Collagen Hydrogel Viscoelasticity Regulates MSC Chondrogenesis in a ROCK-dependent Manner

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2023 Feb 10

PMID 36763657

Authors

Danyang Huang

Yuehong Li

Zihan Ma

Hai Lin

Xiangdong Zhu

Yun Xiao

Xingdong Zhang

Affiliations

Soon will be listed here.

Abstract

Mesenchymal stem cell (MSC) chondrogenesis in three-dimensional (3D) culture involves dynamic changes in cytoskeleton architecture during mesenchymal condensation before morphogenesis. However, the mechanism linking dynamic mechanical properties of matrix to cytoskeletal changes during chondrogenesis remains unclear. Here, we investigated how viscoelasticity, a time-dependent mechanical property of collagen hydrogel, coordinates MSC cytoskeleton changes at different stages of chondrogenesis. The viscoelasticity of collagen hydrogel was modulated by controlling the gelling process without chemical cross-linking. In slower-relaxing hydrogels, although a disordered cortical actin promoted early chondrogenic differentiation, persistent myosin hyperactivation resulted in Rho-associated kinase (ROCK)-dependent apoptosis. Meanwhile, faster-relaxing hydrogels promoted cell-matrix interactions and eventually facilitated long-term chondrogenesis with mitigated myosin hyperactivation and cell apoptosis, similar to the effect of ROCK inhibitors. The current work not only reveals how matrix viscoelasticity coordinates MSC chondrogenesis and survival in a ROCK-dependent manner but also highlights viscoelasticity as a design parameter for biomaterials for chondrogenic 3D culture.

Citing Articles

Design and Fabrication of Viscoelastic Hydrogels as Extracellular Matrix Mimicry for Cell Engineering.

Li Z, Li T, Yang H, Ding M, Chen L, Yu S Chem Bio Eng. 2025; 1(11):916-933.

PMID: 39975568 PMC: 11835267. DOI: 10.1021/cbe.4c00129.

Viscoelastic hydrogel combined with dynamic compression promotes osteogenic differentiation of bone marrow mesenchymal stem cells and bone repair in rats.

Yang C, Cai W, Xiang P, Liu Y, Xu H, Zhang W Regen Biomater. 2025; 12:rbae136.

PMID: 39845143 PMC: 11751691. DOI: 10.1093/rb/rbae136.

Mechanobiology of 3D cell confinement and extracellular crowding.

Da Silva Andre G, Labouesse C Biophys Rev. 2025; 16(6):833-849.

PMID: 39830117 PMC: 11735831. DOI: 10.1007/s12551-024-01244-z.

Cartilaginous microtissues exhibit extreme resilience under compression with size-dependent mechanical properties.

Androulidakis C, Svitina H, Ioannidis K, Dunn A, Papantoniou I Biomaterials. 2025; 317:123074.

PMID: 39799695 PMC: 11850221. DOI: 10.1016/j.biomaterials.2024.123074.

3D Culture of MSCs for Clinical Application.

Gao Q, Cekuc M, Ergul Y, Pius A, Shinohara I, Murayama M Bioengineering (Basel). 2025; 11(12.

PMID: 39768017 PMC: 11726872. DOI: 10.3390/bioengineering11121199.

References

Lou J, Stowers R, Nam S, Xia Y, Chaudhuri O . Stress relaxing hyaluronic acid-collagen hydrogels promote cell spreading, fiber remodeling, and focal adhesion formation in 3D cell culture. Biomaterials. 2017; 154:213-222. DOI: 10.1016/j.biomaterials.2017.11.004. View

Cameron A, Frith J, Cooper-White J . The influence of substrate creep on mesenchymal stem cell behaviour and phenotype. Biomaterials. 2011; 32(26):5979-93. DOI: 10.1016/j.biomaterials.2011.04.003. View

Holmes D, Chapman J, Prockop D, Kadler K . Growing tips of type I collagen fibrils formed in vitro are near-paraboloidal in shape, implying a reciprocal relationship between accretion and diameter. Proc Natl Acad Sci U S A. 1992; 89(20):9855-9. PMC: 50232. DOI: 10.1073/pnas.89.20.9855. View

Leo L, Bridelli M, Polverini E . Insight on collagen self-assembly mechanisms by coupling molecular dynamics and UV spectroscopy techniques. Biophys Chem. 2019; 253:106224. DOI: 10.1016/j.bpc.2019.106224. View

Chaudhuri O, Gu L, Klumpers D, Darnell M, Bencherif S, Weaver J . Hydrogels with tunable stress relaxation regulate stem cell fate and activity. Nat Mater. 2015; 15(3):326-34. PMC: 4767627. DOI: 10.1038/nmat4489. View

Wu Y, Shu J, He C, Li M, Wang Y, Ou W . ROCK inhibitor Y27632 promotes proliferation and diminishes apoptosis of marmoset induced pluripotent stem cells by suppressing expression and activity of caspase 3. Theriogenology. 2015; 85(2):302-14. DOI: 10.1016/j.theriogenology.2015.09.020. View

Li Y, Lam K, Chen A, Zhang W, Chan B . Collagen microencapsulation recapitulates mesenchymal condensation and potentiates chondrogenesis of human mesenchymal stem cells - A matrix-driven in vitro model of early skeletogenesis. Biomaterials. 2019; 213:119210. DOI: 10.1016/j.biomaterials.2019.05.021. View

Lewis D, Tang V, Jain N, Isser A, Xia Z, Gerecht S . Collagen Fiber Architecture Regulates Hypoxic Sarcoma Cell Migration. ACS Biomater Sci Eng. 2021; 4(2):400-409. DOI: 10.1021/acsbiomaterials.7b00056. View

Gadjanski I, Spiller K, Vunjak-Novakovic G . Time-dependent processes in stem cell-based tissue engineering of articular cartilage. Stem Cell Rev Rep. 2011; 8(3):863-81. PMC: 3412929. DOI: 10.1007/s12015-011-9328-5. View

10.

Yang J, Chen X, Yuan T, Yang X, Fan Y, Zhang X . Regulation of the secretion of immunoregulatory factors of mesenchymal stem cells (MSCs) by collagen-based scaffolds during chondrogenesis. Mater Sci Eng C Mater Biol Appl. 2016; 70(Pt 2):983-991. DOI: 10.1016/j.msec.2016.04.096. View

11.

Ray P, Chapman S . Cytoskeletal Reorganization Drives Mesenchymal Condensation and Regulates Downstream Molecular Signaling. PLoS One. 2015; 10(8):e0134702. PMC: 4523177. DOI: 10.1371/journal.pone.0134702. View

12.

Abou Neel E, Bozec L, Knowles J, Syed O, Mudera V, Day R . Collagen--emerging collagen based therapies hit the patient. Adv Drug Deliv Rev. 2012; 65(4):429-56. DOI: 10.1016/j.addr.2012.08.010. View

13.

McBeath R, Pirone D, Nelson C, Bhadriraju K, Chen C . Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell. 2004; 6(4):483-95. DOI: 10.1016/s1534-5807(04)00075-9. View

14.

Quintin S, Gally C, Labouesse M . Epithelial morphogenesis in embryos: asymmetries, motors and brakes. Trends Genet. 2008; 24(5):221-30. DOI: 10.1016/j.tig.2008.02.005. View

15.

Leikin S, Rau D, Parsegian V . Temperature-favoured assembly of collagen is driven by hydrophilic not hydrophobic interactions. Nat Struct Biol. 1995; 2(3):205-10. DOI: 10.1038/nsb0395-205. View

16.

Woods A, Wang G, Beier F . RhoA/ROCK signaling regulates Sox9 expression and actin organization during chondrogenesis. J Biol Chem. 2005; 280(12):11626-34. DOI: 10.1074/jbc.M409158200. View

17.

Foyt D, Taheem D, Ferreira S, Norman M, Petzold J, Jell G . Hypoxia impacts human MSC response to substrate stiffness during chondrogenic differentiation. Acta Biomater. 2019; 89:73-83. PMC: 6481516. DOI: 10.1016/j.actbio.2019.03.002. View

18.

Sebbagh M, Renvoize C, Hamelin J, Riche N, Bertoglio J, Breard J . Caspase-3-mediated cleavage of ROCK I induces MLC phosphorylation and apoptotic membrane blebbing. Nat Cell Biol. 2001; 3(4):346-52. DOI: 10.1038/35070019. View

19.

Vining K, Mooney D . Mechanical forces direct stem cell behaviour in development and regeneration. Nat Rev Mol Cell Biol. 2017; 18(12):728-742. PMC: 5803560. DOI: 10.1038/nrm.2017.108. View

20.

Xu J, Wang W, Ludeman M, Cheng K, Hayami T, Lotz J . Chondrogenic differentiation of human mesenchymal stem cells in three-dimensional alginate gels. Tissue Eng Part A. 2008; 14(5):667-80. DOI: 10.1089/tea.2007.0272. View