Comparison of cytotoxicity effects induced by four different types of nanoparticles in human corneal and conjunctival epithelial cells.
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ABSTRACT:
SUBMITTER: Li X
PROVIDER: S-EPMC8742118 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Integrity of the cornea, the most anterior part of the eye is indispensable for vision. 45 million individuals are bilaterally blind and another 135 millions have severely impaired vision in both eyes because of loss of corneal transparency; treatments range from local medications to corneal transplants and more recently to stem cell therapy. The corneal epithelium is a squamous epithelium that is constantly renewing with a vertical turnover of seven to fourteen days in many mammals3. Identification of slow cycling cells (label-retaining cells or LRCs) in the limbus of the mouse has led to the notion that the limbus is the niche for the stem cells responsible for the long-term renewal of the cornea4; hence, the corneal epithelium is supposedly renewed by cells generated at and migrating from the limbus, in striking opposition to other squamous epithelia in which each resident stem cell has in charge a limited area of epithelium. Here, we show that the corneal epithelium of the mouse can be serially transplanted, is self-maintained and contains oligopotent stem cells with the capacity to generate goblet cells if provided with a conjunctival environment. In addition, the entire ocular surface of the pig, including the cornea, contains oligopotent stem cells (holoclones) with the capacity to generate individual colonies of corneal and conjunctival cells; hence, the limbus is not the only niche for corneal stem cells and corneal renewal is not different from other squamous epithelia. Keywords: Cell type comparison
2008-08-29 | GSE12604 | GEO
Project description:Mucin glycoproteins on the ocular surface are rich in O-glycans and have important roles in the protection from physical, chemical and microbial impact. In this work, we have cultured human corneal and conjunctival epithelial cells to examine the glycosyltransferase activities that synthesize the O-glycans of mucins. The results indicate that ocular surface epithelial cells have active enzymes that synthesize O-glycans with sialylated core 1, Galβ1-3GalNAcα, and core 2, GlcNAcβ1-6(Galβ1-3)GalNAcα structures which corresponds to previous structural studies. Eye cells also have enzymes that synthesize complex N-glycans that are found on mucins. Results from treatment of eye cells with TNFα suggest that epithelial O-glycosylation changes in a dynamic fashion during inflammatory stimuli of the eye surface.
| S-EPMC6311131 | biostudies-literature
Project description:Polydopamine has acquired great relevance in the field of nanomedicine due to its physicochemical properties. Previously, it has been reported that nanoparticles synthetized from this polymer are able to decrease the viability of breast and colon tumor cells. In addition, it is well known that the size of therapeutic particles plays an essential role in their effect. As a consequence, the influence of this parameter on the cytotoxicity of polydopamine nanoparticles was studied in this work. For this purpose, polydopamine nanoparticles with three different diameters (115, 200 and 420 nm) were synthetized and characterized. Their effect on the viability of distinct sorts of human carcinomas (breast, colon, liver and lung) and stromal cells was investigated, as well as the possible mechanisms that could be responsible for such cytotoxicity. Moreover, polydopamine nanoparticles were also loaded with doxorubicin and the therapeutic action of the resulting nanosystem was analyzed. As a result, it was demonstrated that a smaller nanoparticle size is related to a more enhanced antiproliferative activity, which may be a consequence of polydopamine's affinity for iron ions. Smaller nanoparticles would be able to adsorb more lysosomal Fe3+ and, when they are loaded with doxorubicin, a synergistic effect can be achieved.
| S-EPMC6896006 | biostudies-literature
Project description:Corneal wound healing involves communication between the different cell types that constitute the three cellular layers of the cornea (epithelium, stroma and endothelium), a process ensured in part by a category of extracellular vesicles called exosomes. In the present study, we isolated exosomes released by primary cultured human corneal epithelial cells (hCECs), corneal fibroblasts (hCFs) and corneal endothelial cells (hCEnCs) and determined whether they have wound healing characteristics of their own and to which point they modify the genetic and proteomic pattern of these cell types. Exosomes released by all three cell types significantly accelerated wound closure of scratch-wounded hCECs in vitro compared to controls (without exosomes). Profiling of activated kinases revealed that exosomes from human corneal cells caused the activation of signal transduction mediators that belong to the HSP27, STAT, β-catenin, GSK-3β and p38 pathways. Most of all, data from gene profiling analyses indicated that exosomes, irrespective of their cellular origin, alter a restricted subset of genes that are completely different between each targeted cell type (hCECs, hCFS, hCEnCs). Analysis of the genes specifically differentially regulated for a given cell-type in the microarray data using the Ingenuity Pathway Analysis (IPA) software revealed that the mean gene expression profile of hCECs cultured in the presence of exosomes would likely promote cell proliferation and migration whereas it would reduce differentiation when compared to control cells. Collectively, our findings represent a conceptual advance in understanding the mechanisms of corneal wound repair that may ultimately open new avenues for the development of novel therapeutic approaches to improve closure of corneal wounds.
| S-EPMC9602716 | biostudies-literature
Project description:To determine global mRNA expression levels in corneal and conjunctival epithelia and identify transcripts that exhibit preferential tissue expression.cDNA samples derived from human conjunctival and corneal epithelia were hybridized in three independent experiments to a commercial oligonucleotide array representing more than 22,000 transcripts. The resultant signal intensities and microarray software transcript present/absent calls were used in conjunction with the local pooled error (LPE) statistical method to identify transcripts that are preferentially or exclusively expressed in one of the two tissues at significant levels (expression >1% of the beta-actin level). EASE (Expression Analysis Systematic Explorer software) was used to identify biological systems comparatively overrepresented in either epithelium. Immuno-, and cytohistochemistry was performed to validate or expand on selected results of interest.The analysis identified 332 preferential and 93 exclusive significant corneal epithelial transcripts. The corresponding numbers of conjunctival epithelium transcripts were 592 and 211, respectively. The overrepresented biological processes in the cornea were related to cell adhesion and oxiredox equilibria and cytoprotection activities. In the conjunctiva, the biological processes that were most prominent were related to innate immunity and melanogenesis. Immunohistochemistry for antigen-presenting cells and melanocytes was consistent with these gene signatures. The transcript comparison identified a substantial number of genes that have either not been identified previously or are not known to be highly expressed in these two epithelia, including testican-1, ECM1, formin, CRTAC1, and NQO1 in the cornea and, in the conjunctiva, sPLA(2)-IIA, lipocalin 2, IGFBP3, multiple MCH class II proteins, and the Na-Pi cotransporter type IIb.Comparative gene expression profiling leads to the identification of many biological processes and previously unknown genes that are potentially active in the function of corneal and conjunctival epithelia.
| S-EPMC2909883 | biostudies-literature
Project description:Human conjunctival cell lines are useful tools for modeling ocular surface disease and evaluation of ocular drugs and cosmetics. However, gene expression in these cells may not be comparable to primary cultured cells, raising doubts that they could be used as a substitute. We aimed to ascertain the similarities of global gene expression between commonly used cell lines and primary cells using a microarray approach. The Affymetrix U133A chip (>22,000 genes) was used to investigate conjunctival tissue (CT), primary conjunctival epithelial cells (PCEC), two conjunctival epithelial cell lines (IOBA-NHC and ChWK), and HCEC-T, a human corneal epithelial cell line (control). Using principal component analysis, the PCEC profile was clustered more closely to conjunctival tissue than either of the two cell lines. Certain extracellular matrix genes were differentially upregulated in CT compared to PCEC, suggesting presence of fibroblasts in addition to epithelial cells in CT. Overall, 67.3% (95% CI: 66.7-67.9) of transcripts in IOBA-NHC were within 1.5-fold of the corresponding transcripts in PCEC, but only 62.2% (95% CI: 61.5-62.9) in the case of ChWK. In HCEC-T, the proportion was only 58.8% (95% CI 58.1-59.4), suggesting less resemblance to PCEC than the conjunctival epithelial cell lines. The IOBA-NHC profile was more similar to PCEC than ChWK, for all genes and genes concerned with membrane association, communication, development, and regulation of metabolism, especially protein and nucleic acid metabolism. The correlation of normalized gene expression levels was high between either the IOBA-NHC or ChWK and PCEC for genes concerned with cell defense, viral life cycle, antigen presentation, antioxidation, or ubiquitin ligation. In order to evaluate the functional significance of the altered gene expression in IOBA-NHC cells, we evaluated a few proteins important for epithelial differentiation or defense, corresponding to the transcripts for S100A9, TGM2, and TLR4. Protein levels of S100A9 and TGM2 were indeed raised, and TLR4 decreased, in IOBA-NHC compared to PCEC. Gene expression in conjunctival cell lines differs from primary cells, but the profile varies according to functional gene categories. Depending on the methodology of proposed studies, if there is limited availability of PCEC, NHC-IOBA may be more suitable than ChWK, but even then, epithelial differentiation and innate immunity functions in NHC-IOBA may differ from primary cells.
| S-EPMC6042047 | biostudies-literature
Project description:PurposeOcular surface mucins and glycocalyx are critical for providing ocular hydration as well lubrication and repelling pathogens or allergens. Elevated levels of tear proinflammatory cytokines in dry eye may have detrimental effect on mucins and glycocalyx. The present study tested the effect of proinflammatory cytokines IL-6, TNF-α, and IFN-γ on membrane-tethered mucins expression, glycocalyx, and viability of ocular surface epithelial cells.MethodsStratified cultures of human corneal and conjunctival epithelial cells were exposed to different concentrations of IL-6, TNF-α, and IFN-γ for 24 hours. The mucins gene and protein expressions were quantified by real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). The glycocalyx was imaged using confocal microscopy after staining with Alexa 488-conjugated wheat germ agglutinin lectin. Apoptotic and necrotic cell death was quantified using flow cytometry.ResultsIL-6, TNF-α, and IFN-γ treatment resulted in a significant increase in mucins (MUC)1 and MUC4 gene and protein expression in human corneal epithelial cells but caused no significant changes in the levels of these mucins in conjunctival epithelial cells. Further, these cytokines decreased MUC16 expression in both corneal and conjunctival epithelial cells. Moreover, no notable change in glycocalyx or apoptotic cell death in corneal and conjunctival epithelial cells was noted with any of the tested cytokines, but IL-6 and TNF-α exposure increased necrotic cell death in corneal and conjunctival epithelial cells, respectively.ConclusionsOur results demonstrate that proinflammatory cytokines have differential effects on human corneal and conjunctival epithelial cell mucins expression, but do not cause any damage to ocular surface epithelial cell glycocalyx.
| S-EPMC8212448 | biostudies-literature
Project description:Purpose:Report a case of corneal melt in a patient with conjunctival intraepithelial neoplasia (CIN) treated with topical interferon (IFN) alpha-2B. Observations:An 89-year-old man presented with gelatinous paralimbal lesions of the left eye extending onto the cornea with corneal neovascularization extending 5-6 clock hours. Nasally there was mild absence of the terminal vascular loops of the limbal palisades of Vogt and conjunctivalization. Diffuse punctate epithelial erosions were noted. The corneal graft displayed subepithelial and stromal edema. Anterior segment optical coherence tomography detected hyperreflectivity, sectional thickened epithelium, and abrupt transitions from normal to abnormal tissue. The patient was treated with excision of the corneal and conjunctival lesions with cryotherapy to the conjunctival borders. Excisional biopsy revealed CIN Grade 3 and carcinoma in situ of the cornea. Topical IFN alpha-2B four times daily was initiated postoperatively. Two months later, a central epithelial defect developed. The cornea progressively thinned and corneal melt ensued. The patient had several risk factors for corneal melt including neurotrophic cornea, early limbal stem cell deficiency, history of cryotherapy, keratoconjunctivitis sicca, and chronic use of glaucoma medications and steroid medications. Conclusions:Interferon alpha-2B is an effective first line treatment for CIN with few side effects. It's side effects include punctate epithelial erosions, conjunctival hyperemia, and follicular conjunctivitis. We report a case of pre-existing keratoconjunctivitis sicca, early limbal stem cell deficiency, neurotrophic cornea, and newly diagnosed CIN Grade 3; it was treated with surgical excision, cryotherapy, and topical IFN alpha-2b with development of corneal melt 2 months later. Caution should be taken when using interferon alpha -2b in patients with pre-existing keratoconjunctivitis sicca, neurotrophic cornea, or limbal stem cell deficiency as it could exacerbate these conditions resulting in corneal melt.
| S-EPMC7225610 | biostudies-literature
Project description:Purpose:The present study highlighted the value of anterior segment optical coherence tomography (AS-OCT) for different types of corneal foreign bodies in humans. Methods:This study was a prospective observational study. The patients included were divided into two groups. If the patients were directly diagnosed based on eye injury history and slit-lamp examination, then they were assigned to Group A. Otherwise, the patients were assigned to Group B. We compared and described the characteristics of the corneal foreign body in both groups using AS-OCT. Results:From October 2017 to January 2020, 36 eyes of 36 patients (9 females and 27 males) with a mean age of 37.8?±?11.7 years were included in the study. Patients in Group A were the majority and accounted for 72.2% (26/36). High signals on AS-OCT images were the main constituent and accounted for 92.3% (24/26) in Group A and 70.0% (7/10) in Group B. Most of the patients in Group A, 96.2% (25/26), had clear boundaries. A blurred boundary was observed in 70.0% (7/10) of the patients in Group B. The foreign bodies on AS-OCT images had key characteristics of a high signal followed by a central zone shadowing effect and a low signal followed by a marginal zone shadowing effect. Further, all of the lesions could be directly located in Group B, and 92.3% (24/26) of the patients in Group A did not have directly located lesions. Six representative cases are described in detail. Conclusions:AS-OCT is a valuable tool in the diagnosis and management of corneal foreign bodies, especially for unusual corneal foreign body.
| S-EPMC7439195 | biostudies-literature
Project description:Amoebic keratitis causes significant ocular morbidity in contact lens wearers. Current diagnostic methods for amoebic keratitis are insensitive and labor-intensive and have poor turnaround time. We evaluated four laboratory methods for detection of acanthamoebae in clinical specimens. Deidentified, delinked consecutive specimens from patients with suspected amoebic keratitis were assayed for acanthamoebae by direct smear analysis, culture, and PCR using two different primer sets specific for Acanthamoeba ribosomal DNA. The consensus reference standard was considered fulfilled when the results for any two of the four tests were positive, and the outcome measures were sensitivity and specificity. Of 107 specimens assayed over an 18-month period, 20 were positive for acanthamoebae. The sensitivity and specificity of each assay were as follows, respectively: for smear analysis, 55% (95% confidence interval [CI], 33.2 to 76.8%) and 100%; for culture, 73.7% (95% CI, 54.4 to 93.0%) and 100%; for PCR using Nelson primers, 90% (95% CI, 76.9 to 100%) and 90.8% (95% CI, 84.7 to 96.9%); and for PCR using JDP primers, 65% (95% CI, 44.1 to 85.9%) and 100%. Nelson primer PCR demonstrated a single-organism level of analytic sensitivity. The performance characteristics of the assays varied by specimen type, with contact lenses and casings showing the highest rates of detectable acanthamoebae and the highest diagnostic sensitivities for direct smear analysis, culture, and JDP primer PCR, though these results are based on small numbers and should be interpreted cautiously. These findings have important implications for clinicians collecting diagnostic specimens and for diagnostic laboratories, especially in outbreak situations.
| S-EPMC2681827 | biostudies-literature