2013 Fiscal Year Final Research Report
Elucidation of structural function and basic research for clinical application of dentin sialophosphoprotein in porcine pulp
| Project/Area Number |
24890265
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional basic dentistry
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| Research Institution | Tsurumi University |
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Principal Investigator |
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| Project Period (FY) |
2012-08-31 – 2014-03-31
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| Keywords | 歯学 / 再生医学 / 細胞・組織 / タンパク質 / 生理活性物質 |
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| Research Abstract |
We fractionated DPP and DSP along with TGF-b-like activity by ion exchange (IE) chromatography from developing pig molars and measured their alkaline phosphatase (ALP) stimulating activity in human periodontal (HPDL) cells with or without TGF-b receptor inhibitor. We then purified TGF-b-unbound or -bound DPP and DSP by reverse phase high performance liquid chromatography (RP-HPLC) using ALP-HPDL system. The TGF-b isoform bound to DPP and DSP was identified as being TGF-b1 by ELISA and LC-MS/MS analysis. We incubated carrier-free human recombinant TGF-b1 (CF-hTGF-b1) with TGF-b-unbound DPP or DSP and characterized the binding on IE-HPLC using the ALP-HPDL system. DPP and DSP rescued the loss of TGF-b1 activity. Approximately 19% and 10% of the ALP stimulating activities were retained by the binding of TGF-b1 to DPP and DSP, respectively. The type I collagen infrequently bound to CF-hTGFb1. We conclude that both DPP and DSP help retain TGF-b1 activity in porcine dentin.
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