Skip to main content

Advertisement

SpringerLink
Log in

In vitro release of silver ions and expression of osteogenic genes by MC3T3-E1 cell line cultured on nano-hydroxyapatite and silver/strontium-coated titanium plates

  • Original Article
  • Published:
Odontology Aims and scope Submit manuscript

Abstract

Attempts are ongoing to improve the surface properties of dental implants by application of different coatings, aiming to enhance osseointegration, and decrease the adverse effects of titanium and its alloys used in dental implants. Coating of implant surface with hydroxyapatite (HA) is one suggested strategy for this purpose due to its high biocompatibility and similar structure to the adjacent bone. This study aimed to quantify the release of silver ions and expression of osteogenic genes by MC3T3-E1 cells cultured on nano-HA and silver/strontium (Ag/Sr)-coated titanium plates via the electrochemical deposition method. Plates measuring 10 × 10 × 0.9 mm were fabricated from Ti-6Al-4 V alloy, and polished with silicon carbide abrasive papers before electrochemical deposition to create a smooth, mirror-like surface. After applying homogenous nano-HA coatings with/without silver/strontium on the surface of the plates, the composition of coatings was confirmed by energy-dispersive X-ray spectroscopy (EDS), and their morphological properties were analyzed by scanning electron microscopy (SEM). The coated specimens were then immersed in simulated body fluid (SBF), and the concentration of released sliver ions was quantified by spectroscopy at 7–14 days. The MC3T3-E1 osteoblastic cell line was cultured in osteogenic medium for 7–14 days, and after RNA extraction and cDNA synthesis, the expression of runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OPN); osteogenic genes was quantified by polymerase chain reaction (PCR) using SYBR Green Master Mix kit. The expression of genes and the released amount of silver ions were compared between the two groups using the Mann–Whitney U test. The two groups were not significantly different regarding silver ion release at 14 days (P > 0.05). However, silver ion release was significantly higher from nano-HA coatings with silver/strontium at 7 days (P = 0.03). The difference in expression of RUNX2 (P = 0.04), OPN (P = 0.04), and OCN (P = 0.03) genes was also significant between nano-HA coating groups with and without silver/strontium at 7 days, and the expressions were higher in nano-HA with silver/strontium group, but this difference was not significant at 14 days. Addition of silver and strontium to specimens coated with nano-HA increased the release of silver ions within the non-toxic range, and enhanced the expression of osteogenic genes particularly after 7 days.

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

Similar content being viewed by others

Data availability

The datasets obtained and analyzed during the current study are available from the corresponding author upon reasonable request.

Abbreviations

HA:

Hydroxyapatite

Ag/Sr:

Silver/Strontium

EDS:

Energy-Dispersive X-ray Spectroscopy

SEM:

Scanning Electron Microscopy

SBF:

Simulated Body Fluid

RUNX2:

Runt-Related Transcription Factor 2

OCN:

Osteocalcin

OPN:

Osteopontin

PCR:

Polymerase Chain Reaction

References

  1. Ambard AJ, Mueninghoff L. Calcium phosphate cement: Review of mechanical and biological properties. J Prosthodont. 2006;15:321–8.

    Article  Google Scholar 

  2. Brunette DM, Tengvall P, Textor M TP, Textor M, Thomsen P. (2013) Titanium in medicine: material science, surface science, engineering, biological responses and medical applications. Springer Sci Bus Media 13–24.

  3. Bernardi S, Bianchi S, Tomei AR, Continenza MA, Macchiarelli G. Microbiological and SEM-EDS evaluation of titanium surfaces exposed to periodontal gel. In Vitro Study Mater (Basel). 2019;12(9):1448.

    Article  Google Scholar 

  4. Kula Z, Semenov M, Klimek L. Carbon coatings deposited on prosthodontic Ni-Cr alloy. Appl Sci. 2021;11(10):4551.

    Article  Google Scholar 

  5. Prodanov L, Lamers E, Domanski M, Luttge R, Jansen JA, Walboomers XF. The effect of nanometric surface texture on bone contact to titanium implants in rabbit tibia. Biomaterials. 2013;34:2920–7.

    Article  Google Scholar 

  6. Bral A, Mommaerats MY. In vivo bio-functionalization of titanium patient-specific implants with nano-hydroxyapatite and other nano-calcium phosphate coatings: a systematic review. J Cranio-maxillofac Surg. 2016;44:400–12.

    Article  Google Scholar 

  7. Webster TJ, Ejiofor JU. Increased osteoblast adhesion on nano-phase metals: Ti, Ti-6Al-4V, and CoCrMo. Biomaterials. 2004;25:4731–9.

    Article  Google Scholar 

  8. Gopi D, Shinyjoy E, Kavitha L. Synthesis and spectral characterization of silver/magnesium co-substituted hydroxyapatite for biomedical applications. Spectrochim Acta Part A Mol Biomol Spectrosc. 2014;127:286–91.

    Article  Google Scholar 

  9. Lv M, Su S, He Y, Huang Q, Hu W, Li D, Fan C, Lee ST. Long-term antimicrobial effect of silicon nanowires decorated with silver nanoparticles. Adv Mater. 2010;22:5463–7.

    Article  Google Scholar 

  10. Zhou JH, Li B, Lu SM, Zhang L, Han Y. Regulation of osteoblast proliferation and differentiation by interred spacing of Sr-HA nanorods on microporous titania coating. ACS Appl Mater Int. 2013;5:5358–65.

    Article  Google Scholar 

  11. Wong KL, Wong CT, Liu WC, Pan HB, Fong MK, Lam WM, Cheung WL, Tang WM, Chiu KY, Luk KDK, Lu WW. Mechanical properties and in vitro response of strontium-containing hydroxyapatite/polyetherether-ketone composites. Biomaterials. 2009;30:3810–7.

    Article  Google Scholar 

  12. Roy M, Fielding GA, Beyenal H, Bandyopadhyay A, Bose S. Mechanical in vitro antimicrobial, and biological properties of plasma sprayed silver doped hydroxyapatite coating. ACS Appl Mater Interfaces. 2012;4(3):1341–9.

    Article  Google Scholar 

  13. El-Wassefy NA, Reicha FM, Aref NS. Electrochemical deposition of nano hydroxyapatite-zinc coating on titanium metal substrate. Int J Implant Dent. 2017;3:39–50.

    Article  Google Scholar 

  14. Geng Z, Wang R, Zhuo X, Li Z, Huange Y, Ma L, Cuia Z, Zhu S, Liang Y, Liu Y, Bao H, Li X, Huo Q, Liu Z, Yang X. Incorporation of silver and strontium in hydroxyapatite coating on titanium surface for enhanced antibacterial and biological properties. Mater Sci Eng. 2017;71:852–61.

    Article  Google Scholar 

  15. Huang Y, Zhang X, Zhang H, Qiao H, Zhang X, Jia T, Han S, Gao Y, Xiao H, Yang H. Fabrication of silver- and strontium-doped hydroxyapatite/ TiO2 nanotube bilayer coatings for enhancing bactericidal effect and osteoinductivity. Ceramics Int. 2017;43(1):992–1007.

    Article  Google Scholar 

  16. Fu C, Zhang X, Savino K, Gabrys P, Gao Y, Chaimayo W, Miller BL, Yates MZ. Antimicrobial silver hydroxyapatite composite coatings through two-stage electro-chemical synthesis. J Ceramics. 2016;301:13–9.

    Google Scholar 

  17. Liang Y, Li H, Xu J, Li X, Li X, Yan Y, Qi M, Hu M. Strontium coating by electrochemical deposition improves implant osseointegration in osteopenic models. Exp Ther Med. 2015;9:172–6.

    Article  Google Scholar 

  18. Shimazaki T, Miyamoto H, Ando Y, Noda I, Yonekura Y, Kawano S, Miyazaki M, Mawatari M, Hotokebuchi T. In vivo antibacterial and silver-releasing properties of novel thermal sprayed silver-containing hydroxy-apatite coating. J Biomed Mater Res B Appl Biomater. 2010;92(2):386–9.

    Google Scholar 

  19. Chen Y, Zheng X, Xie Y, Ji H, Ding C, Li H, Dai K. Silver release from silver-containing hydroxyapatite coatings. Surf Coat Technol. 2010;205:1892–6.

    Article  Google Scholar 

  20. Fu C, Savino K, Gabrys P, Zeng A, Guan B, Olvera D, et al. Hydroxyapatite thin films with giant electrical polarization. Chem Mater. 2015;27:1164–71.

    Article  Google Scholar 

  21. Bartmanski M, Cieslik B, Glodowska J, Kalka P, Pawloski L, Pieper M, Zielingski A. Electrophoretic deposition (EPD) of nanohydroxyapatite- nano-silver coatings on Ti13Zr13Nb alloy. Ceramics Int. 2017;43:11820–9.

    Article  Google Scholar 

  22. Erdem U, Turkoz MB. Silver release of Ag (I) doped hydroxyapatite: In vitro study. Microsc Res Tech. 2019;82(7):961–71.

    Article  Google Scholar 

  23. Mirzaee M, Vaezi M, Palizdar Y. Synthesis and characterization of silver doped hydroxyapatite nano-composite coatings and evaluation of their antibacterial and corrosion resistance properties in simulated body fluid. Mater Sci Eng C. 2016;69:675–84.

    Article  Google Scholar 

  24. Zhu Z-Y, Zhang F-Q, Zheng X-B. Study on the slow release of silver ion from silver containing antibacterial HA coating material. Shanghai Kou Qiang Yi Xue. 2009;18(1):66–8.

    Google Scholar 

  25. Fielding GA, Roy M, Bandyopadhyay A, Bose S. Antibacterial and biological characteristics of silver containing and strontium doped plasma sprayed hydroxyl-apatite coatings. Acta Biomater. 2012;8:3144–52.

    Article  Google Scholar 

  26. Geng Z, Cui ZD, Li ZY, Zhu SL, Liang YQ, Liu YD. Strontium incorporation to optimize the antibacterial and biological characteristics of silver-substituted hydroxyl-apatite coating. Mater Sci Eng C. 2016;58:467–77.

    Article  Google Scholar 

  27. Guo X, Gough JE, Xiao P, Liu J, Shen Z. Fabrication of nanostructured hydroxyapatite and analysis of human osteoblastic cellular response. J Biomed Mater Res A. 2007;82:1022–32.

    Article  Google Scholar 

  28. Matsumoto N, Sato K, Yoshida K, Hashimoto K, Toda Y. Preparation and characterization of β-tricalcium phosphate co-doped with monovalent and divalent antibacterial metal ions. Acta Biomater. 2009;5:3157–64.

    Article  Google Scholar 

  29. Song WH, Ryu HS, Hong SH. Antibacterial properties of Ag (or Pt)-containing calcium phosphate coatings formed by micro-arc oxidation. J Biomed Mater Res. 2009;88A:246–54.

    Article  Google Scholar 

  30. Hata K, Kokubo T, Nakamura T, Yamamuro T. Growth of a bonelike apatite layer on a substrate by a biomimetic process. J Am Ceram Soc. 1995;78(4):1049–53.

    Article  Google Scholar 

  31. Bonnelye E, Chabadel A, Saltel F, Jurdic P. Dual effect of strontium ranelate: stimulation of osteoblast differentiation and inhibition of osteoclast formation and resorption in vitro. Bone. 2008;42:129–38.

    Article  Google Scholar 

  32. Ni GX, Yao ZP, Huang GT, Liu WG, Lu WW. The effect of strontium incorporation in hydroxyapatite on osteoblasts in vitro. J Mater Sci Mater Med. 2011;22:961–7.

    Article  Google Scholar 

  33. Choudhary S, Halbout P, Alander C, Raisz L, Pilbeam C. Strontium ranelate promotes osteoblastic differentiation and mineralization of murine bone marrow stromal cells: involvement of prostaglandins. J Bone Miner Res. 2007;22:1002–10.

    Article  Google Scholar 

  34. Gotoh Y, Hiraiwa K, Nagayama M. In vitro mineralization of osteoblastic cells derived from human bone. Bone Miner. 1990;8(3):239–50.

    Article  Google Scholar 

  35. Stein GS, Lian JB, Owen TA. Relationship of cell growth to the regulation of tissue-specific gene expression during osteoblast differentiation. FASEB J. 1990;4:3111–23.

    Article  Google Scholar 

  36. Capuccini C, Torricelli P, Sima F. Strontium-substituted hydroxyapatite coatings synthesized by pulsed-laser deposition: in vitro osteoblast and osteoclast response. Acta Biomater. 2008;4:1885–93.

    Article  Google Scholar 

  37. Caverzasio J, Thouverey C. Activation of FGF receptors is a new mechanism by which strontium ranelate induces osteoblastic cell growth. Cell Physiol Biochem. 2011;27:243–50.

    Article  Google Scholar 

  38. Baier M, Staudt P, Klein R. Strontium enhances osseointegration of calcium phosphate cement: a histomorphometric pilot study in ovariectomized rats. J Orthop Surg Res. 2013;8:16.

    Article  Google Scholar 

  39. Pan HB, Zhao XL, Zhang X. Strontium borate glass: potential biomaterial for bone regeneration. J R Soc Interface. 2010;7:1025–31.

    Article  Google Scholar 

  40. Xuejiao Z, Bingbing W, Lifei M, Lei X, Hao Y, Yichao L, Saisai W, Haixia Q, He L, Jingpin L, Yong H. Chemical stability, antibacterial and osteogenic activities study of strontium-silver co-substituted fluorohydroxyapatite nanopillars: a potential multifunctional biological coating. Ceram Int. 2020;17:27758–73.

    Google Scholar 

  41. Bernardi S, Macchiarelli G, Bianchi S. Autologous Materials in Regenerative Dentistry: Harvested Bone, Platelet Concentrates and Dentin Derivates. Molecules. 2020;25(22):5330.

    Article  Google Scholar 

  42. Bianchi S, Mancini L, Torge D, Cristiano L, Mattei A, Varvara G, Macchiarelli G, Marchetti E, Bernardi S. Bio-Morphological Reaction of Human Periodontal Ligament Fibroblasts to Different Types of Dentinal Derivates: In Vitro Study. International Journal of Molecular Sciences. 2021; 22(16):8681.

Download references

Acknowledgements

The authors gratefully acknowledge the financial support for this work that was provided by Research Institute for Dental Sciences, Shahid Beheshti University of Medical Sciences.

Funding

This research was supported by Research Institute for Dental Sciences, Shahid Beheshti University of Medical Sciences.

Author information

Authors and Affiliations

  1. Department of Periodontics, School of Dentistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Ardeshir Lafzi & Azadeh Esmaeil Nejad

  2. Dental Research Center, School of Dentistry, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Maryam Rezai Rad

  3. Community Oral Health Department, School of Dentistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Mahshid Namdari

  4. Department of Periodontics, School of Dentistry, Iran University of Medical Sciences, Tehran, Iran

    Tohid Sabetmoghaddam

Authors
  1. Ardeshir Lafzi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Azadeh Esmaeil Nejad
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maryam Rezai Rad
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mahshid Namdari
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tohid Sabetmoghaddam
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

A.L. and A.E.N. conceived the study; M.R.R. and T.S. performed the experiments; M.N. analyzed the data; A.L. and T.S. wrote the manuscript. All authors have read and approved the final manuscript.

Corresponding author

Correspondence to Tohid Sabetmoghaddam.

Ethics declarations

Conflict of interests

The authors have declared that No competing interests exist.

Ethics approval and consent to participate

This study was approved by the Research Institute of Dental Sciences (Shahid Beheshti University of Medical Sciences) Ethics Committee (approval number 161).

Consent for publication

Not applicable.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lafzi, A., Esmaeil Nejad, A., Rezai Rad, M. et al. In vitro release of silver ions and expression of osteogenic genes by MC3T3-E1 cell line cultured on nano-hydroxyapatite and silver/strontium-coated titanium plates. Odontology 111, 33–40 (2023). https://doi.org/10.1007/s10266-022-00747-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10266-022-00747-z

Keywords

Search

Navigation