Substrate Stiffness of Bone Microenvironment Controls Functions of Pre-Osteoblasts and Fibroblasts In Vitro
Source abstract
Abstract
The formation of bone in a bone defect is accomplished by osteoblasts, while the over activation of fibroblasts promotes fibrosis. However, it is not clear how the extracellular matrix stiffness of the bone-regeneration microenvironment affects the function of osteoblasts and fibroblasts. This study aim to investigate the effect of bone-regeneration microenvironment stiffness on cell adhesion, cell proliferation, cell differentiation, synthesizing matrix ability and its potential mechanisms in mechanotransduction, in pre-osteoblasts and fibroblasts. Polyacrylamide substrates mimicking the matrix stiffness of different stages of the bone-healing process (15 kPa, mimic granulation tissue; 35 kPa, mimic osteoid; 150 kPa, mimic calcified bone matrix) were prepared. Mouse pre-osteoblasts MC3T3-E1 and mouse fibroblasts NIH3T3 were plated on three types of substrates, respectively. There were significant differences in the adhesion of pre-osteoblasts and fibroblasts on different polyacrylamide substrates. Runx2 expression increased with increasing substrate stiffness in pre-osteoblasts, while no statistical differences were found in the Acta2 expression in fibroblasts on three substrates. OPN expression in pre-osteoblasts, as well as Fn1 and Col1a1 expression in fibroblasts, decreased with increasing stiffness. The difference between the cell traction force generated by pre-osteoblasts and fibroblasts on substrates was also found. Our results indicated that substrate stiffness is a potent regulator of pre-osteoblasts and fibroblasts with the ability of promoting osteogenic differentiation of pre-osteoblasts, while having no effect on myofibroblast differentiation of fibroblasts.
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Research context
OpenAlex topics: Bone Tissue Engineering Materials · Cellular Mechanics and Interactions · Bone Metabolism and Diseases
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
Department of Prosthodontics, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, Beijing Key Laboratory of Digital Stomatology, Research Center of Engineering and Technology for Computerized Dentistry Ministry of Health, NMPA Key Laboratory for Dental Materials, Central Laboratory, Peking University School and Hospital of Stomatology, No. 22, Zhongguancun South Avenue, Haidian District, Beijing 100081, China
Department of Implantology, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, No. 22, Zhongguancun South Avenue, Haidian District, Beijing 100081, China
Department of Geriatric Dentistry, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, No. 22, Zhongguancun South Avenue, Haidian District, Beijing 100081, China
Institute of Biomechanics and Medical Engineering, School of Aerospace Engineering, Tsinghua University, Beijing 100084, China
Funding
National Key R&D Program of China · Beijing Natural Science Foundation · National Program for Multidisciplinary Cooperative Treatment on Major Diseases
References (53)
Show source-provided references
1. Mechanisms of bone development and repair10.1038/s41580-020-00279-w ↗
2. The decisive early phase of bone regeneration10.1038/s41584-022-00887-0 ↗
3. Barrier Membrane in Regenerative Therapy: A Narrative Review10.3390/membranes12050444 ↗
4. Osteoblast/fibroblast coculture derived bioactive ECM with unique matrisome profile facilitates bone regeneration10.1016/j.bioactmat.2020.06.017 ↗
5. Fracture healing: Mechanisms and interventions10.1038/nrrheum.2014.164 ↗
6. The healing potential of the periosteum molecular aspects10.1016/j.injury.2005.07.030 ↗
7. Substrate Stiffness Combined with Hepatocyte Growth Factor Modulates Endothelial Cell Behavior10.1021/acs.biomac.6b00318 ↗
8. Glossary of Oral and Maxillofacial Implants10.11607/jomi.2017.4.gomi ↗
9. Bone physiological microenvironment and healing mechanism: Basis for future bone-tissue engineering scaffolds10.1016/j.bioactmat.2021.03.043 ↗
10. The perivascular origin of pathological fibroblasts10.1172/JCI93558 ↗
11. Direct contribution of skeletal muscle mesenchymal progenitors to bone repair10.1038/s41467-021-22842-5 ↗
12. Overview of the fracture healing cascade10.1016/j.injury.2005.07.027 ↗
13. Bone remodeling during fracture repair: The cellular picture10.1016/j.semcdb.2008.07.004 ↗
14. Building strong bones: Molecular regulation of the osteoblast lineage10.1038/nrm3254 ↗
15. Topography-Supported Nanoarchitectonics of Hybrid Scaffold for Systematically Modulated Bone Regeneration and Remodeling10.1002/adfm.202206863 ↗
16. Fibronectin at select sites binds multiple growth factors and enhances their activity: Expansion of the collaborative ECM-GF paradigm10.1038/jid.2013.484 ↗
17. Stem Cell Differentiation is Regulated by Extracellular Matrix Mechanics10.1152/physiol.00026.2017 ↗
18. Materials as stem cell regulators10.1038/nmat3937 ↗
19. Mechanism of regulation of stem cell differentiation by matrix stiffness10.1186/s13287-015-0083-4 ↗
20. Effect of matrix stiffness on osteoblast functionalization10.1111/cpr.12338 ↗
21. Combined role for YAP-TEAD and YAP-RUNX2 signalling in substrate-stiffness regulation of cardiac fibroblast proliferation10.1016/j.bbamcr.2022.119329 ↗
22. Extracellular matrix type modulates mechanotransduction of stem cells10.1016/j.actbio.2019.06.048 ↗
23. Integrin ligands at a glance10.1242/jcs.03098 ↗
24. Cell-extracellular matrix mechanotransduction in 3D10.1038/s41580-023-00583-1 ↗
25. Intrinsic extracellular matrix properties regulate stem cell differentiation10.1016/j.jbiomech.2009.09.009 ↗
26. Integrin activation and internalization on soft ECM as a mechanism of induction of stem cell differentiation by ECM elasticity10.1073/pnas.1106467108 ↗
27. Rigidity sensing and adaptation through regulation of integrin types10.1038/nmat3960 ↗
28. High-resolution traction force microscopy10.1016/B978-0-12-420138-5.00020-3 ↗
29. Preparation of hydrogel substrates with tunable mechanical properties10.1002/0471143030.cb1016s47 ↗
30. An extended relationship for the characterization of Young’s modulus and Poisson’s ratio of tunable polyacrylamide gels
31. Ghiasi, M.S., Chen, J.E., Rodriguez, E.K., Vaziri, A., and Nazarian, A. (2019). Computational modeling of human bone fracture healing affected by different conditions of initial healing stage. BMC Musculoskelet. Disord., 20.10.1186/s12891-019-2854-z ↗
32. Matrix elasticity directs stem cell lineage specification10.1016/j.cell.2006.06.044 ↗
33. The Bone Extracellular Matrix in Bone Formation and Regeneration10.3389/fphar.2020.00757 ↗
34. Substrate stiffness regulates the differentiation profile and functions of osteoclasts via cytoskeletal arrangement10.1111/cpr.13172 ↗
35. Olivares-Navarrete, R., Lee, E.M., Smith, K., Hyzy, S.L., Doroudi, M., Williams, J.K., Gall, K., Boyan, B.D., and Schwartz, Z. (2017). Substrate Stiffness Controls Osteoblastic and Chondrocytic Differentiation of Mesenchymal Stem Cells without Exogenous Stimuli. PLoS ONE, 12.10.1371/journal.pone.0170312 ↗
36. The role of α-smooth muscle actin in fibroblast-mediated matrix contraction and remodeling10.1016/j.bbadis.2016.11.006 ↗
37. Mobilizing endogenous stem cells for repair and regeneration: Are we there yet?10.1016/j.stem.2012.05.004 ↗
38. Extracellular matrix stiffness dictates Wnt expression through integrin pathway10.1038/srep20395 ↗
39. Neurite branching on deformable substrates10.1097/00001756-200212200-00007 ↗
40. Integrin expression and function on human osteoblast-like cells10.1359/jbmr.1997.12.8.1189 ↗
41. Human gingival fibroblast integrin subunit expression on titanium implant surfaces10.1902/jop.2005.76.10.1743 ↗
42. Matrix stiffness regulation of integrin-mediated mechanotransduction during osteogenic differentiation of human mesenchymal stem cells10.1002/jbmr.278 ↗
43. Integrin endocytosis on elastic substrates mediates mechanosensing10.1016/j.jbiomech.2016.05.024 ↗
44. Quantitative Analyses of Dynamic Features of Fibroblasts on Different Protein-Coated Compliant Substrates10.1021/acsbiomaterials.6b00598 ↗
45. The effect of substrate stiffness, thickness, and cross-linking density on osteogenic cell behavior10.1016/j.bpj.2015.02.022 ↗
46. Effect of substrate stiffness on proliferation and differentiation of periodontal ligament stem cells10.1111/cpr.12478 ↗
47. Force loading explains spatial sensing of ligands by cells10.1038/nature24662 ↗
48. Periostin and matrix stiffness combine to regulate myofibroblast differentiation and fibronectin synthesis during palatal healing10.1016/j.matbio.2020.07.002 ↗
49. Fibronectin synthesis, but not alpha-smooth muscle expression, is regulated by periostin in gingival healing through FAK/JNK signaling10.1038/s41598-018-35805-6 ↗
50. An overview of substrate stiffness guided cellular response and its applications in tissue regeneration10.1016/j.bioactmat.2021.12.005 ↗
51. Multi-level molecular clutches in motile cell processes10.1016/j.tcb.2009.07.001 ↗
52. Force Triggers YAP Nuclear Entry by Regulating Transport across Nuclear Pores10.1016/j.cell.2017.10.008 ↗
53. Screening of rat mesenchymal stem cell behaviour on polydimethylsiloxane stiffness gradients10.1016/j.actbio.2011.09.030 ↗