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J Adv Prosthodont. 2015 Dec;7(6):496-505. doi: 10.4047/jap.2015.7.6.496. Epub 2015 Dec 30.

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

Author information

1
Department of Biomaterials & Prosthodontics, Kyung Hee University Hospital at Gangdong, Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul, Republic of Korea.
2
Department of Dentistry, Graduate School of Dentistry, Kyung Hee University, Seoul, Republic of Korea.
3
ED Dental Clinic, Seoul, Republic of Korea.
4
Department of Oral and Maxillofacial Surgery, Kyung Hee University Hospital at Gangdong, Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul, Republic of Korea.
5
Department of Prosthodontics, National Health Insurance Medical Center Ilsan Hospital, Goyang, Gyeonggi, Republic of Korea.
6
Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul, Republic of Korea.

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 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 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.

KEYWORDS:

Fibronectin conjugation; Gene expression; Microgrooves; Osteoblast differentiation; Titanium

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