An advanced in vitro model to assess glaucoma onset

S.C. Sacc; S. Tirendi; S. Scarji; M. Fassalacqua; F. Oddone; C.K. Traverso; S. Vernazza; A.M. Bassi

doi:10.14573/altex.1909262

An advanced in vitro model to assess glaucoma onset

S.C. Sacc, S. Tirendi, S. Scarji, M. Fassalacqua, F. Oddone, C.K. Traverso, S. Vernazza, A.M. Bassi

Fondazione G.B. Bietti

Research output: Contribution to journal › Article › peer-review

Abstract

Glaucoma is the second leading cause of blindness worldwide. Currently, glaucoma treatments aim to lower intraocular pressure by decreasing aqueous humor production or increasing aqueous humor outflow through pharmacological approaches or trabeculectomy. The lack of an effective cure requires new therapeutic strategies. We compared the biological responses of a three-dimensional trabecular meshwork model with or without perfusion bioreactor technology to better understand the early molecular changes induced by prolonged oxidative stress conditions induced by repeated daily peroxide exposure. We used standard 3D cultures of trabecular meshwork cells in Matrigel cultured under either static and dynamic conditions for one week. We studied changes in F-actin expression and organization in the cells, cellular metabolic activity, proinflammatory gene expression, expression of pro- and anti-apoptotic proteins, PARP-1 cleavage, and NFKB activation in the model. We demonstrate that the dynamic conditions improve the adaptive behavior of 3D trabecular meshwork cultures to chronic oxidative stress via offsetting pathway activation. © 2020 ALTEX Edition. All rights reserved.

Original language	English
Pages (from-to)	265-274
Number of pages	10
Journal	Altex
Volume	37
Issue number	2
DOIs	https://doi.org/10.14573/altex.1909262
Publication status	Published - 2020

Keywords

caspase 3
collagen type 1
collagen type 1 alpha 1
complementary DNA
cytochrome c
gelatinase B
immunoglobulin enhancer binding protein
interleukin 1alpha
interleukin 1beta
interleukin 6
interstitial collagenase
matrigel
nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase 1
peroxide
protein Bax
protein bcl 2
protein p53
second mitochondrial activator of caspase
stromelysin
synaptotagmin I
transforming growth factor beta2
tumor necrosis factor
ubiquitin C
unclassified drug
ubiquitin
antiapoptotic activity
apoptosis assay
Article
chronic stress
gene expression
glaucoma
human
human cell
immunoreactivity
in vitro study
metabolic activity assay
oxidative stress
polyacrylamide gel electrophoresis
protein expression
real time polymerase chain reaction
resazurin assay
RNA extraction
three dimensional cell culture
trabecular meshwork
Western blotting
adaptive behavior
cell culture technique
cell growth
confocal microscopy
controlled study
cytokine production
down regulation
flow cytometry
genetic transfection
signal transduction
upregulation

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10.14573/altex.1909262

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85082979653&doi=10.14573%2faltex.1909262&partnerID=40&md5=cb93fb65d3ce90b9f4d8b986e1feaf32

Cite this

@article{3fd336e992304c0f853dd55b6f292e01,

title = "An advanced in vitro model to assess glaucoma onset",

abstract = "Glaucoma is the second leading cause of blindness worldwide. Currently, glaucoma treatments aim to lower intraocular pressure by decreasing aqueous humor production or increasing aqueous humor outflow through pharmacological approaches or trabeculectomy. The lack of an effective cure requires new therapeutic strategies. We compared the biological responses of a three-dimensional trabecular meshwork model with or without perfusion bioreactor technology to better understand the early molecular changes induced by prolonged oxidative stress conditions induced by repeated daily peroxide exposure. We used standard 3D cultures of trabecular meshwork cells in Matrigel cultured under either static and dynamic conditions for one week. We studied changes in F-actin expression and organization in the cells, cellular metabolic activity, proinflammatory gene expression, expression of pro- and anti-apoptotic proteins, PARP-1 cleavage, and NFKB activation in the model. We demonstrate that the dynamic conditions improve the adaptive behavior of 3D trabecular meshwork cultures to chronic oxidative stress via offsetting pathway activation. {\textcopyright} 2020 ALTEX Edition. All rights reserved.",

keywords = "caspase 3, collagen type 1, collagen type 1 alpha 1, complementary DNA, cytochrome c, gelatinase B, immunoglobulin enhancer binding protein, interleukin 1alpha, interleukin 1beta, interleukin 6, interstitial collagenase, matrigel, nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase 1, peroxide, protein Bax, protein bcl 2, protein p53, second mitochondrial activator of caspase, stromelysin, synaptotagmin I, transforming growth factor beta2, tumor necrosis factor, ubiquitin C, unclassified drug, ubiquitin, antiapoptotic activity, apoptosis assay, Article, chronic stress, gene expression, glaucoma, human, human cell, immunoreactivity, in vitro study, metabolic activity assay, oxidative stress, polyacrylamide gel electrophoresis, protein expression, real time polymerase chain reaction, resazurin assay, RNA extraction, three dimensional cell culture, trabecular meshwork, Western blotting, adaptive behavior, cell culture technique, cell growth, confocal microscopy, controlled study, cytokine production, down regulation, flow cytometry, genetic transfection, signal transduction, upregulation",

author = "S.C. Sacc and S. Tirendi and S. Scarji and M. Fassalacqua and F. Oddone and C.K. Traverso and S. Vernazza and A.M. Bassi",

note = "Cited By :3 Export Date: 2 February 2021 CODEN: ALTEE Correspondence Address: Vernazza, S.; Department of Experimental Medicine, Italy Chemicals/CAS: caspase 3, 169592-56-7; cytochrome c, 9007-43-6, 9064-84-0; gelatinase B, 146480-36-6; interstitial collagenase, 9001-12-1; matrigel, 119978-18-6; peroxide, 14915-07-2; protein bcl 2, 219306-68-0; stromelysin, 79955-99-0; ubiquitin C, 151821-62-4; ubiquitin, 60267-61-0 References: Aarskog, N., Vedeler, C., Real-time quantitative polymerase chain reaction (2000) Hum Genet, 107, pp. 494-498; Acott, T. S., Kelley, M., Extracellular matrix in the trabecular meshwork (2008) Eye Res, 86, pp. 543-561. , 3; Ahluwalia, A., Allometric scaling in-vitro (2017) Sci Rep, 7, p. 42113; Benton, G., Arnaoutova, I., George, J., Maui- gel: From discovery and ECM mimicry to assays and mod- els for cancer research (2014) Adv Drug Del/v Rev, 79-80, pp. 3-18; Berger, E., Paczia, N., Millifluidic culture improves human midbrain organoid vitality and differentiation (2018) Lab Chip, 18, pp. 3172-3183. , Magliaro. C; Boardman., K. C., Aryal, A. M., Miller., W. M., Actin re-distribution in response to hydrogen peroxide in airway epithelial cells (2004) JCell .Physiol, 199 (5), pp. 7-66; Roubeix, C., Kessal, K., Effect of benzalkonium chloride on trabecular meshwork cells in a new in vitro 3D trabecular meshwork model for glaucoma (2017) Toxicol In Vitro, 41, pp. 21-29. , Bouchemi. M; Bouhenni, P.. A., Dunmire, 3., Sewell, A., Animal models of glaucoma (2012) J Biomed Biotechnol, 2012, p. 692609; Brancato, V., Gioiella, F., 3D breast cancer microtissue reveals the role of tumor microenvironment on the transport and efficacy of free-doxorubicin in vitro (2018) Acta Biomater, 75, pp. 200-212. , Imparato. 0; Burgoyne, C. F., The non-human primate experimental glaucoma model (2015) Eye Res, 141, pp. 57-73; Elmore, S., Apoptosis: Areview of programmed cell death (2007) ToxicolPathol35, 495-5 16; de-la-Torre, A., Experimental models of glaucoma: A powerful translational tool for the future development of new therapies for glaucoma in humans 1/2A review of the literature (2019) Med/dna (Ma), 55, p. 280. , Evangelho. K., Mastronardi. C. A. and; Fuchshofer, R., Modulation of extracellular matrix turnover in the trabecular meshwork (2009) Erp Eye Res, 88, pp. 683-688. , and Tamm. E. R; Giusti, S., La, Marca. M., A novel dual-flow bioreactor simulates increased fluorescein permeability in epithelial tissue barriers (2014) Biotechnol J, 9, pp. 1175-1184. , Sbrana. T; Gonzalez, J. M., Hamm-Alvarez, . S., Tan, 3. C. H., Analyzing live cellularity in the human trabecular meshwork (2013) Invest Opthalmol fir SeE, 54, pp. 1039-1047; Hughes, C. S., Postovit, L. M., Lajoie, G. A., Math- gel: A complex protein mixture required for optimal growth of cell culture (2010) Proteomics, 10, pp. 1886-1890; Ishikawa, M., Yoshitomi, T., Zorumski, C. F., Experimentally induced mammalian models of glaucoma (2015) Biomed ResInt2OlS, p. 281214. , lO.1155/2015/281214; Izzotti, A., Longobardi, S. C.., Sensitivity of ocular anterior chamber tissues to oxidative damage and its relevance to the pathogenesis of glaucoma (2009) Invest Ophthalmol V/s SeE, 50, pp. 5251-5258. , Sacc 1/2 M. et al. lO.1l67/iovs.09-387l; Dynamics of H202 availability to ARPE- 19 cultures in models of oxidative stress (2010) Free Radic Blot Med, 48, pp. 1064-1070. , Kaczara. P., Sarna. T. and Burke, 3. M; Keller, K. E., Bhattacharya, S. K., Consensus recommendations for trabecular meshwork cell isola tion, characterization and culture (2018) ELtp Eye Res, 171, pp. 164-173. , Borr 1/2s, T; Kim, H., Inhibitory effect of astaxanthin on oxidative stress-induced mitochondrial dysfunction 1/2 A mini-review (2018) Nutrients, 10, p. 1137. , Kim. S. H. and; Kim, Y. W., Park, K. H., Exogenous influences on intraocular pressure (2019) Br J Ophthalmol, 2018, p. 313381; Kohen, N. T., Little, L. E., Healy, K. E., Characterization of Matrigel interfaces during defined human embryonic stem cell culture (2009) Bzoznterphases, 4, pp. 69-79; Adcock, I. M., Towards a 2P-century roadmap for biomedical research and drug discovery: Consensus report and recommendations (2017) Drug Discov Today, 22, pp. 327-339. , Busquet. F Langley. 0. R; Li, O., Lee, C., Agrahari, V., In vivo measurement of trabecular meshwork stifibess in a corticosteroid-induced ocular hypertensive mouse model (2019) Proc NatlAcadSci USA, 116, pp. 1714-1722. , lO.l073/pnas.l814889ll6; Lv, X., Liu, S., Hu, Z.-W., Autophagy-inducing natural compounds: A treasure resource for developing therapeutics against tissue fibrosis (2017) JAsian Nat Prod Res, 19, pp. 101-108; Yousaf, S., Khan, M. I., Polymorphisms in matrix metalloproteinases MIvIP1 and MMP9 are associated with primary open-angle and angle closure glaucoma in a Pakistani population (2013) Mol V/s, 19, pp. 441-447. , Micheal. S; Osmond, M., Bernier, S. M., Pantcheva, M. B., Collagen and collagen-chondroitin sulfate scaffolds with uniaxially aligned pores for the biomimetic, three dimensional culture of trabecular meshwork cells (2017) Biotechnol Bioeng, 114, pp. 915-923; Reissig, K., Schonfeld, P., Repeated H202 exposure drives cell cycle progression in an in vitro model of ulcerative colitis (2013) J Cell Mol Med, 17, pp. 1619-1631. , Poehlmann. A; Seydel, A., Camoirano, A., Oxidative damage and autophagy in the human trabecular meshwork as related with ageing (2014) PLoS One, 9, p. e98106. , Pulliero. A); Izzotti, A., Focus on molecular events in the anterior chamber leading to glaucoma (2014) Cell Mol Lfe SeE, 71, pp. 2197-2218. , Sacc 1/2, S. C. and; Gandolfi, S., Bagnis, A., From DNA damage to functional changes of the trabecular meshwork in aging and glaucoma (2016) Ageing Res Rev, 29, pp. 26-41. , Sacc 1/2, S. C., (a); Gandolfi, S., Bagnis, A., The outflow pathway: A tissue with morphological and functional unity (2016) J Cell Physiol, 231, pp. 1876-1893. , Sacc 1/2, S. C., (b); Corazza, P., Substances of interest that support glaucoma therapy (2019) Nutrients, 11, p. 239. , Sacc 1/2, S. C., Gandoffi. S. et al; Singh, D., Srivastava, S. K., Chaudhuri, T. K., Multifaceted role of matrix metalloproteinases (MMPs) (2015) Front Mol Biosci, 2, p. 19; Deutscher, M. P., Efficient mammalian protein synthesis requires an intact F-actin system (1997) JBIo! Chem, 272, pp. 24980-24986. , Stapulionis. R., Kolli. S. and; Struebing, F. L., What animal models can tell us about glaucoma (2015) ProgMo!Biol Traits! Sd, 134, pp. 365-380. , and Geisert. E. E; Ripandelli, O., Potential regulatory molecules in the human trabecular meshwork of patients with glaucoma: Immunohistochemical profile of a number of inflammatory cytokines (2015) Mo! Med Rep, 11, pp. 1384-1390. , Pacella. E Taurone. S; Li, X., Wong, T. Y., Global prevalence of glaucoma and projections of glaucoma burden through 2040: A systematic review and meta-analysis (2014) Ophthalmology, 121, pp. 2081-2090. , Tham. Y.-C., l0.10l6/j.ophtha.2014.05.013; Ucciferri, N., Aliluwalia, A., Allomelric scaling and cell ratios in multi-organ in vitro models of human metabolism (2014) Front Bloeng Biotechno, 2, p. 74. , Sbrana. T. and; TNFR2: A novel target for cancer immunotherapy (2017) Trends Mo! Med, 23 (103), pp. 7-1046. , Vanamee. E. S. and Faustman. D. L; De Preter, K., Pattyn, F., Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes (2002) Genome Biol, 3. , researchoO34. 1; Vernazza, S., Tirendi, S., Scarfi, S., 2D- and 3D-cultures of human trabecular meshwork cells: A preliminary assessment of an in vitro model for glaucoma study (2019) PLoS One, 14, p. e0221942; Waduthanthri, K. D., He, Y., Montemagno, C., An injectable peptide hydrogel for reconstruction of the human trabecular meshwork (2019) Acta Biomater, 100, pp. 244-254; Wang, J., Hanis, A., Prendes, M. A., Targeting transforming growth factor-n signaling in primary open- angle glaucoma (2017) J Glaucoma, 26, pp. 390-395; Wang, N., Chintala, S. K., Fini, lvi. E., Activation of a tissue-specific stress response in the aqueous outflow pathway of the eye defines the glaucoma disease phenotype (2001) Nat Med, 7, pp. 304-309; Xiang, Y., Li, B., Li, G.-G., Effects of endothelin-1 on the cytoskeleton protein F-actin of human trabecular meshwork cells in vitro (2010) mt j Ophthamo, 3, pp. 61-63; Zahir, N., Weaver, V. M., Death in the third dimension: Apoptosis regulation and tissue architecture (2004) Curr Opin GenetDev, 14, pp. 71-80; Zhao, J., Wang, S., Zhong, W., Oxidative stress in the trabecular meshwork (Review) (2016) mt J Mo! Med, 38, pp. 995-1002",

year = "2020",

doi = "10.14573/altex.1909262",

language = "English",

volume = "37",

pages = "265--274",

journal = "Altex",

issn = "1868-596X",

publisher = "ALTEX Edition",

number = "2",

}

TY - JOUR

T1 - An advanced in vitro model to assess glaucoma onset

AU - Sacc, S.C.

AU - Tirendi, S.

AU - Scarji, S.

AU - Fassalacqua, M.

AU - Oddone, F.

AU - Traverso, C.K.

AU - Vernazza, S.

AU - Bassi, A.M.

N1 - Cited By :3 Export Date: 2 February 2021 CODEN: ALTEE Correspondence Address: Vernazza, S.; Department of Experimental Medicine, Italy Chemicals/CAS: caspase 3, 169592-56-7; cytochrome c, 9007-43-6, 9064-84-0; gelatinase B, 146480-36-6; interstitial collagenase, 9001-12-1; matrigel, 119978-18-6; peroxide, 14915-07-2; protein bcl 2, 219306-68-0; stromelysin, 79955-99-0; ubiquitin C, 151821-62-4; ubiquitin, 60267-61-0 References: Aarskog, N., Vedeler, C., Real-time quantitative polymerase chain reaction (2000) Hum Genet, 107, pp. 494-498; Acott, T. S., Kelley, M., Extracellular matrix in the trabecular meshwork (2008) Eye Res, 86, pp. 543-561. , 3; Ahluwalia, A., Allometric scaling in-vitro (2017) Sci Rep, 7, p. 42113; Benton, G., Arnaoutova, I., George, J., Maui- gel: From discovery and ECM mimicry to assays and mod- els for cancer research (2014) Adv Drug Del/v Rev, 79-80, pp. 3-18; Berger, E., Paczia, N., Millifluidic culture improves human midbrain organoid vitality and differentiation (2018) Lab Chip, 18, pp. 3172-3183. , Magliaro. C; Boardman., K. C., Aryal, A. M., Miller., W. M., Actin re-distribution in response to hydrogen peroxide in airway epithelial cells (2004) JCell .Physiol, 199 (5), pp. 7-66; Roubeix, C., Kessal, K., Effect of benzalkonium chloride on trabecular meshwork cells in a new in vitro 3D trabecular meshwork model for glaucoma (2017) Toxicol In Vitro, 41, pp. 21-29. , Bouchemi. M; Bouhenni, P.. A., Dunmire, 3., Sewell, A., Animal models of glaucoma (2012) J Biomed Biotechnol, 2012, p. 692609; Brancato, V., Gioiella, F., 3D breast cancer microtissue reveals the role of tumor microenvironment on the transport and efficacy of free-doxorubicin in vitro (2018) Acta Biomater, 75, pp. 200-212. , Imparato. 0; Burgoyne, C. F., The non-human primate experimental glaucoma model (2015) Eye Res, 141, pp. 57-73; Elmore, S., Apoptosis: Areview of programmed cell death (2007) ToxicolPathol35, 495-5 16; de-la-Torre, A., Experimental models of glaucoma: A powerful translational tool for the future development of new therapies for glaucoma in humans 1/2A review of the literature (2019) Med/dna (Ma), 55, p. 280. , Evangelho. K., Mastronardi. C. A. and; Fuchshofer, R., Modulation of extracellular matrix turnover in the trabecular meshwork (2009) Erp Eye Res, 88, pp. 683-688. , and Tamm. E. R; Giusti, S., La, Marca. M., A novel dual-flow bioreactor simulates increased fluorescein permeability in epithelial tissue barriers (2014) Biotechnol J, 9, pp. 1175-1184. , Sbrana. T; Gonzalez, J. M., Hamm-Alvarez, . S., Tan, 3. C. H., Analyzing live cellularity in the human trabecular meshwork (2013) Invest Opthalmol fir SeE, 54, pp. 1039-1047; Hughes, C. S., Postovit, L. M., Lajoie, G. A., Math- gel: A complex protein mixture required for optimal growth of cell culture (2010) Proteomics, 10, pp. 1886-1890; Ishikawa, M., Yoshitomi, T., Zorumski, C. F., Experimentally induced mammalian models of glaucoma (2015) Biomed ResInt2OlS, p. 281214. , lO.1155/2015/281214; Izzotti, A., Longobardi, S. C.., Sensitivity of ocular anterior chamber tissues to oxidative damage and its relevance to the pathogenesis of glaucoma (2009) Invest Ophthalmol V/s SeE, 50, pp. 5251-5258. , Sacc 1/2 M. et al. lO.1l67/iovs.09-387l; Dynamics of H202 availability to ARPE- 19 cultures in models of oxidative stress (2010) Free Radic Blot Med, 48, pp. 1064-1070. , Kaczara. P., Sarna. T. and Burke, 3. M; Keller, K. E., Bhattacharya, S. K., Consensus recommendations for trabecular meshwork cell isola tion, characterization and culture (2018) ELtp Eye Res, 171, pp. 164-173. , Borr 1/2s, T; Kim, H., Inhibitory effect of astaxanthin on oxidative stress-induced mitochondrial dysfunction 1/2 A mini-review (2018) Nutrients, 10, p. 1137. , Kim. S. H. and; Kim, Y. W., Park, K. H., Exogenous influences on intraocular pressure (2019) Br J Ophthalmol, 2018, p. 313381; Kohen, N. T., Little, L. E., Healy, K. E., Characterization of Matrigel interfaces during defined human embryonic stem cell culture (2009) Bzoznterphases, 4, pp. 69-79; Adcock, I. M., Towards a 2P-century roadmap for biomedical research and drug discovery: Consensus report and recommendations (2017) Drug Discov Today, 22, pp. 327-339. , Busquet. F Langley. 0. R; Li, O., Lee, C., Agrahari, V., In vivo measurement of trabecular meshwork stifibess in a corticosteroid-induced ocular hypertensive mouse model (2019) Proc NatlAcadSci USA, 116, pp. 1714-1722. , lO.l073/pnas.l814889ll6; Lv, X., Liu, S., Hu, Z.-W., Autophagy-inducing natural compounds: A treasure resource for developing therapeutics against tissue fibrosis (2017) JAsian Nat Prod Res, 19, pp. 101-108; Yousaf, S., Khan, M. I., Polymorphisms in matrix metalloproteinases MIvIP1 and MMP9 are associated with primary open-angle and angle closure glaucoma in a Pakistani population (2013) Mol V/s, 19, pp. 441-447. , Micheal. S; Osmond, M., Bernier, S. M., Pantcheva, M. B., Collagen and collagen-chondroitin sulfate scaffolds with uniaxially aligned pores for the biomimetic, three dimensional culture of trabecular meshwork cells (2017) Biotechnol Bioeng, 114, pp. 915-923; Reissig, K., Schonfeld, P., Repeated H202 exposure drives cell cycle progression in an in vitro model of ulcerative colitis (2013) J Cell Mol Med, 17, pp. 1619-1631. , Poehlmann. A; Seydel, A., Camoirano, A., Oxidative damage and autophagy in the human trabecular meshwork as related with ageing (2014) PLoS One, 9, p. e98106. , Pulliero. A); Izzotti, A., Focus on molecular events in the anterior chamber leading to glaucoma (2014) Cell Mol Lfe SeE, 71, pp. 2197-2218. , Sacc 1/2, S. C. and; Gandolfi, S., Bagnis, A., From DNA damage to functional changes of the trabecular meshwork in aging and glaucoma (2016) Ageing Res Rev, 29, pp. 26-41. , Sacc 1/2, S. C., (a); Gandolfi, S., Bagnis, A., The outflow pathway: A tissue with morphological and functional unity (2016) J Cell Physiol, 231, pp. 1876-1893. , Sacc 1/2, S. C., (b); Corazza, P., Substances of interest that support glaucoma therapy (2019) Nutrients, 11, p. 239. , Sacc 1/2, S. C., Gandoffi. S. et al; Singh, D., Srivastava, S. K., Chaudhuri, T. K., Multifaceted role of matrix metalloproteinases (MMPs) (2015) Front Mol Biosci, 2, p. 19; Deutscher, M. P., Efficient mammalian protein synthesis requires an intact F-actin system (1997) JBIo! Chem, 272, pp. 24980-24986. , Stapulionis. R., Kolli. S. and; Struebing, F. L., What animal models can tell us about glaucoma (2015) ProgMo!Biol Traits! Sd, 134, pp. 365-380. , and Geisert. E. E; Ripandelli, O., Potential regulatory molecules in the human trabecular meshwork of patients with glaucoma: Immunohistochemical profile of a number of inflammatory cytokines (2015) Mo! Med Rep, 11, pp. 1384-1390. , Pacella. E Taurone. S; Li, X., Wong, T. Y., Global prevalence of glaucoma and projections of glaucoma burden through 2040: A systematic review and meta-analysis (2014) Ophthalmology, 121, pp. 2081-2090. , Tham. Y.-C., l0.10l6/j.ophtha.2014.05.013; Ucciferri, N., Aliluwalia, A., Allomelric scaling and cell ratios in multi-organ in vitro models of human metabolism (2014) Front Bloeng Biotechno, 2, p. 74. , Sbrana. T. and; TNFR2: A novel target for cancer immunotherapy (2017) Trends Mo! Med, 23 (103), pp. 7-1046. , Vanamee. E. S. and Faustman. D. L; De Preter, K., Pattyn, F., Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes (2002) Genome Biol, 3. , researchoO34. 1; Vernazza, S., Tirendi, S., Scarfi, S., 2D- and 3D-cultures of human trabecular meshwork cells: A preliminary assessment of an in vitro model for glaucoma study (2019) PLoS One, 14, p. e0221942; Waduthanthri, K. D., He, Y., Montemagno, C., An injectable peptide hydrogel for reconstruction of the human trabecular meshwork (2019) Acta Biomater, 100, pp. 244-254; Wang, J., Hanis, A., Prendes, M. A., Targeting transforming growth factor-n signaling in primary open- angle glaucoma (2017) J Glaucoma, 26, pp. 390-395; Wang, N., Chintala, S. K., Fini, lvi. E., Activation of a tissue-specific stress response in the aqueous outflow pathway of the eye defines the glaucoma disease phenotype (2001) Nat Med, 7, pp. 304-309; Xiang, Y., Li, B., Li, G.-G., Effects of endothelin-1 on the cytoskeleton protein F-actin of human trabecular meshwork cells in vitro (2010) mt j Ophthamo, 3, pp. 61-63; Zahir, N., Weaver, V. M., Death in the third dimension: Apoptosis regulation and tissue architecture (2004) Curr Opin GenetDev, 14, pp. 71-80; Zhao, J., Wang, S., Zhong, W., Oxidative stress in the trabecular meshwork (Review) (2016) mt J Mo! Med, 38, pp. 995-1002

PY - 2020

Y1 - 2020

N2 - Glaucoma is the second leading cause of blindness worldwide. Currently, glaucoma treatments aim to lower intraocular pressure by decreasing aqueous humor production or increasing aqueous humor outflow through pharmacological approaches or trabeculectomy. The lack of an effective cure requires new therapeutic strategies. We compared the biological responses of a three-dimensional trabecular meshwork model with or without perfusion bioreactor technology to better understand the early molecular changes induced by prolonged oxidative stress conditions induced by repeated daily peroxide exposure. We used standard 3D cultures of trabecular meshwork cells in Matrigel cultured under either static and dynamic conditions for one week. We studied changes in F-actin expression and organization in the cells, cellular metabolic activity, proinflammatory gene expression, expression of pro- and anti-apoptotic proteins, PARP-1 cleavage, and NFKB activation in the model. We demonstrate that the dynamic conditions improve the adaptive behavior of 3D trabecular meshwork cultures to chronic oxidative stress via offsetting pathway activation. © 2020 ALTEX Edition. All rights reserved.

AB - Glaucoma is the second leading cause of blindness worldwide. Currently, glaucoma treatments aim to lower intraocular pressure by decreasing aqueous humor production or increasing aqueous humor outflow through pharmacological approaches or trabeculectomy. The lack of an effective cure requires new therapeutic strategies. We compared the biological responses of a three-dimensional trabecular meshwork model with or without perfusion bioreactor technology to better understand the early molecular changes induced by prolonged oxidative stress conditions induced by repeated daily peroxide exposure. We used standard 3D cultures of trabecular meshwork cells in Matrigel cultured under either static and dynamic conditions for one week. We studied changes in F-actin expression and organization in the cells, cellular metabolic activity, proinflammatory gene expression, expression of pro- and anti-apoptotic proteins, PARP-1 cleavage, and NFKB activation in the model. We demonstrate that the dynamic conditions improve the adaptive behavior of 3D trabecular meshwork cultures to chronic oxidative stress via offsetting pathway activation. © 2020 ALTEX Edition. All rights reserved.

KW - caspase 3

KW - collagen type 1

KW - collagen type 1 alpha 1

KW - complementary DNA

KW - cytochrome c

KW - gelatinase B

KW - immunoglobulin enhancer binding protein

KW - interleukin 1alpha

KW - interleukin 1beta

KW - interleukin 6

KW - interstitial collagenase

KW - matrigel

KW - nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase 1

KW - peroxide

KW - protein Bax

KW - protein bcl 2

KW - protein p53

KW - second mitochondrial activator of caspase

KW - stromelysin

KW - synaptotagmin I

KW - transforming growth factor beta2

KW - tumor necrosis factor

KW - ubiquitin C

KW - unclassified drug

KW - ubiquitin

KW - antiapoptotic activity

KW - apoptosis assay

KW - Article

KW - chronic stress

KW - gene expression

KW - glaucoma

KW - human

KW - human cell

KW - immunoreactivity

KW - in vitro study

KW - metabolic activity assay

KW - oxidative stress

KW - polyacrylamide gel electrophoresis

KW - protein expression

KW - real time polymerase chain reaction

KW - resazurin assay

KW - RNA extraction

KW - three dimensional cell culture

KW - trabecular meshwork

KW - Western blotting

KW - adaptive behavior

KW - cell culture technique

KW - cell growth

KW - confocal microscopy

KW - controlled study

KW - cytokine production

KW - down regulation

KW - flow cytometry

KW - genetic transfection

KW - signal transduction

KW - upregulation

U2 - 10.14573/altex.1909262

DO - 10.14573/altex.1909262

M3 - Article

VL - 37

SP - 265

EP - 274

JO - Altex

JF - Altex

SN - 1868-596X

IS - 2

ER -

An advanced in vitro model to assess glaucoma onset

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