Nerve Growth Factor-Binding Engineered Silk Films Promote Neuronal Attachment and Neurite Outgrowth

Lizzy A. Baker; Lingling Xu; Farideh Badichi Akher; Madelaine K. Robertson; Lauren Pugsley-DeBruyn; Chloe Xiaoyi Ma; Xiang Qin Liu; John P. Frampton; Jan K. Rainey

doi:10.1002/adfm.202205178

Nerve Growth Factor-Binding Engineered Silk Films Promote Neuronal Attachment and Neurite Outgrowth

Lizzy A. Baker, Lingling Xu, Farideh Badichi Akher, Madelaine K. Robertson, Lauren Pugsley-DeBruyn, Chloe Xiaoyi Ma, Xiang Qin Liu, John P. Frampton, Jan K. Rainey

Medicine

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

6 Citations (Scopus)

Abstract

Spider silks have outstanding potential as biomaterials due to their sought-after mechanical properties and low immunogenicity. The toughest spider silk is aciniform silk, which is used by spiders to wrap prey and produce egg sacs. A variety of recombinant aciniform silk constructs are now been developed, including hybrid silks with domains from multiple spider silk proteins fused together. In this study, an engineered aciniform silk construct, termed N_BSilk, fused both N- and C-terminally to heptapeptide motifs that bind the neurotrophic factor and nerve growth factor-β (NGF) is introduced. N_BSilk is shown to be amenable to casting into robust films that remain intact while sequestering and maintaining bioactive NGF at the film surface for at least 7 days. These films support cell survival while enhancing differentiation and neurite density and outgrowth in neuron-like PC12 cells, with elevation of signaling through both the mitogen-activated protein kinase (MAPK) and protein kinase B (AKT) signaling pathways. Strikingly, preloading of N_BSilk films with NGF enhances neuritogenesis even over conditions where cells are grown with NGF supplementation in the culture medium. N_BSilk scaffolds, thus, warrant future development and evaluation as biomaterials for nerve regeneration.

Original language	English
Journal	Advanced Functional Materials
DOIs	https://doi.org/10.1002/adfm.202205178
Publication status	Accepted/In press - 2022

Bibliographical note

Funding Information:
This work was supported by funds from the Natural Sciences and Engineering Research Council of Canada (J.P.F., RGPIN/04298‐2016; X‐Q.L., RGPIN/06083‐2020; J.K.R., RGPIN/05907‐2017; J.K.R, J.P.F., and X‐Q.L. RTI/000030‐2020), the New Frontiers in Research Fund (J.P.F and J.K.R, NFRFE/2018‐00356), the Canada Research Chairs Program (J.P.F.), the Canada Foundation for Innovation (CFI – J.P.F., Project #33533; J.K.R., Project #15719), and Dalhousie University's Student Employment Program (J.K.R.). Advanced computing resources were provided by Compute Canada, the organization responsible for digital research infrastructure in Canada, and ACENET, the regional partner in Atlantic Canada. ACENET was funded by the CFI, the provinces of New Brunswick, Newfoundland & Labrador, Nova Scotia and Prince Edward Island, as well as the Atlantic Canada Opportunities Agency. L.A.B. was supported by scholarships from the Canadian Institutes of Health Research and from Research Nova Scotia (formerly the Nova Scotia Health Research Foundation). F.B.A. was a trainee in the Cancer Research Training Program of the Beatrice Hunter Cancer Research Institute, with funds provided by the Dalhousie Medical Research Foundation's Rosetti Endowment. Finally, the authors wish to acknowledge Dalhousie University that is located in Mi'kma'ki, the ancestral and unceded territory of the Mi'kmaq.

Publisher Copyright:
© 2022 Wiley-VCH GmbH.

ASJC Scopus Subject Areas

General Chemistry
General Materials Science
Condensed Matter Physics

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10.1002/adfm.202205178

Cite this

@article{de58582bc1c74ef2b1493b82635e7d51,

title = "Nerve Growth Factor-Binding Engineered Silk Films Promote Neuronal Attachment and Neurite Outgrowth",

abstract = "Spider silks have outstanding potential as biomaterials due to their sought-after mechanical properties and low immunogenicity. The toughest spider silk is aciniform silk, which is used by spiders to wrap prey and produce egg sacs. A variety of recombinant aciniform silk constructs are now been developed, including hybrid silks with domains from multiple spider silk proteins fused together. In this study, an engineered aciniform silk construct, termed NBSilk, fused both N- and C-terminally to heptapeptide motifs that bind the neurotrophic factor and nerve growth factor-β (NGF) is introduced. NBSilk is shown to be amenable to casting into robust films that remain intact while sequestering and maintaining bioactive NGF at the film surface for at least 7 days. These films support cell survival while enhancing differentiation and neurite density and outgrowth in neuron-like PC12 cells, with elevation of signaling through both the mitogen-activated protein kinase (MAPK) and protein kinase B (AKT) signaling pathways. Strikingly, preloading of NBSilk films with NGF enhances neuritogenesis even over conditions where cells are grown with NGF supplementation in the culture medium. NBSilk scaffolds, thus, warrant future development and evaluation as biomaterials for nerve regeneration.",

author = "Baker, {Lizzy A.} and Lingling Xu and {Badichi Akher}, Farideh and Robertson, {Madelaine K.} and Lauren Pugsley-DeBruyn and Ma, {Chloe Xiaoyi} and Liu, {Xiang Qin} and Frampton, {John P.} and Rainey, {Jan K.}",

note = "Funding Information: This work was supported by funds from the Natural Sciences and Engineering Research Council of Canada (J.P.F., RGPIN/04298‐2016; X‐Q.L., RGPIN/06083‐2020; J.K.R., RGPIN/05907‐2017; J.K.R, J.P.F., and X‐Q.L. RTI/000030‐2020), the New Frontiers in Research Fund (J.P.F and J.K.R, NFRFE/2018‐00356), the Canada Research Chairs Program (J.P.F.), the Canada Foundation for Innovation (CFI – J.P.F., Project #33533; J.K.R., Project #15719), and Dalhousie University's Student Employment Program (J.K.R.). Advanced computing resources were provided by Compute Canada, the organization responsible for digital research infrastructure in Canada, and ACENET, the regional partner in Atlantic Canada. ACENET was funded by the CFI, the provinces of New Brunswick, Newfoundland & Labrador, Nova Scotia and Prince Edward Island, as well as the Atlantic Canada Opportunities Agency. L.A.B. was supported by scholarships from the Canadian Institutes of Health Research and from Research Nova Scotia (formerly the Nova Scotia Health Research Foundation). F.B.A. was a trainee in the Cancer Research Training Program of the Beatrice Hunter Cancer Research Institute, with funds provided by the Dalhousie Medical Research Foundation's Rosetti Endowment. Finally, the authors wish to acknowledge Dalhousie University that is located in Mi'kma'ki, the ancestral and unceded territory of the Mi'kmaq. Publisher Copyright: {\textcopyright} 2022 Wiley-VCH GmbH.",

year = "2022",

doi = "10.1002/adfm.202205178",

language = "English",

journal = "Advanced Functional Materials",

issn = "1616-301X",

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AU - Baker, Lizzy A.

AU - Xu, Lingling

AU - Badichi Akher, Farideh

AU - Robertson, Madelaine K.

AU - Pugsley-DeBruyn, Lauren

AU - Ma, Chloe Xiaoyi

AU - Liu, Xiang Qin

AU - Frampton, John P.

AU - Rainey, Jan K.

N1 - Funding Information: This work was supported by funds from the Natural Sciences and Engineering Research Council of Canada (J.P.F., RGPIN/04298‐2016; X‐Q.L., RGPIN/06083‐2020; J.K.R., RGPIN/05907‐2017; J.K.R, J.P.F., and X‐Q.L. RTI/000030‐2020), the New Frontiers in Research Fund (J.P.F and J.K.R, NFRFE/2018‐00356), the Canada Research Chairs Program (J.P.F.), the Canada Foundation for Innovation (CFI – J.P.F., Project #33533; J.K.R., Project #15719), and Dalhousie University's Student Employment Program (J.K.R.). Advanced computing resources were provided by Compute Canada, the organization responsible for digital research infrastructure in Canada, and ACENET, the regional partner in Atlantic Canada. ACENET was funded by the CFI, the provinces of New Brunswick, Newfoundland & Labrador, Nova Scotia and Prince Edward Island, as well as the Atlantic Canada Opportunities Agency. L.A.B. was supported by scholarships from the Canadian Institutes of Health Research and from Research Nova Scotia (formerly the Nova Scotia Health Research Foundation). F.B.A. was a trainee in the Cancer Research Training Program of the Beatrice Hunter Cancer Research Institute, with funds provided by the Dalhousie Medical Research Foundation's Rosetti Endowment. Finally, the authors wish to acknowledge Dalhousie University that is located in Mi'kma'ki, the ancestral and unceded territory of the Mi'kmaq. Publisher Copyright: © 2022 Wiley-VCH GmbH.

PY - 2022

Y1 - 2022

N2 - Spider silks have outstanding potential as biomaterials due to their sought-after mechanical properties and low immunogenicity. The toughest spider silk is aciniform silk, which is used by spiders to wrap prey and produce egg sacs. A variety of recombinant aciniform silk constructs are now been developed, including hybrid silks with domains from multiple spider silk proteins fused together. In this study, an engineered aciniform silk construct, termed NBSilk, fused both N- and C-terminally to heptapeptide motifs that bind the neurotrophic factor and nerve growth factor-β (NGF) is introduced. NBSilk is shown to be amenable to casting into robust films that remain intact while sequestering and maintaining bioactive NGF at the film surface for at least 7 days. These films support cell survival while enhancing differentiation and neurite density and outgrowth in neuron-like PC12 cells, with elevation of signaling through both the mitogen-activated protein kinase (MAPK) and protein kinase B (AKT) signaling pathways. Strikingly, preloading of NBSilk films with NGF enhances neuritogenesis even over conditions where cells are grown with NGF supplementation in the culture medium. NBSilk scaffolds, thus, warrant future development and evaluation as biomaterials for nerve regeneration.

AB - Spider silks have outstanding potential as biomaterials due to their sought-after mechanical properties and low immunogenicity. The toughest spider silk is aciniform silk, which is used by spiders to wrap prey and produce egg sacs. A variety of recombinant aciniform silk constructs are now been developed, including hybrid silks with domains from multiple spider silk proteins fused together. In this study, an engineered aciniform silk construct, termed NBSilk, fused both N- and C-terminally to heptapeptide motifs that bind the neurotrophic factor and nerve growth factor-β (NGF) is introduced. NBSilk is shown to be amenable to casting into robust films that remain intact while sequestering and maintaining bioactive NGF at the film surface for at least 7 days. These films support cell survival while enhancing differentiation and neurite density and outgrowth in neuron-like PC12 cells, with elevation of signaling through both the mitogen-activated protein kinase (MAPK) and protein kinase B (AKT) signaling pathways. Strikingly, preloading of NBSilk films with NGF enhances neuritogenesis even over conditions where cells are grown with NGF supplementation in the culture medium. NBSilk scaffolds, thus, warrant future development and evaluation as biomaterials for nerve regeneration.

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Nerve Growth Factor-Binding Engineered Silk Films Promote Neuronal Attachment and Neurite Outgrowth

Abstract

Bibliographical note

ASJC Scopus Subject Areas

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