF-1 was quantitated by qRT-PCR. Exposure to hp-lysate (also containing polyP) increased cell migration already at day 1. Even stronger was the effect of polyP. This effect was blocked by a mast cell serine protease. The formation of cell multilayers was enhanced by hp-lysate or even more by polyP. ESEM revealed continuous cell junctions and prominent microvilli on the surface of adjacent cells exposed to polyP; those structures were only rarely seen in the controls. The hp-lysate and, more potently, polyP increased the proliferation of hCECs, as well as SDF-1 expression. The findings indicate the potential usefulness of the natural polymer, polyP, for topical treatment of corneal epithelial defects. Future studies are directed to develop suitable formulations of polyP, such as biomimetic polyP nano/microparticles showing an adjustable release kinetics.

Advanced Technology IOL Cataract Surgery Optimized for Pre-existing Corneal Disease

Discuss the management of ocular surface disease for favorable outcomes with advanced technology IOLs. Ocular surface disease poses a challenge for favorable outcomes with advanced technology IOLs, especially given current patient expectations that glasses-free outcomes are guaranteed. Long-established and recent techniques for optimizing the ocular surface prior to cataract surgery, combined with education and careful selection of IOLs can help meet patient expectations. Careful measurement, planning, and treatment of ocular surface disorders can enable more patients to benefit from advanced technology IOLs.

The use of autologous serum eye drops for the treatment of ocular surface disorders

ObjectivesTo investigate the use of autologous serum (AS) eye drops in patients with ocular surface disorders who were refractory to conventional treatments.MethodsA retrospective cohort study was conducted at a tertiary...

Amniotic membrane use for management of corneal limbal stem cell deficiency

The current article reviews the most recent surgical techniques for management of corneal limbal stem cell deficiency (LSCD) using amniotic membrane tissue. Early successes with amniotic membrane transplantation (AMT) for the treatment of ocular surface disorders have encouraged clinicians to investigate new applications. The use of AMT as a temporary patch in emergency cases in which LSCD may develop has considerably improved the prognosis of these patients. Amniotic membrane does not have stem cells of its own, but it supports regeneration of limbal epithelial stem cells (LESCs). Similarly, the combination of AMT with classic surgical techniques has enhanced the surgical success rates in most case series. Furthermore, based on its advantageous properties as a cell carrier, new applications to support in-vivo and ex-vivo cell expansion have been reported recently. LSCD constitutes a general indication for AMT. Based on the clinical scenario, AMT may be performed alone to support regeneration of LESCs, in combination with other surgical techniques, or even supporting the in-vivo or ex-vivo expansion of LESCs.

Conjunctival flap cover surgery: 10-year review

Background: To evaluate the conjunctival flap cover surgery for the treatment of ocular surface disorders. Methods: Retrospective uncontrolled study was performed. A total of 253 eyes of 253 patients who had received conjunctival flap cover surgery during a 10-year period were included. The indications, symptoms and complications experienced by patients undergoing partial or total conjunctival flap cover surgery were reviewed. Results: In all of these cases, satisfactory anatomic and functional outcomes were achieved. The BCVA of 224 eyes out of 253 patients (88.5%) were no worse than that of preoperative and the BCVA of 29 eyes out of 253 patients (11.5%) decreased postoperatively. No eye pain, foreign body sensation and other discomforts presented in the last visit. The surgical indications was therapeutic for 138 out of 253 patients (54.5%), aesthetic for 63 eyes out of 253 patients (24.9%), tectonic for 45 eyes out of 253 patients (17.8%), and analgesic for 7 eyes out of 253 patients (2.8%). The primary diagnose of therapeutic indications is refractory corneal ulcer for 135 eyes out of 253 patients (53.4%). The main diagnose of aesthetic indications is atrophy of eyeball for 45 eyes out of 253 patients (17.8%) and corneal leukoma for 18 eyes out of 253 patients (7.1%). The principal diagnose of tectonic indications is corneal perforation for 43 eyes out of 253 patients (17%). The main intraoperative complications is buttonhole formation for 8 eyes out of 253 patients (3.2%), which can be handled by simple suture. The most severe complications postoperatively was conjunctival flaps dissolving and corneal perforation for 3 eyes out of 253 patients (1.2%), and corneal transplantation had to be performed to avoid enucleation. In addition, 4 eyes out of 253 patients (1.6%) presented cystic flap and two of them received surgical excision. Conclusions: Conjunctival flap cover surgery is an underused technique. Its primary indication is refractory corneal ulcer and corneal perforation, and its second indication is aesthetic with poor visual potential coexisting ocular surface diseases. It represents an interesting alternative to more mutilating surgeries.