Experimental eye research最新文献

{"title":"Proteome of pericytes from retinal vasculature of diabetic donor eyes.","authors":"Sharmila Rajendran, Angayarkanni Narayansamy, Radha Annamalai, Lawrence D Cruze, Kaviarasan Kuppan","doi":"10.1016/j.exer.2024.110178","DOIUrl":"10.1016/j.exer.2024.110178","url":null,"abstract":"<p><p>Retinal pericytes (PCs) are contractile microvascular smooth muscle cells that wrap around the endothelial cells (ECs) maintaining intact retinal vasculature (RV) with a 1:1 ratio. Microvascular complications like diabetic retinopathy (DR) due to chronic diabetes causes apoptotic loss of PCs followed by diminished vessel stability, EC apoptosis, and ischemia, leading to retinal angiogenesis, and eventually severe vision loss. This study aimed to analyze the proteins in PCs isolated from the RV of diabetic human donor eyes and compare them with remaining mixed population (MP) of retinal vascular cells. PCs and MP proteomes were analyzed using semi-quantitative proteomics. Proteins were extracted, quantified, and analyzed in duplicate using LC-MS/MS on a Tandem mass spectrometer. Overall, 42 PC and 27 MP proteins, with 19 shared proteins, were identified. Functional enrichment analysis indicated that PC proteins share common biological processes, such as negative regulation of fibrinolysis and vLDL particle remodeling, nitric oxide transport, phospholipid efflux, positive control over the clearance of apoptotic cells, chondrocyte proliferation, lipoprotein lipase activity, and oxidative stress-induced intrinsic atrophic signaling pathways. In the fold enrichment analysis, the PC proteins were associated with cholesterol metabolism, Complement and coagulant, ECM-receptor interaction, longevity regulating pathway, Peroxisome proliferator-activated receptors (PPAR), focal adhesion and PI3 Akt signaling pathways. Among the PC proteins, vitronectin, gelsolin, hornerin, apolipoprotein A1, C3, H, and complement Factors C3, C4, and C9 were identified as the most highly ranked proteins in diabetes. The identified unique proteins of retinal PC could prove beneficial as a therapeutic target in the management of DR.</p>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":" ","pages":"110178"},"PeriodicalIF":3.0,"publicationDate":"2024-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142695322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel three-dimensional method for detailed analysis of RGC central projections under acute ocular hypertension","authors":"Wenhan Lu ,&nbsp;Yu Wang ,&nbsp;Wei Hu ,&nbsp;Xinyi Lin ,&nbsp;Xiaoyu Tong ,&nbsp;Yi Tian ,&nbsp;Yuning Chen ,&nbsp;Yicong Wang ,&nbsp;Yan Xiao ,&nbsp;Hongfang Yang ,&nbsp;Yi Feng ,&nbsp;Xinghuai Sun","doi":"10.1016/j.exer.2024.110157","DOIUrl":"10.1016/j.exer.2024.110157","url":null,"abstract":"<div><div>Normal perception of visual information relies not only on the quantity and quality of retinal ganglion cells (RGCs), but also on the integrity of the visual pathway, within which RGC central projection predominates. However, the exact changes of RGC central projection under particular pathological conditions remain to be elucidated. Here, we report a whole-brain clearing method modified from iDISCO for 3D visualization of RGC central projection. The CTB-labeled RGC central projection was visualized three-dimensionally with minimized both fluorescence quenching and the time taken. For observation of RGC axonal degeneration pattern under pathological conditions, we took acute ocular hypertension (AOH) as an example. Mice were intracamerally irrigated, and fluorescent signal in brain subregions where RGC axons projected to were quantified. The novel methodology is well-applied for rapid clearing and observation of RGC central projection in C57BL/6J, showing damaged RGC central projection on the AOH side and the most statistically significant degeneration in the superior colliculi (SC). Detailed analysis also revealed a distinct injury pattern among lateral geniculate nuclei (LGN) subregions, with the parvocellular part of the pregeniculate nuclei (PrGPC) being more vulnerable compared with the magnocellular part (PrGMC). The intracranial retrograde labeling of RGC subgroups based on brain damage variation showed PrGPC-projecting RGCs (<em>Plgn</em> RGC) being smaller than PrGMC-projecting RGCs (<em>Mlgn</em> RGC) in size and less in number, yet more vulnerable in terms of degeneration under AOH. Our data revealed the methodology for visualizing selective neuronal vulnerability under AOH, and in the meantime provided novel approach for future mechanisms exploration regarding RGC degeneration.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"250 ","pages":"Article 110157"},"PeriodicalIF":3.0,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142686461","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TGF-β1-induced apoptosis in retinal endothelial cells is implicated in retinal vein occlusion","authors":"Fengyu Chen ,&nbsp;Qi Wang ,&nbsp;Yujin Li ,&nbsp;Fen Li ,&nbsp;Lin Zhang ,&nbsp;Xuezhong Gu","doi":"10.1016/j.exer.2024.110168","DOIUrl":"10.1016/j.exer.2024.110168","url":null,"abstract":"<div><div>Retinal vein occlusion (RVO) is a serious vascular condition that impairs vision due to retinal endothelial cell injury and apoptosis. This study aimed to identify key molecular pathways and therapeutic targets involved in RVO pathogenesis. Transcriptomic analysis of the retinal tissues from a mouse RVO model was performed to identify differentially expressed genes and co-expression modules associated with RVO. Protein-protein interaction network analysis pinpointed putative hub genes. <em>In vitro</em> experiments using human retinal microvascular endothelial cells (HRMECs) validated the involvement of identified genes/pathways in apoptosis induced by oxygen-glucose deprivation/reperfusion (OGD/R) and UV exposure. Gene expression was assessed by RT-qPCR, while protein levels and phosphorylation were measured by ELISA and Western blotting. Apoptosis was evaluated using flow cytometry, and reactive oxygen species (ROS) were quantified using a fluorescence-based assay. A total of 392 genes were identified as putatively involved in RVO-associated apoptosis, enriched in MAPK, TGF-β and other signaling pathways. Among top hub genes, TGF-β1 emerged as a central regulator whose expression and signaling (pSmad2/3) increased after OGD/R induction or UV exposure in HRMECs. TGF-β1-induced HRMEC apoptosis was mediated by p38/JNK activation. Similar effects were observed for OGD/R and UV triggering TGF-β1-dependent p38/JNK signaling and apoptosis. Pharmacological inhibition of TGF-β signaling attenuated the apoptotic and oxidative stress responses induced by OGD/R and UV exposure. This study elucidates TGF-β1 as a crucial mediator of retinal endothelial injury through p38/JNK-induced apoptosis, suggesting TGF-β1 pathway inhibition as a potential therapeutic strategy for RVO.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"250 ","pages":"Article 110168"},"PeriodicalIF":3.0,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142692849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tracking astrocyte polarization in the retina in retinopathy of prematurity","authors":"Xiaoxiao Feng,&nbsp;Liwei Zhang,&nbsp;Kangwei Jiao,&nbsp;Yunqing Li,&nbsp;Min Wu,&nbsp;Yu Xie,&nbsp;Libo Xiao","doi":"10.1016/j.exer.2024.110170","DOIUrl":"10.1016/j.exer.2024.110170","url":null,"abstract":"<div><div>Astrocyte patterns affect the normal development of the retinal vascular network in retinopathy of prematurity (ROP), which is associated with VEGF secretion. However, the role of the astrocyte polarization in this process remains unknown. Therefore, this study aimed to track the status of A1/A2 reactive astrocytes in the retinas of the oxygen-induced retinopathy (OIR) model and their association with VEGF expression. The C57BL/6 mouse OIR model was constructed to characterize the pathological changes in ROP. Immunofluorescence of iB4 and GFAP staining was performed to observe changes in the vascular network and astrocyte pattern at different time points (P0, P7, P12, P17, and P21). C3-labeled A1 reactive and S100A10-labeled A2 reactive astrocytes and VEGF were also observed. The pattern of GFAP-labeled astrocyte was altered concurrently with the iB4-positive vascular network during OIR. Astrocyte activity was significantly weakened at P12 and significantly enhanced at P17. Notably, the number of C3-labeled A1 reactive astrocytes was significantly increased at P12, decreased at P17, and normalized at P21 in OIR models. S100A10-labeled A2 reactive astrocytes were significantly increased at P17 but did not change significantly at P12 or P17. VEGF levels were decreased at P7-P12 and increased at P12-P17. The expression pattern of VEGF was opposite to that of C3-labeled A1 reactive astrocytes and identical to that of S100A10-labeled A2 reactive astrocytes. In conclusion, the astrocyte pattern and vascular network exhibited similar changes during the OIR process, and the periods of vaso-obliteration and neo-vascularization display an abnormal activation in A1-and A2-reactive astrocytes.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"250 ","pages":"Article 110170"},"PeriodicalIF":3.0,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142692850","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Choroidal macrophages in homeostasis, aging and age-related macular degeneration","authors":"Adnan H. Khan ,&nbsp;Kelly Mulfaul","doi":"10.1016/j.exer.2024.110159","DOIUrl":"10.1016/j.exer.2024.110159","url":null,"abstract":"<div><div>With increasing age, the optimal functioning of the choroid is essential for efficient removal of waste products formed from photoreceptor renewal. A decline in regulatory elements of the immune system, termed immunosenescence, and the failure of para-inflammation to restore tissue homeostasis can result in the progression of healthy aging to sight-threatening inflammation of the choroid. Macrophages are uniquely situated between the innate and adaptive immune systems, with a high capacity for phagocytosis, recognition of complement components, as well as antigen presentation. In this review, we provide an overview of macrophages and their properties in the healthy choroid and cover the impact of aging, immunosenescence and inflammaging on the function of choroidal macrophages. We will discuss the impact of age on macrophage phenotype and behaviour in the pathophysiology of age-related macular degeneration.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"250 ","pages":"Article 110159"},"PeriodicalIF":3.0,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142692846","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Conditioned media from dental pulp stem cells to counteract age-related macular degeneration","authors":"Giulia Carozza ,&nbsp;Darin Zerti ,&nbsp;Fanny Pulcini ,&nbsp;Loreto Lancia ,&nbsp;Simona Delle Monache ,&nbsp;Vincenzo Mattei ,&nbsp;Rita Maccarone","doi":"10.1016/j.exer.2024.110167","DOIUrl":"10.1016/j.exer.2024.110167","url":null,"abstract":"<div><h3>Purpose</h3><div>Age-related macular degeneration (AMD) is the leading cause of blindness in the elderly. To date, there are no effective therapies to counteract AMD towards the most severe stages characterised by a progressive loss of photoreceptors triggered by retinal pigmented epithelium dysfunction. Given their easy source and their high proliferative potential, Dental Pulp Stem Cells (DPSCs) are considered promising for regenerative medicine. The main advantage of DPSCs is related to their paracrine immunosuppressive and immunoregulatory abilities, including the capability to promote regeneration of damaged tissues. Recent studies demonstrated the therapeutic potential of DPSCs-conditioned media (CM) in neurodegenerative diseases. In addition, we have already shown a differential expression of some growth factors and cytokines in CM derived from DPSCs cultured in hypoxia and normoxia conditions.</div></div><div><h3>Aim</h3><div>In this study we evaluated the capability of DPSCs-CM to counteract retinal degeneration in an animal model of AMD. DPSCs-CM were intravitreally injected the day before the exposure of albino rats to high intensity light (LD).</div></div><div><h3>Results</h3><div>We evaluated the retinal function, and we performed morphological and molecular analysis a week after the LD, in accordance with the well-established protocol of our light damage model. DPSCs-CM obtained from hypoxia (HYPO-CM) or normoxia (NORM-CM), were able to preserve the retinal function, to reduce the damaged area and to counteract the upregulation of key factors involved in retinal degeneration, like FGF-2. Furthermore, we demonstrated that neither conditioned media modified inflammatory activation, as shown by both microglia activation and GFAP upregulation, but <em>in vitro</em> studies demonstrated a significant effect of both CM to counteract oxidative stress, one of the main causes of AMD.</div></div><div><h3>Conclusion</h3><div>Taken together, our study demonstrated that NORM-CM and HYPO-CM, albeit with a different chemical composition, could represent eligible candidates to counteract retinal degeneration in an animal model of AMD. Further studies are needed to obtain conditioned media with the best performance in term of retinal protection.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"250 ","pages":"Article 110167"},"PeriodicalIF":3.0,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142686386","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The effect of immobilisation strategies on the ability of peptoids to reduce the adhesion of P. aeruginosa strains to contact lenses","authors":"Manjulatha Sara ,&nbsp;Sudip Chakraborty ,&nbsp;Renxun Chen ,&nbsp;Dennis Palms ,&nbsp;Georgio Katsifis ,&nbsp;Zhongyan Li ,&nbsp;Syamak Farajikhah ,&nbsp;Vinod Massedupally ,&nbsp;Alex Hui ,&nbsp;Edgar H.H. Wong ,&nbsp;Naresh Kumar ,&nbsp;Krasimir Vasilev ,&nbsp;David Mackenzie ,&nbsp;Linda Losurdo ,&nbsp;Farida Dehghani ,&nbsp;Havard Jenssen ,&nbsp;Kristian Sorensen ,&nbsp;Jennifer S. Lin ,&nbsp;Annelise E. Barron ,&nbsp;Mark Willcox","doi":"10.1016/j.exer.2024.110149","DOIUrl":"10.1016/j.exer.2024.110149","url":null,"abstract":"<div><h3>Aim</h3><div>Previous studies have demonstrated that contact lenses coated with the antimicrobial cationic peptide Mel4, a derivative of melimine, can reduce the occurrence of keratitis. However, the antimicrobial activity of Mel4 weakened over time due to its susceptibility to proteolytic degradation. Oligo-<em>N</em>-substituted glycine peptoids such as TM5 and TM18 possess antimicrobial properties and are resistant to proteolytic breakdown. This study focused on exploring methods for covalently attaching these peptoids to contact lenses to enhance their durability and performance <em>in vitro</em>.</div></div><div><h3>Methods</h3><div>The peptoids TM5 and TM18 were covalently attached to etafilcon lenses via carbodiimide chemistry (EDC/NHS), oxazoline plasma, and plasma ion immersion implantation (PIII). The lenses were analysed using X-ray photoelectron spectroscopy (XPS), surface charge, and hydrophobicity. Inhibition of adhesion of multidrug-resistant <em>Pseudomonas aeruginosa</em> and cytotoxicity on corneal epithelial cells were evaluated. The impact of moist heat sterilization on activity was also assessed.</div></div><div><h3>Results</h3><div>XPS confirmed peptoid binding to lenses. Peptoid coatings slightly increased contact angles (≤23°) without affecting overall charge. Peptoids, bound via carbodiimide, inhibited <em>P. aeruginosa</em> adhesion by over 5 log10 CFU per lens, outperforming melimine, which required six times the concentration for a 3 log10 reduction. Peptoids attached via oxazoline or PIII reduced adhesion by &gt; 5 log10 CFU. All covalent methods significantly reduced bacterial adhesion compared to untreated lenses (<em>P</em> &lt; 0.0001). Peptoid-bound lenses were non-toxic to corneal epithelial cells. Sterilization did not affect carbodiimide-treated lenses but reduced the activity of oxazoline and PIII surfaces by 1–2 log10 CFU.</div></div><div><h3>Conclusion</h3><div>Peptoids TM5 and TM18 effectively reduced <em>P. aeruginosa</em> adhesion on lenses, with carbodiimide-bound surfaces retaining activity post-sterilization, showing promise for the development of antimicrobial contact lenses.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"250 ","pages":"Article 110149"},"PeriodicalIF":3.0,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142686463","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Increased glucose concentration modifies TGF-β1 and NFκB signaling pathways in aniridia limbal fibroblasts, in vitro","authors":"Zhen Li ,&nbsp;Tanja Stachon ,&nbsp;Sabrina Häcker ,&nbsp;Fabian N. Fries ,&nbsp;Ning Chai ,&nbsp;Berthold Seitz ,&nbsp;Lei Shi ,&nbsp;Shao-Lun Hsu ,&nbsp;Shuailin Li ,&nbsp;Shanhe Liu ,&nbsp;Maryam Amini ,&nbsp;Shweta Suiwal ,&nbsp;Nóra Szentmáry","doi":"10.1016/j.exer.2024.110163","DOIUrl":"10.1016/j.exer.2024.110163","url":null,"abstract":"<div><div>To determine the impact of increased glucose concentration on gene expression of primary healthy human limbal fibroblasts (LFCs) and congenital aniridia human limbal fibroblasts (AN-LFCs), <em>in vitro</em>.</div><div>LFCs (n = 8) and AN-LFCs (n = 8) were isolated and cultured in serum containing DMEM, including either normal glucose (17.5 mM) or increased glucose (70 mM) concentration for 48h or 72h, respectively<strong>.</strong> mRNA and protein expression of transforming growth factor beta 1 (TGF-β1), alpha-smooth muscle actin (ACTA)2A1, SMAD 2/3, hypoxia markers such as nuclear factor kappa B (NFκB), inducible nitric oxide synthase (iNOS), hypoxia-inducible factor 1-alpha (HIF-1ɑ), oxidative stress markers such as nuclear factor erythroid 2-related factor 2 (Nrf2) and Catalase (CAT) were analyzed using qPCR and Western blot.</div><div>In 70 mM glucose concentration medium for 48 h, <em>TGF-β1</em> mRNA expression was significantly lower (p = 0.001, p &lt; 0.001), <em>Nrf2</em> (p = 0.001, p = 0.001) and <em>CAT</em> (p = 0.001, p = 0.001) mRNA expression was significantly higher in LFCs and AN-LFCs, than using 17.5 mM glucose concentration medium. In addition, in 70 mM glucose concentration medium for 48 h, <em>SMAD 2</em>, <em>SMAD 3</em>, <em>NFκB</em>, <em>HIF-1ɑ</em> mRNA expression was significantly lower in AN-LFCs, than in 17.5 mM glucose concentration medium (p = 0.003, p = 0.002, p = 0.008, p = 0.020). At this time-point in 70 mM glucose concentration medium, at protein level, TGF-β1, SMAD2/3 and NFκB were significantly lower in AN-LFCs, than in 17.5 mM glucose concentration medium (p = 0.041, p = 0.002, p = 0.012).</div><div>In 70 mM glucose concentration medium for 72h, <em>TGF-β1</em> was significantly higher (p &lt; 0.001, p &lt; 0.001) and <em>Nrf2</em> (p = 0.001, p = 0.001) and <em>CAT</em> (p &lt; 0.001, p &lt; 0.001) mRNA were significantly lower in LFCs and AN-LFCs, than in 17.5 mM glucose concentration medium. At this time-point, in 70 mM glucose concentration medium, <em>NFκB</em> mRNA was significantly higher (p &lt; 0.001) in LFCs<strong>,</strong> than in 17.5 mM glucose concentration DMEM medium. In 70 mM glucose concentration medium for 72 h, TGF-β1 and NFκB protein were significantly lower in AN-LFCs, than in 17.5 mM glucose concentration medium (p &lt; 0.001, p &lt; 0.001).</div><div>Our study confirmed that high glucose concentration has an impact on TGF-β1 and NFκB signaling both in AN-LFCs and LFCs. These findings highlight that prolonged exposure to high glucose levels may contribute to cellular stress and dysfunction in LFCs and AN-LFCs.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"250 ","pages":"Article 110163"},"PeriodicalIF":3.0,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142692848","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Macromolecular crowding agent dependent extracellular matrix deposition and growth factor retention in human corneal fibroblast cultures","authors":"Mehmet Gurdal,&nbsp;Dimitrios I. Zeugolis","doi":"10.1016/j.exer.2024.110162","DOIUrl":"10.1016/j.exer.2024.110162","url":null,"abstract":"<div><div>The major obstacle in the commercialisation and clinical translation of tissue engineered medicines is the required for the development of implantable tissue surrogates prolonged <em>in vitro</em> culture. Macromolecular crowding (MMC) enhances and accelerates extracellular matrix (ECM) deposition, thus offering an opportunity to bridge the gap between research and development in tissue engineered substitutes. However, the optimal MMC agent is still elusive. Herein, we first assessed the biophysical properties of the most widely used MMC agents [κλ carrageenan (κλ CR), λ carrageenan (λ CR) and Ficoll™ cocktail (FC)] and then assessed their effect in basic cell function, ECM deposition and growth factor retention in human corneal fibroblast (hCF) cultures. Dynamic light scattering analysis revealed that both CR macromolecules had significantly lower and higher zeta potential and hydrodynamic radius, respectively, than the FC. None of the MMC agents affected hCF morphology and all induced similar hCF viability, proliferation and metabolic activity. Electrophoresis and immunofluorescence analyses made apparent that at day 10 (longest time point assessed), the FC brought about the highest fibronectin and collagen types I, III, IV, V and VI deposition. Deposited ECM pattern analysis showed that at day 10, the FC induced the lowest lacunarity and normalised end points and the highest fractal dimension and % high density matrix. Further immunofluorescence analysis revealed no significant differences between the groups in vimentin, aldehyde dehydrogenase 3 family member A1, keratocan, paired box protein 6 and α-smooth muscle actin. Importantly, at day 10, the FC resulted in the highest growth factor retention (20 molecules). Our data clearly illustrate a MMC agent dependent cell response, with the FC having the highest positive effect in hCF cultures.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"250 ","pages":"Article 110162"},"PeriodicalIF":3.0,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142686462","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A modified calculation formula for meibomian gland grading","authors":"Yang Liu ,&nbsp;Yaoyao Ren ,&nbsp;Wenjing Li ,&nbsp;Wei Liu ,&nbsp;Min Ke","doi":"10.1016/j.exer.2024.110166","DOIUrl":"10.1016/j.exer.2024.110166","url":null,"abstract":"<div><div>This study aimed to establish a modified calculation formula for the grading of meibomian glands. Meibography images from 102 participants by different examiners on separate machines on two consecutive days were analyzed, quantified and compared side-by-side. Measure and calculate the ratio of the MGs area to the whole eyelid area and the ratio of the MGs to the corneal base area. Our findings demonstrate that there were significant differences in the ratio of the meibomian gland area to the whole eyelid area between two measurements, but not in the ratio of the meibomian gland area to the corneal base area. The measurement of the eyelid area showed bigger variations and poorer repeatability than the meibomian gland area and the corneal base area. As such, the ratio of the meibomian gland area to the corneal base area is a more stable indicator for the grading of meibomian glands over multiple measurement.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"250 ","pages":"Article 110166"},"PeriodicalIF":3.0,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142686457","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}