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.

Activating an adaptive immune response from a hydrogel scaffold imparts regenerative wound healing

Abstract

Microporous annealed particle (MAP) scaffolds are flowable, in situ crosslinked, microporous scaffolds composed of microgel building blocks and were previously shown to accelerate wound healing. To promote more extensive tissue ingrowth before scaffold degradation, we aimed to slow MAP degradation by switching the chirality of the crosslinking peptides from l- to d-amino acids. Unexpectedly, despite showing the predicted slower enzymatic degradation in vitro, d-peptide crosslinked MAP hydrogel (d-MAP) hastened material degradation in vivo and imparted significant tissue regeneration to healed cutaneous wounds, including increased tensile strength and hair neogenesis. MAP scaffolds recruit IL-33 type 2 myeloid cells, which is amplified in the presence of d-peptides. Remarkably, d-MAP elicited significant antigen-specific immunity against the d-chiral peptides, and an intact adaptive immune system was required for the hydrogel-induced skin regeneration. These findings demonstrate that the generation of an adaptive immune response from a biomaterial is sufficient to induce cutaneous regenerative healing despite faster scaffold degradation.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Fig. 1: d-MAP hydrogel degradation is enhanced in wounds of SKH1 hairless mice.
Fig. 2: d-MAP hydrogel induces neogenesis of hair follicles in full-thickness skin wounds in B6 mice.
Fig. 3: Peptide recognition by pattern recognition receptors is not required for myeloid cell recruitment.
Fig. 4: d-MAP induces antibody responses and the recruitment of myeloid cells via adaptive immunity.
Fig. 5: d-MAP requires an intact adaptive immunity to induce hair follicle neogenesis.
Fig. 6: d-MAP changes the wound fate from scar formation to regeneration by type 2 immune activation.

Data availability

The data that support the findings of this study are available from the corresponding authors upon reasonable request. Source data are provided with this paper.

References

  1. 1.

    Griffin, D. R., Weaver, W. M., Scumpia, P., Di Carlo, D. & Segura, T. Accelerated wound healing by injectable microporous gel scaffolds assembled from annealed building blocks. Nat. Mater. 14, 737–744 (2015).

    CAS  Google Scholar 

  2. 2.

    Nih, L. R., Sideris, E., Carmichael, S. T. & Segura, T. Injection of microporous annealing particle (MAP) hydrogels in the stroke cavity reduces gliosis and inflammation and promotes NPC migration to the lesion. Adv. Mater. 29, 1606471 (2017).

    Google Scholar 

  3. 3.

    Xu, Q. et al. Injectable hyperbranched poly(β-amino ester) hydrogels with on-demand degradation profiles to match wound healing processes. Chem. Sci. 9, 2179–2187 (2018).

    CAS  Google Scholar 

  4. 4.

    Zhu, S., Nih, L., Carmichael, S. T., Lu, Y. & Segura, T. Enzyme-responsive delivery of multiple proteins with spatiotemporal control. Adv. Mater. 27, 3620–3625 (2015).

    CAS  Google Scholar 

  5. 5.

    Sela, M. & Zisman, E. Different roles of d-amino acids in immune phenomena. FASEB J. 11, 449–456 (1997).

    CAS  Google Scholar 

  6. 6.

    Doloff, J. C. et al. Colony stimulating factor-1 receptor is a central component of the foreign body response to biomaterial implants in rodents and non-human primates. Nat. Mater. 16, 671–680 (2017).

    CAS  Google Scholar 

  7. 7.

    Mishra, P. K. et al. Sterile particle-induced inflammation is mediated by macrophages releasing IL-33 through a Bruton’s tyrosine kinase-dependent pathway. Nat. Mater. 18, 289–297 (2019).

    CAS  Google Scholar 

  8. 8.

    Chung, L., Maestas, D. R., Housseau, F. & Elisseeff, J. H. Key players in the immune response to biomaterial scaffolds for regenerative medicine. Adv. Drug Deliv. Rev. 114, 184–192 (2017).

    CAS  Google Scholar 

  9. 9.

    Sadtler, K. et al. Developing a pro-regenerative biomaterial scaffold microenvironment requires T helper 2 cells. Science 352, 366–370 (2016).

    CAS  Google Scholar 

  10. 10.

    Galiano, R. D., Michaels, J., Dobryansky, M., Levine, J. P. & Gurtner, G. C. Quantitative and reproducible murine model of excisional wound healing. Wound Repair Regen 12, 485–492 (2004).

    Google Scholar 

  11. 11.

    Ito, M. et al. Wnt-dependent de novo hair follicle regeneration in adult mouse skin after wounding. Nature 447, 316–320 (2007).

    CAS  Google Scholar 

  12. 12.

    Seifert, A. W. et al. Skin shedding and tissue regeneration in African spiny mice (Acomys). Nature 489, 561–565 (2012).

    CAS  Google Scholar 

  13. 13.

    Nelson, A. M. et al. dsRNA released by tissue damage activates TLR3 to drive skin regeneration. Cell Stem Cell 17, 139–151 (2015).

    CAS  Google Scholar 

  14. 14.

    Guerrero-Juarez, C. F. et al. Wound regeneration deficit in rats correlates with low morphogenetic potential and distinct transcriptome profile of epidermis. J. Invest. Dermatol. 138, 1409–1419 (2018).

    CAS  Google Scholar 

  15. 15.

    Marshall, C. D. et al. Sanativo wound healing product does not accelerate reepithelialization in a mouse cutaneous wound healing model. Plast. Reconstr. Surg. 139, 343–352 (2017).

    CAS  Google Scholar 

  16. 16.

    Lim, C. H. et al. Hedgehog stimulates hair follicle neogenesis by creating inductive dermis during murine skin wound healing. Nat. Commun. 9, 4903 (2018).

    Google Scholar 

  17. 17.

    Carlson, M. A. & Chakkalakal, D. Tensile properties of the murine ventral vertical midline incision. PLoS ONE 6, e24212 (2011).

    CAS  Google Scholar 

  18. 18.

    Plikus, M. V. et al. Regeneration of fat cells from myofibroblasts during wound healing. Science 355, 748–752 (2017).

    CAS  Google Scholar 

  19. 19.

    Guerrero-Juarez, C. F. et al. Single-cell analysis reveals fibroblast heterogeneity and myeloid-derived adipocyte progenitors in murine skin wounds. Nat. Commun. 10, 650 (2019).

    CAS  Google Scholar 

  20. 20.

    Warren, K. S. A functional classification of granulomatous inflammation. Ann. NY Acad. Sci. 278, 7–18 (1976).

    CAS  Google Scholar 

  21. 21.

    Chensue, S. W. et al. Cytokine responses during mycobacterial and schistosomal antigen-induced pulmonary granuloma formation. Production of Th1 and Th2 cytokines and relative contribution of tumor necrosis factor. Am. J. Pathol. 145, 1105–1113 (1994).

    CAS  Google Scholar 

  22. 22.

    Wills-Karp, M. et al. Trefoil factor 2 rapidly induces interleukin 33 to promote type 2 immunity during allergic asthma and hookworm infection. J. Exp. Med. 209, 607–622 (2012).

    CAS  Google Scholar 

  23. 23.

    Hardman, C. S., Panova, V. & McKenzie, A. N. J. IL-33 citrine reporter mice reveal the temporal and spatial expression of IL-33 during allergic lung inflammation. Eur. J. Immunol. 43, 488–498 (2013).

    CAS  Google Scholar 

  24. 24.

    de Kouchkovsky, D. A., Ghosh, S. & Rothlin, C. V. Induction of sterile type 2 inflammation. Nat. Mater. 18, 193–194 (2019).

    Google Scholar 

  25. 25.

    Koh, J. et al. Enhanced in vivo delivery of stem cells using microporous annealed particle scaffolds. Small 15, e1903147 (2019).

    Google Scholar 

  26. 26.

    Purbey, P. K. et al. Defined sensing mechanisms and signaling pathways contribute to the global inflammatory gene expression output elicited by ionizing radiation. Immunity 47, 421–434 (2017).

    CAS  Google Scholar 

  27. 27.

    Scumpia, P. O. et al. Opposing roles of Toll-like receptor and cytosolic DNA-STING signaling pathways for Staphylococcus aureus cutaneous host defense. PLoS Pathog. 13, e1006496 (2017).

    Google Scholar 

  28. 28.

    Tong, A.-J. et al. A stringent systems approach uncovers gene-specific mechanisms regulating inflammation. Cell 165, 165–179 (2016).

    CAS  Google Scholar 

  29. 29.

    Kim, S. D. et al. The agonists of formyl peptide receptors prevent development of severe sepsis after microbial infection. J. Immunol. 185, 4302–4310 (2010).

    CAS  Google Scholar 

  30. 30.

    Kang, H. K. et al. The synthetic peptide Trp-Lys-Tyr-Met-Val-d-Met inhibits human monocyte-derived dendritic cell maturation via formyl peptide receptor and formyl peptide receptor-like 2. J. Immunol. 175, 685–692 (2005).

    CAS  Google Scholar 

  31. 31.

    Schepetkin, I. A. et al. 3-(1 H-indol-3-yl)-2-[3-(4-nitrophenyl)ureido]propanamide enantiomers with human formyl-peptide receptor agonist activity: molecular modeling of chiral recognition by FPR2. Biochem. Pharmacol. 85, 404–416 (2013).

    CAS  Google Scholar 

  32. 32.

    Zisman, E., Dayan, M., Sela, M. & Mozes, E. Ia-antigen–T-cell interactions for a thymus-independent antigen composed of d amino acids. Proc. Natl Acad. Sci. USA 90, 994–998 (1993).

    CAS  Google Scholar 

  33. 33.

    Cernysiov, V., Gerasimcik, N., Mauricas, M. & Girkontaite, I. Regulation of T-cell-independent and T-cell-dependent antibody production by circadian rhythm and melatonin. Int. Immunol. 22, 25–34 (2010).

    CAS  Google Scholar 

  34. 34.

    Honda, S. et al. Enhanced humoral immune responses against T-independent antigens in Fc alpha/muR-deficient mice. Proc. Natl Acad. Sci. USA 106, 11230–11235 (2009).

    CAS  Google Scholar 

  35. 35.

    Mongini, P. K., Stein, K. E. & Paul, W. E. T cell regulation of IgG subclass antibody production in response to T-independent antigens. J. Exp. Med. 153, 1–12 (1981).

    CAS  Google Scholar 

  36. 36.

    Weinstein, J. S. et al. Maintenance of anti-Sm/RNP autoantibody production by plasma cells residing in ectopic lymphoid tissue and bone marrow memory B cells. J. Immunol. 190, 3916–3927 (2013).

    CAS  Google Scholar 

  37. 37.

    Germann, T. et al. Interleukin-12 profoundly up-regulates the synthesis of antigen-specific complement-fixing IgG2a, IgG2b and IgG3 antibody subclasses in vivo. Eur. J. Immunol. 25, 823–829 (1995).

    CAS  Google Scholar 

  38. 38.

    Boehler, R. M., Graham, J. G. & Shea, L. D. Tissue engineering tools for modulation of the immune response. BioTechniques 51, 239–254 (2011).

    CAS  Google Scholar 

  39. 39.

    Song, J. et al. A mouse model for the human pathogen Salmonella typhi. Cell Host Microbe 8, 369–376 (2010).

    CAS  Google Scholar 

  40. 40.

    Park, C. G. et al. Extended release of perioperative immunotherapy prevents tumor recurrence and eliminates metastases. Sci. Transl. Med. 10, eaar1916 (2018).

    Google Scholar 

  41. 41.

    Kim, J. et al. Injectable, spontaneously assembling, inorganic scaffolds modulate immune cells in vivo and increase vaccine efficacy. Nat. Biotechnol. 33, 64–72 (2015).

    CAS  Google Scholar 

  42. 42.

    Ramirez-Carrozzi, V. R. et al. A unifying model for the selective regulation of inducible transcription by CpG islands and nucleosome remodeling. Cell 138, 114–128 (2009).

    CAS  Google Scholar 

Download references

Acknowledgements

We thank the National Institutes of Health F32EB018713-01A1 (D.R.G.), T32-GM008042 (M.M.A.), T32AR071307 (M.M.A), U01AR073159 (M.V.P.), R01NS094599 (T.S.), R01HL110592 (T.S.), R03AR073940 (P.O.S.), K08AR066545 (P.O.S.), Pew Charitable Trust (M.V.P.), LEO Foundation (M.V.P.), the National Science Foundation grant DMS1763272, Simons Foundation Grant (594598, QN) (M.V.P.), and the Presidential Early Career Award for Scientists and Engineers (N00014-16-1-2997) (D.D.) for funding. We thank S. C. Lesher-Perez and M. Bogumil for their assistance with MATLAB coding. We thank Y. Liu for assistance with running the endotoxin texts. We also thank the Advanced Light Microscopy and Spectroscopy at California NanoSystems Institute and Electron Microscopy Core Laboratory of the Brain Research Institute at UCLA and, particularly, for the significant help of M. Cilluffo.

Author information

Affiliations

Authors

Contributions

D.R.G., P.O.S. and T.S. conceived the experiments. D.R.G., W.M.W., E.S., M.M.A. and J.K. carried out the microfluidic design and fabrication, and D.D.C. oversaw the microfluidic design and fabrication. D.R.G., M.M.A., C.-H.K., W.M.W, J.S.W., A.C.F., E.S., A.R., V.R. and P.O.S. performed the experiments. D.R.G., M.M.A., J.S.W., A.R., M.V.P., T.S. and P.O.S. analysed and interpreted the data. D.R.G., M.M.A., P.O.S. and T.S. wrote the manuscript and all the authors discussed the results and contributed to writing portions of the manuscript and editing the manuscript. D.R.G. and M.M.A. contributed equally to this work. The co-principal investigators are P.O.S. and T.S.

Corresponding authors

Correspondence to Tatiana Segura or Philip O. Scumpia.

Ethics declarations

Competing interests

D.R.G., W.M.W., D.D.C., T.S., and P.O.S. have a financial interest in Tempo Therapeutics, which aims to commercialize MAP technology.

Additional information

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

Supplementary information

Supplementary Information

Supplementary Figs. 1–5 and Supplementary Notes.

Reporting Summary

Source data

Source Data Fig. 1

Raw data for gel degradation and wound characterization.

Source Data Fig. 2

Figure 2 replicate images.

Source Data Fig. 3

Source data for cell infiltration and gene expression.

Source Data Fig. 4

Source data for antibody production and cell infiltration.

Source Data Fig. 5

Source data for wound characterization.

Source Data Supplementary Fig 2

Assessment of wound size.

Source Data Supplementary Fig 3

Quantification of F4/80+CD11b+ macrophages in the edge of L-MAP and D-MAP implant.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Griffin, D.R., Archang, M.M., Kuan, CH. et al. Activating an adaptive immune response from a hydrogel scaffold imparts regenerative wound healing. Nat. Mater. 20, 560–569 (2021). https://doi.org/10.1038/s41563-020-00844-w

Download citation

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing