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Inorganic Polyphosphate: a Possible Stimulant of Bone Formation

Y. Hacchou1,2, T. Uematsu1,2,*, O. Ueda3, Y. Usui4, S. Uematsu4, M. Takahashi1, T. Uchihashi1, Y. Kawazoe5, T. Shiba5, S. Kurihara4, M. Yamaoka1, and K. Furusawa1,2

1 Department of Oral and Maxillofacial Surgery,
2 Institute for Oral Science,
3 Department of Oral Microbiology, and
4 Department of Orthodontics, Matsumoto Dental University School of Dentistry, 1780 Gobara Hirooka, Shiojiri, Nagano 399-0874, Japan; and
5 Regenetiss Inc., 1-5-17, Akabane, Okaya, Nagano 394-0002, Japan


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  		Figure 1. Expression levels of osteoblast-related genes by real-time polymerase chain-reaction and alkaline phosphatase activity in MC 3T3-E1 cells. Cells were cultured in differentiation medium with 1 mM Poly(P) or without 1 mM Poly(P) (Control) for 35 days. Relative mRNA levels of OPN (A), OCN (B), Col I{alpha}(C), OPG (D), and ALP activity (E)—without treatment (open circles) and following treatment with 1 mM Poly(P) (closed circles)—are shown. Error bars represent the mean ± SD of three independent analyses.

 

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  		Figure 2. Influence of Poly(P) on calcification and decalcification with in vitro analysis. MC 3T3-E1 cells were cultured with 1 mM Poly(P)75 or 1 mM NaPO4 for 25 days and stained by the von Kossa method. Arrow represents nodule formation (A). After co-culture of mouse calvarial osteoblasts and bone marrow macrophages, a pit formation assay in the cultivation with the dentin slice was performed. Dentin decalcified by osteoclasts was stained by Mayer’s hematoxylin (B) and counted microscopically (C). Bar: 5 µm. Error bars represent the mean ± SD of three independent analyses.

 

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  		Figure 3. Histological and immunohistochemical analyses of periodontal tissue generation. (A) HE staining and immunostaining for ALP in the tissue samples given 1 mM CMC or 1 mM CMC + 1 mM Poly(P)75, with the periodontal regeneration mice model, 2 wks following surgery. Arrow: reference notch. Arrowhead: ALP-positive osteoblasts. Bar: 50 µm. (B) Comparison of bone regeneration area 1–3 wks after surgery. CMC: Carboxymethylcellulose. Error bars represent the mean ± SD of five independent analyses.

 




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