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Neurotrophic keratopathy (NK) is a degenerative corneal disease produced by different factors, including infection, trauma, and neurogenesis, that lead to trigeminal nerve damage and impaired corneal sensitivity. Extensive epithelial breakdown, impaired corneal epithelial healing and corneal ulceration, stromal melting, and perforation are main NK features. The proliferation of the corneal epithelium is endogenously regulated by a balance between adrenergic cAMP-dependent and cholinergic cGMP-dependent pathways. A careful balance of epitheliotropic neuromediators and neurotrophic factors expressed by corneal nerves and epithelial cells, respectively, is required to maintain corneal homeostasis. Even in its early stages, NK can cause reduced vision secondary to epithelial disturbance. Diagnosing NK is challenging, requiring the acquisition of a thorough clinical history and a comprehensive neurological and ophthalmic examination. Following suspicion of a clinical NK diagnosis, corneal sensitivity must be assessed qualitatively with the wisp of the cotton-tipped applicator and quantitatively through Cochet-Bonnet esthesiometry (CBE). A myriad of therapies is used for NK, and new, more specific modalities are being developed and investigated. Medical treatment with topical recombinant human nerve growth factor and surgical treatment through corneal neurotization are promising therapies aiming to target NK pathophysiology. Coexistent ocular surface disorders must be managed concomitantly to improve its prognosis. This review describes the up-to-date knowledge of the molecular basis regarding the pathogenesis of NK, and the novel target-specific therapeutic approaches based on this molecular mechanism.

Purpose The objective was to evaluate the clinical results obtained from the use of immunosafe plasma rich in growth factors (isPRGF) in the treatment of patients with cicatrizing conjunctivitis (CC) who had not responded to the usual therapy. Patients and methods This is a retrospective study that included patients diagnosed with CC, in whom isPRGF was used in different phases (I: eye drops; II: eye drops and injectable; III: eye drops, injectable and surgical treatment) to achieve control of the inflammation. As a clinical follow-up of the patients, the better corrected visual acuity (BCVA), degree of inflammation (measured from 1 to 4), the severity of the CC, Schirmer I test, IOP and TBUT were analyzed. The adverse events were also evaluated. Results Ten eyes (6 patients) were evaluated, 50% corresponded to Stevens-Johnson Syndrome and 50% to ocular mucous membrane pemphigoid. The mean age was 59.7 ± 16.5 (39-80) years, and 50% were women. Fifty per cent of the cases were initially considered severe CC, and 10% of the cases (one eye of one patient) were considered severe CC at the end of the treatment (p = 0.046). The initial degree of inflammation was 2 in 4 eyes, 3 in two eyes, and 4 in 4 eyes, and final inflammation degree was 1 in all cases (p = 0.004). Twenty per cent of the cases achieved stability in Phase I of the treatment with immunosafe PRGF, 70% with both Phases I and II, and only one case underwent Phase III to achieve stability. The IOP improved significantly (p = 0.027) though the BCVA, TBUT and Schirmer I test showed no significant changes. The follow-up time was 23.1 ± 6.7 (13.6-30.3) months. No adverse effects were reported. Conclusion Treatment with PRGF technology in its injectable and topical immunosafe formulations may be a novel alternative for the treatment of patients with CC, given its complement activity modulating effect, as well as its anti-inflammatory, antifibrotic and regenerative properties.

Purpose: To report the clinical findings of a patient who presented with an atypical bilateral fungal retinitis that was established by retinochoroidal biopsy. Methods: Case report. Results: A 56-year-old systemically healthy man presented with progressive visual loss in his left eye for 3 weeks. Visual acuity was 20/40 in the left eye, and 20/20 in the right eye and fundus examination showed macular retinal pigmented epithelium changes in his left eye. Over the following four months, his lesions progressed to serpiginous-like widespread retinal pigmented epithelium atrophy and his visual acuity decreased to 20/100, but no signs of ocular inflammation were found. Treatment with oral corticoids, valganciclovir and trimethoprim/sulfamethoxazole showed no efficacy. Blood analysis and cultures, laboratory investigations, and imaging tests were carried out looking for infectious and inflammatory diseases, but all tests were negative. Two months later, the patient presented with the same kind of lesions in the other eye (right eye), so he was subjected to retinochoroidal biopsy. Histopathological examination of specimen revealed the presence of intraretinal and choroidal fungal hyphae. Oral voriconazole was initiated achieving clinical remission, but no visual improvement was obtained. The source of the infection remains unknown since all tests results were negative. However, his profession as brewmaster might be related to the origin of the infection. Conclusion: Diagnosis of intraocular fungal infection can be challenging. Retinochoroidal biopsy may be useful to establish the diagnosis in those atypical cases with nonrevealing workup and inflammation localized to the retina.

Previous studies have reported that heparan sulfate proteoglycans (HSPGs) promote amyloid-beta peptide and tau fibrillization in Alzheimer disease (AD) and provide resistance against proteolytic breakdown. We compared the expression levels of 17 HSPG core proteins in 18 AD cases and 6 controls. RT-PCR was used to analyze transcription levels. Immunohistochemistry was performed to localize HSPGs in the brain tissue. We detected expression of all HSPG genes investigated. SDC1, GPC3, and CD44v3 showed the lowest levels of expression, while SDC3 and GPC1 showed the highest. Remarkably, SDC4 and SRGN were overexpressed in most of the areas analyzed. Immunohistochemistry revealed the presence of both SDC4 and SRGN mostly associated with tau and amyloid-β pathology throughout the AD brains. In conclusion, in view of the involvement of HSPGs in AD pathology, especially SDC4 and SRGN, there would seem to be a relationship between the regulation of core protein expression and the pathological features suggesting HSPGs are potential inducers of the disease.

[Purpose]: To describe the association between Sars-CoV-2 infection and small fiber neuropathy in the cornea identified by in vivo corneal confocal microscopy., [Methods]: Twenty-three patients who had overcome COVID-19 were recruited to this observational retrospective study. Forty-six uninfected volunteers were also recruited and studied as a control group. All subjects were examined under in vivo confocal microscopy to obtain images of corneal subbasal nerve fibers in order to study the presence of neuroma-like structures, axonal beadings and dendritic cells. The Ocular Surface Disease Index (OSDI) questionnaire and Schirmer tear test were used as indicators of Dry Eye Disease (DED) and ocular surface pathology., [Results]: Twenty-one patients (91.31%) presented alterations of the corneal subbasal plexus and corneal tissue consistent with small fiber neuropathy. Images from healthy subjects did not indicate significant nerve fiber or corneal tissue damage. Eight patients reported increased sensations of ocular dryness after COVID-19 infection and had positive DED indicators. Beaded axons were found in 82.60% of cases, mainly in patients reporting ocular irritation symptoms. Neuroma-like images were found in 65.22% patients, more frequently in those with OSDI scores >13. Dendritic cells were found in 69.56% of patients and were more frequent in younger asymptomatic patients. The presence of morphological alterations in patients up to 10 months after recovering from Sars-CoV-2 infection points to the chronic nature of the neuropathy., [Conclusions]: Sars-CoV-2 infection may be inducing small fiber neuropathy in the ocular surface, sharing symptomatology and morphological landmarks with DED and diabetic neuropathy.

Aim To evaluate whether the Visian Implantable Collamer Lens with a central port (V4c ICL®; STAAR Surgical, Switzerland) affects the retinal nerve fibre layer (RNFL), macula and optic nerve head (ONH) measurements obtained by optical coherence tomography (OCT), and Heidelberg Retina Tomography (HRT). Methods This prospective study included myopic patients undergoing V4c ICL® implantation. RNFL thickness, macular thickness, ganglion cell analysis (GCA) and ONH main parameters were evaluated with RTVue OCT (Optovue Inc., USA) and Cirrus-HD OCT (Carl Zeiss Meditec, USA). ONH variables were also analysed with HRT-3 (Heidelberg Engineering, Germany). All measurements were performed before and 1 week and 12 months after the surgery. Results 31 eyes of 31 patients (mean age 30.1 ± 5.5 years) were included. Comparing with preoperative values, no significant differences in average RNFL thickness were found with RTVue, while a slight increase (4.3 µm) was detected with Cirrus-HD (85.2 ± 10.3 µm, preoperatively) at 1-week postoperatively (89.5 ± 8.3 µm; p Conclusions The implantation of V4c ICL® did not induce a clinically significant impact on the results of the RNFL/ONH analysis with OCT and HRT.

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.

Background This study presents an experiment regarding the introduction of gamification strategies in occupational therapy courses. Based on previous studies, the objective is to adapt the idea of recreational escape rooms to educational environments of health sciences like occupational therapy to increase student motivation and promote game-based learning and key skills, such as teamwork. Methods Computer software was created for a collaborative escape room which allows on-line simultaneous play of up to 24–30 students. It was tested three times in an occupational therapy degree program with 75 students and it was based on two different subjects, although it can be adapted to others. The escape room was evaluated using feedback surveys and comparing students’ performances before and after the game. Descriptive exploratory statistical analysis was performed using SPSS version 24. Results An appropriate use of educational escape rooms can have significant positive impacts on students’ engagement and learning. Students were found to prefer using gamification tools in their learning. Their degrees of satisfaction exceeded their expectations. Conclusions Educational escape rooms may have a positive impact on students’ motivation and a statistically significant improvement of test scores after playing was found. Comments from the feedback surveys were used to improve successive versions of the software and design of the game. Trial registration T.F.G. n° 2020.038 (Research Ethics Committee of the Principality of Asturias).

Introduction: The use of blood derivatives and especially Plasma rich in growth factors (PRGF), for regenerative purposes has been a common trend along the last decades in the field of oral surgery, dermatology, orthopedics, and more recently in ophthalmology.Areas covered: PRGF is a type of platelet-rich plasma that is being explored for the treatment of ocular injuries. The present review article highlights 50 ophthalmology-related publications about the application of PRGF in the treatment of acute and chronic pathologies in ophthalmology as well as most relevant challenges and future prospects.Expert opinion: PRGF technology provides a wide range of formulations that can be used therapeutically in many different acute and chronic ocular pathologies. In addition to eye drops enriched with autologous growth factors, PRGF enables the preparation of both immunologically safe and fibrin-based formulations. Recent advances in the field have promoted PRGF storage for 12 months under freezing conditions, its daily use for 7 days at room temperature and the freeze-dried formulation. The thermally treated immunosafe formulation has shown promising clinical results for the treatment of several diseases such as Sjogren syndrome, graft versus host disease or cicatrizing conjunctivitis. In addition, several fibrin formulations have been preclinically evaluated and clinically incorporated as an adjuvant to ocular surface or glaucoma surgeries, dermal fat graft procedures, limbal stem cell expansion and retinal surgeries. The present review explores the latest scientific and clinical data, current challenges, and main prospects of this technology for the treatment of several ocular injuries.

Among several requirements for the manufacture of Advanced Therapy Medicinal Products (ATMP) are: following the guidelines of a pharmaceutical quality system, complying with Good Manufacturing Practice (GMP) and access to a cleanroom fulfilling strict environmental conditions (Class A work area and Class B environment). This makes ATMP expensive. Moreover, the production of many of these therapeutic products may also be unprofitable, as in most cases their use is limited to a few patients and to a single batch per manufacturing unit. To reduce costs, ATMP may be produced in a scaled-down system isolated from the external environment (isolator), allowing for placement of this facility in a Class D environment, which is much more permissive and less costly. In this work, we confirm that it is possible to manufacture bioengineered corneal epithelium inside an isolator while fulfilling all the safety assurance standards at an affordable cost for patients. This small-scale ultra-clean working environment complies with GMP guidelines and could be a solution for the high costs associated with conventional cleanroom ATMP production.

This study aimed to investigate the use of Plasma Rich in Growth Factors (PRGF) associated with tissue ReGeneraTing Agent (RGTA) drops for the treatment of noninfectious corneal ulcers. RGTA treatment was applied (one drop every two days), however, if ulcer closure was not achieved, PRGF eye drops treatment was added (four times/day). The time taken to reach the ulcer closure, the Best Corrected Visual Acuity (BCVA), intraocular pressure (IOP), Visual Analog Scale (VAS, in terms of frequency and severity of symptoms), and Ocular Surface Disease Index (OSDI) were evaluated. Seventy-four patients (79 eyes) were included, and the mean age was 56.8 ± 17.3 years. The neurotrophic corneal ulcer was the most frequent disorder (n = 27, 34.2%), mainly for herpes virus (n = 15, 19.0%). The time of PRGF eye drops treatment associated with the RGTA matrix was 4.2 ± 2.2 (1.5–9.0) months, and the follow-up period was 44.9 ± 31.5 months. The ulcer closure was achieved in 76 eyes (96.2%). BCVA, VAS and OSDI improved from the baseline (p &lt, 0.001), and IOP remained unchanged (p = 0.665). RGTA and PRGF in noninfectious ulcers were effective and could be a therapeutic alternative for this type of corneal disease.

Purpose To describe the case of a patient with a recurrent large myopic macular hole (MH), who was successfully treated with a plasma rich in growth factors (PRGF) membrane. Case Report A 71-year-old patient presented a myopic MH in his right eye. One month later, pars plana vitrectomy with internal limiting membrane (ILM) peeling was performed, achieving closure of the MH. Three months later a recurrence was observed (700 µm) without visual acuity (VA) improvement. A new surgery was carried out placing an autologous PRGF-membrane into the MH, and performing a fluid-gas exchange at the end of the surgery. After two months of follow-up, the MH was completely closed, obtaining the anatomic recovery of the foveal depression, and improving the VA to 0.1. No adverse reactions were associated with the use of PRGF and there were no new recurrences of the MH in a follow-up period of six months. Conclusion The use of PRGF-membrane can be used as adjunctive therapy in the surgical repair of recurrent large myopic MHs, thus improving anatomic and visual outcomes.

Resumen Recientemente se ha planteado que las alteraciones de las capas de la retina podrian ser un biomarcador de determinados trastornos mentales, al derivar esta de la misma capa embrionaria que el cerebro y estar conectada con este a traves del nervio optico. El objeto del presente articulo es ofrecer una revision sistematica de la literatura y una sintesis tematica sobre el estado actual de las alteraciones de las capas de la retina identificadas mediante tomografia de coherencia optica en los pacientes con esquizofrenia, trastorno bipolar y depresion mayor. Para ello se realizo una busqueda sistematica de la literatura, la lectura critica de los articulos seleccionados y la sintesis tematica de los resultados. Los pacientes con esquizofrenia son los que presentan mas alteraciones, seguidos de los pacientes con trastorno bipolar, siendo muy escasos los hallazgos en la depresion. La capa de fibras nerviosas de la retina es la capa retiniana con mas alteraciones en la esquizofrenia y en el trastorno bipolar, mientras que ningun estudio en depresion mayor encontro alteraciones en ella. De los parametros clinicos, la duracion de la enfermedad correlaciona significativa e inversamente con el grosor de las distintas capas en todos los trastornos. A la hora de interpretar estos datos es necesario tener en cuenta las limitaciones y diferencias de los estudios, especialmente el tiempo medio de evolucion de los trastornos. Dado que este era muy diferente entre los 3 trastornos (mas del doble en el caso de la esquizofrenia respecto a la depresion mayor), las diferencias en los resultados encontrados podrian deberse mas al efecto del tiempo de evolucion que al trastorno en si. En conclusion, los hallazgos de la tomografia de coherencia optica son esperanzadores, ya que podrian proporcionar biomarcadores de la neurodegeneracion y/o neuroprogresion tanto de la esquizofrenia como del trastorno bipolar.

A new in vitro human corneal epithelial model (QobuR) obtained from normal limbal tissue has been developed to study ocular irritancy of different ophthalmic compounded drugs. Phenotypical characterization and trans-epithelial electrical resistance (TEER) of QobuR revealed essential similarities compared with a native human cornea, displaying functional markers and TEER values near 1500 Ωcm2 at day 7th of cellular differentiation. Using this model, ocular irritancy and barrier integrity alterations were evaluated using MTT reaction and variations in TEER. We found that some of the Non-Irritant products evaluated still damage the corneal epithelial integrity and current protocols for ocular irritancy should therefore include a barrier integrity evaluation. Moreover, in order to comprehensively evaluate corneal permeability of the active ingredients, we propose the use of QobuR as an all-in-one alternative method for evaluating ocular irritancy, barrier disruptions and permeability rates of topically applied ocular drugs to improve current in vitro drug testing procedures.

Age-related macular degeneration (AMD) causes the degeneration of photoreceptors and retinal cells leading to vision loss in older subjects. Among possible exogenous risk factors, it has been recently proposed that long-term exposure to blue light could aggravate the course of AMD. In the search for therapeutic options, plasma rich in growth factors (PRGF) has been shown to enhance cell antioxidant pathways and protect photoreceptors against the harm produced by blue light, although its mechanism of action remains unknown. One possible mechanism, autophagy, is one of the most conservative cell renewal systems used in eukaryotes to destroy cellular components that have been damaged by some kind of insult. The oxidative stress of exposure to blue light is known to induce cell autophagy. In this study, we examined the combined effects on autophagy of blue light and PRGF in a retinal cell line, ARPE19. In response to treatment with both PRGF and blue light, we detected the modulated expression of autophagy markers such as NF-kB, p62/sqstm1, Atg5, LC3 and Beclin1, and inflammatory markers such as IL1B and IL18. Our findings suggest that PRGF promotes cell autophagy in response to exposure to blue light.

To evaluate the efficacy and safety of plasma rich in growth factors (PRGF) in photorefractive keratectomy (PRK) versus Mitomycin C (MMC). This is a comparative, longitudinal and retrospective case-control study (MMC vs PRGF), in patients with a spherical correction from −0.25 to −8.00 D and cylinder correction from −0.25 to −3.00. The uncorrected distance visual acuity (UDVA), refractive efficacy and safety indices, and changes in endothelial cell density were evaluated. The predictability was assessed with the postoperative manifest spherical equivalent. Forty-four patients (72 eyes) were treated with MMC and twenty-five patients (45 eyes) with PRGF. The final UDVA (LogMar) in MMC was 0.029 ± 0.065 and in PRGF it was 0.028 ± 0.048 (p = 0.383). The efficacy index for MMC was 0.98 ± 0.10 and 1.10 ± 0.46 for patients treated with PRGF (p = 0.062). The safety index for MMC was 1.03 ± 0.11 and 1.12 ± 0.46 (p = 0.158) for PRGF group. The change percentage of endothelial cell density was 0.9 ± 11.6 for MMC and 4.3 ± 13.1 for PRGF (p = 0.593). The predictability for MMC was 92.1% and for the PRGF was 91.9% (p = 0.976). Hyperemia, eye pain and superficial keratitis were observed in 11.1% of the MMC group; no adverse events were observed with the PRGF. The use of PRGF in PRK surgery is as effective as MMC. The PRGF shows a better safety profile than MMC for its intraoperative use in PRK.

Purpose To analyze whether plasma rich in growth factors (PRGFs) eye drops preserve their activity and biological properties after storage for 9 and 12 months at -20°C, and at 4°C, and at room temperature (RT) for 3 and 7 days in comparison to fresh samples (t0). Methods PRGF eye drops were obtained from 6 healthy donors. Then, they were stored for 9 and 12 months at -20°C. At each time, different PRGF eye drops samples were thawed and maintained at RT or at 4°C for 3 and 7 days. Platelet-derived growth factor-AB, epidermal growth factor, transforming growth factor-β1, vascular endothelial growth factor, angiopoietin-1, and thrombospondin-1 were analyzed at each time and temperature of storage. In addition, the pH level, the microbial contamination, and the proliferative potential on primary human corneal stromal fibroblasts human keratocytes of each obtained PRGF eye drops were also evaluated. Results All growth factor levels were preserved at each time and storage condition. No differences were observed on the human keratocytes proliferation after treatment with PRGF eye drops at any studied time or temperature. No microbial contamination was observed in any of the PRGF eye drops. Finally, the pH levels increased significantly after 9 and 12 months of storage at -20°C compared with t0. Conclusions PRGF eye drops can be stored for up to 12 months without reduction of the main growth factors and proteins and without any microbial contamination. Furthermore, the biological activity of the PRGF eye drops is maintained after storing for 3 and 7 days at 4°C or at RT.

To restore corneal transparency and vision loss after an injury on the ocular surface, the use of human stem cells from different origins has been recently proposed. Mesenchymal stem cells (MSCs) seem to be an appropriate adult source of autologous stem cells due to their accessibility, high proliferation rate, and multipotent capacity. In this work, we developed a simple culture system to prepare a graft based on a fibrin membrane seeded with human MSCs. A commercial kit, PRGF Endoret®, was used to prepare both, the growth factors used as culture media supplement and the fibrin membrane grafts. Adipose-derived MSCs (Ad-MSCs) were expanded, characterised by flow cytometry and their multilineage differentiation potential confirmed by inducing adipogenesis, osteogenesis and chondrogenesis. Ad-MSCs seeded on the fibrin membranes were grafted onto athymic mice showing good biocompatibility with no adverse reactions observed during the follow up period. These findings support the assumption that a system in which all the biological components (cells, grow factors and carrier) are autologous, could potentially be used for future ex vivo expansion of Ad-MSCs to treat ocular conditions such as an inflammatory milieu, traumatic scars and loss of the regenerative capacity of the corneal epithelium that compromise the quality of vision.

The ocular surface flora perform an important role in the defense mechanisms of the ocular surface system. Its regulation of the immunological activity and the barrier effect against pathogen invasion are remarkable. Composition of the flora differs according to the methods of investigation, because the microbiome, composed of the genetic material of bacteria, fungi, viruses, protozoa, and eukaryotes on the ocular surface, differs from the microbiota, which are the community of microorganisms that colonize the ocular surface. The observed composition of the ocular surface flora depends on harvesting and examining methods, whether with traditional culture or with more refined genetic analysis based on rRNA and DNA sequencing. Environment, diet, sex, and age influence the microbial flora composition, thus complicating the analysis of the baseline status. Moreover, potentially pathogenic organisms can affect its composition, as do various disorders, including chronic inflammation, and therapies applied to the ocular surface. A better understanding of the composition and function of microbial communities at the ocular surface could bring new insights and clarify the epidemiology and pathology of ocular surface dynamics in health and disease. The purpose of this review is to provide an up-to-date overview of knowledge about this topic.

Purpose: The purpose of this study was to analyze the biological content and activity of freeze-dried plasma rich in growth factors eye drops after their storage at 4°C and at room temperature for 3 months with respect to fresh samples (time 0). Methods: Plasma rich in growth factors was obtained after blood centrifugation from three healthy donors. After platelet activation, the obtained plasma rich in growth factors eye drops were lyophilized alone or in combination with lyoprotectant (trehalose), then they were stored for 3 months at room temperature or at 4°C. Several growth factors were analyzed at each storage time and condition. Furthermore, the proliferative and migratory potential of freeze-dried plasma rich in growth factors eye drops kept for 3 months at different temperature conditions was evaluated on primary human keratocytes. Results: The different growth factors analyzed maintained their levels at each time and storage condition. Freeze-dried plasma rich in growth factors eye drops stored at room temperature or 4°C for 3 months showed no significant differences on the proliferative activity of keratocytes in comparison with fresh samples. However, the number of migratory human keratocytes increased significantly after treatment with lyophilized plasma rich in growth factors eye drops kept for 3 months compared to those obtained at time 0. No significant differences were observed between the freeze-dried plasma rich in growth factors eye drops whether mixed or not with lyoprotectant. Conclusion: Freeze-dried plasma rich in growth factors eye drops preserve the main growth factors and their biological activity after storage at room temperature or 4°C for up to 3 months. Lyophilized plasma rich in growth factors eye drops conserve their biological features even without the use of lyoprotectants for at least 3 months.

Background and importance Sutures to replace tissue adhesives have enhanced importance. However, commercialised drugs are allogenic, synthetic and expensive, increasing surgery costs. Aim and objectives To produce an autologous tissue adhesive (ATA) easily compounded in ophthalmological surgery. To show evidences of the safe and effectiveness of the ATA in preclinical studies. Material and methods To produce 4 mL of ATA based on a fibrinogen (FC) and thrombin concentrate (TC) (proportion1:1), 20 mL of donor blood plasma were precipitated with protamine to prepare FC, and then 20 mL of plasma were precipitated with acetic acid to obtain a TC in a buffer (CaCl2, NaHCO3, NaCl). Drug was conditioned in two 2 mL syringes for topical ophthalmic administration by mixing with a needle. The in vitro toxicity of the drug was studied in a human corneal epithelial model (described as QobuR), to evaluate the grade of irritation after 30 min of exposition time.1 Pterygium surgery was performed in four eyes of white New Zealand rabbits, using ATA to fix a frontal conjunctival autograft (4×5 mm) into the temporal bulbar conjunctive. The grafted eyes were evaluated in vivo by clinical evaluation for 14–28 days and ex vivo by histology. Results ATA produced from each donor showed a mean of 18.0 g/L of fibrinogen and 1500 UI/mL of thrombin. ATA instantly produced homogeneous clots when it was mixed with a needle. Three in vitro studies of four ATA showed non-irritation due to high survival cell viabilities (>80%). Good preclinical results were found: 20 mm2 autograft could be fixed successfully. Time for complete tissue adhesion was minimal (3–5 min). Inflammation and adverse events were absent in all cases. The prospective clinical evaluation was positive for follow-up in all cases and included integration and vascularisation of the grafts. Histology supported the in vivo evidence. Staining of the autograft section showed inner vessels and the regeneration of the surrounding conjunctive tissue. Conclusion and relevance It is possible to compound an ATA easily from whole blood, in a hospital pharmacy, for ophthalmological surgery, where the necessary volume is very low. This ATA was safe and effective, supported by our preclinical studies. This ATA could allow the possibility of replacing the suture in surgery with a low cost drug. References and/or acknowledgements 1. https://www.ncbi.nlm.nih.gov/pubmed/30690064 No conflict of interest.

Background This study analysed the effectiveness of plasma rich in growth factors (PRGF) in reducing the oxidative stress induced by blue light exposition on retinal pigment epithelial (RPE) cells. Methods Blood from six healthy donors was collected to obtain the PRGF. Retinal pigment epithelium (ARPE-19) cells were exposed to blue light. Then, cells were incubated with PRGF or with control for 24 and 48 hours maintaining exposure to blue light. The cytoprotective effect of PRGF on ARPE cells was evaluated by measuring the cell viability, the reactive oxygen species (ROS) production and the expression of different proteins such as heme oxygenase 1 (HO-1), catalase (CAT), superoxide dismutase (SOD-1), apoptosis-inducing factor (AIF), pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor (VEGF). Results The cell viability increased significantly at 24 and 48 hours after PRGF treatment compared to the control group. ROS synthesis was significantly reduced in PRGF-treated cells with respect to control. Furthermore, the levels of HO-1, SOD-1 and AIF were significantly reduced after PRGF treatment at both times of treatment. However, CAT levels were only significantly reduced after PRGF treatment at 48 hours. The high expression of VEGF by RPE cells exposed to blue light was only counterbalanced in the PRGF group by increasing the expression of PEDF in comparison to the control group. Conclusion The present results show that PRGF treatment reduces the cytotoxic effects induced in RPE cells exposed to an oxidative stress environment. Furthermore, PRGF treatment preserves the mitochondrial activity and cell viability of RPE cells subjected to an oxidative stress.

AimTo investigate the difference in adhesion and rebubbling rate between eye bank and surgeon prepared Descemet membrane endothelial keratoplasty (DMEK) tissues.MethodsLaboratory and clinical retrospective comparative interventional case series. Research corneal tissues were obtained for laboratory investigation. The clinical study involved patients with endothelial dysfunction who underwent DMEK surgery and tamponade with air. Tissues were stripped using a standard DMEK stripping technique (SCUBA) and shipped as prestripped or loaded in a 2.2 intra-ocular lens cartridge with endothelium facing inwards (preloaded) before transporting from the eye bank to the surgeon. For surgeon prepared tissues, all the grafts were stripped in the theatre and transplanted or stripped in the laboratory and tested immediately. Adhesion force and elastic modulus were measured in the centre and mid-periphery in a laboratory ex vivo investigation using atomic force microscopy, while rebubbling rates were recorded in the clinical study.ResultsThere was no difference in endothelial cell viability between surgeon or eye bank prepared tissue. Surgeon-stripped DMEK grafts in the laboratory investigation showed significantly higher elastic modulus and adhesion force compared to prestripped and preloaded tissues (pConclusionsDecreased adhesion forces and elastic modulus in eye bank prepared tissues may contribute to increased rebubbling rates.

Oxidative stress has a strong impact on the development of retinal diseases such as age-related macular degeneration (AMD). Plasma rich in growth factors (PRGF) is a novel therapeutic approach in ophthalmological pathologies. The aim of this study was to analyze the antioxidant effect of PRGF in retinal epithelial cells (EPR) in in vitro and ex vivo retinal phototoxicity models. In vitro analyses were performed on ARPE19 human cell line. Viability and mitochondrial status were assessed in order to test the primary effects of PRGF. GSH level, and protein and gene expression of the main antioxidant pathway (Keap1, Nrf2, GCL, HO-1, and NQO1) were also studied. Ex vivo analyses were performed on rat RPE, and HO-1 and Nrf2 gene and protein expression were evaluated. The results show that PRGF reduces light insult by stimulating the cell response against oxidative damage and modulates the antioxidant pathway. We conclude that PRGF&rsquo, s protective effect could prove useful as a new therapy for treating neurodegenerative disorders such as AMD.

Purpose To investigate whether plasma rich in growth factors (PRGF) eye drops maintain their biological potential after a freeze drying process. The addition of a lyoprotectant like trehalose was also evaluated. Methods Blood from three healthy donors was collected to obtain eye drops by PRGF technology. The resultant eye drops were divided in four groups: PRGF, freeze-dried PRGF (PRGF lyo), and PRGF lyophilized mixed with 2,5% trehalose (PRGF lyo+2.5T) or 5% trehalose (PRGF lyof+5T). Chemical and biological characteristics were evaluated. Photorefractive keratectomy was performed on C57BL/6 mice which were divided in three treatment groups: control, PRGF, and PRGF lyo. Corneal wound healing and haze formation were evaluated macroscopically. Eyes were collected at 1, 2, 3, and 7 days after surgery, and were processed for histologic studies. Results The pH values of PRGF samples increased significantly after the lyophilization process. Osmolarity levels increased significantly in PRGF samples mixed with trehalose in comparison with PRGF samples without protectants. The freeze drying process maintained growth factors levels as well as the biological properties of PRGF eye drops even without the use of lyoprotectants. PRGF lyo treatment significantly decreased the re-epithelialization time and haze formation in photorefractive keratectomy-treated corneas regarding PRGF and control groups. Furthermore, the PRGF lyo group significantly decreased the number of smooth muscle actin-positive cells in comparison with the control group at each time of the study and at days 2 and 3 in the PRGF group. Conclusions The freeze drying process preserves the protein and growth factor content as well as the biological properties of PRGF eye drops, even without the use of protectants. Freeze-dried PRGF eye drops accelerate corneal tissue regeneration after photorefractive keratectomy in comparison with the control group. Translational Relevance Our study shows the feasibility to preserve the biological capability of PRGF eye drops as freeze-dried formulation, avoiding the addition of protectants., Resumen (Foreign Language Abstract) Objetivo Investigar si el colirio de plasma rico en factores de crecimiento (PRGF) mantiene su potencial biológico después de un proceso de liofilización. También se evaluó la adición de un lioprotector como la trehalosa. Métodos Se recogió sangre de tres donantes sanos para obtener colirio mediante la tecnología PRGF. El colirio resultante se dividió en cuatro grupos: PRGF, PRGF liofilizado (PRGF lyo) y PRGF liofilizado mezclado con trehalosa al 2,5% (PRGF lyo + 2.5T) o trehalosa al 5% (PRGF lyof + 5T). Se evaluaron las características químicas y biológicas de cada uno de los colirios. Se realizó una queratectomía fotorrefractiva (PRK) en ratones C57BL/6 que se dividieron en tres grupos de tratamiento: Control, PRGF y PRGF lyo. La curación de la herida corneal y la formación de “haze” se evaluaron macroscópicamente. Los ojos se recolectaron en los días 1, 2, 3 y 7 después de la cirugía, y se procesaron para la realización de estudios histológicos. Resultados Los valores de pH de las muestras de PRGF aumentaron significativamente después del proceso de liofilización. Los niveles de osmolaridad aumentaron significativamente en las muestras de PRGF mezcladas con trehalosa en comparación con las muestras de PRGF sin protectores. El proceso de liofilización mantuvo los niveles de factores de crecimiento, así como las propiedades biológicas de los colirios de PRGF, incluso sin el uso de lioprotectores. El tratamiento con PRGF lyo redujo significativamente el tiempo de reepitelización y la formación de “haze” en las córneas tratadas con PRK con respecto al grupo de control y PRGF. Además, el grupo PRGF lyo redujo significativamente el número de células SMA positivas en comparación con el grupo control en cada tiempo de estudio y en los días 2 y 3 con respecto al grupo de PRGF. Conclusiones el proceso de liofilización conserva el contenido de proteínas y factores de crecimiento, así como las propiedades biológicas del colirio de PRGF, incluso sin el uso de protectores. El colirio de PRGF liofilizado acelera la regeneración del tejido corneal después de la PRK en comparación con el grupo de control. Relevancia Traslacional nuestro estudio muestra la viabilidad de preservar la capacidad biológica del colirio de PRGF como formulación liofilizada evitando la adición de protectores.

Purpose: The aim was to report on the visual outcomes and safety of myopic LASIK performed in patients with corneas of central thickness below average( Methods: This is a retrospective cohort study conducted at a private practice setting on Mexican Hispanic patients who underwent myopic LASIK between January 2014 and January 2015. An analysis of records of patients >18 years-old with previous normal topography, stable refraction, corrected visual acuity ≥ 20/20 (Snellen), Central Corneal Thickness (CCT) < 540μm and at least 12 months follow up after surgery was conducted. The main outcome measures were standard visual outcomes (efficacy, safety, refractive stability) and Percent Tissue Altered (PTA) analysis was conducted. Results: A total of 51 patients (102 eyes) were included; 56% (n=57) were female. The mean age was 26.52 ± 8.06 (range 18-55 years) with a mean follow up of 13.9 ± 1.2 months. Preoperative CCT was 515.44 ± 17.87μm (range 452- 540μm), with a mean refractive spherical equivalent (SEQ): -4.08 ± 2.17 D (range -0.75 to -9.75 D), and mean refractive cylinder: -1.44 ± 1.29 D (range 0.00 to -6.00 D). Mean predictability of postoperative SEQ was -0.20 ± 0.40 D (range -1.25 to +1.25). Postoperative SEQ was ±0.50 D in 71%, ±1.00 D in 93% of the eyes. Postoperative uncorrected distance visual acuity was ≥20/20 in 78% and ≥20/25 in 95%. One line of CDVA was lost in 3% of the eyes, no eyes lost ≥2 lines. No ectasia cases were observed during follow-up. Conclusion: LASIK surgery in Mexican Hispanic patients with thinner than “normal” corneas (

Purpose: To evaluate the efficacy and safety of plasma rich in growth factors eye drops for the treatment of corneal and ocular surface disorders in patients with graft versus host disease. Methods: This retrospective and longitudinal study included graft versus host disease patients with ocular disorders. The resolution of corneal ulcers (area and density staining) was evaluated as primary outcome. Best corrected visual acuity, intraocular pressure, tear film breakup time, Schirmer test, ocular surface disease index, and visual analog score were evaluated as secondary outcomes. All variables were analyzed before and after plasma rich in growth factors treatment. The safety of plasma rich in growth factors treatment was also assessed. Results: Twelve patients (23 eyes) with ocular graft versus host disease were evaluated. Statistically significant improvement in the area (75.7%) and density (73.3%) of the corneal staining, in best corrected visual acuity (74.7%), in ocular surface disease index scale (75.4%), visual analog score frequency (81.4%) and visual analog score severity (81.9%), and an increase of 3.8 s in tear film breakup time and 6 mm in Schirmer test was observed after plasma rich in growth factors treatment (p Conclusion: Immunosafe plasma rich in growth factors eye drops for the treatment of patients with ocular graft versus host disease could be safe and effective, showing a high rate of corneal ulcer resolution and dry eye disease control. Plasma rich in growth factors eye drops may help to maintain corneal stability and prevent it against higher ocular complications.

The use of filters in ophthalmology is not new, but usually they operate by just blocking light of a particular wavelength. In contrast, photoluminescence filters are theoretically able to provide an altered wavelength of light to reach the retina, an approach which potentially can be exploited for therapeutic purposes. Our working hypothesis is that providing a means to deliver a reduced amount of harmful UV-blue light and an increased amount of beneficial red-near infrared light to retinal neuronal mitochondria will improve the health of the tissue and so enhance the health of the retina. The purpose of this communication is a) to briefly review the evidence behind our working hypothesis, b) to propose and describe luminescent spectral conversion filters that might be used to non-invasively achieve our aims related to our hypothesis, and c] to provide some preliminary experimental data to show that such luminescent filters coated on a glass substrate can convert UV-blue light to sufficient red light to enhance cell survival in cultured cells.

Corneal nerves are key components of the physiological system that controls ocular surface homeostasis. The cornea is primarily innervated by the ophthalmic branch of the trigeminal nerves (cranial nerve V), which distend bilaterally from the pons. The nasociliary branch (afferent) of the ophthalmic nerve is sensory for cornea, eyelid and conjunctiva. These nerve fibres play a role in sensing temperature, chemical and mechanical stimuli, and pain, whereas, branches of the facial nerve (cranial nerve VII) contain motor nerves that control blinking and autonomic (sympathetic and a paucity of parasympathetic) fibres that stimulate tear production and secretion via feedback loops between the ocular surface, lacrimal glands and brain. Disruption of these nerves with interruption of neural feedback loops between the ocular surface and lacrimal glands can lead to corneal diseases such as dry eye disease (DED) and neurotrophic keratopathy (NK). Inversely, hypersensitivity of the nerve fibres and/or dysregulation of pain-controlling nervous centres may lead to neuropathic pain. Recently, medications that specifically target regeneration of corneal nerves have started to become available - and considering the high prevalence of diseases associated with corneal nerve dysfunction, these agents promise to fulfil a hitherto important unmet need. In this review, we explore the physiology of corneal nerves, the pathology of corneal nerve diseases and how these relate to neuropathic pain, NK and DED. We also discuss what novel treatments may be useful against diseases involving corneal nerves.

Ronald M Sánchez-Avila,1 Jesus Merayo-Lloves,1 Maria Laura Fernández,1,2 Luis Alberto Rodríguez-Gutiérrez,1 Pedro Pablo Rodríguez-Calvo,1 Andres Fernández-Vega Cueto,1 Francisco Muruzabal,3,4 Gorka Orive,3–6 Eduardo Anitua3,4 1University Institute Fernández-Vega, Ophthalmological Research Foundation, University of Oviedo, Oviedo, Spain; 2Quilmes Eye Center, Buenos Aires, Argentina; 3University Institute for Regenerative Medicine and Oral Implantology (UIRMI), University of the Basque Country, Eduardo Anitua Foundation, Vitoria, Spain; 4Biotechnology Institute (BTI), Vitoria, Spain; 5Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country, Vitoria, Spain; 6Networking Biomedical Research Center on Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, Vitoria, Spain Purpose: To evaluate the efficacy and safety of plasma rich in growth factors (PRGF) eye drops in patients with glaucoma with secondary ocular surface disorders (OSDs) due to surgeries and topical hypotensive drugs use. Materials and methods: A retrospective case-series study design was used including six patients (eight eyes) diagnosed with glaucoma who received surgical (nonpenetrating deep sclerectomy and/or trabeculectomy) and medical treatments (hypotensive eye drops) to control intraocular pressure (IOP) and who developed secondary OSDs, unresponsive to conventional treatments. Patients were treated with PRGF eye drops (four times a day). Outcome measures were ocular surface disease index (OSDI), best-corrected visual acuity (BCVA, in logarithm of the minimum angle of resolution), visual analog scale (VAS), frequency and severity of symptoms, and IOP. The safety of the treatment was also evaluated. Results: Six patients (seven eyes with open-angle glaucoma and one eye with uveitic glaucoma) treated with PRGF eye drops were evaluated. Mean age was 71 years (SD=7.2, range 58–79 years). Five were female and one was male. The mean treatment time was 21.8 weeks (SD=9.0, range 12–36 weeks). The mean time to reach closure of the corneal ulcer was 14.5 (SD=5.5) weeks. A statistical significant reduction in OSDI scale (50.6%), VAS frequency (53.1%), VAS severity (42.0%), and a 41.8% improvement in BCVA were observed (p

In the present study mechanical damage to the corneal endothelium was induced by elevation of intraocular pressure (IOP, 140 mmHg, 60 min) to one eye of rats, delivered either in complete darkness or in the presence of red light (16.5 W/m 2 , 3000 lx, 625–635 nm). IOP raised in the dark revealed the endothelium to be damaged as staining for the gap junction protein ZO-1 was irregular in appearance with some cells displaced in position or lost to leave gaps or holes. This damage was clearly attenuated when red light was focused through the pupil during the insult of raised IOP. Moreover, staining of endothelium with JC-1 dye showed mitochondria to be activated by both elevated IOP and red light but the activation of mitochondria persisted longer for red light. We interpret this finding to suggest that raised IOP causes apoptosis of endothelial cells and that their mitochondria are activated in the initial stages of the process. In contrast, red light activates mitochondria to induce a protective mechanism to counteract the negative influence of raised IOP on endothelial cells. Evidence is provided to support this notion by the finding that red light stimulates mitochondrial cytochrome oxidase IV (COX IV). Moreover, mitochondria in corneal endothelial cell cultures are activated by red light, revealed by staining with JC-1, that results in an increased rate of proliferation and are also able to counteract toxic insults (sodium azide or cobalt chloride) to the cultures. The present studies therefore show that a non-toxic level of red light attenuates damage to the corneal endothelium both in situ and in vitro through action on COX IV located in mitochondria that results in an enhancement of a cell's survival mechanisms. The study provides proof of principle for the non-invasive use of red-light therapy to attenuate any dysfunctions associated with the corneal endothelium and so preserve maximum visual acuity.

Purpose The objective of this study was to determine the efficacy and safety of topical tacrolimus compounded in the Pharmacy Service for the treatment of subepithelial corneal infiltrates (SEIs) secondary to adenoviral keratoconjunctivitis. Methods This retrospective study included patients who had been dispensed topical tacrolimus for the treatment of SEIs during the previous year. Patients were treated with tacrolimus 0.03% eye drops twice daily or tacrolimus 0.02% ointment once daily. The following data were recorded: length of treatment, visual acuity before and after treatment, intraocular pressure before, during, and at the end of treatment, previous treatments, and the presence of SEIs after treatment. The subjective symptoms of the patients were also assessed. Results Fifty-five patients (85 eyes) were included, 54.5% with bilateral involvement. A total of 31 (36.5%) eyes were treated with tacrolimus ointment and 54 eyes (63.5%) with tacrolimus eye drops. The median length of treatment was 185 days (p25-75: 93.5-426), and the mean follow-up duration was 363 days (p25-75: 148-540). In 62.35% of the eyes, the SEIs were reduced in number and size, and in 31.76%, they were eliminated. The patients had better visual acuity after treatment with highly statistically significant differences. Tolerance was good overall, being better in the eye drops group. Conclusions Topical tacrolimus, compounded in the pharmacy, seems to be an effective and safe alternative for the treatment of SEIs secondary to adenovirus keratoconjunctivitis.

Experimental studies in animals, as well as observational and intervention studies in humans, seem to support the premise that the development of juvenile myopia is promoted by a combination of the effect of genetic and environmental factors, with a complex interaction between them. The very rapid increase in myopia rates in some parts of the world, such as Southeast Asia, supports a significant environmental effect. Several lines of evidence suggest that humans might respond to various external factors, such as increased activity in near vision, increased educational pressure, decreased exposure to sunlight outdoors, dietary changes (including increased intake of carbohydrates), as well as low light levels indoors. All these factors could be associated with a higher prevalence of myopia.

BACKGROUND Heparan sulfate proteoglycans (HSPGs) promote amyloid-β peptide and tau fibrillization in Alzheimer's disease (AD) and provide resistance against proteolytic breakdown. Heparanase (HPSE) is the only enzyme that cleaves heparan sulfate (HS). Heparanase 2 (HPSE2) lacks HS-degrading activity, although it is able to interact with HS with high affinity. OBJECTIVE To analyze HPSE and HPSE2 expressions at different stages of AD. METHODS RT-PCR was used to analyze transcription levels of both heparanases at different stages of AD, and immunohistochemistry was performed to localize each one in different parts of the brain. RESULTS Both proteins appeared overexpressed at different stages of AD. Immunohistochemistry indicated that the presence of the heparanases was related to AD pathology, with intracellular deposits found in degenerated neurons. At the extracellular level, HPSE was observed only in neuritic plaques with a fragmented core, while HPSE2 appeared in those with compact cores as well. CONCLUSION Given the involvement of HSPGs in AD pathology, there would seem to be a relationship between the regulation of heparanase expression, the features of the disease, and a possible therapeutic alternative.

BACKGROUND & AIMS Minimal hepatic encephalopathy (mHE) has been shown to affect daily functioning, quality of life, driving and overall mortality. However, little is known about treating or diagnosing early impairments in mHE. We studied one of its precipitating factors, portal hypertension which is driving the inflammatory process behind mHE. The purpose was to describe an indirect diagnostic method able to detect the pathology at early stages based on the study of the vascularization and mast cells conjunctival hyperplasia as secondary inflammatory response associated to portal hypertension. Finally, we correlated the presence of histological changes in the eye in mHE with deficits in behavioral task acquisition. METHODS Rats were trained on a stimulus-response task and a spatial working memory task using the Morris water maze. Two groups of animals were used: a SHAM (sham-operated) group (n=10) and a portal hypertension (HT) group (n=10). The triple portal vein ligation method was used to create an animal model of mHE. Latencies to reach the platform, number of glial fibrillary acidic protein-immunoreactive astrocytes (GFAP-IR), mast cell expression and presence/absence of blood and lymphatic vessels were examined. RESULTS There were differences in stimulus-response behavioral performance, with a deficit in the acquisition in the HT group. However, no differences between groups were found on the spatial working memory task. At the same time, differences between groups were found in the GFAP-IR density, which was lower in the HT group, and in the number of mast cells and the presence of vessels, which were higher in the HT group. CONCLUSIONS In this study, we provide the first preliminary insight into the validity of exploring the eye as a possible tool to assess the diagnosis of mHE conditions.

Defining the corrosive properties of chemical products generally involves the use of animal models for human health safety assessment. However, a few alternatives to animal experimentation are currently internationally accepted in order to reduce animal suffering. One of these alternatives makes use of in vitro reconstructed human epidermis (RhE) models and predicts corrosive potential based on the evaluation of cell viability after topical exposure. These models rely on its similarity to human skin, both functional and histological, and are currently worldwide marketed by a few private companies. In this manuscript, we describe the fundamentals of the production of a Do It Yourself (DIY) RhE model, and the operating procedures for the assessment of skin corrosion based on the guidelines proposed for the development of new alternative methods for skin corrosion. Our results indicate that the DIY-RhE model resembles the anatomy of the normal human epidermis as seen by immunohistochemical analysis. Moreover, barrier properties of DIY-RhE were assessed by the measure of Transepithelial Electrical Resistance. Applicability of DIY-RhE for the assessment of skin corrosion was evaluated by measuring cell viability after topical exposure of twelve reference chemicals for 3 and 60 min. Predictive performance resulted in 100% sensitivity, 100% specificity and 100% accuracy matching current requirements for new RhE models proposed for the discrimination of corrosives and non-corrosives.

To evaluate the endothelial cell loss in patients with iris-claw phakic lenses (Artisan®) in a long-term follow-up. We analyzed the medical records of patients who had undergone iris-claw phakic lens implantation and who had at least 5 years of follow-up. We included 67 eyes with myopic errors (follow-up 9.6 ± 3.0 years) and 10 eyes with mixed astigmatism or hyperopic errors (follow-up 8.8 ± 2.5 years). The mean total endothelial density loss at the last follow-up visit was 18.5% ± 17.0% and 10.5% ± 12.3%, respectively. 29.9% of the eyes in the myopic group and 20% in the hyperopic group lost more than 25% of the preoperative endothelial cell density. During the postoperative follow-up period, 60.8% of the eyes in the myopic group and 40% of the eyes in the hyperopic group lost a higher percentage than the expected physiological loss. Two eyes in the myopic group (3.0%) had a final cell density of less than 1200 cells/mm2. None of the variables studied had a statistically significant association with the percentage of annual endothelial loss in the postoperative period. Three phakic lenses were explanted: two by cataract and one by cataract and severe decrease of the endothelial density (862 cells/mm2). There is a significant endothelial cell loss in a low percentage of the eyes with Artisan® lenses in the long term, and it can decrease to critical levels. Periodic endothelial density evaluations are required for these patients. The selection criteria of surgical candidates could be reevaluated.

Purpose To develop and characterize a new type of plasma rich in growth factors (PRGF) membrane for patients in which immune system is involved in the disease etiology. Methods Blood from three healthy donors was collected to obtain the different fibrin membranes by PRGF technology. PRGF obtained volumes were activated and divided into two groups: PRGF membrane (mPRGF) obtained after incubation at 37 °C for 30 min (control); and is-mPRGF: mPRGF obtained after incubation for 30 min at 56 °C. The concentration of several growth factors, proteins, immunoglobulin E and the complement activity was determined in the different mPRGF. The proliferative potential of heat-inactivated mPRGF were assayed on keratocytes (HK) and conjunctival fibroblasts (HConF). In addition, morphological and physical features of the inactivated mPRGF were evaluated in contrast to the control mPRGF. Results Heat-inactivation of the mPRGF preserves the content of most of the growth factors involved in the ocular wound healing while reducing drastically the content of IgE and the complement activity. The heat-inactivated mPRGF conserve the morphological and physical characteristics of the fibrin meshwork in comparison with the control mPRGF. Furthermore, no significant differences were found in the biological activity of the control mPRGF regarding the heat-inactivated mPRGF (is-mPRGF) in any of both ocular cell types evaluated. Conclusions The heat-inactivation of the PRGF membranes (is-mPRGF) reduces drastically the content of IgE and complement activity while preserving the content of most of the proteins and morphogens involved in ocular wound healing. Furthermore, the morphological and physical features of the immunosafe mPRGF were also preserved after heat-inactivation.

Resumen El uso de los preparados ricos en plaquetas ha experimentado un aumento significativo en los ultimos anos debido a su papel en la reparacion y regeneracion tisular. El objetivo del presente estudio es recopilar la evidencia disponible respecto a la aplicacion de plasma rico en factores de crecimiento y sus variantes sobre la superficie ocular: el efecto de los factores de crecimiento derivados de plaquetas, las implicaciones de los distintos metodos de preparacion, los estudios publicados en patologias de la superficie ocular, asi como sus contraindicaciones y reacciones adversas. Pese al uso generalizado de los preparados de plaquetas, no existe un consenso sobre el metodo de preparacion mas adecuado, variando las concentraciones de factores de crecimiento segun el sistema empleado. Estos preparados se han utilizado en el tratamiento de enfermedades de la superficie ocular como del ojo seco o los defectos epiteliales persistentes, entre otras, con un perfil adecuado de eficacia y seguridad, aunque son necesarios mas estudios para su posicionamiento terapeutico respecto a las alternativas actualmente disponibles.

Ocular graft versus host disease (oGVHD) is part of a systemic inflammatory disease that usually affects ocular surface tissues manifesting as a dry eye syndrome. Current treatments provide unsatisfactory results. Blood-derived products, like plasma rich in growth factors (PRGF) emerge as a potential therapy for this disease. The purpose of this study was to evaluate the tissue regeneration and anti-inflammatory capability of PRGF, an autologous platelet enriched plasma eye-drop, compared to autologous serum (AS) obtained from oGVHD patients on ocular surface cells cultured in a pro-inflammatory environment. PRGF and AS were obtained from four GVHD patients. Cell proliferation and inflammation markers, intercellular adhesion molecule-1 (ICAM-1) and cyclooxygenase-2 (COX-2), were measured in corneal and conjunctival fibroblastic cells cultured under pro-inflammatory conditions and after treatment with PRGF or AS eye drops. Moreover, cell proliferation increased after treatment with PRGF and AS, though this enhancement in the case of keratocytes was significantly higher with PRGF. PRGF eye drops showed a significant reduction of both inflammatory markers with respect to the initial inflammatory situation and to the AS treatment. Our results concluded that PRGF exerts more potent regenerative and anti-inflammatory effects than autologous serum on ocular surface fibroblasts treated with pro-inflammatory IL-1β and TNFα.

The use of platelet-rich preparations has experienced a significant increase in recent years due to its role in tissue-repair and regeneration. The aim of this study is to examine the available evidence regarding the application of plasma rich in growth factors, and its variations, on the ocular surface. A review is also presented on the effects of platelet-derived growth factors, the implications of the preparation methods, and the existing literature on the safety and efficacy of these therapies in ocular surface diseases. Despite the widespread use of platelet preparations there is no consensus on the most appropriate preparation method, and growth factors concentration vary with different systems. These preparations have been used in the treatment of ocular surface diseases, such as dry eye or persistent epithelial defects, among others, with good safety and efficacy profiles, but further studies are needed to compare to the currently available alternatives.

Purpose: The purpose of this study was to examine the effect of plasma rich in growth factors (PRGFs) under blue light conditions in an in vivo model of retinal degeneration. Methods: Male Wistar rats were exposed to dark/blue light conditions for 9 days. On day 7, right eyes were injected with saline and left eyes with PRGF. Electroretinography (ERG) and intraocular pressure (IoP) measurements were performed before and after the experiment. After sacrifice, retinal samples were collected. Hematoxylin and eosin staining was performed to analyze the structure of retinal sections. Immunofluorescence for brain-specific homeobox/POU domain protein 3A (Brn3a), choline acetyltransferase (ChAT), rhodopsin, heme oxygenase-1 (HO-1), and glial fibrillary acidic protein (GFAP) was performed to study the retinal conditions. Results: Retinal signaling measured by ERG was reduced by blue light and recovered with PRGF, however, IoP measurements did not show significant differences among treatments. Blue light reduced the expression for Brn3a, ChAT, and rhodopsin. Treatment with PRGF showed a recovery in their expressions. HO-1 and GFAP results showed that blue light increased their expression but the use of PRGF reduced the effect of light. Conclusions: Blue light causes retinal degeneration. PRGF mitigated the injury, restoring the functionality of these cells and maintaining the tissue integrity.

Biological membranes are currently used in Ophthalmology in order to treat different ocular disorders. These membranes have different properties such as cellular biocompatibility and promoting wound healing. Moreover, intrinsic antimicrobial properties could also be desirable because it would allow their use reducing the risk of infections. Graphene and its derivatives are promising biomaterials that already proved their bactericidal effect. However, their clinical use is limited due to the controversial results regarding their toxicity. In this work, we have developed and characterized a reduced graphene oxide membrane (rGOM) for its use in ocular Regenerative Medicine, and studied its in vitro and in vivo biocompatibility and genotoxicity with different types of human ocular cells. We proved that rGOM allowed the growth of different ocular cells without inducing in vitro or in vivo cytotoxicity or genotoxicity in the short-term. These results indicate that rGOM may be a promising candidate in Regenerative Medicine for the treatment of different ocular pathologies., This work was supported by Consejería de Economía y Empleo del Principado de Asturias: [grant number: IDE/2014/000737 and IDE/2015/000889].

It has recently been suggested that alterations of the layers of the retina could be a biomarker of specific mental disorders since they originate in the same embryonic layer as the brain and both are interconnected through the optic nerve. The purpose of this article is to offer a systematic review of the literature and a thematic synthesis on the current state of the alterations of the retina layers identified by optical coherence tomography in patients with schizophrenia, bipolar disorder and major depression. For this purpose, we performed a bibliographic search, a systematic review of the studies and a thematic synthesis of the reported findings. Patients with schizophrenia have more abnormal findings followed by patients with bipolar disorder, with very few findings in depression. The nerve fiber layer is the retinal layer with more abnormal findings both in schizophrenia and in bipolar disorder, while no study in major depression found alterations in it. Of the clinical parameters, the duration of the illness correlates significantly and inversely with the thickness of the different layers in all disorders. When interpreting these data, it is necessary to take into account the limitations and differences of the studies, especially the mean length of the disorders. Given that this was very different among the 3 disorders (more than doubled in the case of schizophrenia respect to major depression), the differences in the results found could be due more to the effect of the length of illness than to the disorder itself. In summary, optical coherence tomography findings are promising, since they could provide biomarkers of neurodegeneration and/or neuroprogression of both schizophrenia and bipolar disorder.

Purpose The main objective of this study is to compare the protein expression of human keratocytes treated with Plasma rich in growth factors (PRGF) or autologous serum (AS) and previously induced to myofibroblast by TGF-beta 1 treatment. Methods Blood from healthy donor was collected and processed to obtain AS and PRGF eye drops. Blood derivates were aliquoted and stored at-80 degrees C until use. Keratocyte cells were pretreated for 60 minutes with 2.5 ng/ml TGF-beta 1. After that, cells were treated with PRGF, AS or with TGF-beta 1 (control). To characterize the proteins deregulated after PRGF and AS treatment, a proteomic approach that combines 1D-SDS-PAGE approach followed by LC-MS/MS was carried out. Results Results show a catalogue of key proteins in close contact with a myofibroblastic differentiated phenotype in AS treated-cells, whereas PRGF-treated cells show attenuation on this phenotype. The number of proteins downregulated after PRGF treatment or upregulated in AS-treated cells suggest a close relationship between AS-treated cells and cytoskeletal functions. On the other hand, proteins upregulated after PRGF-treatment or downregulated in AS-treated cells reveal a greater association with processes such as protein synthesis, proliferation and cellular motility. Conclusion This proteomic analysis helps to understand the molecular events underlying AS and PRGF-driven tissue regeneration processes, providing new evidence that comes along with the modulation of TGF-beta 1 activity and the reversion of the myofibroblastic phenotype by PRGF. This study was fully supported by BTI Biotechnology Institute, a dental implant company that investigates in the fields of oral implantology and PRGF-Endoret technology. MF and FM received a salary as scientists from BTI Biotechnology Institute. EA is the Scientific Director and president of BTI Biotechnology Institute. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Purpose: To compare the clinical and histological outcomes after intrastromal corneal ring segment (ICRS) implantation with and without plasma rich in growth factors (PRGF) in an experimental animal model. Materials and Methods: First, the toxicity of PRGF was tested in hen’s keratocyte cultures. Then, an animal model with 18 hens was randomly divided into 2 groups. In the first group, one ICRS was implanted in each eye (ICRS group). In the second group, the ICRS was firstly immersed 30 min in PRGF-Endoret solution, then implanted and, finally, PRGF-Endoret was inoculated into the channel (PRGF-ICRS group). Animals of each group were also separated into 3 groups regarding the time they were sacrificed, and corneal tissue was fixed for histological analysis at 2, 7 and 30 days. Cell death was detected by terminal uridine nick end labelling (TUNEL) assay. Proliferation was labelled by 5-bromo-2-deoxyuridine (BrdU) incorporation and myofibroblast differentiation by alpha-smooth muscle actin (αSMA) immunodetection. Clinical examination, analyzing epithelial wound closure, deposits and stromal haze, was carried out at the different study times. Results: No toxic effect was observed by PRGF in hen stromal cell cultures. Clinically, in PRGF-ICRS corneas at 7 days, there were more deposits with higher intensity than in ICRS group. Histologically, at day 2 there was less epithelial damage over the segment in the PRGF-ICRS group, corneal oedema around the segment disappeared earlier and, at day 7, there was also double the number of cells around the segment than in the ICRS group displaying different morphologies. The number of TUNEL-positive cells was statistically higher in the PRGF-ICRS group at 7 and 30 days, and the number of BrdU-positive cells was statistically higher at all analyzed times. However, there were no differences in the number of αSMA-positive cells at 30 days between both groups. Conclusions: The ICRS immersion in PRGF-Endoret prior and after to its corneal implantation, in an experimental animal model, enhances clinical deposits and histological cell turnover without increasing myofibroblast differentiation reducing stromal wound-healing time after surgery.