Repopulation of decellularized retinas with hiPSC-derived retinal pigment epithelial and ocular progenitor cells shows cell engraftment, organization and differentiation

The retinal extracellular matrix (ECM) provides architectural support, adhesion and signal guidance that controls retinal development. Decellularization of the ECM affords great potential to tissue engineering; however, how structural retinal ECM affects in vitro development, differentiation and maturation of ocular cells remains to be elucidated. Here, mouse and porcine retinas were decellularized and the protein profile analyzed. Acellular retinal ECM (arECM) scaffolds were then repopulated with human iPSC-derived retinal pigment epithelial (RPE) cells or ocular progenitor cells (OPC) to assess their integration, proliferation and organization. 3837 and 2612 unique proteins were identified in mouse and porcine arECM, respectively, of which 93 and 116 proteins belong to the matrisome. GO analysis shows that matrisome-related proteins were associated with the extracellular region and cell junction and KEGG pathways related to signalling transduction, nervous and endocrine systems and cell junctions were enriched. Interestingly, mouse and porcine arECMs were successfully repopulated with both RPE and OPC, the latter exhibiting cell lineage-specific clusters. Retinal cells organized into different layers containing well-defined areas with pigmented cells, photoreceptors, Müller glia, astrocytes, and ganglion cells, whereas in other areas, conjunctival/limbal, corneal and lens cells re-arranged in cell-specific self-organized areas. In conclusion, our results demonstrated that decellularization of both mouse and porcine retinas retains common native ECM components that upon cell repopulation could guide similar ocular cell adhesion, migration and organization

A review of current management of vitreomacular traction and macular hole

The paper presents a review of the sequence of events of posterior vitreous detachment (PVD), vitreomacular adhesion (VMA), vitreomacular traction (VMT), and macular hole (MH) from their pathophysiological aspects, clinical features, diagnostic implications, and current management strategies. A treatment algorithm to be used in clinical practice in patients with VMA, VMT, and MH based on the presence of symptoms, visual acuity, associated epiretinal membrane, and width of the vitreous attachment is presented. Observation, pharmacologic vitreolysis with ocriplasmin, and surgical treatment are positioned as treatment options in the different steps of the therapeutic algorithm, with clear indications of the paths to be followed according to the initial presenting manifestations and the patient's clinical course

Repopulation of decellularized retinas with hiPSC-derived retinal pigment epithelial and ocular progenitor cells shows cell engraftment, organization and differentiation

Decellularization; Ocular progenitors; RetinaDescelularización; Progenitores oculares; RetinaDescel·lularització; Progenitors oculars; RetinaThe retinal extracellular matrix (ECM) provides architectural support, adhesion and signal guidance that controls retinal development. Decellularization of the ECM affords great potential to tissue engineering; however, how structural retinal ECM affects in vitro development, differentiation and maturation of ocular cells remains to be elucidated. Here, mouse and porcine retinas were decellularized and the protein profile analyzed. Acellular retinal ECM (arECM) scaffolds were then repopulated with human iPSC-derived retinal pigment epithelial (RPE) cells or ocular progenitor cells (OPC) to assess their integration, proliferation and organization. 3837 and 2612 unique proteins were identified in mouse and porcine arECM, respectively, of which 93 and 116 proteins belong to the matrisome. GO analysis shows that matrisome-related proteins were associated with the extracellular region and cell junction and KEGG pathways related to signalling transduction, nervous and endocrine systems and cell junctions were enriched. Interestingly, mouse and porcine arECMs were successfully repopulated with both RPE and OPC, the latter exhibiting cell lineage-specific clusters. Retinal cells organized into different layers containing well-defined areas with pigmented cells, photoreceptors, Müller glia, astrocytes, and ganglion cells, whereas in other areas, conjunctival/limbal, corneal and lens cells re-arranged in cell-specific self-organized areas. In conclusion, our results demonstrated that decellularization of both mouse and porcine retinas retains common native ECM components that upon cell repopulation could guide similar ocular cell adhesion, migration and organization.This work was supported by La Marató de TV3 Foundation (484/C/2012); ERA-NET EuroNanoMed III-AC19/00080/ISCIII (CELLUX); Instituto de Salud Carlos III (CA18/00045 and PI18/00219); and the European Social Fund, the Ministerio de Ciencia, Innovación y Universidades, which is part of the Agencia Estatal de Investigación (PTA2018-016371-I). A.D. was supported by PT13/0001/0041 PRB2-ISCIII-SGEFI-FEDER-PE I+D+i 2013–2016 and ISCIII-FEDER RETICS (Oftared; RD16/0008). We thank the CERCA Programme/Generalitat de Catalunya for institutional support

Sudden hearing loss: National survey in Spain

This is the peer reviewed version of the following article: Acta Otorrinolaringológica 67.2 (2016): 59-65, which has been published in final form at http://dx.doi.org/10.1016/j.otorri.2015.03.003The objective of our study was to identify the diagnostic and therapeutic approaches in the different ENT Departments of Spain with respect to sudden deafness. We wanted to establish a basis to help to create a new nation-wide consensus, unifying treatment, diagnostic and follow-up criteria for this disease. Methods: We carried out an anonymous Internet survey, addressing Spanish ENT doctors nation-wide (n = 2,029), gathering in 33 questions different aspects about diagnostic criteria, additional tests, treatment procedures and prognostic factors in sudden deafness, according to the different protocols and experience of the participants in the survey. Results: A total of 293 Spanish ENT doctors (14%) took part anonymously. In relation to diagnostic criteria, is the most noteworthy was the requisite of a confirmed neurosensorial loss (91.1%) followed by "initiated in less than three days" (75%) and 3 consecutive frequencies affected (76.4%). More than half of the participants requested an MRI of the IAC/CPA (68.7%) and 88.2% used gadolinium in this test. The prognostic factor most frequently considered was delay in commencement of treatment onset (84.8%).As far as treatment of primary cases, most of the responders agreed on the use of corticosteroids (99.7%). Oral administration was the most widely used (66%), followed by intravenous (29.6%) and intratympanic (1.4%) administration. Ninety-two percent had not had any major complications with systemic steroids. Intratympanic treatments were used by 70% of responders for rescue in failure. Conclusions: In Spain there is currently a significant disparity of concepts regarding the diagnosis of sudden deafness, and more agreement as to using steroids as their treatment. This highlights the need to implement measures to promote a better approach, which would be homogeneous and consensual, to this condition.Introducción: El objetivo de nuestro estudio es identificar las actitudes diagnósticas y terapéuticas que se llevan a cabo en los diferentes servicios de otorrinolaringología (tanto del ámbito privado como público) en España con respecto a la sordera súbita. Esto permitirá establecer una base que ayude a generar un nuevo consenso a nivel nacional, unificando criterios para el tratamiento, diagnóstico y seguimiento de esta patología. Material y métodos: Se realizó una encuesta anónima por Internet, dirigida a otorrinolaringólogos españoles a nivel nacional (n = 2.029 especialistas afiliados a la SEORL), recopilando en 33 preguntas diferentes aspectos en relación a los criterios diagnósticos, pruebas complementarias, pautas de tratamiento y factores pronósticos en la sordera súbita, según los diferentes protocolos instaurados y experiencia de los participantes en la encuesta. Resultados: Participaron 293 otorrinolaringólogos españoles de forma anónima (14% del total). Respecto a criterios diagnósticos, destaca el requerimiento de confirmar una hipoacusia neurosensorial (91,1%), de inicio en menos de tres días (75%) y afectación de tres frecuencias consecutivas (76,4%). Más de la mitad de los participantes solicitan resonancia magnética de CAI/APC (68,7%), y el 88,2% utiliza contraste con gadolinio en esta prueba. El factor pronóstico que se consideró con mayor frecuencia en la encuesta fue la demora hasta inicio del tratamiento con un 84,8%. Respecto al tratamiento empleado en casos primarios, la gran mayoría de los encuestados (99,7%) coinciden en la administración de corticoides. La vía oral es la más utilizada (66%), seguida de la administración intravenosa (29,6%), e intratimpánica (1,4%). El 92% no han tenido complicaciones mayores con el tratamiento corticoide sistémico. La vía intratimpánica es empleada en un 70% como rescate en fracasos Conclusiones: En España, existe actualmente una importante disparidad en el uso de los medios diagnósticos en la sordera súbita y un mayor acuerdo en el uso de corticoides como su tratamiento. Sería necesario implementar medidas que permitan un mejor abordaje, homogéneo y consensuado de esta patologí

Early and intermediate age-related macular degeneration: update and clinical review

Age-related macular degeneration (AMD) is the leading cause of irreversible central vision loss in developed countries. With the aging of population, AMD will become globally an increasingly important and prevalent disease worldwide. It is a complex disease whose etiology is associated with both genetic and environmental risk factors. An extensive decline in the quality of life and progressive need of daily living assistance resulting from AMD among those most severely affected highlights the essential role of preventive strategies, particularly advising patients to quit smoking. In addition, maintaining a healthy diet, controlling other risk factors (such as hypertension, obesity, and atherosclerosis), and the use of nutritional supplements (antioxidants) are recommendable. Genetic testing may be especially important in patients with a family history of AMD. Recently, unifying criteria for the clinical classification of AMD, defining no apparent aging changes; normal aging changes; and early, intermediate, and late AMD stages, are of value in predicting AMD risk of progression and in establishing recommendations for the diagnosis, therapeutic approach, and follow-up of patients. The present review is focused on early and intermediate AMD and presents a description of the clinical characteristics and ophthalmological findings for these stages, together with algorithms for the diagnosis and management of patients, which are easily applicable in daily clinical practice

Management of wet age-related macular degeneration in Spain: challenges for treat and extend implementation in routine clinical practice

Purpose: to ascertain wet AMD (wAMD) management patterns in Spain. Methods: a two-round Delphi study conducted through a questionnaire-based survey designed from literature review and validated by an independent Steering Committee. Results: forty-nine retina specialists experienced in wAMD participated by answering the two-round study questionnaire. Retina specialists are the main responsible for wAMD diagnosis and monitoring, including visits and associated procedures, with a median time per visit of 15 minutes. Standard treatment strategies are based on anti-VEGF administration, including standard loading dose administration followed by maintenance with aflibercept or ranibizumab (81% of patients). Although treat and extend (T&E) dosing strategy is considered as optimal for wAMD management (78% of the panelists), the main routine healthcare limitations (i.e., visits overload, reduced staff, short visit time, coordination issues, lack of facilities) conduct to self-defined "flexible" strategies, based on T&E and pro-re-nata (PRN) protocols. Conclusion: proactive treatment patterns (T&E) are the preferred ones by the retina specialists in Spain. However, their proper implementation is difficult due to healthcare resource limitations, as well as organisation and logistic issues. The use of anti-VEGF agents with longer duration of action could facilitate the use of strict T&E approaches according to routine clinical practices

All-trans retinoic acid modulates pigmentation, neuroretinal maturation, and corneal transparency in human multiocular organoids

Ácido transretinoico; Retina; Células madreÀcid transreitnoic; Retina; Cèl·lules mareAll-trans retinoic acid; Retina; Stem cellsBackground All-trans retinoic acid (ATRA) plays an essential role during human eye development, being temporally and spatially adjusted to create gradient concentrations that guide embryonic anterior and posterior axis formation of the eye. Perturbations in ATRA signaling can result in severe ocular developmental diseases. Although it is known that ATRA is essential for correct eye formation, how ATRA influences the different ocular tissues during the embryonic development of the human eye is still not well studied. Here, we investigated the effects of ATRA on the differentiation and the maturation of human ocular tissues using an in vitro model of human-induced pluripotent stem cells-derived multiocular organoids. Methods Multiocular organoids, consisting of the retina, retinal pigment epithelium (RPE), and cornea, were cultured in a medium containing low (500 nM) or high (10 µM) ATRA concentrations for 60 or 90 days. Furthermore, retinal organoids were cultured with taurine and T3 to further study photoreceptor modulation during maturation. Histology, immunochemistry, qPCR, and western blot were used to study gene and protein differential expression between groups. Results High ATRA levels promote the transparency of corneal organoids and the neuroretinal development in retinal organoids. However, the same high ATRA levels decreased the pigmentation levels of RPE organoids and, in long-term cultures, inhibited the maturation of photoreceptors. By contrast, low ATRA levels enhanced the pigmentation of RPE organoids, induced the opacity of corneal organoids—due to an increase in collagen type IV in the stroma— and allowed the maturation of photoreceptors in retinal organoids. Moreover, T3 promoted rod photoreceptor maturation, whereas taurine promoted red/green cone photoreceptors. Conclusion ATRA can modulate corneal epithelial integrity and transparency, photoreceptor development and maturation, and the pigmentation of RPE cells in a dose-dependent manner. These experiments revealed the high relevance of ATRA during ocular tissue development and its use as a potential new strategy to better modulate the development and maturation of ocular tissue through temporal and spatial control of ATRA signaling.This research project was funded by a grant from ERA-NET EuroNanoMed III-ISCIII (AC19/00080) (CELLUX), and Instituto de Salud Carlos III (PI18/00219). A.D. was supported by ISCIII-FEDER RETICS (Oftared; RD16/0008). H.IM. was supported by EuroNanoMed III-ISCIII (AC19/00080)

Management of wet age-related macular degeneration in Spain: challenges for treat and extend implementation in routine clinical practice

Age-related macular degeneration; Ophthalmology services; Anti-VEGFDegeneración macular relacionada con la edad; Servicios de oftalmología; Anti-VEGFDegeneració macular relacionada amb l’edat; Serveis d’oftalmologia; Anti-VEGFPurpose. To ascertain wet AMD (wAMD) management patterns in Spain. Methods. A two-round Delphi study conducted through a questionnaire-based survey designed from literature review and validated by an independent Steering Committee. Results. Forty-nine retina specialists experienced in wAMD participated by answering the two-round study questionnaire. Retina specialists are the main responsible for wAMD diagnosis and monitoring, including visits and associated procedures, with a median time per visit of 15 minutes. Standard treatment strategies are based on anti-VEGF administration, including standard loading dose administration followed by maintenance with aflibercept or ranibizumab (81% of patients). Although treat and extend (T&E) dosing strategy is considered as optimal for wAMD management (78% of the panelists), the main routine healthcare limitations (i.e., visits overload, reduced staff, short visit time, coordination issues, lack of facilities) conduct to self-defined “flexible” strategies, based on T&E and pro-re-nata (PRN) protocols. Conclusion. Proactive treatment patterns (T&E) are the preferred ones by the retina specialists in Spain. However, their proper implementation is difficult due to healthcare resource limitations, as well as organisation and logistic issues. The use of anti-VEGF agents with longer duration of action could facilitate the use of strict T&E approaches according to routine clinical practices

Prevalence and implications of the presence of intraocular silicone oil droplets in patients treated with intravitreal injections of anti-VEGF

Intraocular pressure; Macular diseases; Silicone dropletsPresión intraocular; Enfermedades maculares; Gotas de siliconaPressió intraocular; Malalties maculars; Gotes de siliconaBackground To determine the percentage of patients who have silicone droplets in the vitreous after treatment with different anti-Vascular Endothelial Growth Factor (anti-VEGF) intravitreal injections (IVI) and how symptomatic they are. Methods One hundred fifty-two eyes of 140 patients who had at least received an IVI were recruited for this study. Data collection included the number and type of IVI (aflibercept, ranibizumab and bevacizumab) and the follow-up time. A complete ophthalmologic examination was carried out and patients were classified in four groups according to the amount of silicone droplets found in dilated fundoscopy (nonexistent, scarce, moderate and abundant). Measurement of intraocular pressure (IOP) was also carried out. An interview was conducted to report the presence and intensity of the symptomatology. Results Silicone oil droplets were reported in 109 eyes (71.7%). A positive correlation was found between the number of IVIs received and the quantity of droplets found, especially when aflibercept was used. Posterior vitreous detachment (PVD) was present in 65.8% of the patients, showing a positive correlation with the number of bubbles. Regarding the symptomatology, 60 eyes (39.5%) had floaters and the disturbance was reported to be 4 out of 10. The group with a moderate amount of silicone droplets had the highest percentage of floaters (60%). No statistical differences in the IOP were found between groups, although the group with abundant droplets had a higher mean IOP. Conclusion A high prevalence of silicone droplets in vitreous of patients who undergo IVI treatment was found. It appears to have little impact on symptomatology and rise of IOP

Fundus flavimaculatus-like in myotonic dystrophy: a case report

Informe d'un cas; Fundus flavimaculatus; Distròfia miotònicaReporte de un caso; Fundus flavimaculatus; Distrofia miotónicaCase report; Fundus flavimaculatus; Myotonic dystrophyBackground Myotonic dystrophy is an inherited disease characterized by progressive muscle weakness and myotonia. It is a multisystemic disorder that affects different parts of the body, including the eye. Dysfunction of ocular muscles, ptosis and cataract are the most common ophthalmologic manifestations, but it can also present with pigmentary changes in the retina. This report presents and discusses an unusual case of a pigmented pattern dystrophy simulating a fundus flavimaculatus in a patient with myotonic dystrophy. Case presentation We present a case of a woman with a history of myotonic dystrophy and complaints of progressive vision loss who presented bilateral retinal pigmentary changes in posterior pole and midperiphery. The characteristics and distribution of pigmented deposits, as well as ancillary tests, showed a retinal phenotype compatible with a multifocal pattern dystrophy or a fundus flavimaculatus. Conclusions There are a few publications about retinal disorders in patients with myotonic dystrophy. When macular area is affected it tends to adopt a patterned-shape defined as butterfly dystrophy or reticular dystrophy. To our knowledge, this is the first report of a patient with myotonic dystrophy and multifocal pattern dystrophy or fundus flavimaculatus