Sustained Neurotrophin Release from Protein Nanoparticles Mediated by Matrix Metalloproteinases Induces the Alignment and Differentiation of Nerve Cells
The spatial and temporal availability of cytokines, and the microenvironments this creates, is critical to tissue development and homeostasis. Creating concentration gradients in vitro using soluble proteins is challenging as they do not provide a self-sustainable source. To mimic the sustained cyto...
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| Format: | Article |
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MDPI AG
2019-09-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/9/10/510 |
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| author | Yuka Matsuzaki Rina Maruta Keiko Takaki Eiji Kotani Yasuko Kato Ryoichi Yoshimura Yasuhisa Endo Ciara Whitty Christian Pernstich Raj Gandhi Michael Jones Hajime Mori |
| author_facet | Yuka Matsuzaki Rina Maruta Keiko Takaki Eiji Kotani Yasuko Kato Ryoichi Yoshimura Yasuhisa Endo Ciara Whitty Christian Pernstich Raj Gandhi Michael Jones Hajime Mori |
| author_sort | Yuka Matsuzaki |
| collection | DOAJ |
| description | The spatial and temporal availability of cytokines, and the microenvironments this creates, is critical to tissue development and homeostasis. Creating concentration gradients in vitro using soluble proteins is challenging as they do not provide a self-sustainable source. To mimic the sustained cytokine secretion seen in vivo from the extracellular matrix (ECM), we encapsulated a cargo protein into insect virus-derived proteins to form nanoparticle co-crystals and studied the release of this cargo protein mediated by matrix metalloproteinase-2 (MMP-2) and MMP-8. Specifically, when nerve growth factor (NGF), a neurotrophin, was encapsulated into nanoparticles, its release was promoted by MMPs secreted by a PC12 neuronal cell line. When these NGF nanoparticles were spotted onto a cover slip to create a uniform circular field, movement and alignment of PC12 cells via their extended axons along the periphery of the NGF nanoparticle field was observed. Neural cell differentiation was confirmed by the expression of specific markers of tau, neurofilament, and GAP-43. Connections between the extended axons and the growth cones were also observed, and expression of connexin 43 was consistent with the formation of gap junctions. Extensions and connection of very fine filopodia occurred between growth cones. Our studies indicate that crystalline protein nanoparticles can be utilized to generate a highly stable cytokine gradient microenvironment that regulates the alignment and differentiation of nerve cells. This technique greatly simplifies the creation of protein concentration gradients and may lead to therapies for neuronal injuries and disease. |
| first_indexed | 2024-12-11T21:19:55Z |
| format | Article |
| id | doaj.art-2163037a146d44baa5ede03fb9200469 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-12-11T21:19:55Z |
| publishDate | 2019-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-2163037a146d44baa5ede03fb92004692022-12-22T00:50:29ZengMDPI AGBiomolecules2218-273X2019-09-0191051010.3390/biom9100510biom9100510Sustained Neurotrophin Release from Protein Nanoparticles Mediated by Matrix Metalloproteinases Induces the Alignment and Differentiation of Nerve CellsYuka Matsuzaki0Rina Maruta1Keiko Takaki2Eiji Kotani3Yasuko Kato4Ryoichi Yoshimura5Yasuhisa Endo6Ciara Whitty7Christian Pernstich8Raj Gandhi9Michael Jones10Hajime Mori11Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, JapanDepartment of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, JapanDepartment of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, JapanDepartment of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, JapanDepartment of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, JapanDepartment of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, JapanDepartment of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, JapanCell Guidance Systems, Maia Building, Babraham Research Campus, Cambridge, CB22 3AT, UKCell Guidance Systems, Maia Building, Babraham Research Campus, Cambridge, CB22 3AT, UKCell Guidance Systems, Maia Building, Babraham Research Campus, Cambridge, CB22 3AT, UKCell Guidance Systems, Maia Building, Babraham Research Campus, Cambridge, CB22 3AT, UKDepartment of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, JapanThe spatial and temporal availability of cytokines, and the microenvironments this creates, is critical to tissue development and homeostasis. Creating concentration gradients in vitro using soluble proteins is challenging as they do not provide a self-sustainable source. To mimic the sustained cytokine secretion seen in vivo from the extracellular matrix (ECM), we encapsulated a cargo protein into insect virus-derived proteins to form nanoparticle co-crystals and studied the release of this cargo protein mediated by matrix metalloproteinase-2 (MMP-2) and MMP-8. Specifically, when nerve growth factor (NGF), a neurotrophin, was encapsulated into nanoparticles, its release was promoted by MMPs secreted by a PC12 neuronal cell line. When these NGF nanoparticles were spotted onto a cover slip to create a uniform circular field, movement and alignment of PC12 cells via their extended axons along the periphery of the NGF nanoparticle field was observed. Neural cell differentiation was confirmed by the expression of specific markers of tau, neurofilament, and GAP-43. Connections between the extended axons and the growth cones were also observed, and expression of connexin 43 was consistent with the formation of gap junctions. Extensions and connection of very fine filopodia occurred between growth cones. Our studies indicate that crystalline protein nanoparticles can be utilized to generate a highly stable cytokine gradient microenvironment that regulates the alignment and differentiation of nerve cells. This technique greatly simplifies the creation of protein concentration gradients and may lead to therapies for neuronal injuries and disease.https://www.mdpi.com/2218-273X/9/10/510extracellular matrixmatrix metalloproteinaseneurotrophinNGFPC12 cellspolyhedraprotein nanoparticles |
| spellingShingle | Yuka Matsuzaki Rina Maruta Keiko Takaki Eiji Kotani Yasuko Kato Ryoichi Yoshimura Yasuhisa Endo Ciara Whitty Christian Pernstich Raj Gandhi Michael Jones Hajime Mori Sustained Neurotrophin Release from Protein Nanoparticles Mediated by Matrix Metalloproteinases Induces the Alignment and Differentiation of Nerve Cells Biomolecules extracellular matrix matrix metalloproteinase neurotrophin NGF PC12 cells polyhedra protein nanoparticles |
| title | Sustained Neurotrophin Release from Protein Nanoparticles Mediated by Matrix Metalloproteinases Induces the Alignment and Differentiation of Nerve Cells |
| title_full | Sustained Neurotrophin Release from Protein Nanoparticles Mediated by Matrix Metalloproteinases Induces the Alignment and Differentiation of Nerve Cells |
| title_fullStr | Sustained Neurotrophin Release from Protein Nanoparticles Mediated by Matrix Metalloproteinases Induces the Alignment and Differentiation of Nerve Cells |
| title_full_unstemmed | Sustained Neurotrophin Release from Protein Nanoparticles Mediated by Matrix Metalloproteinases Induces the Alignment and Differentiation of Nerve Cells |
| title_short | Sustained Neurotrophin Release from Protein Nanoparticles Mediated by Matrix Metalloproteinases Induces the Alignment and Differentiation of Nerve Cells |
| title_sort | sustained neurotrophin release from protein nanoparticles mediated by matrix metalloproteinases induces the alignment and differentiation of nerve cells |
| topic | extracellular matrix matrix metalloproteinase neurotrophin NGF PC12 cells polyhedra protein nanoparticles |
| url | https://www.mdpi.com/2218-273X/9/10/510 |
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