Background: The ex vivo model represented by mouse retinal explants in culture is a useful experimental model to investigate the molecular mechanism involved in neurovascular diseases such as diabetic retinopathy (DR). It ensures an experimental overview with more complete respect to isolate cells and reduce problems in terms of accessibility and management with respect to in vivo model. In particular, it allows the evaluation of the relationship between retinal cells in response to the typical stressors involved in DR pathogenesis.
Methods: Ex vivo retinal fragments derived from 3- to 5-week-old C57BL/6J mice. In particular, after dissection, the retina is cut into 4 separate fragments and transferred onto inserts placed with ganglion cells up. Once in culture, the explants could be treated in stress conditions typical of DR. In particular, this study protocol describes the procedure for the preparation and the culture of retinal explants with specific metabolic stressors such as high glucose (HG), advanced glycation end product (AGE), and oxidative stress (OS). In the end, this paper provides the protocols to perform molecular analyses in order to evaluate the response of retinal explants to stress and/or neuroprotective treatments.
Discussion: The cultured retinal explants represent an ex vivo experimental model to investigate the molecular mechanisms involved in neurovascular diseases such as DR. Moreover, they could be useful to test the effect of neuroprotective compounds in response to metabolic stressors in a fewer time respect to an in vivo model. In conclusion, retinal explants in culture represent a valuable experimental model to conduct further studies to better understand the pathophysiology of DR.
Background: To investigate the outcome of cataract surgery in patients with legal blindness defined as best-corrected visual acuity (BCVA) of 20/1,000 or lower and to determine factors influencing the visual outcome in these patients.
Methods: Medical records of 68 eyes of 62 patients diagnosed with legal blindness and underwent cataract surgery were reviewed. The study population was divided into 3 groups based on types of cataracts (Group A: posterior subcapsular cataract, Group B: mature or brunescent cataract, and Group C: cataract combined with other ocular diseases). Data including demographics, predisposing factors, BCVA before and 6 months after surgery and post-operative complications were collected and analyzed.
Results: Mean preoperative logMAR BCVA was 1.88±0.24, 2.24±0.26 and 1.96±0.31 in Groups A, B and C, respectively (P=0.003). The postoperative BCVA was 0.49±0.35, 0.51±0.47 and 0.90±0.53 in Groups A, B and C, respectively (p=0.003). Significant improvement in BCVA was shown in all 3 groups (P<0.001 in Groups A and B and, P=0.001 in Group C). There was significant difference in the amount of visual improvement among the 3 groups, P<0.001). Although there was no significant difference in the amount of visual improvement between group A and C (P=0.379), significantly higher visual improvement was achieved in group B compared with group A (P=0.012) and C (P=0.001).
Conclusions: Cataract surgery should be encouraged for patients with legal blindness, even in the presence of other ocular disease. Significant visual recovery was observed in all the groups, particularly in those with mature or brunescent cataract.
Background: The complexity of the glaucoma pathophysiology is directly reflected on its experimental modeling for studies about pathological mechanisms and treatment approaches. Currently, a variety of in vivo models are available for the study of glaucoma, although they do not reach an exact reproduction of all aspects characterizing the human glaucoma. Therefore, a comprehensive view of disease onset, progression and treatment efficacy can only be obtained by the integration of outcomes deriving from different experimental models.
Methods: The present article summary experimental procedures and analytical methodologies related with two experimental models of glaucoma belonging to the classes of induced intraocular pressure (IOP)-elevation and genetic models, methyl cellulose (MCE)-induced ocular hypertension and DBA/2J mouse strain. Point-by-point protocols are reported with a particular focus on the critical point for the realization of each model. Moreover, typical strength and drawbacks of each model are described in order to critically handle the outcomes deriving from each model.
Discussion: This paper provides a guideline for the realization, analysis and expected outcomes of two models allowing to study IOP-driven neurodegenerative mechanisms rather than IOP-independent neurodegeneration. The complementary information from these models could enhance the analysis of glaucomatous phenomena from different points of view potentiating the basic and translational study of glaucoma.
