Screening of Self-Assembling of Collagen IV Fragments into Stable Structures Potentially Useful in Regenerative Medicine
The aim of the research was to check whether it is possible to use fragments of type IV collagen to obtain, as a result of self-assembling, stable spatial structures that could be used to prepare new materials useful in regenerative medicine. Collagen IV fragments were obtained by using DMT/NMM/TosO...
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| Format: | Article |
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MDPI AG
2021-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/24/13584 |
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| author | Marcin Kolasa Grzegorz Galita Ireneusz Majsterek Ewa Kucharska Katarzyna Czerczak Joanna Wasko Angelika Becht Justyna Fraczyk Anna Gajda Lukasz Pietrzak Lukasz Szymanski Agnieszka Krakowiak Zbigniew Draczynski Beata Kolesinska |
| author_facet | Marcin Kolasa Grzegorz Galita Ireneusz Majsterek Ewa Kucharska Katarzyna Czerczak Joanna Wasko Angelika Becht Justyna Fraczyk Anna Gajda Lukasz Pietrzak Lukasz Szymanski Agnieszka Krakowiak Zbigniew Draczynski Beata Kolesinska |
| author_sort | Marcin Kolasa |
| collection | DOAJ |
| description | The aim of the research was to check whether it is possible to use fragments of type IV collagen to obtain, as a result of self-assembling, stable spatial structures that could be used to prepare new materials useful in regenerative medicine. Collagen IV fragments were obtained by using DMT/NMM/TosO<sup>−</sup> as a coupling reagent. The ability to self-organize and form stable spatial structures was tested by the CD method and microscopic techniques. Biological studies covered: resazurin assay (cytotoxicity assessment) on BJ, BJ-5TA and C2C12 cell lines; an alkaline version of the comet assay (genotoxicity), Biolegend Legendplex human inflammation panel 1 assay (SC cell lines, assessment of the inflammation activity) and MTT test to determine the cytotoxicity of the porous materials based on collagen IV fragments. It was found that out of the pool of 37 fragments (peptides <b>1</b>–<b>33</b> and <b>2.1</b>–<b>2.4</b>) reconstructing the outer sphere of collagen IV, nine fragments (peptides: <b>2</b>, <b>4</b>, <b>5</b>, <b>6</b>, <b>14</b>, <b>15</b>, <b>25</b>, <b>26</b> and <b>30</b>), as a result of self-assembling, form structures mimicking the structure of the triple helix of native collagens. The stability of spatial structures formed as a result of self-organization at temperatures of 4 °C, 20 °C, and 40 °C was found. The application of the MST method allowed us to determine the K<sub>d</sub> of binding of selected fragments of collagen IV to ITGα1β1. The stability of the spatial structures of selected peptides made it possible to obtain porous materials based on their equimolar mixture. The formation of the porous materials was found for cross-linked structures and the material stabilized only by weak interactions. All tested peptides are non-cytotoxic against all tested cell lines. Selected peptides also showed no genotoxicity and no induction of immune system responses. Research on the use of porous materials based on fragments of type IV collagen, able to form stable spatial structures as scaffolds useful in regenerative medicine, will be continued. |
| first_indexed | 2024-03-10T03:54:49Z |
| format | Article |
| id | doaj.art-9a14a48f361f45e8bca227edf3de6991 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T03:54:49Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-9a14a48f361f45e8bca227edf3de69912023-11-23T08:48:34ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-12-0122241358410.3390/ijms222413584Screening of Self-Assembling of Collagen IV Fragments into Stable Structures Potentially Useful in Regenerative MedicineMarcin Kolasa0Grzegorz Galita1Ireneusz Majsterek2Ewa Kucharska3Katarzyna Czerczak4Joanna Wasko5Angelika Becht6Justyna Fraczyk7Anna Gajda8Lukasz Pietrzak9Lukasz Szymanski10Agnieszka Krakowiak11Zbigniew Draczynski12Beata Kolesinska13General Command of the Polish Armed Forces, Medical Division, Zwirki i Wigury 103/105, 00-912 Warsaw, PolandDepartment of Clinical Chemistry and Biochemistry, Medical University of Lodz, Narutowicza 60, 90-136 Lodz, PolandDepartment of Clinical Chemistry and Biochemistry, Medical University of Lodz, Narutowicza 60, 90-136 Lodz, PolandDepartment Geriatrics and Social Work, Jesuit University Ignatianum in Cracow, Kopernika 26, 31-501 Krakow, PolandInstitute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, PolandInstitute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, PolandInstitute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, PolandInstitute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, PolandInstitute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, PolandInstitute of Mechatronics and Information Systems, Faculty of Electrical, Electronic, Computer and Control Engineering, Lodz University of Technology, Stefanowskiego 18/22, 90-924 Lodz, PolandInstitute of Mechatronics and Information Systems, Faculty of Electrical, Electronic, Computer and Control Engineering, Lodz University of Technology, Stefanowskiego 18/22, 90-924 Lodz, PolandCentre of Molecular and Macromolecular Studies Polish Academy of Sciences, Department of Bioorganic Chemistry, Sienkiewicza 112, 90-363 Lodz, PolandInstitute of Material Sciences of Textiles and Polymer Composites, Faculty of Material Technologies and Textile Design, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, PolandInstitute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego 116, 90-924 Lodz, PolandThe aim of the research was to check whether it is possible to use fragments of type IV collagen to obtain, as a result of self-assembling, stable spatial structures that could be used to prepare new materials useful in regenerative medicine. Collagen IV fragments were obtained by using DMT/NMM/TosO<sup>−</sup> as a coupling reagent. The ability to self-organize and form stable spatial structures was tested by the CD method and microscopic techniques. Biological studies covered: resazurin assay (cytotoxicity assessment) on BJ, BJ-5TA and C2C12 cell lines; an alkaline version of the comet assay (genotoxicity), Biolegend Legendplex human inflammation panel 1 assay (SC cell lines, assessment of the inflammation activity) and MTT test to determine the cytotoxicity of the porous materials based on collagen IV fragments. It was found that out of the pool of 37 fragments (peptides <b>1</b>–<b>33</b> and <b>2.1</b>–<b>2.4</b>) reconstructing the outer sphere of collagen IV, nine fragments (peptides: <b>2</b>, <b>4</b>, <b>5</b>, <b>6</b>, <b>14</b>, <b>15</b>, <b>25</b>, <b>26</b> and <b>30</b>), as a result of self-assembling, form structures mimicking the structure of the triple helix of native collagens. The stability of spatial structures formed as a result of self-organization at temperatures of 4 °C, 20 °C, and 40 °C was found. The application of the MST method allowed us to determine the K<sub>d</sub> of binding of selected fragments of collagen IV to ITGα1β1. The stability of the spatial structures of selected peptides made it possible to obtain porous materials based on their equimolar mixture. The formation of the porous materials was found for cross-linked structures and the material stabilized only by weak interactions. All tested peptides are non-cytotoxic against all tested cell lines. Selected peptides also showed no genotoxicity and no induction of immune system responses. Research on the use of porous materials based on fragments of type IV collagen, able to form stable spatial structures as scaffolds useful in regenerative medicine, will be continued.https://www.mdpi.com/1422-0067/22/24/13584collagen IVbasement membraneself-assemblingability to mimic native collagen structuresporous materialsolid phase peptide synthesis |
| spellingShingle | Marcin Kolasa Grzegorz Galita Ireneusz Majsterek Ewa Kucharska Katarzyna Czerczak Joanna Wasko Angelika Becht Justyna Fraczyk Anna Gajda Lukasz Pietrzak Lukasz Szymanski Agnieszka Krakowiak Zbigniew Draczynski Beata Kolesinska Screening of Self-Assembling of Collagen IV Fragments into Stable Structures Potentially Useful in Regenerative Medicine International Journal of Molecular Sciences collagen IV basement membrane self-assembling ability to mimic native collagen structures porous material solid phase peptide synthesis |
| title | Screening of Self-Assembling of Collagen IV Fragments into Stable Structures Potentially Useful in Regenerative Medicine |
| title_full | Screening of Self-Assembling of Collagen IV Fragments into Stable Structures Potentially Useful in Regenerative Medicine |
| title_fullStr | Screening of Self-Assembling of Collagen IV Fragments into Stable Structures Potentially Useful in Regenerative Medicine |
| title_full_unstemmed | Screening of Self-Assembling of Collagen IV Fragments into Stable Structures Potentially Useful in Regenerative Medicine |
| title_short | Screening of Self-Assembling of Collagen IV Fragments into Stable Structures Potentially Useful in Regenerative Medicine |
| title_sort | screening of self assembling of collagen iv fragments into stable structures potentially useful in regenerative medicine |
| topic | collagen IV basement membrane self-assembling ability to mimic native collagen structures porous material solid phase peptide synthesis |
| url | https://www.mdpi.com/1422-0067/22/24/13584 |
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