Hydrogel crosslinking modulates macrophages, fibroblasts, and their communication, during wound healing

Sergei Butenko; Raji R Nagalla; Christian F Guerrero-Juarez; Francesco Palomba; Li-Mor David; Ronald Q Nguyen; Denise Gay; Axel A Almet; Michelle A Digman; Qing Nie; Philip O Scumpia; Maksim V Plikus; Wendy F Liu

doi:10.1038/s41467-024-50072-y

Hydrogel crosslinking modulates macrophages, fibroblasts, and their communication, during wound healing

Nat Commun. 2024 Aug 9;15(1):6820. doi: 10.1038/s41467-024-50072-y.

Authors

Sergei Butenko^#^{1

2}, Raji R Nagalla^#¹, Christian F Guerrero-Juarez³, Francesco Palomba¹, Li-Mor David¹, Ronald Q Nguyen¹, Denise Gay², Axel A Almet^{4

5}, Michelle A Digman^{1

6}, Qing Nie^{4

5

7}, Philip O Scumpia^{8

9

10}, Maksim V Plikus^{2

4

11}, Wendy F Liu^{12

13

14

15

16}

Affiliations

¹ Department of Biomedical Engineering, University of California Irvine, Irvine, CA, USA.
² Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, USA.
³ Carle Illinois College of Medicine, University of Illinois at Urbana-Champaign, Urbana-Champaign, IL, USA.
⁴ NSF-Simons Center for Multiscale Cell Fate Research, University of California, Irvine, Irvine, CA, USA.
⁵ Department of Mathematics, University of California, Irvine, Irvine, CA, USA.
⁶ Laboratory of Fluorescence Dynamics, The Henry Samueli School of Engineering, University of California, Irvine, CA, USA.
⁷ Center for Complex Biological Systems, University of California Irvine, Irvine, CA, USA.
⁸ Division of Dermatology, Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA.
⁹ Jonsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA.
¹⁰ Department of Dermatology, Veterans Administration Greater Los Angeles Healthcare System, Los Angeles, CA, USA.
¹¹ Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA, USA.
¹² Department of Biomedical Engineering, University of California Irvine, Irvine, CA, USA. wendy.liu@uci.edu.
¹³ UCI Edwards Lifesciences Foundation Cardiovascular Innovation and Research Center, University of California Irvine, Irvine, CA, USA. wendy.liu@uci.edu.
¹⁴ Institute for Immunology, University of California, Irvine, Irvine, CA, USA. wendy.liu@uci.edu.
¹⁵ Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA, USA. wendy.liu@uci.edu.
¹⁶ Department of Chemical and Biomolecular Engineering, University of California Irvine, Irvine, CA, USA. wendy.liu@uci.edu.

^# Contributed equally.

Abstract

Biomaterial wound dressings, such as hydrogels, interact with host cells to regulate tissue repair. This study investigates how crosslinking of gelatin-based hydrogels influences immune and stromal cell behavior and wound healing in female mice. We observe that softer, lightly crosslinked hydrogels promote greater cellular infiltration and result in smaller scars compared to stiffer, heavily crosslinked hydrogels. Using single-cell RNA sequencing, we further show that heavily crosslinked hydrogels increase inflammation and lead to the formation of a distinct macrophage subpopulation exhibiting signs of oxidative activity and cell fusion. Conversely, lightly crosslinked hydrogels are more readily taken up by macrophages and integrated within the tissue. The physical properties differentially affect macrophage and fibroblast interactions, with heavily crosslinked hydrogels promoting pro-fibrotic fibroblast activity that drives macrophage fusion through RANKL signaling. These findings suggest that tuning the physical properties of hydrogels can guide cellular responses and improve healing, offering insights for designing better biomaterials for wound treatment.

MeSH terms

Animals
Biocompatible Materials / chemistry
Cell Communication / drug effects
Cross-Linking Reagents / chemistry
Female
Fibroblasts* / drug effects
Fibroblasts* / metabolism
Gelatin / chemistry
Hydrogels* / chemistry
Inflammation / metabolism
Inflammation / pathology
Macrophages* / drug effects
Macrophages* / metabolism
Mice
Mice, Inbred C57BL
RANK Ligand / metabolism
Wound Healing* / drug effects

Substances

Hydrogels
Biocompatible Materials
RANK Ligand
Cross-Linking Reagents
Gelatin
Tnfsf11 protein, mouse

Abstract

MeSH terms

Substances

Grants and funding