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[PMID]:28400915
[Au] Autor:Cejka C; Kossl J; Hermankova B; Holan V; Cejkova J
[Ad] Endereço:Institute of Experimental Medicine, Czech Academy of Sciences, 14220 Prague 4, Czech Republic.
[Ti] Título:Molecular Hydrogen Effectively Heals Alkali-Injured Cornea via Suppression of Oxidative Stress.
[So] Source:Oxid Med Cell Longev;2017:8906027, 2017.
[Is] ISSN:1942-0994
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The aim of this study was to examine the effect of molecular hydrogen (H ) on the healing of alkali-injured cornea. The effects of the solution of H in phosphate buffered saline (PBS) or PBS alone topically applied on the alkali-injured rabbit cornea with 0.25 M NaOH were investigated using immunohistochemical and biochemical methods. Central corneal thickness taken as an index of corneal hydration was measured with an ultrasonic pachymeter. Results show that irrigation of the damaged eyes with H solution immediately after the injury and then within next five days renewed corneal transparency lost after the injury and reduced corneal hydration increased after the injury to physiological levels within ten days after the injury. In contrast, in injured corneas treated with PBS, the transparency of damaged corneas remained lost and corneal hydration elevated. Later results-on day 20 after the injury-showed that in alkali-injured corneas treated with H solution the expression of proinflammatory cytokines, peroxynitrite, detected by nitrotyrosine residues (NT), and malondialdehyde (MDA) expressions were very low or absent compared to PBS treated injured corneas, where NT and MDA expressions were present. In conclusion, H solution favorably influenced corneal healing after alkali injury via suppression of oxidative stress.
[Mh] Termos MeSH primário: Lesões da Córnea/etiologia
Hidrogênio/farmacologia
Estresse Oxidativo/efeitos dos fármacos
Hidróxido de Sódio/toxicidade
[Mh] Termos MeSH secundário: Actinas/metabolismo
Animais
Córnea/metabolismo
Córnea/patologia
Lesões da Córnea/metabolismo
Lesões da Córnea/patologia
Citocinas/metabolismo
Modelos Animais de Doenças
Feminino
Expressão Gênica/efeitos dos fármacos
Imuno-Histoquímica
Interleucina-1beta/genética
Interleucina-1beta/metabolismo
Queratina-12/metabolismo
Queratina-3/metabolismo
Malondialdeído/metabolismo
Ácido Peroxinitroso/metabolismo
Coelhos
Fator A de Crescimento do Endotélio Vascular/genética
Fator A de Crescimento do Endotélio Vascular/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Actins); 0 (Cytokines); 0 (Interleukin-1beta); 0 (Keratin-12); 0 (Keratin-3); 0 (Vascular Endothelial Growth Factor A); 14691-52-2 (Peroxynitrous Acid); 4Y8F71G49Q (Malondialdehyde); 55X04QC32I (Sodium Hydroxide); 7YNJ3PO35Z (Hydrogen)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170504
[Lr] Data última revisão:
170504
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170413
[St] Status:MEDLINE
[do] DOI:10.1155/2017/8906027


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[PMID]:27250558
[Au] Autor:Mathan JJ; Ismail S; McGhee JJ; McGhee CN; Sherwin T
[Ad] Endereço:Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, The University of Auckland, Private Bag 92019, Auckland, 1010, New Zealand.
[Ti] Título:Sphere-forming cells from peripheral cornea demonstrate the ability to repopulate the ocular surface.
[So] Source:Stem Cell Res Ther;7(1):81, 2016 Jun 01.
[Is] ISSN:1757-6512
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: The limbus forms the outer rim of the cornea at the corneoscleral junction and harbours a population of stem cells for corneal maintenance. Injuries to the limbus, through disease or accidents such as chemical injuries or burns, may lead to significant visual impairment due to depletion of the native stem cells of the tissue. METHODS: Sphere-forming cells were isolated from peripheral cornea for potential use as transplantable elements for limbal stem cell repopulation and limbal reconstruction. Immunocytochemistry, live cell imaging and quantitative PCR were used to characterize spheres and elucidate activity post implantation into human cadaveric corneal tissue. RESULTS: Spheres stained positively for stem cell markers ∆NP63α, ABCG2 and ABCB5 as well as the basal limbal marker and putative niche marker, notch 1. In addition, spheres also stained positively for markers of corneal cells, vimentin, keratin 3, keratocan and laminin, indicating a heterogeneous mix of stromal and epithelial-origin cells. Upon implantation into decellularized corneoscleral tissue, 3D, polarized and radially orientated cell migration with cell proliferation was observed. Cells migrated out from the spheres and repopulated the entire corneal surface over 14 days. Post-implantation analysis revealed qualitative evidence of stem, stromal and epithelial cell markers while quantitative PCR showed a quantitative reduction in keratocan and laminin expression indicative of an enhanced progenitor cell response. Proliferation, quantified by PCNA expression, significantly increased at 4 days subsequently followed by a decrease at day 7 post implantation. CONCLUSION: These observations suggest great promise for the potential of peripheral corneal spheres as transplantable units for corneal repair, targeting ocular surface regeneration and stem cell repopulation.
[Mh] Termos MeSH primário: Epitélio Anterior/citologia
Limbo da Córnea/citologia
Esferoides Celulares/transplante
Células-Tronco/citologia
[Mh] Termos MeSH secundário: Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética
Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo
Biomarcadores/metabolismo
Cadáver
Diferenciação Celular
Movimento Celular
Proliferação Celular
Epitélio Anterior/metabolismo
Expressão Gênica
Seres Humanos
Queratina-3/genética
Queratina-3/metabolismo
Laminina/genética
Laminina/metabolismo
Limbo da Córnea/metabolismo
Proteínas de Neoplasias/genética
Proteínas de Neoplasias/metabolismo
Antígeno Nuclear de Célula em Proliferação/genética
Antígeno Nuclear de Célula em Proliferação/metabolismo
Proteoglicanas/genética
Proteoglicanas/metabolismo
Receptor Notch1/genética
Receptor Notch1/metabolismo
Esferoides Celulares/citologia
Esferoides Celulares/metabolismo
Transplante de Células-Tronco
Células-Tronco/metabolismo
Técnicas de Cultura de Tecidos
Fatores de Transcrição/genética
Fatores de Transcrição/metabolismo
Proteínas Supressoras de Tumor/genética
Proteínas Supressoras de Tumor/metabolismo
Vimentina/genética
Vimentina/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (ABCB5 protein, human); 0 (ABCG2 protein, human); 0 (ATP Binding Cassette Transporter, Sub-Family G, Member 2); 0 (ATP-Binding Cassette, Sub-Family B, Member 1); 0 (Biomarkers); 0 (KERA protein, human); 0 (KRT3 protein, human); 0 (Keratin-3); 0 (Laminin); 0 (NOTCH1 protein, human); 0 (Neoplasm Proteins); 0 (Proliferating Cell Nuclear Antigen); 0 (Proteoglycans); 0 (Receptor, Notch1); 0 (TP63 protein, human); 0 (Transcription Factors); 0 (Tumor Suppressor Proteins); 0 (Vimentin)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160603
[St] Status:MEDLINE
[do] DOI:10.1186/s13287-016-0339-7


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[PMID]:26937166
[Au] Autor:Selver OB; Durak I; Gürdal M; Baysal K; Ates H; Ozbek Z; Wang Z; Wu A; Wolosin JM
[Ad] Endereço:Department of Ophtalmology, Dokuz Eylul University School of Medicine, Izmir, Turkey.
[Ti] Título:Corneal recovery in a rabbit limbal stem cell deficiency model by autologous grafts of tertiary outgrowths from cultivated limbal biopsy explants.
[So] Source:Mol Vis;22:138-49, 2016.
[Is] ISSN:1090-0535
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:PURPOSE: To determine the corneal regenerative capacity of sequentially generated primary, secondary, and tertiary limbal explant outgrowths in a limbal stem cell deficiency (LSCD) surgical model. METHODS: Two-millimeter-long limbal shallow biopsies were surgically excised from the upper quadrant of the right eye of rabbits and set on preserved amniotic membrane for explant culture. After the generation of primary outgrowth, the biopsies were sequentially transferred to new amniotic membrane to generate secondary and then tertiary outgrowths. Eighteen rabbits were subjected to a 360° limbal peritomy extending into the scleral zone and combined with superficial keratectomy of the corneal periphery and thorough mechanical debridement of the central cornea in their left eye. Right eye outgrowths, six of each generation, were engrafted on the ocular surface. Clinical outcomes (neovascularization, corneal clarity, and corneal fluorescein staining) were graded after 6 months. Post-mortem corneas were compared with histology, immunochemistry for p63 and Krt3, ABCG2-dependent dye exclusion, and capacity for outgrowths in explant culture. RESULTS: Immunohistology and western blot of the outgrowths for p63 and Krt3 indicated no differences in expression between the primary and tertiary outgrowths for these two markers of growth and differentiation. Clinically, all rabbits treated with amniotic membrane alone developed severe LSCD. Most rabbits grafted with cell outgrowths from all three outgrowth generations achieved stable (>6 months) recovery of the ocular surface. There were partial failures of grafts performed with two secondary and tertiary outgrowths. However, Kruskal-Wallis statistical analysis of the clinical scores yielded no significant difference between the three groups (p=0.524). Histology showed full anatomic recovery of grafts made with primary and tertiary outgrowths. Krt3 and p63 expression throughout the whole limbal corneal epithelium with primary or tertiary outgrowths was not distinguishable from each other. The percentage of dye-excluding cells present within this zone and the capacity of the explant epithelial outgrowth of the regenerated peripheral corneal zone were also on par with those of the donor corneas. The Krt3-negative cells that characterize the basal epithelial layer of the normal limbus could not be found in any regenerated cornea from the primary to tertiary outgrowths. CONCLUSIONS: Our results demonstrate that in rabbits post-primary explant outgrowths retain the capacity for LSCD recovery found in primary explants.
[Mh] Termos MeSH primário: Córnea/fisiologia
Doenças da Córnea/terapia
Modelos Animais de Doenças
Epitélio Anterior/citologia
Limbo da Córnea/patologia
Transplante de Células-Tronco
Células-Tronco/patologia
[Mh] Termos MeSH secundário: Âmnio
Animais
Biópsia
Western Blotting
Técnicas de Cultura de Células
Doenças da Córnea/fisiopatologia
Epitélio Anterior/metabolismo
Citometria de Fluxo
Seres Humanos
Imuno-Histoquímica
Queratina-3/metabolismo
Coelhos
Recuperação de Função Fisiológica/fisiologia
Regeneração/fisiologia
Tecidos Suporte
Fatores de Transcrição/metabolismo
Transplante Autólogo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Keratin-3); 0 (Transcription Factors)
[Em] Mês de entrada:1610
[Cu] Atualização por classe:161230
[Lr] Data última revisão:
161230
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160304
[St] Status:MEDLINE


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[PMID]:26908015
[Au] Autor:Hazra S; Nandi S; Naskar D; Guha R; Chowdhury S; Pradhan N; Kundu SC; Konar A
[Ad] Endereço:Department of Veterinary Surgery &Radiology, West Bengal University of Animal &Fishery Sciences, Kolkata-700037, West Bengal, India.
[Ti] Título:Non-mulberry Silk Fibroin Biomaterial for Corneal Regeneration.
[So] Source:Sci Rep;6:21840, 2016 Feb 24.
[Is] ISSN:2045-2322
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:PURPOSE: Successful repair of a damaged corneal surface is a great challenge and may require the use of a scaffold that supports cell growth and differentiation. Amniotic membrane is currently used for this purpose, in spite of its limitations. A thin transparent silk fibroin film from non-mulberry Antheraea mylitta (Am) has been developed which offers to be a promising alternative. The silk scaffolds provide sufficient rigidity for easy handling, the scaffolds support the sprouting, migration, attachment and growth of epithelial cells and keratocytes from rat corneal explants; the cells form a cell sheet, preserve their phenotypes, express cytokeratin3 and vimentin respectively. The films also support growth of limbal stem cell evidenced by expression of ABCG2. The cell growth on the silk film and the amniotic membrane is comparable. The implanted film within the rabbit cornea remains transparent, stable. The clinical examination as well as histology shows absence of any inflammatory response or neovascularization. The corneal surface integrity is maintained; tear formation, intraocular pressure and electroretinography of implanted eyes show no adverse changes. The silk fibroin film from non-mulberry silk worms may be a worthy candidate for use as a corneal scaffold.
[Mh] Termos MeSH primário: Materiais Biocompatíveis/farmacologia
Córnea/fisiologia
Fibroínas/farmacologia
Regeneração/efeitos dos fármacos
[Mh] Termos MeSH secundário: Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo
Âmnio/transplante
Animais
Materiais Biocompatíveis/química
Adesão Celular/efeitos dos fármacos
Movimento Celular/efeitos dos fármacos
Proliferação Celular/efeitos dos fármacos
Células Cultivadas
Córnea/patologia
Córnea/ultraestrutura
Ceratócitos da Córnea/citologia
Ceratócitos da Córnea/efeitos dos fármacos
Ceratócitos da Córnea/metabolismo
Eletrorretinografia
Fibroínas/química
Pressão Intraocular/fisiologia
Queratina-3/metabolismo
Microscopia Eletrônica de Varredura
Microscopia de Fluorescência
Mariposas/metabolismo
Coelhos
Ratos
Ratos Sprague-Dawley
Refratometria
Vimentina/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (ATP Binding Cassette Transporter, Sub-Family G, Member 2); 0 (Biocompatible Materials); 0 (Keratin-3); 0 (Vimentin); 9007-76-5 (Fibroins)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:170220
[Lr] Data última revisão:
170220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160225
[St] Status:MEDLINE
[do] DOI:10.1038/srep21840


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[PMID]:26899008
[Au] Autor:Sasamoto Y; Hayashi R; Park SJ; Saito-Adachi M; Suzuki Y; Kawasaki S; Quantock AJ; Nakai K; Tsujikawa M; Nishida K
[Ad] Endereço:Department of Ophthalmology, Osaka University Medical School, Suita, Osaka 565-0871, Japan.
[Ti] Título:PAX6 Isoforms, along with Reprogramming Factors, Differentially Regulate the Induction of Cornea-specific Genes.
[So] Source:Sci Rep;6:20807, 2016 Feb 22.
[Is] ISSN:2045-2322
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:PAX6 is the key transcription factor involved in eye development in humans, but the differential functions of the two PAX6 isoforms, isoform-a and isoform-b, are largely unknown. To reveal their function in the corneal epithelium, PAX6 isoforms, along with reprogramming factors, were transduced into human non-ocular epithelial cells. Herein, we show that the two PAX6 isoforms differentially and cooperatively regulate the expression of genes specific to the structure and functions of the corneal epithelium, particularly keratin 3 (KRT3) and keratin 12 (KRT12). PAX6 isoform-a induced KRT3 expression by targeting its upstream region. KLF4 enhanced this induction. A combination of PAX6 isoform-b, KLF4, and OCT4 induced KRT12 expression. These new findings will contribute to furthering the understanding of the molecular basis of the corneal epithelium specific phenotype.
[Mh] Termos MeSH primário: Olho/crescimento & desenvolvimento
Queratina-12/biossíntese
Queratina-3/biossíntese
Fatores de Transcrição Kruppel-Like/biossíntese
Fator 3 de Transcrição de Octâmero/biossíntese
Fator de Transcrição PAX6/genética
[Mh] Termos MeSH secundário: Linhagem Celular
Células Epiteliais/metabolismo
Epitélio Anterior/crescimento & desenvolvimento
Epitélio Anterior/metabolismo
Olho/metabolismo
Regulação da Expressão Gênica no Desenvolvimento
Seres Humanos
Queratina-12/genética
Queratina-3/genética
Fatores de Transcrição Kruppel-Like/genética
Fator 3 de Transcrição de Octâmero/genética
Fator de Transcrição PAX6/biossíntese
Isoformas de Proteínas/biossíntese
Isoformas de Proteínas/genética
Transdução Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (GKLF protein); 0 (KRT12 protein, human); 0 (KRT3 protein, human); 0 (Keratin-12); 0 (Keratin-3); 0 (Kruppel-Like Transcription Factors); 0 (Octamer Transcription Factor-3); 0 (PAX6 Transcription Factor); 0 (PAX6 protein, human); 0 (POU5F1 protein, human); 0 (Protein Isoforms)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:170922
[Lr] Data última revisão:
170922
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160223
[St] Status:MEDLINE
[do] DOI:10.1038/srep20807


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[PMID]:26758872
[Au] Autor:Allen EH; Courtney DG; Atkinson SD; Moore JE; Mairs L; Poulsen ET; Schiroli D; Maurizi E; Cole C; Hickerson RP; James J; Murgatroyd H; Smith FJ; MacEwen C; Enghild JJ; Nesbit MA; Leslie Pedrioli DM; McLean WH; Moore CB
[Ad] Endereço:School of Biomedical Sciences, University of Ulster, Coleraine BT52 1SA, Northern Ireland, UK, Centre for Dermatology and Genetic Medicine, Division of Biological Chemistry and Drug Discovery, School of Life Sciences, University of Dundee, Scotland DD1 5EH, UK.
[Ti] Título:Keratin 12 missense mutation induces the unfolded protein response and apoptosis in Meesmann epithelial corneal dystrophy.
[So] Source:Hum Mol Genet;25(6):1176-91, 2016 Mar 15.
[Is] ISSN:1460-2083
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Meesmann epithelial corneal dystrophy (MECD) is a rare autosomal dominant disorder caused by dominant-negative mutations within the KRT3 or KRT12 genes, which encode the cytoskeletal protein keratins K3 and K12, respectively. To investigate the pathomechanism of this disease, we generated and phenotypically characterized a novel knock-in humanized mouse model carrying the severe, MECD-associated, K12-Leu132Pro mutation. Although no overt changes in corneal opacity were detected by slit-lamp examination, the corneas of homozygous mutant mice exhibited histological and ultrastructural epithelial cell fragility phenotypes. An altered keratin expression profile was observed in the cornea of mutant mice, confirmed by western blot, RNA-seq and quantitative real-time polymerase chain reaction. Mass spectrometry (MS) and immunohistochemistry demonstrated a similarly altered keratin profile in corneal tissue from a K12-Leu132Pro MECD patient. The K12-Leu132Pro mutation results in cytoplasmic keratin aggregates. RNA-seq analysis revealed increased chaperone gene expression, and apoptotic unfolded protein response (UPR) markers, CHOP and Caspase 12, were also increased in the MECD mice. Corneal epithelial cell apoptosis was increased 17-fold in the mutant cornea, compared with the wild-type (P < 0.001). This elevation of UPR marker expression was also observed in the human MECD cornea. This is the first reporting of a mouse model for MECD that recapitulates the human disease and is a valuable resource in understanding the pathomechanism of the disease. Although the most severe phenotype is observed in the homozygous mice, this model will still provide a test-bed for therapies not only for corneal dystrophies but also for other keratinopathies caused by similar mutations.
[Mh] Termos MeSH primário: Distrofia Corneana Epitelial Juvenil de Meesmann/genética
Queratina-12/genética
Queratina-3/genética
Mutação de Sentido Incorreto
[Mh] Termos MeSH secundário: Adulto
Animais
Apoptose/genética
Modelos Animais de Doenças
Éxons
Feminino
Heterozigoto
Seres Humanos
Camundongos
Camundongos Transgênicos
Mutação
Linhagem
Resposta a Proteínas não Dobradas
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (KRT12 protein, human); 0 (KRT3 protein, human); 0 (Keratin-12); 0 (Keratin-3)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:170922
[Lr] Data última revisão:
170922
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160114
[St] Status:MEDLINE
[do] DOI:10.1093/hmg/ddw001


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[PMID]:25897888
[Au] Autor:Baradaran-Rafii A; Biazar E; Heidari-keshel S
[Ad] Endereço:a Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences , Tehran , Iran .
[Ti] Título:Cellular Response of Limbal Stem Cells on Polycaprolactone Nanofibrous Scaffolds for Ocular Epithelial Regeneration.
[So] Source:Curr Eye Res;41(3):326-33, 2016.
[Is] ISSN:1460-2202
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:PURPOSE: The aim of this study was to develop nanofibrous polycaprolactone (PCL) substrate for limbal stem cell (LSC) expansion that can serve as a potential alternative substrate to replace human amniotic membrane (AM). MATERIALS AND METHODS: The human limbus stem cell was used to evaluate the biocompatibility of substrates (nanofibrous scaffold and, human AM) based on their phenotypic profile, viability, proliferation and attachment ability. RESULTS: Biocompatibility results indicated that the all substrates were highly biocompatible, as LSCs could favorably attach and proliferate on the nanofibrous surface. Microscopic figures showed that the human LSCs were firmly anchored to the substrates and were able to retain a normal corneal stem cell phenotype. Microscopic analyses illustrated that cells infiltrated the nanofibers and successfully formed a three-dimensional corneal epithelium, which was viable for two weeks. Immunocytochemistry (ICC) and real time-PCR results revealed no change in the expression profile of LECs grown on nanofibrous substrate when compared to those grown on human AM. CONCLUSION: In addition, electrospun nanofibrous PCL substrate provides not only a milieu supporting LSCs expansion, but also serve as a useful alternative carrier for ocular surface tissue engineering and could be used as an alternative substrate to AM.
[Mh] Termos MeSH primário: Epitélio Anterior/fisiologia
Limbo da Córnea/citologia
Poliésteres
Regeneração/fisiologia
Células-Tronco/citologia
Tecidos Suporte
[Mh] Termos MeSH secundário: Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética
Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo
Materiais Biocompatíveis
Biomarcadores/metabolismo
Adesão Celular/fisiologia
Proliferação Celular/fisiologia
Sobrevivência Celular/fisiologia
Citometria de Fluxo
Expressão Gênica
Seres Humanos
Queratina-12/genética
Queratina-12/metabolismo
Queratina-3/genética
Queratina-3/metabolismo
Limbo da Córnea/metabolismo
Microscopia Eletrônica de Varredura
Nanofibras
Proteínas de Neoplasias/genética
Proteínas de Neoplasias/metabolismo
Reação em Cadeia da Polimerase em Tempo Real
Células-Tronco/metabolismo
Fatores de Transcrição/genética
Fatores de Transcrição/metabolismo
Proteínas Supressoras de Tumor/genética
Proteínas Supressoras de Tumor/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (ABCG2 protein, human); 0 (ATP Binding Cassette Transporter, Sub-Family G, Member 2); 0 (Biocompatible Materials); 0 (Biomarkers); 0 (KRT12 protein, human); 0 (KRT3 protein, human); 0 (Keratin-12); 0 (Keratin-3); 0 (Neoplasm Proteins); 0 (Polyesters); 0 (TP63 protein, human); 0 (Transcription Factors); 0 (Tumor Suppressor Proteins); 24980-41-4 (polycaprolactone)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:161230
[Lr] Data última revisão:
161230
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150422
[St] Status:MEDLINE
[do] DOI:10.3109/02713683.2015.1019004


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[PMID]:25803495
[Au] Autor:López-García JS; García-Lozano I; Rivas L; Ramírez N; Méndez MT; Raposo R
[Ad] Endereço:a Ophthalmology Service, Hospital Cruz Roja , Madrid , Spain .
[Ti] Título:Stability of Growth Factors in Autologous Serum Eyedrops After Long-Term Storage.
[So] Source:Curr Eye Res;41(3):292-8, 2016.
[Is] ISSN:1460-2202
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:PURPOSE: The purpose of this study is to assess the stability of the growth factors (GF) in autologous serum eyedrops under different storage conditions. METHODS: The concentration of epidermal growth factor (EGF), transforming growth factor-ß (TGF-ß1), platelet-derived growth factor AB (PDGF-AB), and albumin was measured in fresh and defrosted samples of autologous serum under different storage conditions. The fresh and defrosted samples were cooled at 4 °C, and they were studied immediately after preparation, or after defrosting, and after 1, 2, 3, and 4 weeks. The concentration of GF was also assessed after 1, 3, 6, and 9 months at -20 °C. We also investigated how the different storage conditions influence the biological effects of autologous serum on conjunctival and corneal cell cultures. RESULTS: The concentration of EGF, TGF-ß1, PDGF-AB, and albumin remained stable over the 4 weeks at 4 °C, both in fresh and in defrosted samples. Likewise, no statistically significant differences were found between the GF concentration in fresh samples and after 1, 3, 6, and 9 months of freezing at -20 °C. Moreover, no differences were found on the cell proliferation and differentiation between cultured cells with fresh or defrosted samples after 4 weeks at 4 °C or after 1, 3, 6, or 9 months at -20 °C. CONCLUSIONS: Long-term storage of autologous serum eyedrops at -20 °C does not affect the concentration of GF, simplifies clinical logistics, and reduces the frequency of blood extractions from the patients.
[Mh] Termos MeSH primário: Albuminas/metabolismo
Fator de Crescimento Epidérmico/sangue
Soluções Oftálmicas/química
Fator de Crescimento Derivado de Plaquetas/metabolismo
Soro/química
Fator de Crescimento Transformador beta1/sangue
[Mh] Termos MeSH secundário: Adulto
Diferenciação Celular/fisiologia
Proliferação Celular/fisiologia
Células Cultivadas
Túnica Conjuntiva/citologia
Túnica Conjuntiva/efeitos dos fármacos
Túnica Conjuntiva/metabolismo
Criopreservação
Estabilidade de Medicamentos
Armazenamento de Medicamentos
Ensaio de Imunoadsorção Enzimática
Voluntários Saudáveis
Seres Humanos
Queratina-19/metabolismo
Queratina-3/metabolismo
Limbo da Córnea/citologia
Limbo da Córnea/efeitos dos fármacos
Limbo da Córnea/metabolismo
Meia-Idade
Soluções Oftálmicas/farmacologia
Soro/fisiologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Albumins); 0 (Keratin-19); 0 (Keratin-3); 0 (Ophthalmic Solutions); 0 (Platelet-Derived Growth Factor); 0 (Transforming Growth Factor beta1); 0 (platelet-derived growth factor AB); 62229-50-9 (Epidermal Growth Factor)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:161230
[Lr] Data última revisão:
161230
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150325
[St] Status:MEDLINE
[do] DOI:10.3109/02713683.2015.1016180


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[PMID]:26788030
[Au] Autor:Chen JL; Lin BR; Gee KM; Gee JA; Chung DW; Frausto RF; Deng SX; Aldave AJ
[Ad] Endereço:Stein Eye Institute, David Geffen School of Medicine at UCLA, Los Angeles, CA.
[Ti] Título:Identification of presumed pathogenic KRT3 and KRT12 gene mutations associated with Meesmann corneal dystrophy.
[So] Source:Mol Vis;21:1378-86, 2015.
[Is] ISSN:1090-0535
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:PURPOSE: To report potentially pathogenic mutations in the keratin 3 (KRT3) and keratin 12 (KRT12) genes in two individuals with clinically diagnosed Meesmann corneal dystrophy (MECD). METHODS: Slit-lamp examination was performed on the probands and available family members to identify characteristic features of MECD. After informed consent was obtained, saliva samples were obtained as a source of genomic DNA, and screening of KRT3 and KRT12 was performed. Potentially pathogenic variants were screened for in 200 control chromosomes. PolyPhen-2, SIFT, and PANTHER were used to predict the functional impact of identified variants. Short tandem repeat genotyping was performed to confirm paternity. RESULTS: Slit-lamp examination of the first proband demonstrated bilateral, diffusely distributed, clear epithelial microcysts, consistent with MECD. Screening of KRT3 revealed a heterozygous missense variant in exon 1, c.250C>T (p.(Arg84Trp)), which has a minor allele frequency of 0.0076 and was not identified in 200 control chromosomes. In silico analysis with PolyPhen-2 and PANTHER predicted the variant to be damaging to protein function; however, SIFT analysis predicted tolerance of the variant. The second proband demonstrated bilateral, diffusely distributed epithelial opacities that appeared gray-white on direct illumination and translucent on retroillumination. Neither parent demonstrated corneal opacities. Screening of KRT12 revealed a novel heterozygous insertion/deletion variant in exon 6, c.1288_1293delinsAGCCCT (p.(Arg430_Arg431delinsSerPro)). This variant was not present in either of the proband's parents or in 200 control chromosomes and was predicted to be damaging by PolyPhen-2, PANTHER, and SIFT. Haplotype analysis confirmed paternity of the second proband, indicating that the variant arose de novo. CONCLUSIONS: We present a novel KRT12 mutation, representing the first de novo mutation and the first indel in KRT12 associated with MECD. In addition, we report a variant of uncertain significance in KRT3 in an individual with MECD. Although the potential pathogenicity of this variant is unknown, it is the first variant affecting the head domain of K3 to be reported in an individual with MECD and suggests that disease-causing variants associated with MECD may not be restricted to primary sequence alterations of either the helix-initiation or helix-termination motifs of K3 and K12.
[Mh] Termos MeSH primário: Distrofia Corneana Epitelial Juvenil de Meesmann/genética
Queratina-12/genética
Queratina-3/genética
Mutação
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Substituição de Aminoácidos
Sequência de Bases
Criança
Distrofia Corneana Epitelial Juvenil de Meesmann/patologia
Análise Mutacional de DNA
Feminino
Heterozigoto
Seres Humanos
Mutação INDEL
Queratina-12/química
Queratina-3/química
Masculino
Meia-Idade
Mutação de Sentido Incorreto
Linhagem
Polimorfismo de Nucleotídeo Único
[Pt] Tipo de publicação:CASE REPORTS; JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (KRT12 protein, human); 0 (KRT3 protein, human); 0 (Keratin-12); 0 (Keratin-3)
[Em] Mês de entrada:1609
[Cu] Atualização por classe:161019
[Lr] Data última revisão:
161019
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160121
[St] Status:MEDLINE


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[PMID]:26297801
[Au] Autor:Grieve K; Ghoubay D; Georgeon C; Thouvenin O; Bouheraoua N; Paques M; Borderie VM
[Ad] Endereço:Quinze Vingts National Ophthalmology Hospital, 28 rue de Charonne, 75012 Paris, France; Vision Institute, UPMC Université Paris 06, UMR_S 968/INSERM, U968/CHNO des XV-XX/CNRS, UMR_7210, France. Electronic address: kategrieve@gmail.com.
[Ti] Título:Three-dimensional structure of the mammalian limbal stem cell niche.
[So] Source:Exp Eye Res;140:75-84, 2015 Nov.
[Is] ISSN:1096-0007
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Although the existence of the limbal stem cell (LSC) niche is accepted, precise knowledge of its three-dimensional (3D) architecture remains incomplete. The LSC niche was explored on freshly excised and organ-cultured corneoscleral rims from human donors (n = 47), pigs (n = 15) and mice (n = 27) with full-field optical coherence microscopy (FFOCM). Limbal crypt features were detected in 90% of organ-cultured human corneoscleral rims, extending between the palisades of Vogt as radially oriented rectangular (74% of eyes) and/or rounded (23% of eyes) forms, often branching off to, or becoming interconnected by, sub-scleral radially or circumferentially oriented crypts (in 56% of eyes). Mean crypt volume represented 16% of sampled limbal volume on the vertical axis and 8% on the horizontal axis. In pigs, palisades were finer and crypts wider with relatively uniform distribution around the eye, and radial orientation, connecting to numerous narrow criss-crossing invaginations beneath the scleral surface. In mice, only a circumferential limbal trough was detected. Mean crypt volume represented 13% of sampled limbal volume in humans and 9% in pigs. FFOCM combined with fluorescence, and confocal fluorescence microscopy, showed presence of p63-α+ cells and cytokeratin-3+ cells in the limbal crypts. To assess colony forming efficiency (CFE), limbal epithelial cells were cultured at low density with mitomycin-arrested 3T3 feeders. CFE increased with limbal crypt volume and was not significantly decreased in organ-cultured cornea, despite degradation of the epithelial roof, suggesting that stem cells remain protected at the base of crypts during organ culture. CFE in human samples was significantly greater than in pig, and CFE in mouse was zero. Crypt architecture in the three species appears associated with eye exposure to light. LSC density increased with percentage limbal volume occupied by crypts.
[Mh] Termos MeSH primário: Epitélio Anterior/citologia
Limbo da Córnea/citologia
Nicho de Células-Tronco/fisiologia
Células-Tronco/citologia
[Mh] Termos MeSH secundário: Adulto
Idoso
Idoso de 80 Anos ou mais
Animais
Biomarcadores/metabolismo
Contagem de Células
Epitélio Anterior/metabolismo
Feminino
Seres Humanos
Imagem Tridimensional
Queratina-3/metabolismo
Limbo da Córnea/metabolismo
Masculino
Camundongos
Camundongos Endogâmicos C57BL
Microscopia Confocal
Meia-Idade
Técnicas de Cultura de Órgãos
Células-Tronco/metabolismo
Suínos
Tomografia de Coerência Óptica
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Biomarkers); 0 (Keratin-3)
[Em] Mês de entrada:1602
[Cu] Atualização por classe:170801
[Lr] Data última revisão:
170801
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150823
[St] Status:MEDLINE



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