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. 2016 Nov 13;9(3):132-142.
doi: 10.11138/orl/2016.9.3.132. eCollection 2016 Jul-Sep.

Quantitive evaluation of dentin sialoprotein (DSP) using microbeads - a potential early marker of root resorption

L Lombardo  1 , F Carinci  2 , M Martini  1 , D Gemmati  3 , M Nardone  4 , G Siciliani  1
Affiliations

Quantitive evaluation of dentin sialoprotein (DSP) using microbeads - a potential early marker of root resorption

L Lombardo et al. Oral Implantol (Rome). .

Abstract

Purpose: This study had the aim of comparing two different methods of analysing dentin sialoprotein (DSP) in the gingival crevicular fluid (GCF): the conventional eLISA approach and a new method involving the use of magnetic micro-beads coated with an antibody specific for DSP prior to eLISA analysis.

Materials and methods: GCF was taken from six patients following twelve weeks of orthodontic treatment using paper strips inserted into the mesial and distal sulci of the upper incisors, and analysed using both methods.

Results: Statistical analysis of the results using the Mann-Whitney non-parametric test showed that the micro-bead approach conferred more reliability and less variability on the conventional eLISA approach. Furthermore, this method, for the first time, enables the quantification of the DSP in the sample in ng/μl.

Conclusions: The innovative micro-bead/eLISA approach proposed provides a reliable means of quantifying the DSP in the GCF.

Keywords: marker; resorption; root; sialoprotein; tooth.

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Figures

Figure 1
Figure 1
The sterile paper strip is inserted into the gingival sulcus to a depth of 1–2 mm and left in place for 1 minute.
Figure 2
Figure 2
Centrifuge tubes containing the paper GCF-collection strips during the laboratory analysis.
Figure 3
Figure 3
MagSi micro-beads (MagnaMedics®) coated with protein A or G and bound to specific antibodies (i.e., DSP-goat polyclonal IgG), which selectively bind to the target protein (i.e., DSP).
Figure 4
Figure 4
Calibration curve showing the behaviour of the purified protein and antibody.
Figure 5
Figure 5
Graph of eLISA test results obtained with the mean of the measurements for the six patients at each site using the micro-beads method.
Figure 6
Figure 6
Graph of eLISA test results obtained with the mean of the measurements for the six patients at each site using the eLISA-only method.
Figure 7
Figure 7
Results of the non-parametric Mann-Whitney test.
See this image and copyright information in PMC

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