The clinical need for maxillofacial reconstruction requires a deeper understanding of site-specific skeletal stem cell biology. The periosteum serves as a critical reservoir of adult skeletal stem progenitor cells (SSPCs) that navigate microscale stiffness gradients during bone healing. However, whether periosteal SSPCs from distinct skeletal sites exhibit divergent mechanosensitive responses remains poorly understood. This study compared the phenotypic characteristics and mechanotactic behaviors of mouse periosteal SSPCs (passage 3) from mandible (m-pSSPCs) and long bone (l-pSSPCs). Furthermore, we evaluated cell migration and adhesion using our previously reported flat PDMS substrate with periodic subsurface stiffness patterns (20-µm or 50-µm periods; stiffness differentials of ~0.3 MPa and ~0.5 MPa, respectively). While both cell types exhibited similar basal morphologies, flowcytometry and western blot analysis revealed a significantly higher proportion of osteogenic progenitors in m-pSSPCs compared to l-pSSPCs. Notably, m-pSSPCs displayed scale-dependent directional migration, morphological elongation and alignment exclusively on 50-µm patterns, whereas l-pSSPCs remained unresponsive across all scales. Transcriptomic profiling identified a marked enrichment of calcium-channel genes in m-pSSPCs, with Trpv2 significantly upregulated. Selective Trpv2 silencing abolished the characteristic polarization and alignment of m-pSSPCs on 50-μm stiffness patterns. Our findings demonstrate that m-pSSPCs possess a unique, Trpv2-dependent sensitivity to microscale mechanical cues, revealing site-specific mechanosensing programs in skeletal progenitors. These results highlight the necessity of considering tissue-specific cellular programs when designing “instructive” biomaterials for craniofacial regeneration.
- Open Access
- Article
Mandibular Periosteal Cells Exhibit Trpv2-Dependent Mechanosensitivity to Microscale Stiffness Patterns
- Qianying Lin 1,†,
- Conghui Zhang 1,†,
- Kaijie Zhang 2,
- Siyu Jin 1,
- Longjian Xue 2,*,
- Wei Ji 1,3,*
Author Information
Received: 01 Apr 2026 | Revised: 18 May 2026 | Accepted: 26 May 2026 | Published: 16 Jun 2026
Abstract
Keywords
mechanosensitivity | periosteal cells | skeletal stem progenitor cells | microscale stiffness | Trpv2
References
- 1.
Kumar, B.P.; Venkatesh, V.; Kumar, K.A.J.; et al. Mandibular Reconstruction: Overview. J. Maxillofac. Oral Surg. 2015, 15, 425–441. https://doi.org/10.1007/s12663-015-0766-5.
- 2.
Momoh, A.O.; Yu, P.; Skoracki, R.J.; et al. A Prospective Cohort Study of Fibula Free Flap Donor-Site Morbidity in 157 Consecutive Patients. Plast. Reconstr. Surg. 2011, 128, 714–720. https://doi.org/10.1097/PRS.0b013e318221dc2a.
- 3.
Dimitriou, R.; Mataliotakis, G.I.; Angoules, A.G.; et al. Complications following autologous bone graft harvesting from the iliac crest and using the RIA: A systematic review. Injury 2011, 42, S3-S15. https://doi.org/10.1016/j.injury.2011.06.015.
- 4.
Wu, J.; Min, A.; Wang, W.; et al. Trends in the incidence, prevalence and years lived with disability of facial fracture at global, regional and national levels from 1990 to 2017. PeerJ 2021, 9, e10693. https://doi.org/10.7717/peerj.10693.
- 5.
Bahney, C.S.; Zondervan, R.L.; Allison, P.; et al. Cellular biology of fracture healing. J. Orthop. Res. 2018, 37, 35–50. https://doi.org/10.1002/jor.24170.
- 6.
Colnot, C. Skeletal Cell Fate Decisions Within Periosteum and Bone Marrow During Bone Regeneration. J. Bone Miner. Res. 2009, 24, 274–282. https://doi.org/10.1359/jbmr.081003.
- 7.
Ortinau, L.C.; Wang, H.; Lei, K.; et al. Identification of Functionally Distinct Mx1+αSMA+ Periosteal Skeletal Stem Cells. Cell Stem Cell 2019, 25, 784–796.e5. https://doi.org/10.1016/j.stem.2019.11.003.
- 8.
Liu, R.; Jiao, Y.-R.; Huang, M.; et al. Mechanosensitive protein polycystin-1 promotes periosteal stem/progenitor cells osteochondral differentiation in fracture healing. Theranostics 2024, 14, 2544–2559. https://doi.org/10.7150/thno.93269.
- 9.
Dong, G.; Wang, J.; Chen, Z.; et al. Regulatory effects of stress release from decellularized periosteum on proliferation, migration, and osteogenic differentiation of periosteum-derived cells. Biomater. Sci. 2024, 12, 3360–3373. https://doi.org/10.1039/d4bm00358f.
- 10.
Ghiasi, M.S.; Chen, J.; Vaziri, A.; et al. Bone fracture healing in mechanobiological modeling: A review of principles and methods. Bone Rep. 2017, 6, 87–100. https://doi.org/10.1016/j.bonr.2017.03.002.
- 11.
Fu, X.; Liu, G.; Halim, A.; et al. Mesenchymal Stem Cell Migration and Tissue Repair. Cells 2019, 8, 784. https://doi.org/10.3390/cells8080784.
- 12.
Lim, R.; Banerjee, A.; Biswas, R.; et al. Mechanotransduction through adhesion molecules: Emerging roles in regulating the stem cell niche. Front. Cell Dev. Biol. 2022, 10, 966662. https://doi.org/10.3389/fcell.2022.966662.
- 13.
Putra, V.D.L.; Kilian, K.A.; Knothe Tate, M.L. Biomechanical, biophysical and biochemical modulators of cytoskeletal remodelling and emergent stem cell lineage commitment. Commun. Biol. 2023, 6, 75. https://doi.org/10.1038/s42003-022-04320-w.
- 14.
Shellard, A.; Mayor, R. Collective durotaxis along a self-generated stiffness gradient in vivo. Nature 2021, 600, 690–694. https://doi.org/10.1038/s41586-021-04210-x.
- 15.
Lewis, K.J. Osteocyte calcium signaling—A potential translator of mechanical load to mechanobiology. Bone 2021, 153, 116136. https://doi.org/10.1016/j.bone.2021.116136.
- 16.
Qin, L.; He, T.; Chen, S.; et al. Roles of mechanosensitive channel Piezo1/2 proteins in skeleton and other tissues. Bone Res. 2021, 9, 44. https://doi.org/10.1038/s41413-021-00168-8.
- 17.
Leucht, P.; Kim, J.-B.; Amasha, R.; et al. Embryonic origin and Hox status determine progenitor cell fate during adult bone regeneration. Development 2008, 135, 2845–2854. https://doi.org/10.1242/dev.023788.
- 18.
Siyu, J.; Conghui, Z.; Wei, J. Single-cell Sequencing Resolves Heterogeneity in Mandible and Limb Bud Development. J. Oral Sci. Res. 2025, 41, 849–855. https://doi.org/10.13701/j.cnki.kqyxyj.2025.10.004.
- 19.
Soares, A.P.; Fischer, H.; Aydin, S.; et al. Uncovering the unique characteristics of the mandible to improve clinical approaches to mandibular regeneration. Front. Physiol. 2023, 14. https://doi.org/10.3389/fphys.2023.1152301.
- 20.
Zhu, D.; Tong, X.; Trinh, P.; et al. Mimicking Cartilage Tissue Zonal Organization by Engineering Tissue-Scale Gradient Hydrogels as 3D Cell Niche. Tissue Eng. Part A 2017, 24, 1–10. https://doi.org/10.1089/ten.tea.2016.0453.
- 21.
Zhang, S.; Zhu, J.; Jin, S.; et al. Jawbone periosteum-derived cells with high osteogenic potential controlled by R-spondin 3. FASEB J. 2024, 38, e70079. https://doi.org/10.1096/fj.202400988RR.
- 22.
Chan, C.K.; Seo, E.Y.; Chen, J.Y.; et al. Identification and Specification of the Mouse Skeletal Stem Cell. Cell 2015, 160, 285–298. https://doi.org/10.1016/j.cell.2014.12.002.
- 23.
Lin, Q.; Zhang, K.; Jin, S.; et al. Microfabrication of Substrates with Microscale Stiffness Gradients to Guide Bone Marrow Stromal Cell Migration. J. Vis. Exp. 2025, 225, e69473. https://doi.org/10.3791/69473.
- 24.
Gorelik, R.; Gautreau, A. Quantitative and unbiased analysis of directional persistence in cell migration. Nat. Protoc. 2014, 9, 1931–1943. https://doi.org/10.1038/nprot.2014.131.
- 25.
Horzum, U.; Ozdil, B.; Pesen-Okvur, D. Step-by-step quantitative analysis of focal adhesions. MethodsX 2014, 1, 56–59. https://doi.org/10.1016/j.mex.2014.06.004.
- 26.
Langmead, B.; Salzberg, S.L. Fast gapped-read alignment with Bowtie 2. Nat. Methods 2012, 9, 357–359. https://doi.org/10.1038/nmeth.1923.
- 27.
Li, B.; Dewey, C.N. RSEM: Accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinform. 2011, 12, 323. https://doi.org/10.1186/1471-2105-12-323.
- 28.
Love, M.I.; Huber, W.; Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014, 15, 550. https://doi.org/10.1186/s13059-014-0550-8.
- 29.
Ambrosi, T.H.; Taheri, S.; Chen, K.; et al. Human skeletal development and regeneration are shaped by functional diversity of stem cells across skeletal sites. Cell Stem Cell 2025, 32, 811–823.e11. https://doi.org/10.1016/j.stem.2025.02.013.
- 30.
Pham, J.T.; Xue, L.; del Campo, A.; et al. Guiding cell migration with microscale stiffness patterns and undulated surfaces. Acta Biomater. 2016, 38, 106–115. https://doi.org/10.1016/j.actbio.2016.04.031.
- 31.
Nikkhah, M.; Edalat, F.; Manoucheri, S.; et al. Engineering microscale topographies to control the cell–substrate interface. Biomaterials 2012, 33, 5230–5246. https://doi.org/10.1016/j.biomaterials.2012.03.079.
- 32.
Aegerter, C.M.; Wehner, T.; Steiner, M.; et al. Prediction of the Time Course of Callus Stiffness as a Function of Mechanical Parameters in Experimental Rat Fracture Healing Studies—A Numerical Study. PLoS ONE 2014, 9, e115695. https://doi.org/10.1371/journal.pone.0115695.
- 33.
Miga, M.I.; Weis, J.A.; Granero-Molto, F.; et al. Quantifying Mechanical Properties in a Murine Fracture Healing System Using an Inverse Geometric Nonlinear Elasticity Modeling Framework. In International Symposium on Biomedical Simulation; Springer: Berlin/Heidelberg, Germany, 2010.
- 34.
Panzetta, V.; Fusco, S.; Netti, P.A. Cell mechanosensing is regulated by substrate strain energy rather than stiffness. Proc. Natl. Acad. Sci.USA 2019, 116, 22004–22013. https://doi.org/10.1073/pnas.1904660116.
- 35.
Prager-Khoutorsky, M.; Lichtenstein, A.; Krishnan, R.; et al. Fibroblast polarization is a matrix-rigidity-dependent process controlled by focal adhesion mechanosensing. Nat. Cell Biol. 2011, 13, 1457–1465. https://doi.org/10.1038/ncb2370.
- 36.
Aghaloo, T.L.; Chaichanasakul, T.; Bezouglaia, O.; et al. Osteogenic Potential of Mandibular vs. Long-bone Marrow Stromal Cells. J. Dent. Res. 2010, 89, 1293–1298. https://doi.org/10.1177/0022034510378427.
- 37.
Ichikawa, Y.; Watahiki, J.; Nampo, T.; et al. Differences in the developmental origins of the periosteum may influence bone healing. J. Periodontal Res. 2014, 50, 468–478. https://doi.org/10.1111/jre.12229.
- 38.
Li, L.; Liu, Y.; Qian, X.; et al. Modulating the phenotype and function of bone marrow-derived macrophages via mandible and femur osteoblasts. Int. Immunopharmacol. 2024, 132. https://doi.org/10.1016/j.intimp.2024.112000.
- 39.
Galea, G.L.; Zein, M.R.; Allen, S.; et al. Making and shaping endochondral and intramembranous bones. Dev. Dyn. 2020, 250, 414–449. https://doi.org/10.1002/dvdy.278.
- 40.
Goodwin, K.; Nelson, C.M. Mechanics of Development. Dev. Cell 2021, 56, 240–250. https://doi.org/10.1016/j.devcel.2020.11.025.
- 41.
Christensen, A.P.; Corey, D.P. TRP channels in mechanosensation: Direct or indirect activation? Nat. Rev. Neurosci. 2007, 8, 510–521. https://doi.org/10.1038/nrn2149.
- 42.
Eijkelkamp, N.; Quick, K.; Wood, J.N. Transient Receptor Potential Channels and Mechanosensation. Annu. Rev. Neurosci. 2013, 36, 519–546. https://doi.org/10.1146/annurev-neuro-062012-170412.
- 43.
Kyprianou, N.; Oulidi, A.; Bokhobza, A.; et al. TRPV2 Mediates Adrenomedullin Stimulation of Prostate and Urothelial Cancer Cell Adhesion, Migration and Invasion. PLoS ONE 2013, 8, e64885. https://doi.org/10.1371/journal.pone.0064885.
- 44.
Schwab, A.; Fabian, A.; Hanley, P.J.; et al. Role of Ion Channels and Transporters in Cell Migration. Physiol. Rev. 2012, 92, 1865–1913. https://doi.org/10.1152/physrev.00018.2011.
- 45.
Wei, C.; Wang, X.; Zheng, M.; et al. Calcium gradients underlying cell migration. Curr. Opin. Cell Biol. 2012, 24, 254–261. https://doi.org/10.1016/j.ceb.2011.12.002.
- 46.
Rocereta, J.A.; Sturhahn, T.; Pumroy, R.A.; et al. Structural insights into TRPV2 modulation by probenecid. Nat. Struct. Mol. Biol. 2025, 32, 1019–1029. https://doi.org/10.1038/s41594-025-01494-9.
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