The Journal of Advanced Prosthodontics

The Korean Academy of Prosthodonitics (대한치과보철학회)
권호 표지
DOI QR코드

DOI QR Code

Effect of microgrooves and fibronectin conjugation on the osteoblast marker gene expression and differentiation

  • Park, Su-Jung (Department of Biomaterials & Prosthodontics, Kyung Hee University Hospital at Gangdong, Institute of Oral Biology, School of Dentistry, Kyung Hee University) ;
  • Leesungbok, Richard (Department of Biomaterials & Prosthodontics, Kyung Hee University Hospital at Gangdong, Institute of Oral Biology, School of Dentistry, Kyung Hee University) ;
  • Ahn, Su-Jin (Department of Biomaterials & Prosthodontics, Kyung Hee University Hospital at Gangdong, Institute of Oral Biology, School of Dentistry, Kyung Hee University) ;
  • Im, Byung-Jin (Department of Dentistry, Graduate School of Dentistry, Kyung Hee University) ;
  • Lee, Do Yun (ED Dental Clinic) ;
  • Jee, Yu-Jin (Department of Oral and Maxillofacial Surgery, Kyung Hee University Hospital at Gangdong, Institute of Oral Biology, School of Dentistry, Kyung Hee University) ;
  • Yoon, Joon-Ho (Department of Prosthodontics, National Health Insurance Medical Center Ilsan Hospital) ;
  • Cui, Taixing (Department of Dentistry, Graduate School of Dentistry, Kyung Hee University) ;
  • Lee, Sang Cheon (Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University) ;
  • Lee, Suk Won (Department of Biomaterials & Prosthodontics, Kyung Hee University Hospital at Gangdong, Institute of Oral Biology, School of Dentistry, Kyung Hee University)
  • Received : 2015.11.25
  • Accepted : 2015.12.22
  • Published : 2015.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

PURPOSE. To determine the effect of fibronectin (FN)-conjugated, microgrooved titanium (Ti) on osteoblast differentiation and gene expression in human bone marrow-derived mesenchymal stem cells (MSCs). MATERIALS AND METHODS. Photolithography was used to fabricate the microgrooved Ti, and amine functionalization (silanization) was used to immobilize fibronectin on the titanium surfaces. Osteoblast differentiation and osteoblast marker gene expression were analyzed by means of alkaline phosphatase activity assay, extracellular calcium deposition assay, and quantitative real-time PCR. RESULTS. The conjugation of fibronectin on Ti significantly increased osteoblast differentiation in MSCs compared with non-conjugated Ti substrates. On the extracellular calcium deposition assays of MSCs at 21 days, an approximately two-fold increase in calcium concentration was observed on the etched 60-${\mu}m$-wide/10-${\mu}m$-deep microgrooved surface with fibronectin (E60/10FN) compared with the same surface without fibronectin (E60/10), and a more than four-fold increase in calcium concentration was observed on E60/10FN compared with the non-etched control (NE0) and etched control (E0) surfaces. Through a series of analyses to determine the expression of osteoblast marker genes, a significant increase in all the marker genes except type I collagen ${\alpha}1$ mRNA was seen with E60/10FN more than with any of the other groups, as compared with NE0. CONCLUSION. The FN-conjugated, microgrooved Ti substrate can provide an effective surface to promote osteoblast differentiation and osteoblast marker gene expression in MSCs.

Keywords

References

  1. Lee SW, Kim SY, Lee MH, Lee KW, Leesungbok R, Oh N. Influence of etched microgrooves of uniform dimension on in vitro responses of human gingival fibroblasts. Clin Oral Implants Res 2009;20:458-66. https://doi.org/10.1111/j.1600-0501.2008.01671.x
  2. Lee MH, Oh N, Lee SW, Leesungbok R, Kim SE, Yun YP, Kang JH. Factors influencing osteoblast maturation on microgrooved titanium substrata. Biomaterials 2010;31:3804-15. https://doi.org/10.1016/j.biomaterials.2010.01.117
  3. Park JA, Leesungbok R, Ahn SJ, Lee SW. Effect of etched microgrooves on hydrophilicity of titanium and osteoblast responses: A pilot study. J Adv Prosthodont 2010;2:18-24. https://doi.org/10.4047/jap.2010.2.1.18
  4. Kim SY, Oh N, Lee MH, Kim SE, Leesungbok R, Lee SW. Surface microgrooves and acid etching on titanium substrata alter various cell behaviors of cultured human gingival fibroblasts. Clin Oral Implants Res 2009;20:262-72. https://doi.org/10.1111/j.1600-0501.2008.01652.x
  5. Guida L, Annunziata M, Rocci A, Contaldo M, Rullo R, Oliva A. Biological response of human bone marrow mesenchymal stem cells to fluoride-modified titanium surfaces. Clin Oral Implants Res 2010;21:1234-41. https://doi.org/10.1111/j.1600-0501.2010.01929.x
  6. Lee MH, Kang JH, Lee SW. The significance of differential expression of genes and proteins in human primary cells caused by microgrooved biomaterial substrata. Biomaterials 2012;33:3216-34. https://doi.org/10.1016/j.biomaterials.2012.01.034
  7. Engvall E, Ruoslahti E. Binding of soluble form of fibroblast surface protein, fibronectin, to collagen. Int J Cancer 1977;20:1-5. https://doi.org/10.1002/ijc.2910200102
  8. Dean JW, 3rd, Culbertson KC, D'Angelo AM. Fibronectin and laminin enhance gingival cell attachment to dental implant surfaces in vitro. Int J Oral Maxillofac Implants 1995;10:721-8.
  9. Gallant ND, Michael KE, Garcia AJ. Cell adhesion strengthening: contributions of adhesive area, integrin binding, and focal adhesion assembly. Mol Biol Cell 2005;16:4329-40. https://doi.org/10.1091/mbc.E05-02-0170
  10. Fini M, Savarino L, Nicoli Aldini N, Martin L, Giavaresi G, Rizzi G, Martini D, Ruggeri A, Giunti A, Giardino R. Biomechanical and histomorphometric investigations on two morphologically differing titanium surfaces with and without fluorohydroxyapatite coating: an experimental study in sheep tibiae. Biomaterials 2003;24:3183-92. https://doi.org/10.1016/S0142-9612(03)00164-9
  11. Rezania A, Thomas CH, Healy KE. A probabilistic approach to measure the strength of bone cell adhesion to chemically modified surfaces. Ann Biomed Eng 1997;25:190-203. https://doi.org/10.1007/BF02738550
  12. Middleton CA, Pendegrass CJ, Gordon D, Jacob J, Blunn GW. Fibronectin silanized titanium alloy: a bioinductive and durable coating to enhance fibroblast attachment in vitro. J Biomed Mater Res A 2007;83:1032-8.
  13. Pendegrass CJ, Middleton CA, Gordon D, Jacob J, Blunn GW. Measuring the strength of dermal fibroblast attachment to functionalized titanium alloys in vitro. J Biomed Mater Res A 2010;92:1028-37.
  14. Chou L, Firth JD, Uitto VJ, Brunette DM. Substratum surface topography alters cell shape and regulates fibronectin mRNA level, mRNA stability, secretion and assembly in human fibroblasts. J Cell Sci 1995;108 (Pt 4):1563-73.
  15. Lee SW, Kim SY, Rhyu IC, Chung WY, Leesungbok R, Lee KW. Influence of microgroove dimension on cell behavior of human gingival fibroblasts cultured on titanium substrata. Clin Oral Implants Res 2009;20:56-66. https://doi.org/10.1111/j.1600-0501.2008.01597.x
  16. Jaiswal N, Haynesworth SE, Caplan AI, Bruder SP. Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem 1997;64:295-312. https://doi.org/10.1002/(SICI)1097-4644(199702)64:2<295::AID-JCB12>3.0.CO;2-I
  17. Branemark PI, Albrektsson T. Titanium implants permanently penetrating human skin. Scand J Plast Reconstr Surg 1982;16:17-21. https://doi.org/10.3109/02844318209006565
  18. Dalby MJ, McCloy D, Robertson M, Wilkinson CD, Oreffo RO. Osteoprogenitor response to defined topographies with nanoscale depths. Biomaterials 2006;27:1306-15. https://doi.org/10.1016/j.biomaterials.2005.08.028
  19. den Braber ET, de Ruijter JE, Ginsel LA, von Recum AF, Jansen JA. Quantitative analysis of fibroblast morphology on microgrooved surfaces with various groove and ridge dimensions. Biomaterials 1996;17:2037-44. https://doi.org/10.1016/0142-9612(96)00032-4
  20. Li Z, Hassan MQ, Jafferji M, Aqeilan RI, Garzon R, Croce CM, van Wijnen AJ, Stein JL, Stein GS, Lian JB. Biological functions of miR-29b contribute to positive regulation of osteoblast differentiation. J Biol Chem 2009;284:15676-84. https://doi.org/10.1074/jbc.M809787200
  21. Valencia S, Gretzer C, Cooper LF. Surface nanofeature effects on titanium-adherent human mesenchymal stem cells. Int J Oral Maxillofac Implants 2009;24:38-46.
  22. Mendonca G, Mendonca DB, Simoes LG, Araujo AL, Leite ER, Duarte WR, Aragao FJ, Cooper LF. The effects of implant surface nanoscale features on osteoblast-specific gene expression. Biomaterials 2009;30:4053-62. https://doi.org/10.1016/j.biomaterials.2009.04.010
  23. Mendonca G, Mendonca DB, Aragao FJ, Cooper LF. The combination of micron and nanotopography by H(2)SO(4)/ H(2)O(2) treatment and its effects on osteoblast-specific gene expression of hMSCs. J Biomed Mater Res A 2010;94:169-79.
  24. Chau JF, Leong WF, Li B. Signaling pathways governing osteoblast proliferation, differentiation and function. Histol Histopathol 2009;24:1593-606.
  25. Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR, de Crombrugghe B. The novel zinc fingercontaining transcription factor osterix is required for osteoblast differentiation and bone formation. Cell 2002;108:17-29. https://doi.org/10.1016/S0092-8674(01)00622-5
  26. Balloni S, Calvi EM, Damiani F, Bistoni G, Calvitti M, Locci P, Becchetti E, Marinucci L. Effects of titanium surface roughness on mesenchymal stem cell commitment and differentiation signaling. Int J Oral Maxillofac Implants 2009;24:627-35.
  27. Jayaraman M, Meyer U, Buhner M, Joos U, Wiesmann HP. Influence of titanium surfaces on attachment of osteoblastlike cells in vitro. Biomaterials 2004;25:625-31. https://doi.org/10.1016/S0142-9612(03)00571-4
  28. Sato M, Morii E, Komori T, Kawahata H, Sugimoto M, Terai K, Shimizu H, Yasui T, Ogihara H, Yasui N, Ochi T, Kitamura Y, Ito Y, Nomura S. Transcriptional regulation of osteopontin gene in vivo by PEBP2alphaA/CBFA1 and ETS1 in the skeletal tissues. Oncogene 1998;17:1517-25. https://doi.org/10.1038/sj.onc.1202064
  29. Komori T. Regulation of bone development and extracellular matrix protein genes by RUNX2. Cell Tissue Res 2010;339:189-95. https://doi.org/10.1007/s00441-009-0832-8
  30. Ducy P, Starbuck M, Priemel M, Shen J, Pinero G, Geoffroy V, Amling M, Karsenty G. A Cbfa1-dependent genetic pathway controls bone formation beyond embryonic development. Genes Dev 1999;13:1025-36. https://doi.org/10.1101/gad.13.8.1025
  31. Maruyama Z, Yoshida CA, Furuichi T, Amizuka N, Ito M, Fukuyama R, Miyazaki T, Kitaura H, Nakamura K, Fujita T, Kanatani N, Moriishi T, Yamana K, Liu W, Kawaguchi H, Nakamura K, Komori T. Runx2 determines bone maturity and turnover rate in postnatal bone development and is involved in bone loss in estrogen deficiency. Dev Dyn 2007;236:1876-90. https://doi.org/10.1002/dvdy.21187

Cited by

  1. Regulation of human gingival fibroblast gene expression on microgrooves: A DNA microarray study vol.55, pp.4, 2017, https://doi.org/10.4047/jkap.2017.55.4.361
  2. Promotion of osteoblastic differentiation and osteogenic transcription factor expression on a microgroove titanium surface with immobilized fibronectin or bone sialoprotein II vol.11, pp.3, 2015, https://doi.org/10.1088/1748-6041/11/3/035020