Modulation of the mechanical properties of ventricular extracellular matrix hydrogels with a carbodiimide crosslinker and investigation of their cellular compatibility

Kyohei Fujita, Zhonggang Feng, Daisuke Sato, Tadashi Kosawada, Takao Nakamura, Yasuyuki Shiraishi, Mitsuo Umezu

    Research output: Contribution to journalArticle

    Abstract

    Hydrogels made from the cardiac extracellular matrix (ECM) as two-dimensional (2D) or 3D cell-culture substrates have beneficial biochemical effects on the differentiation of stem cells into cardiomyocytes. The mechanical properties of the substrates that match those of the host tissues have been identified as critical biophysical cues for coaxing the tissue-specific differentiation of stem cells. The objectives of the present study are (1) to fabricate hydrogels comprising pure ventricular ECM (vECM), (2) to make the gels possess mechanical properties similar to those of the decellularized ventricular tissue, and (3) to evaluate the cellular compatibility of the hydrogels. In order to achieve these aims, (1) a simplified protocol was developed to produce vECM solution easily and rapidly, (2) N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDAC) was chosen to crosslink the hydrogels made from the vECM solution to enhance their mechanical properties and stabilize the microstructure of the gels, (3) rat embryonic fibroblasts or cardiomyocytes were cultured on these gels to determine the cellular compatibility of the gels. In particular, the nonlinearity and viscoelasticity of the gels were characterized quantitatively using a newly proposed nonlinear Kelvin model. The results showed that EDAC treatment allowed modulation of the mechanical properties of the gels to the same level as those of decellularized ventricular tissue in terms of the equilibrium elasticity and relaxation coefficient. Cell culture confirmed the cellular compatibility of the gels. Furthermore, an empirical relationship between the equilibrium elastic modulus of the gels and the vECM and EDAC concentrations was derived, which is important to tailor the mechanical properties of the gels. Finally, the influence of the mechanical properties of the gels on the behavior of cultured fibroblasts and cardiomyocytes was discussed.

    Original languageEnglish
    Pages (from-to)54-74
    Number of pages21
    JournalAIMS Materials Science
    Volume5
    Issue number1
    DOIs
    Publication statusPublished - 2018 Jan 1

    Fingerprint

    Carbodiimides
    Hydrogels
    Gels
    Modulation
    Mechanical properties
    Military electronic countermeasures
    Tissue
    Fibroblasts
    Stem cells
    Cell culture
    Viscoelasticity
    Substrates
    Rats
    Elasticity
    Elastic moduli

    Keywords

    • Cellular biocompatibility
    • Decellularization
    • Hydrogel
    • Nonlinearity
    • Ventricular extracellular matrix
    • Viscoelasticity

    ASJC Scopus subject areas

    • Materials Science(all)

    Cite this

    Modulation of the mechanical properties of ventricular extracellular matrix hydrogels with a carbodiimide crosslinker and investigation of their cellular compatibility. / Fujita, Kyohei; Feng, Zhonggang; Sato, Daisuke; Kosawada, Tadashi; Nakamura, Takao; Shiraishi, Yasuyuki; Umezu, Mitsuo.

    In: AIMS Materials Science, Vol. 5, No. 1, 01.01.2018, p. 54-74.

    Research output: Contribution to journalArticle

    Fujita, Kyohei ; Feng, Zhonggang ; Sato, Daisuke ; Kosawada, Tadashi ; Nakamura, Takao ; Shiraishi, Yasuyuki ; Umezu, Mitsuo. / Modulation of the mechanical properties of ventricular extracellular matrix hydrogels with a carbodiimide crosslinker and investigation of their cellular compatibility. In: AIMS Materials Science. 2018 ; Vol. 5, No. 1. pp. 54-74.
    @article{2e80e1cec8a84127bb1f4b58dd26dc61,
    title = "Modulation of the mechanical properties of ventricular extracellular matrix hydrogels with a carbodiimide crosslinker and investigation of their cellular compatibility",
    abstract = "Hydrogels made from the cardiac extracellular matrix (ECM) as two-dimensional (2D) or 3D cell-culture substrates have beneficial biochemical effects on the differentiation of stem cells into cardiomyocytes. The mechanical properties of the substrates that match those of the host tissues have been identified as critical biophysical cues for coaxing the tissue-specific differentiation of stem cells. The objectives of the present study are (1) to fabricate hydrogels comprising pure ventricular ECM (vECM), (2) to make the gels possess mechanical properties similar to those of the decellularized ventricular tissue, and (3) to evaluate the cellular compatibility of the hydrogels. In order to achieve these aims, (1) a simplified protocol was developed to produce vECM solution easily and rapidly, (2) N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDAC) was chosen to crosslink the hydrogels made from the vECM solution to enhance their mechanical properties and stabilize the microstructure of the gels, (3) rat embryonic fibroblasts or cardiomyocytes were cultured on these gels to determine the cellular compatibility of the gels. In particular, the nonlinearity and viscoelasticity of the gels were characterized quantitatively using a newly proposed nonlinear Kelvin model. The results showed that EDAC treatment allowed modulation of the mechanical properties of the gels to the same level as those of decellularized ventricular tissue in terms of the equilibrium elasticity and relaxation coefficient. Cell culture confirmed the cellular compatibility of the gels. Furthermore, an empirical relationship between the equilibrium elastic modulus of the gels and the vECM and EDAC concentrations was derived, which is important to tailor the mechanical properties of the gels. Finally, the influence of the mechanical properties of the gels on the behavior of cultured fibroblasts and cardiomyocytes was discussed.",
    keywords = "Cellular biocompatibility, Decellularization, Hydrogel, Nonlinearity, Ventricular extracellular matrix, Viscoelasticity",
    author = "Kyohei Fujita and Zhonggang Feng and Daisuke Sato and Tadashi Kosawada and Takao Nakamura and Yasuyuki Shiraishi and Mitsuo Umezu",
    year = "2018",
    month = "1",
    day = "1",
    doi = "10.3934/matersci.2018.1.54",
    language = "English",
    volume = "5",
    pages = "54--74",
    journal = "AIMS Materials Science",
    issn = "2372-0484",
    publisher = "AIMS Press",
    number = "1",

    }

    TY - JOUR

    T1 - Modulation of the mechanical properties of ventricular extracellular matrix hydrogels with a carbodiimide crosslinker and investigation of their cellular compatibility

    AU - Fujita, Kyohei

    AU - Feng, Zhonggang

    AU - Sato, Daisuke

    AU - Kosawada, Tadashi

    AU - Nakamura, Takao

    AU - Shiraishi, Yasuyuki

    AU - Umezu, Mitsuo

    PY - 2018/1/1

    Y1 - 2018/1/1

    N2 - Hydrogels made from the cardiac extracellular matrix (ECM) as two-dimensional (2D) or 3D cell-culture substrates have beneficial biochemical effects on the differentiation of stem cells into cardiomyocytes. The mechanical properties of the substrates that match those of the host tissues have been identified as critical biophysical cues for coaxing the tissue-specific differentiation of stem cells. The objectives of the present study are (1) to fabricate hydrogels comprising pure ventricular ECM (vECM), (2) to make the gels possess mechanical properties similar to those of the decellularized ventricular tissue, and (3) to evaluate the cellular compatibility of the hydrogels. In order to achieve these aims, (1) a simplified protocol was developed to produce vECM solution easily and rapidly, (2) N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDAC) was chosen to crosslink the hydrogels made from the vECM solution to enhance their mechanical properties and stabilize the microstructure of the gels, (3) rat embryonic fibroblasts or cardiomyocytes were cultured on these gels to determine the cellular compatibility of the gels. In particular, the nonlinearity and viscoelasticity of the gels were characterized quantitatively using a newly proposed nonlinear Kelvin model. The results showed that EDAC treatment allowed modulation of the mechanical properties of the gels to the same level as those of decellularized ventricular tissue in terms of the equilibrium elasticity and relaxation coefficient. Cell culture confirmed the cellular compatibility of the gels. Furthermore, an empirical relationship between the equilibrium elastic modulus of the gels and the vECM and EDAC concentrations was derived, which is important to tailor the mechanical properties of the gels. Finally, the influence of the mechanical properties of the gels on the behavior of cultured fibroblasts and cardiomyocytes was discussed.

    AB - Hydrogels made from the cardiac extracellular matrix (ECM) as two-dimensional (2D) or 3D cell-culture substrates have beneficial biochemical effects on the differentiation of stem cells into cardiomyocytes. The mechanical properties of the substrates that match those of the host tissues have been identified as critical biophysical cues for coaxing the tissue-specific differentiation of stem cells. The objectives of the present study are (1) to fabricate hydrogels comprising pure ventricular ECM (vECM), (2) to make the gels possess mechanical properties similar to those of the decellularized ventricular tissue, and (3) to evaluate the cellular compatibility of the hydrogels. In order to achieve these aims, (1) a simplified protocol was developed to produce vECM solution easily and rapidly, (2) N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDAC) was chosen to crosslink the hydrogels made from the vECM solution to enhance their mechanical properties and stabilize the microstructure of the gels, (3) rat embryonic fibroblasts or cardiomyocytes were cultured on these gels to determine the cellular compatibility of the gels. In particular, the nonlinearity and viscoelasticity of the gels were characterized quantitatively using a newly proposed nonlinear Kelvin model. The results showed that EDAC treatment allowed modulation of the mechanical properties of the gels to the same level as those of decellularized ventricular tissue in terms of the equilibrium elasticity and relaxation coefficient. Cell culture confirmed the cellular compatibility of the gels. Furthermore, an empirical relationship between the equilibrium elastic modulus of the gels and the vECM and EDAC concentrations was derived, which is important to tailor the mechanical properties of the gels. Finally, the influence of the mechanical properties of the gels on the behavior of cultured fibroblasts and cardiomyocytes was discussed.

    KW - Cellular biocompatibility

    KW - Decellularization

    KW - Hydrogel

    KW - Nonlinearity

    KW - Ventricular extracellular matrix

    KW - Viscoelasticity

    UR - http://www.scopus.com/inward/record.url?scp=85042780010&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=85042780010&partnerID=8YFLogxK

    U2 - 10.3934/matersci.2018.1.54

    DO - 10.3934/matersci.2018.1.54

    M3 - Article

    VL - 5

    SP - 54

    EP - 74

    JO - AIMS Materials Science

    JF - AIMS Materials Science

    SN - 2372-0484

    IS - 1

    ER -