Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

The correlations between structure, rheology, and cell growth in peptide-based multicomponent hydrogels

Abstract

Factorial analysis of the interactions between three hydrogel-forming peptides based on three different biological motifs, namely, fibronectin: Fmoc-GFFRGD, collagen: Fmoc-GFFGER, and laminin: Fmoc-DDIKVAV, was conducted through rheology and live cell imaging using L929 fibroblasts. Gels were formed from each of these three peptide gelators alone and in various combinations. Cellular growth was tracked for the first 48 h in time-lapse movies by counting fluorescent nuclei and segmenting the cell area. The correlation between cell growth and the gel structure was characterized by linear regression analysis. While all peptide combinations showed good biocompatibility, the single-component Fmoc-DDIKVAV gel had the most positive effect on cell growth, while Fmoc-GFFRGD was the least biocompatible and had the lowest growth rate and cell area. Linear regression modeling demonstrated possible negative and positive interactions between Fmoc-GFFRGD*Fmoc-DDIKVAV and Fmoc-GFFRGD*Fmoc-GFFGER, respectively. No correlation was observed between gel stiffness and cellular growth. However, an increase in the strain crossover point for the elastic and loss moduli was associated with greater cell proliferation. This may indicate that elastic gels that store the work of cell deformation during cytokinesis are required for proliferation.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  1. Modepalli VN, Rodriguez AL, Li R, Pavuluri S, Nicholas KR, Barrow CJ et al. In vitro response to functionalized self-assembled peptide scaffolds for three-dimensional cell culture. Biopolymers. 2014;102:197–205.

    Article  CAS  Google Scholar 

  2. Panda JJ, Dua R, Mishra A, Mittra B, Chauhan VS. 3D cell growth and proliferation on a RGD functionalized nanofibrillar hydrogel based on a conformationally restricted residue containing dipeptide. ACS Appl Mater Interfaces. 2010;2:2839–48.

    Article  CAS  Google Scholar 

  3. Farrukh A, Ortega F, Fan W, Marichal N, Paez JI, Berninger B et al. Bifunctional hydrogels containing the laminin motif IKVAV promote neurogenesis. Stem Cell Rep. 2017;9:1432–40.

    Article  CAS  Google Scholar 

  4. Paiva Dos Santos B, Garbay B, Pasqua M, Chevron E, Chinoy ZS, Cullin C et al. Production, purification and characterization of an elastin-like polypeptide containing the Ile-Lys-Val-Ala-Val (IKVAV) peptide for tissue engineering applications. J Biotechnol. 2019;298:35–44.

    Article  CAS  Google Scholar 

  5. Merrifield RB. Solid phase peptide synthesis. I. The synthesis of a tetrapeptide. J Am Chem Soc. 1963;85:2149–54.

    Article  CAS  Google Scholar 

  6. Martin AD, Robinson AB, Thordarson P. Biocompatible small peptide super-hydrogelators bearing carbazole functionalities. J Mat Chem B. 2015;3:2277–80.

    Article  CAS  Google Scholar 

  7. Nagy-Smith K, Beltramo PJ, Moore E, Tycko R, Furst EM, Schneider JP. Molecular, local, and network-level basis for the enhanced stiffness of hydrogel networks formed from coassembled racemic peptides: predictions from Pauling and Corey. ACS Cent Sci. 2017;3:586–97.

    Article  CAS  Google Scholar 

  8. Wu A, Isaacs L. Self-sorting: the exception or the rule? J Am Chem Soc. 2003;125:4831–5.

    Article  CAS  Google Scholar 

  9. Li X, Du X, Gao Y, Shi J, Kuang Y, Xu B. Supramolecular hydrogels formed by the conjugates of nucleobases, Arg-Gly-Asp (RGD) peptides, and glucosamine. Soft Matter. 2012;8:7402–7.

    Article  CAS  Google Scholar 

  10. Du X, Zhou J, Guvench O, Sangiorgi FO, Li X, Zhou N et al. Supramolecular assemblies of a conjugate of nucleobase, amino acids, and saccharide act as agonists for proliferation of embryonic stem cells and development of zygotes. Bioconjug Chem. 2014;25:1031–5.

    Article  CAS  Google Scholar 

  11. Raeburn J, Adams DJ. Multicomponent low molecular weight gelators. Chem Commun. 2015;51:5170–80.

    Article  CAS  Google Scholar 

  12. Horgan CC, Rodriguez AL, Li R, Bruggeman KF, Stupka N, Raynes JK et al. Characterisation of minimalist co-assembled fluorenylmethyloxycarbonyl self-assembling peptide systems for presentation of multiple bioactive peptides. Acta Biomat. 2016;38:11–22.

    Article  CAS  Google Scholar 

  13. Zhou M, Ulijn RV, Gough JE. Extracellular matrix formation in self-assembled minimalistic bioactive hydrogels based on aromatic peptide amphiphiles. J Tissue Eng. 2014;5:2041731414531593.

    Article  Google Scholar 

  14. Niece KL, Hartgerink JD, Donners JJ, Stupp SI. Self-assembly combining two bioactive peptide-amphiphile molecules into nanofibers by electrostatic attraction. J Am Chem Soc. 2003;125:7146–7.

    Article  CAS  Google Scholar 

  15. Liyanage W, Vats K, Rajbhandary A, Benoit DS, Nilsson BL. Multicomponent dipeptide hydrogels as extracellular matrix-mimetic scaffolds for cell culture applications. Chem Commun. 2015;51:11260–3.

    Article  CAS  Google Scholar 

  16. Wojciechowski JP, Martin AD, Du EY, Garvey CJ, Nordon RE, Thordarson P. Non-reversible heat-induced gelation of a biocompatible Fmochexapeptide in water. Nanoscale. 2020;12:8262–7.

    Article  CAS  Google Scholar 

  17. Zhou P, Yin B, Zhang R, Xu Z, Liu Y, Yan Y et al. Molecular basis for RGD-containing peptides supporting adhesion and self-renewal of human pluripotent stem cells on synthetic surface. Colloids Surf B Biointerfaces. 2018;171:451–60.

    Article  CAS  Google Scholar 

  18. Choe G, Park J, Jo H, Kim YS, Ahn Y, Lee JY. Studies on the effects of microencapsulated human mesenchymal stem cells in RGD-modified alginate on cardiomyocytes under oxidative stress conditions using in vitro biomimetic co-culture system. Int J Biol Macromol. 2019;123:512–20.

    Article  CAS  Google Scholar 

  19. Munnix IC, Gilio K, Siljander PR, Raynal N, Feijge MA, Hackeng TM et al. Collagen-mimetic peptides mediate flow-dependent thrombus formation by high- or low-affinity binding of integrin alpha2beta1 and glycoprotein VI. J Thromb Haemost. 2008;6:2132–42.

    Article  CAS  Google Scholar 

  20. Farndale RW, Lisman T, Bihan D, Hamaia S, Smerling CS, Pugh N et al. Cell-collagen interactions: the use of peptide Toolkits to investigate collagen-receptor interactions. Biochem Soc Trans. 2008;36:241–50.

    Article  CAS  Google Scholar 

  21. Somaa FA, Wang TY, Niclis JC, Bruggeman KF, Kauhausen JA, Guo H et al. Peptide-based Scaffolds support human cortical progenitor graft integration to reduce atrophy and promote functional repair in a model of stroke. Cell Rep. 2017;20:1964–77.

    Article  CAS  Google Scholar 

  22. Rodrigues T, Kundu B, Silva-Correia J, Kundu SC, Oliveira JM, Reis RL et al. Emerging tumor spheroids technologies for 3D in vitro cancer modeling. Pharm Ther. 2018;184:201–11.

    Article  CAS  Google Scholar 

  23. Engler AJ, Sen S, Sweeney HL, Discher DE. Matrix elasticity directs stem cell lineage specification. Cell 2006;126:677–89.

    Article  CAS  Google Scholar 

  24. Das RK, Gocheva V, Hammink R, Zouani OF, Rowan AE. Stress-stiffening-mediated stem-cell commitment switch in soft responsive hydrogels. Nat Mater. 2016;15:318–25.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank the Australian Research Council (ARC) for a Linkage Grant (LP160100573) to REN and PT, support from the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology (CE140100036) to PT and the Australian Government for a PhD scholarship to EYD.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Robert E. Nordon or Pall Thordarson.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Du, E.Y., Ziaee, F., Wang, L. et al. The correlations between structure, rheology, and cell growth in peptide-based multicomponent hydrogels. Polym J 52, 947–957 (2020). https://doi.org/10.1038/s41428-020-0351-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41428-020-0351-8

This article is cited by

Search

Quick links