Skip to main content
SpringerLink
Log in

Polydopamine-assisted BMP-2 immobilization on titanium surface enhances the osteogenic potential of periodontal ligament stem cells via integrin-mediated cell-matrix adhesion

  • Research Article
  • Published:
Journal of Cell Communication and Signaling Aims and scope

Abstract

A mussel-inspired polydopamine (PDA), resulting from the oxidative polymerization of dopamine, was reported to be an attractive substrate for advancing biomaterial applications. Thus, this study determined the osteoconductive/osteoinductive properties of titanium (Ti) surfaces coated with PDA and the facilitation of the PDA layer to immobilize bone morphogenetic protein-2 (BMP-2) on Ti substrates. The surface chemistry of PDA or PDA/BMP-2-coated Ti was confirmed by contact angle measurement, scanning electron microscopy (SEM), immunofluorescence staining, atomic force microscopy (AFM), and X-ray photoelectron spectroscopy (XPS). We verified the osteogenic potential of periodontal ligament stem cells (PDLSCs) cultured on the PDA or PDA/BMP-2-Ti surfaces. The osteogenic differentiation of the PDLSCs was assessed by measuring alkaline phosphatase (ALP) activity, intracellular calcium levels, as well as by evaluating osteocalcin (OCN), osterix (OSX), and runt-related transcription factor 2 (RUNX2) protein levels. The PDLSCs cultured on PDA/BMP-2-Ti showed the highest osteogenic activity compared with those on the control Ti and PDA-coated Ti surfaces. Moreover, PDLSCs on PDA and PDA/BMP-2-Ti expressed increased levels of integrin β1 and actin molecules compared to cells on control Ti. Blocking integrin β1 significantly decreased the osteogenic activity of PDLSCs on PDA/BMP-2 surfaces. This study suggests that the PDA coating can efficiently encourage the immobilization of BMP-2 on Ti surfaces and that this modified Ti substrate highly enhanced the osteogenic differentiation of PDLSCs by integrin-mediated cell-matrix adhesion mechanisms.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Abrams GA, Goodman SL, Nealey PE, Franco M, Murphy CJ (2000) Nanoscaletopography of the basement membrane underlying the corneal epithelium of therhesus macaque. Cell Tissue Res 299:39–46

    Article  CAS  Google Scholar 

  • Baxter LC, Frauchiger V, Textor M, ap Gwynn I, Richards RG (2002) Fibroblast and osteoblast adhesion and morphology on calcium phosphate surfaces. Eur Cell Mater 4:1–17

    Article  CAS  Google Scholar 

  • Bhang SH, Kwon SH, Lee S, Kim GC, Han AM, Kwon YH, Kim BS (2013) Enhanced neuronal differentiation of pheochromocytoma 12 cells on polydopamine-modified surface. Biochem Biophys Res Commun 430:1294–1300

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Chien HW, Kuo WH, Wang MJ, Tsai SW, Tsai WB (2012) Tunable micropatterned substrates based on poly(dopamine) deposition via microcontact printing. Langmuir 28:5775–5782

    Article  CAS  Google Scholar 

  • Chou L, Firth JD, Uitto VJ, Brunette DM (1995) Substratum surface topography alters cell shape and regulates fibronectin mRNA level, mRNA stability, secretion and assembly in human fibroblasts. J Cell Sci 108:1563–1573

    CAS  PubMed  Google Scholar 

  • Cochran DL, Schenk RK, Lussi A, Higginbottom FL, Buser D (1998) Bone response to unloaded and loaded titanium implants with a sandblasted and acid-etched surface. J Biomed Mater Res 40:1–11

    Article  CAS  Google Scholar 

  • Fourel L, Valat A, Faurobert E, Guillot R, Bourrin-Reynard I, Ren K, Lafanechère L, Planus E, Picart C, Albiges-Rizo C (2016) β3 integrin-mediated spreading induced by matrix-bound BMP-2 controls Smad signaling in a stiffness-independent manner. J Cell Biol 212:693–706

    Article  CAS  Google Scholar 

  • Humphries JD, Byron A, Humphries MJ (2006) Integrin ligands at a glance. J Cell Sci 119:3901–3903

    Article  CAS  Google Scholar 

  • Ivaska J, Heino J (2011) Cooperation between integrins and growth factor receptors in signaling and endocytosis. Annu Rev Cell Dev Biol 27:291–320

    Article  CAS  Google Scholar 

  • Keselowsky BG, Collard DM, Garcia AJ (2005) Integrin binding specificity regulates biomaterial surface chemistry effects on cell differentiation. Proc Natl Acad Sci U S A 102:5953–5957

    Article  CAS  Google Scholar 

  • Kim SY, Lee JK, Park YD, Kang KL, Lee JC, Heo JS (2013) Hesperetin alleviates the inhibitory effects of high glucose on the osteoblastic differentiation of periodontal ligament stem cells. PLoS One 8:e67504

    Article  CAS  Google Scholar 

  • Ko E, Yang K, Shin J, Cho SW (2013) Polydopamine-assisted osteoinductive peptide immobilization of polymer scaffolds for enhanced bone regeneration by human adipose-derived stem cells. Biomacromolecules 14:3202–3213

    Article  CAS  Google Scholar 

  • Kononen M, Hormia M, Kivilahti J, Hautaniemi J, Thesleff I (1992) Effect of surface processing on the attachment, orientation, and proliferation of human gingival fibroblasts on titanium. J Biomed Mater Res 26:1325–1341

    Article  CAS  Google Scholar 

  • Ku SH, Ryu J, Hong SK, Lee H, Park CB (2010) General functionalization route for cell adhesion on non-wetting surfaces. Biomaterials 31:2535–2541

    Article  CAS  Google Scholar 

  • Lai M, Cai K, Zhao L, Chen X, Hou Y, Yang Z (2011) Surface functionalization of TiO2 nanotubes with bone morphogenetic protein 2 and its synergistic effect on the differentiation of mesenchymal stem cells. Biomacromolecules 12:1097–1105

    Article  CAS  Google Scholar 

  • Lee H, Scherer NF, Messersmith PB (2006) Single-molecule mechanics of mussel adhesion. Proc Natl Acad Sci U S A 103:12999–13003

    Article  CAS  Google Scholar 

  • Lee YB, Shin YM, Lee JH, Jun I, Kang JK, Park JC, Shin H (2012) Polydopamine-mediated immobilization of multiple bioactive molecules for the development of functional vascular graft materials. Biomaterials 33:8343–8352

    Article  CAS  Google Scholar 

  • Lee JS, Yi JK, An SY, Heo JS (2014) Increased osteogenic differentiation of periodontal ligament stem cells on Polydopamine film occurs via activation of integrin and PI3K signaling pathways. Cell Physiol Biochem 34:1824–1834

    Article  CAS  Google Scholar 

  • Liu X, Lim JK, Donahue HJ, Dhurjati R, Mastro AM, Vogler EA (2007) Influence of substratumsurface chemistry/energy and topography on the human fetal osteoblastic cell line hFOB 1.19: phenotypic and genotypic responses observed in vitro. Biomaterials 28:4535–4550

    Article  CAS  Google Scholar 

  • Liu YT, Lee TM, Lui TS (2013) Enhanced osteoblastic cell response on zirconia by bio-inspired surface modification. Colloids Surf B Biointerfaces 106:37–45

    Article  CAS  Google Scholar 

  • Luo R, Tang L, Zhong S, Yang Z, Wang J, Weng Y, Tu Q, Jiang C, Huang N (2013) In vitro investigation of enhanced hemocompatibility and endothelial cell proliferation associated with quinone-rich polydopamine coating. ACS Appl Mater Interfaces 5:1704–1714

    Article  CAS  Google Scholar 

  • Lynge ME, Ogaki R, Laursen AO, Lovmand J, Sutherland DS, Städler B (2011) Polydopamine/liposome coatings and their interaction with myoblast cells. ACS Appl Mater Interfaces 3:2142–2147

    Article  CAS  Google Scholar 

  • Margadant C, Sonnenberg A (2010) Integrin-TGF-beta crosstalk in fibrosis, cancer and wound healing. EMBO Rep 11:97–105

    Article  CAS  Google Scholar 

  • Martins A, Pinho ED, Faria S, Pashkuleva I, Marques AP, Reis RL, Neves NM (2009) Surface modification of electrospun polycaprolactone nanofiber meshes by plasma treatment to enhance biological performance. Small 5:1195–1206

    CAS  PubMed  Google Scholar 

  • Oakley C, Brunette DM (1995) Response of single, pairs, and clusters of epithelial cells to substratum topography. Biochem Cell Biol 73:473–489

    Article  CAS  Google Scholar 

  • Oates TW, Valderrama P, Bischof M, Nedir R, Jones A, Simpson J, Toutenburg H, Cochran DL (2007) Enhanced implant stability with a chemically modified SLA surface: a randomized pilot study. Int J Oral Maxillofac Implants 22:755–760

    PubMed  Google Scholar 

  • Park SY, Kim KH, Gwak EH, Rhee SH, Lee JC, Shin SY, Koo KT, Lee YM, Seol YJ (2015) Ex vivo bone morphogenetic protein 2 gene delivery using periodontal ligament stem cells for enhanced re-osseointegration in the regenerative treatment of peri-implantitis. J Biomed Mater Res A 103:38–47

    Article  Google Scholar 

  • Rim NG, Kim SJ, Shin YM, Jun I, Lim DW, Park JH, Shin H (2012) Mussel-inspired surface modification of poly (L-lactide) electrospun fibers for modulation of osteogenic differentiation of human mesenchymal stem cells. Colloids Surf B Biointerfaces 91:189–197

    Article  CAS  Google Scholar 

  • Seo BM, Miura M, Gronthos S, Bartold PM, Batouli S, Brahim J, Young M, Robey PG, Wang CY, Shi S (2004) Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet 364:149–155

    Article  CAS  Google Scholar 

  • Xu FJ, Wang ZH, Yang WT (2010) Surface functionalization of polycaprolactone films via surface-initiated atom transfer radical polymerization for covalently coupling cell-adhesive biomolecules. Biomaterials 31:3139–3147

    Article  CAS  Google Scholar 

  • Yamaguchi A, Komori T, Suda T (2000) Regulation of osteoblast differentiation mediated by bone morphogenetic proteins, hedgehogs, and Cbfa1. Endocr Rev 21:393–411

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (No. 2015R1A2A2A01006490).

Author information

Authors and Affiliations

  1. Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, 26 Kyunghee-daero, Dongdaemun-gu, Seoul, 02447, South Korea

    Jeong Seok Lee & Jung Sun Heo

  2. Department of Orthodontics and Institute of Oral Biosciences, Research Center of Bioactive Materials, Chonbuk National University, Jeonju, 54896, South Korea

    Jeong-Chae Lee

Authors
  1. Jeong Seok Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jeong-Chae Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jung Sun Heo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jung Sun Heo.

Ethics declarations

Conflict of interest

The authors declare no conflict of interest.

Electronic supplementary material

Supplementary figure 1

Osteogenic differentiation of human osteoblast cell line (hFOB 1.19) cultured on PDA-modified surfaces. (A) ALP activity, (B) intracellular calcium levels, and (C) protein levels of osteogenic markers, OCN and OSX were determined after 4 and 7 days of osteogenic induction. The values reported are the means ± S.D. of four independent experiments. *P < 0.05 vs. 4 day-control value, and #P < 0.05 vs. 7-day control value. (PDF 78 kb)

Supplementary figure 2

Effect of BMP-2 neutralization on PDLSCs osteogenesis. Cells were cultured on Ti, PDA, PDA/BMP-2 (100 ng/ml), and PDA/BMP-2 (100 ng/ml) plus neutralizing antibody-Ti substrates for 4 or 7 days, and (A) ALP activity, (B) intracellular calcium levels, (C) real time RT-PCR for OCN, OPN, RUNX2 were then assessed. The values reported are the means ± S.D. of five independent experiments. *P < 0.05 vs. each control value, and #P < 0.05 vs. PDA/BMP-2 (100 ng/ml) value. (PDF 11 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lee, J.S., Lee, JC. & Heo, J.S. Polydopamine-assisted BMP-2 immobilization on titanium surface enhances the osteogenic potential of periodontal ligament stem cells via integrin-mediated cell-matrix adhesion. J. Cell Commun. Signal. 12, 661–672 (2018). https://doi.org/10.1007/s12079-018-0468-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12079-018-0468-0

Keywords

Search

Navigation