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Original Research

Cell responses to two kinds of nanohydroxyapatite with different sizes and crystallinities

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Pages 1239-1250 | Published online: 07 Mar 2012
 

Abstract

Introduction

Hydroxyapatite (HA) is the principal inorganic constituent of human bone. Due to its good biocompatibility and osteoconductivity, all kinds of HA particles were prepared by different methods. Numerous reports demonstrated that the properties of HA affected its biological effects.

Methods

Two kinds of nanohydroxyapatite with different sizes and crystallinities were obtained via a hydrothermal treatment method under different temperatures. It was found that at a temperature of 140°C, a rod-like crystal (n-HA1) with a diameter of 23 ± 5 nm, a length of 47 ± 14 nm, and crystallinity of 85% ± 5% was produced, while at a temperature of 80°C, a rod-like crystal (n-HA2) with a diameter of 16 ± 3 nm, a length of 40 ± 10 nm, and crystallinity of 65% ± 3% was produced. The influence of nanohydroxyapatite size and crystallinity on osteoblast viability was studied by MTT, scanning electron microscopy, and flow cytometry.

Results

n-HA1 gave a better biological response than n-HA2 in promoting cell growth and inhibiting cell apoptosis, and also exhibited much more active cell morphology. Alkaline phosphatase activity for both n-HA2 and n-HA1 was obviously higher than for the control, and no significant difference was found between n-HA1 and n-HA2. The same trend was observed on Western blotting for expression of type I collagen and osteopontin. In addition, it was found by transmission electron microscopy that large quantities of n-HA2 entered into the cell and damaged the cellular morphology. Release of tumor necrosis factor alpha from n-HA2 was markedly higher than from n-HA1, indicating that n-HA2 might trigger a severe inflammatory response.

Conclusion

This work indicates that not all nanohydroxyapatite should be considered a good biomaterial in future clinical applications.

Acknowledgments

This work was supported by the State Key Development Program for Basic Research of China (grant 2007CB936103), Fundamental Research Funds for the Central Universities, Peking University 985 grant, the National Natural Science Foundation of China (30970720), and Key Medical Program of Science and Technology Development of Shanghai (09411954900).

Disclosure

The authors report no conflicts of interest in this work.