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[1]P. Watson, B.L. Hazleman, P. Mc Cluskey, C. Pavesio. The sclera and systemic disorders. Third edition. JP Medical Ltd London, UK (2012)
[2]J. Smyth, J.J. More, S. Shapourifar-Tehrani, D.A. Lee: The effect of 5-fluorouridine, 5-fluoredeoxyuridine, and 5-fluorodeoxyuridine monophosphate on rabbit tenon’s capsule fibroblasts in vitro. J Ocul Pharmacol 7, 329–338 (1991)
[3]E.J. O’Neill, Q. Qin, N.J. Van Bergen, P.P. Connell, S. Vasudevan, M.A. Coote, I.A. Trounce, T.T. Wong, J.G. Crowston: Antifibrotic activity of bevacizumab on human Tenon’s fibroblasts in vitro. Invest Ophtalmol Vis Sci 51, 6524–6532 (2010)
[4]E. De Falco, G. Scafetta, C. Napoletano, R. Puca R, E.M. Vingolo, G. Ragona, O. Iorio, G. Frati: A standardized laboratory and surgical method for in vitro culture isolation and expansion of primary human Tenon’s fibroblasts. Cell and Tissue Bank 14, 277-287 (2013)
[5]L. Choritz, J. Grub, M. Wegner, N. Pfeiffer, H. Thieme: Paclitaxel inhibits growth, migration and collagen production of human Tenon’s fibroblasts-potential use in drugeluiting glaucoma drainage devices. Graefes Arch Clin Exp Ophtalmol 248, 197–206 (2010)
[6]G. Scafetta, E. Tricoli, C. Siciliano, C. Napoletano, R. Puca, E.M. Vingolo, G. Cavallaro, A. Polistena, G. Frati, E. De Falco: Suitability of human Tenon’s fibroblasts as feeder cells for culturing human limbal epithelial stem cells. Stem cell Rev 9, 847-857 (2013)
[7]F. Deng, H. Hu, M. Chen, X. Sun, X. Liu, Z. Dong, Y. Liu, L. Xi, J. Zhuang, J. Ge: Generation of induced pluripotent stem cells from human Tenon’s capsule fibroblasts. Mol Vis 18, 2871-2881 (2012)
[8]G. Gstraunthaler: Alternatives to the use of fetal bovine serum: serum-free cell culture. ALTEX, 20, 275-281 (2003)
[9]R. Ross, E.W. Raines, D.F. Bowen-Pope: The biology of platelet derived growth factor. Cell 46, 155-169 (1986)
[10]C.H. Heldin, A. Wasteson, B. Westermark: Platelet-derived growth factor. Mol. Cell. Endocrinol. 39, 169-187 (1985)
[11]C. Siciliano, M. Ibrahim, G. Scafetta, C. Napoletano, G. Mangino, L. Pierelli, G. Frati, E. De Falco: Optimization of the isolation and expansion method of human mediastinal-adipose tissue derived mesenchymal stem cells with virally inactivated GMP-grade platelet lysate. Cytotechnology 67, 165-174 (2013)
[12]E. Anitua, M. Sánchez, G. Orive, I. Andía: The potential impact of the preparation rich in growth factors (PRGF) in different medical fields. Biomaterials 28, 4551-4560 (2007)
[13]J.L. Alio, M. Abad, A. Artola, J.L. Rodriguez-Prats, S. Pastor, J. Ruiz-Colecha: Use of autologous platelet-rich plasma in the treatment of dormant corneal ulcers. Ophthalmology 114, 1286-1293 (2007)
[14]J.F. Korobelnik, D. Hannouche, N. Belayachi, M. Branger M, J.E. Guez, T. Hoang-Xuan: Autologous platelet concentrate as an adjunct in macular hole healing. Ophthalmology 103, 590-594 (1996)
[15]S. Gehring, H. Hoerauf, H. Laqua, H. Kirchner, H. Klüter: Preparation of autologous platelets for the ophthalmologic treatment of macular holes. Transfusion 39, 144-148 (1999)
[16]J. Wachtlin, C. Jandeck, S. Potthofer, U. Kellner, M.H. Foerster: Long-term results following pars plana vitrectomy with platelet concentrate in pediatric patients with traumatic macular hole. Am J Ophthalmol 136, 197-199 (2003)
[17]B.J. Vote, W.L. Membrey, A.G. Casswell: Autologous platelets for macular hole surgery: the Sussex Eye Hospital experience. Clin Experiment Ophthalmol 32, 472-477 (2004)
[18]W. Geremicca, C. Fonte, S. Vecchio: Blood components for topical use in tissue regeneration: evaluation of corneal lesions treated with platelet lysate and considerations on repair mechanisms. Blood Transfus 8, 107-112 (2010)
[19]S. Werner, R. Grose: Regulation of wound healing by growth factors and cytokines. Physiol Rev 83, 835-870 (2003)
[20]E. Anitua, I. Andıa, B. Ardanza, P. Nurden, A.T. Nurden: Autologous platelets as a source of proteins for healing and tissue regeneration. Thromb Haemost 91, 4-15 (2004)
[21]G.C. Leitner, R. Gruber, J. Neumüller, A. Wagner, P. Kloimstein, P. Höcker, G.F. Körmöczi, C. Buchta: Platelet content and growth factor release in platelet rich plasma: a comparison of four different systems. Vox Sang 91, 135-139 (2006)
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[23]E. Ranzato, F. Boccafoschi, L. Mazzucco, M. Patrone, B. Burlando: Role of ERK1/2 in Platelet Lysate-Driven Endothelial Cell Repair. J Cell Biochem 110, 783-793 (2010)
[24]E. Ranzato, L. Mazzucco, M. Patrone, B. Burlando: Platelet lysate promotes in vitro wound scratch closure of human dermal fibroblasts: different roles of cell calcium, P38, ERK and PI3K/AKT. J Cell Mol Med 13, 2030-2038 (2009)
[25]D. Bastianelli, C. Siciliano, R. Puca, A. Coccia A, C. Murdoch, A. Bordin, G. Mangino, G. Pompilio, A. Calogero, E. De Falco: Influence of egr-1 in cardiac tissue-derived mesenchymal stem cells in response to glucose variations. Biomed Res Int 2014, 254793 (2014)
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[27]A. Coccia, D. Bastianelli, L. Mosca, R. Monticolo R, I. Panuccio, A. Carbone, A. Calogero, E. Lendaro: Extra Virgin Olive Oil Phenols Suppress Migration and Invasion of T24 Human Bladder Cancer Cells Through Modulation of Matrix Metalloproteinase-2. Nutr Cancer 66, 946-54 (2014)
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Frontiers in Bioscience-Elite (FBE) is published by IMR Press from Volume 13 Issue 2 (2021). Previous articles were published by another publisher on a subscription basis, and they are hosted by IMR Press on imrpress.com as a courtesy and upon agreement with Frontiers in Bioscience.
GMP-grade platelet lysate enhances proliferation and migration of tenon fibroblasts
1 Department of Medical-Surgical Science and Biotechnologies, Faculty of Pharmacy and Medicine, University of Rome, Sapienza, C.so della Repubblica 79, 04100 Latina, Italy
2 Center for Life Nano Science, Sapienza, Istituto Italiano di Tecnologia, Rome, Italy
3 Department of Internal Medicine and Medical Specialties, University of Rome, Sapienza, Rome, Italy
4 Department of Ophthalmology, Sapienza University of Rome, Polo Pontino, A. Fiorini Hospital, Via Firenze, 04019 Terracina
5 Immunohematology and Transfusion Medicine, San Camillo Forlanini Hospital, Rome, Italy
6 Department of Experimental Medicine, University of Rome, Sapienza, Rome, Italy
7 IRCCS Neuromed, Località Camerelle, 86077 Pozzilli, Italy
*Author to whom correspondence should be addressed.
Abstract
Tenon’s fibroblasts (TFs), widely employed as in vitro model for many ophthalmological studies, are routinely cultured with FBS. Platelet Lysate (PL), a hemoderivate enriched with growth factors and cytokines has been largely tested in several clinical applications and as substitute of FBS in culture. Here, we investigate whether PL can exert biological effects on TF populations similarly to other cell types. Results show that PL significantly enhances cell proliferation and migration vs. FBS, without influencing cell size/granularity. Upregulation of EGF, VEGF, KDR, MMP2-9, FAK mRNA levels also occurs and phosphorylation of AKT but not of ERK1/2 is significantly enhanced. The inhibition of the PI3kinase/AKT pathway with the specific inhibitor wortmannin, decreases PL-induced cell migration but not proliferation. Condition supernatants containing PL show increased bioavailability of Nitric Oxide and reduced levels of 8-Iso-PGF2-alpha, correlating with cell proliferation and migration. Pro-angiogenic/inflammatory soluble factors (GRO, Angiogenin, EGF, I-309, PARC) are exclusively or greater expressed in media containing PL than FBS. GMP-grade PL preparations positively influence in vitro biological effects of TFs representing a suitable and safer alternative to FBS.
Keywords
- Tenon Fibroblasts
- Platelet Lysate
- GMP-grade
- Growth Factors
- Cytokines
- AKT
- ERK1/2
- Oxidative Stress
References
- [1] P. Watson, B.L. Hazleman, P. Mc Cluskey, C. Pavesio. The sclera and systemic disorders. Third edition. JP Medical Ltd London, UK (2012)
- [2] J. Smyth, J.J. More, S. Shapourifar-Tehrani, D.A. Lee: The effect of 5-fluorouridine, 5-fluoredeoxyuridine, and 5-fluorodeoxyuridine monophosphate on rabbit tenon’s capsule fibroblasts in vitro. J Ocul Pharmacol 7, 329–338 (1991)
- [3] E.J. O’Neill, Q. Qin, N.J. Van Bergen, P.P. Connell, S. Vasudevan, M.A. Coote, I.A. Trounce, T.T. Wong, J.G. Crowston: Antifibrotic activity of bevacizumab on human Tenon’s fibroblasts in vitro. Invest Ophtalmol Vis Sci 51, 6524–6532 (2010)
- [4] E. De Falco, G. Scafetta, C. Napoletano, R. Puca R, E.M. Vingolo, G. Ragona, O. Iorio, G. Frati: A standardized laboratory and surgical method for in vitro culture isolation and expansion of primary human Tenon’s fibroblasts. Cell and Tissue Bank 14, 277-287 (2013)
- [5] L. Choritz, J. Grub, M. Wegner, N. Pfeiffer, H. Thieme: Paclitaxel inhibits growth, migration and collagen production of human Tenon’s fibroblasts-potential use in drugeluiting glaucoma drainage devices. Graefes Arch Clin Exp Ophtalmol 248, 197–206 (2010)
- [6] G. Scafetta, E. Tricoli, C. Siciliano, C. Napoletano, R. Puca, E.M. Vingolo, G. Cavallaro, A. Polistena, G. Frati, E. De Falco: Suitability of human Tenon’s fibroblasts as feeder cells for culturing human limbal epithelial stem cells. Stem cell Rev 9, 847-857 (2013)
- [7] F. Deng, H. Hu, M. Chen, X. Sun, X. Liu, Z. Dong, Y. Liu, L. Xi, J. Zhuang, J. Ge: Generation of induced pluripotent stem cells from human Tenon’s capsule fibroblasts. Mol Vis 18, 2871-2881 (2012)
- [8] G. Gstraunthaler: Alternatives to the use of fetal bovine serum: serum-free cell culture. ALTEX, 20, 275-281 (2003)
- [9] R. Ross, E.W. Raines, D.F. Bowen-Pope: The biology of platelet derived growth factor. Cell 46, 155-169 (1986)
- [10] C.H. Heldin, A. Wasteson, B. Westermark: Platelet-derived growth factor. Mol. Cell. Endocrinol. 39, 169-187 (1985)
- [11] C. Siciliano, M. Ibrahim, G. Scafetta, C. Napoletano, G. Mangino, L. Pierelli, G. Frati, E. De Falco: Optimization of the isolation and expansion method of human mediastinal-adipose tissue derived mesenchymal stem cells with virally inactivated GMP-grade platelet lysate. Cytotechnology 67, 165-174 (2013)
- [12] E. Anitua, M. Sánchez, G. Orive, I. Andía: The potential impact of the preparation rich in growth factors (PRGF) in different medical fields. Biomaterials 28, 4551-4560 (2007)
- [13] J.L. Alio, M. Abad, A. Artola, J.L. Rodriguez-Prats, S. Pastor, J. Ruiz-Colecha: Use of autologous platelet-rich plasma in the treatment of dormant corneal ulcers. Ophthalmology 114, 1286-1293 (2007)
- [14] J.F. Korobelnik, D. Hannouche, N. Belayachi, M. Branger M, J.E. Guez, T. Hoang-Xuan: Autologous platelet concentrate as an adjunct in macular hole healing. Ophthalmology 103, 590-594 (1996)
- [15] S. Gehring, H. Hoerauf, H. Laqua, H. Kirchner, H. Klüter: Preparation of autologous platelets for the ophthalmologic treatment of macular holes. Transfusion 39, 144-148 (1999)
- [16] J. Wachtlin, C. Jandeck, S. Potthofer, U. Kellner, M.H. Foerster: Long-term results following pars plana vitrectomy with platelet concentrate in pediatric patients with traumatic macular hole. Am J Ophthalmol 136, 197-199 (2003)
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- [19] S. Werner, R. Grose: Regulation of wound healing by growth factors and cytokines. Physiol Rev 83, 835-870 (2003)
- [20] E. Anitua, I. Andıa, B. Ardanza, P. Nurden, A.T. Nurden: Autologous platelets as a source of proteins for healing and tissue regeneration. Thromb Haemost 91, 4-15 (2004)
- [21] G.C. Leitner, R. Gruber, J. Neumüller, A. Wagner, P. Kloimstein, P. Höcker, G.F. Körmöczi, C. Buchta: Platelet content and growth factor release in platelet rich plasma: a comparison of four different systems. Vox Sang 91, 135-139 (2006)
- [22] V. Mirabet, P. Solves, M.D. Miñana, A. Encabo, F. Carbonell-Uberos, A. Blanquer, R. Roig: Human platelet lysate enhances the proliferative activity of cultured human fibroblast-like cells from different tissues. Cell Tissue Bank 9, 1-10 (2008)
- [23] E. Ranzato, F. Boccafoschi, L. Mazzucco, M. Patrone, B. Burlando: Role of ERK1/2 in Platelet Lysate-Driven Endothelial Cell Repair. J Cell Biochem 110, 783-793 (2010)
- [24] E. Ranzato, L. Mazzucco, M. Patrone, B. Burlando: Platelet lysate promotes in vitro wound scratch closure of human dermal fibroblasts: different roles of cell calcium, P38, ERK and PI3K/AKT. J Cell Mol Med 13, 2030-2038 (2009)
- [25] D. Bastianelli, C. Siciliano, R. Puca, A. Coccia A, C. Murdoch, A. Bordin, G. Mangino, G. Pompilio, A. Calogero, E. De Falco: Influence of egr-1 in cardiac tissue-derived mesenchymal stem cells in response to glucose variations. Biomed Res Int 2014, 254793 (2014)
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- [27] A. Coccia, D. Bastianelli, L. Mosca, R. Monticolo R, I. Panuccio, A. Carbone, A. Calogero, E. Lendaro: Extra Virgin Olive Oil Phenols Suppress Migration and Invasion of T24 Human Bladder Cancer Cells Through Modulation of Matrix Metalloproteinase-2. Nutr Cancer 66, 946-54 (2014)
- [28] L. Pacini, E. De Falco, M. Di Bari, A. Coccia, C. Siciliano, D. Ponti, A.L. Pastore, V. Petrozza, A. Carbone, A.M. Tata, A. Calogero: M2 muscarinic receptors inhibit cell proliferation and migration in urothelial bladder cancer cells. Cancer Biol Ther 15, 1489-1498 (2014)
- [29] B.A. Naaijkens, H.W.M. Niessen, H.J. Prins, P.A.J. Krijnen, T.J. Kokhuis, N. de Jong, V.W. van Hinsbergh, O. Kamp, M.N. Helder, R.J. Musters, A. van Dijk, L.J. Juffermans: Human platelet lysate as a fetal bovine serum substitute improves human adipose-derived stromal cell culture for future cardiac repair applications. Cell Tissue Res 348, 119-130 (2012)
- [30] B.J. Tripathi, P.S. Kwait, R.C. Tripathi: Corneal growth factors: a new generation of ophthalmic pharmaceuticals. Cornea 9, 2-9 (1990)
- [31] G. Schultz, N. Chegini, M. Grant, P. Khaw, S. MacKay: Effects of growth factors on corneal wound healing. Acta Ophthalmol Suppl 102, 60-66 (1992)
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- [34] V. Sing, F.L. Barbosa, A.A. Torricelli, M.R. Santhiago MR, S.E. Wilson: Transforming growth factor β and platelet-derived growth factor modulation of myofibroblast development from corneal fibroblasts in vitro. Exp Eye Res 120, 152-160 (2014)
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