Micropatterned nanolayers immobilized with nerve growth factor for neurite formation of PC12 cells

Seong Min Kim,1,2 Masashi Ueki,1 Xueli Ren,3 Jun Akimoto,1 Yasuyuki Sakai,2 Yoshihiro Ito1,3 1Nano Medical Engineering Laboratory, RIKEN Cluster for Pioneering Research, Wako, Saitama 351-0198, Japan; 2Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo 113-8656, Japa...

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Autores principales:	Kim SM, Ueki M, Ren X, Akimoto J, Sakai Y, Ito Y
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2019
Materias:	Nerve growth factor Photoreactive gelatin Micropatterned immobilization PC12 cell Neurite outgrowth Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/c1658d02b9684b00bd14fcdc3e9592a1
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spelling	oai:doaj.org-article:c1658d02b9684b00bd14fcdc3e9592a12021-12-02T04:09:02ZMicropatterned nanolayers immobilized with nerve growth factor for neurite formation of PC12 cells1178-2013https://doaj.org/article/c1658d02b9684b00bd14fcdc3e9592a12019-09-01T00:00:00Zhttps://www.dovepress.com/micropatterned-nanolayers-immobilized-with-nerve-growth-factor-for-neu-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Seong Min Kim,1,2 Masashi Ueki,1 Xueli Ren,3 Jun Akimoto,1 Yasuyuki Sakai,2 Yoshihiro Ito1,3 1Nano Medical Engineering Laboratory, RIKEN Cluster for Pioneering Research, Wako, Saitama 351-0198, Japan; 2Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo 113-8656, Japan; 3Emergent Bioengineering Materials Research Team, RIKEN Center for Emergent Matter Science, Wako, Saitama 351-0198, JapanCorrespondence: Yoshihiro ItoNano Medical Engineering Laboratory, RIKEN Cluster for Pioneering Research, 2-1 Hirosawa, Wako, Saitama 351-0198, JapanTel +81 48 467 5809Fax +81 48 467 9300Email y-ito@riken.jpBackground: Nerve regeneration is important for the treatment of degenerative diseases and neurons injured by accidents. Nerve growth factor (NGF) has been previously conjugated to materials for promotion of neurogenesis.Materials and methods: Photoreactive gelatin was prepared by chemical coupling of gelatin with azidobenzoic acid (P-gel), and then NGF was immobilized on substrates in the presence or absence of micropatterned photomasks. UV irradiation induced crosslinking reactions of P-gel with itself, NGF, and the plate for immobilization.Results: By adjustment of the P-gel concentration, the nanometer-order height of micropatterns was controlled. NGF was quantitatively immobilized with increasing amounts of P-gel. Immobilized NGF induced neurite outgrowth of PC12 cells, a cell line derived from a pheochromocytoma of the rat adrenal medulla, at the same level as soluble NGF. The immobilized NGF showed higher thermal stability than the soluble NGF and was repeatedly used without loss of biological activity. The 3D structure (height of the formed micropattern) regulated the behavior of neurite guidance. As a result, the orientation of neurites was regulated by the stripe pattern width.Conclusion: The micropattern-immobilized NGF nanolayer biochemically and topologically regulated neurite formation.Keywords: nerve growth factor, photoreactive gelatin, micropatterned immobilization, PC12 cell, neurite outgrowthKim SMUeki MRen XAkimoto JSakai YIto YDove Medical PressarticleNerve growth factorPhotoreactive gelatinMicropatterned immobilizationPC12 cellNeurite outgrowthMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 14, Pp 7683-7694 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Nerve growth factor Photoreactive gelatin Micropatterned immobilization PC12 cell Neurite outgrowth Medicine (General) R5-920
spellingShingle	Nerve growth factor Photoreactive gelatin Micropatterned immobilization PC12 cell Neurite outgrowth Medicine (General) R5-920 Kim SM Ueki M Ren X Akimoto J Sakai Y Ito Y Micropatterned nanolayers immobilized with nerve growth factor for neurite formation of PC12 cells
description	Seong Min Kim,1,2 Masashi Ueki,1 Xueli Ren,3 Jun Akimoto,1 Yasuyuki Sakai,2 Yoshihiro Ito1,3 1Nano Medical Engineering Laboratory, RIKEN Cluster for Pioneering Research, Wako, Saitama 351-0198, Japan; 2Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo 113-8656, Japan; 3Emergent Bioengineering Materials Research Team, RIKEN Center for Emergent Matter Science, Wako, Saitama 351-0198, JapanCorrespondence: Yoshihiro ItoNano Medical Engineering Laboratory, RIKEN Cluster for Pioneering Research, 2-1 Hirosawa, Wako, Saitama 351-0198, JapanTel +81 48 467 5809Fax +81 48 467 9300Email y-ito@riken.jpBackground: Nerve regeneration is important for the treatment of degenerative diseases and neurons injured by accidents. Nerve growth factor (NGF) has been previously conjugated to materials for promotion of neurogenesis.Materials and methods: Photoreactive gelatin was prepared by chemical coupling of gelatin with azidobenzoic acid (P-gel), and then NGF was immobilized on substrates in the presence or absence of micropatterned photomasks. UV irradiation induced crosslinking reactions of P-gel with itself, NGF, and the plate for immobilization.Results: By adjustment of the P-gel concentration, the nanometer-order height of micropatterns was controlled. NGF was quantitatively immobilized with increasing amounts of P-gel. Immobilized NGF induced neurite outgrowth of PC12 cells, a cell line derived from a pheochromocytoma of the rat adrenal medulla, at the same level as soluble NGF. The immobilized NGF showed higher thermal stability than the soluble NGF and was repeatedly used without loss of biological activity. The 3D structure (height of the formed micropattern) regulated the behavior of neurite guidance. As a result, the orientation of neurites was regulated by the stripe pattern width.Conclusion: The micropattern-immobilized NGF nanolayer biochemically and topologically regulated neurite formation.Keywords: nerve growth factor, photoreactive gelatin, micropatterned immobilization, PC12 cell, neurite outgrowth
format	article
author	Kim SM Ueki M Ren X Akimoto J Sakai Y Ito Y
author_facet	Kim SM Ueki M Ren X Akimoto J Sakai Y Ito Y
author_sort	Kim SM
title	Micropatterned nanolayers immobilized with nerve growth factor for neurite formation of PC12 cells
title_short	Micropatterned nanolayers immobilized with nerve growth factor for neurite formation of PC12 cells
title_full	Micropatterned nanolayers immobilized with nerve growth factor for neurite formation of PC12 cells
title_fullStr	Micropatterned nanolayers immobilized with nerve growth factor for neurite formation of PC12 cells
title_full_unstemmed	Micropatterned nanolayers immobilized with nerve growth factor for neurite formation of PC12 cells
title_sort	micropatterned nanolayers immobilized with nerve growth factor for neurite formation of pc12 cells
publisher	Dove Medical Press
publishDate	2019
url	https://doaj.org/article/c1658d02b9684b00bd14fcdc3e9592a1
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Micropatterned nanolayers immobilized with nerve growth factor for neurite formation of PC12 cells

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