본문 바로가기
KERD
논문KCI 등재

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

2

조회

0

다운로드

1

인용

0

참고문헌

키워드

초록

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-μm-wide/10-μm-deep microgrooved surface with fibronectin (E60/10FN) compared with the same surface without fibronectin (E60/10), and a more than fourfoldincrease 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 α1 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.

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-μm-wide/10-μm-deep microgrooved surface with fibronectin (E60/10FN) compared with the same surface without fibronectin (E60/10), and a more than fourfoldincrease 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 α1 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.

참고문헌 (31)

  1. Lee SW, "Influence of etched microgrooves of uniform dimension on in vitro responses of human gingival fibroblasts", Clin Oral Implants Res, 20, pp.458~466 (2009)
  2. Lee MH, "Factors influencing osteoblast maturation on microgrooved titanium substrata", Biomaterials, 31, pp.3804~3815 (2010)
  3. 박정애, "Effect of etched microgrooves on hydrophilicity of titanium and osteoblast responses: A pilot study", The Journal of Advanced of Prosthodontics, 2(1), pp.18~24 (2010), 대한치과보철학회
  4. Kim SY, "Surface microgrooves and acid etching on titanium substrata alter various cell behaviors of cultured human gingival fibroblasts", Clin Oral Implants Res, 20, pp.262~272 (2009)
  5. Guida L, "Biological response of human bone marrow mesenchymal stem cells to fluoride-modified titanium surfaces", Clin Oral Implants Res, 21, pp.1234~1241 (2010)
  6. Lee MH, "The significance of differential expression of genes and proteins in human primary cells caused by microgrooved biomaterial substrata", Biomaterials, 33, pp.3216~3234 (2012)
  7. Engvall E, "Binding of soluble form of fibroblast surface protein, fibronectin, to collagen", Int J Cancer, 20, pp.1~5 (1977)
  8. Dean JW, "Fibronectin and laminin enhance gingival cell attachment to dental implant surfaces in vitro", Int J Oral Maxillofac Implants, 10, pp.721~728 (1995)
  9. Gallant ND, "Cell adhesion strengthening:contributions of adhesive area, integrin binding, and focal adhesion assembly", Mol Biol Cell, 16, pp.4329~4340 (2005)
  10. Fini M, "Biomechanical and histomorphometric investigations on two morphologically differing titanium surfaces with and without fluorohydroxyapatite coating: an experimental study in sheep tibiae", Biomaterials, 24, pp.3183~3192 (2003)
  11. Rezania A, "A probabilistic approach to measure the strength of bone cell adhesion to chemically modified surfaces", Ann Biomed Eng, 25, pp.190~203 (1997)
  12. Middleton CA, "Fibronectin silanized titanium alloy: a bioinductive and durable coating to enhance fibroblast attachment in vitro", J Biomed Mater Res A, 83, pp.1032~1038 (2007)
  13. Pendegrass CJ, "Measuring the strength of dermal fibroblast attachment to functionalized titanium alloys in vitro", J Biomed Mater Res A, 92, pp.1028~1037 (2010)
  14. Chou L, "Substratum surface topography alters cell shape and regulates fibronectin mRNA level, mRNA stability, secretion and assembly in human fibroblasts", J Cell Sci, 108(Pt 4), pp.1563~1573 (1995)
  15. Lee SW, "Influence of microgroove dimension on cell behavior of human gingival fibroblasts cultured on titanium substrata", Clin Oral Implants Res, 20, pp.56~66 (2009)
  16. Jaiswal N, "Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro", J Cell Biochem, 64, pp.295~312 (1997)
  17. Branemark PI, "Titanium implants permanently penetrating human skin", Scand J Plast Reconstr Surg, 16, pp.17~21 (1982)
  18. Dalby MJ, "Osteoprogenitor response to defined topographies with nanoscale depths", Biomaterials, 27, pp.1306~1315 (2006)
  19. den Braber ET, "Quantitative analysis of fibroblast morphology on microgrooved surfaces with various groove and ridge dimensions", Biomaterials, 17, pp.2037~2044 (1996)
  20. Li Z, "Biological functions of miR-29b contribute to positive regulation of osteoblast differentiation", J Biol Chem, 284, pp.15676~15684 (2009)
  21. Valencia S, "Surface nanofeature effects on titanium-adherent human mesenchymal stem cells", Int J Oral Maxillofac Implants, 24, pp.38~46 (2009)
  22. Mendonca G, "The effects of implant surface nanoscale features on osteoblast-specific gene expression", Biomaterials, 30, pp.4053~4062 (2009)
  23. Mendonca G, "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, 2010, pp.94, 169~179
  24. Chau JF, "Signaling pathways governing osteoblast proliferation, differentiation and function", Histol Histopathol, 24, pp.1593~1606 (2009)
  25. Nakashima K, "The novel zinc fingercontaining transcription factor osterix is required for osteoblast differentiation and bone formation", Cell, 108, pp.17~29 (2002)
  26. Balloni S, "Effects of titanium surface roughness on mesenchymal stem cell commitment and differentiation signaling", Int J Oral Maxillofac Implants, 24, pp.627~635 (2009)
  27. Jayaraman M, "Influence of titanium surfaces on attachment of osteoblastlike cells in vitro", Biomaterials, 25, pp.625~631 (2004)
  28. Sato M, "Transcriptional regulation of osteopontin gene in vivo by PEBP2alphaA", CBFA1 and ETS1 in the skeletal tissues, 1998, pp.17, 1517~1525
  29. Komori T, "Regulation of bone development and extracellular matrix protein genes by RUNX2", Cell Tissue Res, 339, pp.189~195 (2010)
  30. Ducy P, "A Cbfa1-dependent genetic pathway controls bone formation beyond embryonic development", Genes Dev, 13, pp.1025~1036 (1999)
  31. Maruyama Z, "Runx2 determines bone maturity and turnover rate in postnatal bone development and is involved in bone loss in estrogen deficiency", Dev Dyn, 236, pp.1876~1890 (2007)

관련 논문AI추천

Effect of etched microgrooves on hydrophilicity of titanium and osteoblast responses: A pilot study박정애, 안수진, 이석원, 이성복대한치과보철학회 · The Journal of Advanced of Prosthodontics2010 · 제2권 제1호Heat or radiofrequency plasma glow discharge treatment of a titanium alloy stimulates osteoblast gene expression in the MC3T3 osteoprogenitor cell lineBruce E. Rapuano, Daniel E. MacDonald, Kyle Hackshaw대한치주과학회 · Journal of Periodontal & Implant Science2012 · 제42권 제3호Effect of implant surface microtopography by hydroxyapatite grit-blasting on adhesion, proliferation, and differentiation of osteoblast-like cell line, MG-63김수경, 박성재, 배상범, 송승일, 엄태관대한구강악안면외과학회 · 대한구강악안면외과학회지2011 · 제37권 제3호Comparison of alkaline phosphatase activity of MC3T3-E1 cells cultured on different Ti surfaces: modified sandblasted with large grit and acidetched (MSLA), laser-treated, and laser and acid-treated Ti surfaces김수남, 이인제, 조성암대한치과보철학회 · The Journal of Advanced of Prosthodontics2016 · 제8권 제3호Response of osteoblast-like cells cultured on zirconia to bone morphogenetic protein-2구영, 구기태, 김경화, 김태일, 류인철, 설양조, 이용무, 한승희, 한중석대한치주과학회 · Journal of Periodontal & Implant Science2011 · 제41권 제5호Implant surface treatments affect gene expression of Runx2, osteogenic key marker나영, 곽재영, 김성균, 허성주대한치과보철학회 · The Journal of Advanced of Prosthodontics2009 · 제1권 제2호

이 간행물의 다른 논문 (11건)