Effect of dopamine on the activity of matrix metalloproteinases and degradation of dentin collagen / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the inhibition effect of dopamine on the activity of matrix metalloproteinases (MMP) and the effect of dopamine on degradation of dentin collagen for its potential use in caries treatment and dentin adhesive.</p><p><b>METHODS</b>In the experiment of MMP activity test, 2.0 g/L dopamine + 1.0 g/L highly purified collagenase type VIII from Clostridium histolyticum served as the experimental group, and deionized water + 1.0 g/L highly purified collagenase type VIII from Clostridium histolyticum served as the negative control group, and 2% chlorhexidine + 1.0 g/L highly purified collagenase type VIII from Clostridium histolyticum served as the positive control group, and the mixture volume ratio of the two ingredients in every group was 1:9. After 15 minutes, the enzyme activity of each sample was tested by MMP activity colerimetric quantitative detection kits, and the test was repeated 5 times in each group. In the experiment of collagen degradation, the dentin slices were demineralized with 37% phosphoric acid for 1 min. In sequence, 2 dentin slices were used to observe the morphology, and the remaining 30 dentine slices were randomly divided into three groups (n = 10) according to random number table: the negative control ones were stored in 100 µl deionized water and 900 µl collagenase (7 days, 37 °C), the positive control ones were stored in 100 µl chlorhexidine and 900 µl collagenase (7 days, 37 °C) and the experimental specimens were stored in 100 µl dopamine and 900 µl collagenase (7 days, 37 °C). The degraded collagen was investigated by assaying hydroxyproline. The framework of collagen was evaluated with field emission scanning electron microscope (FE-SEM).</p><p><b>RESULTS</b>The statistical results of completely random design ANOVA showed that the MMP activity and the amount of degraded collagen of the negative control group [(0.089 ± 0.011) µmol · min⁻¹ · mg⁻¹ and (2 837 ± 201) µg/cm²] were significantly higher than those of the positive control group [(0.038 ± 0.006) µmol · min⁻¹ · mg⁻¹ and (1 288 ± 172) µg/cm²] and the experimental group [(0.030 ± 0.009) µmol · min⁻¹ · mg⁻¹ and (1 389 ± 255) µg/cm²] (P < 0.05). SEM observation indicated that the structural integrity of the collagen network on dentin still existed in experiment samples and positive control groups, however, collagen fibrils were destructed and the structural integrity disappeared in the negative control groups.</p><p><b>CONCLUSIONS</b>Dopamine may inhibit MMP activity and reduce the amount of degraded collagen.</p>

Design and evaluation of a kind of biomimetic peptides of dentin matrix protein-1 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To design a kind of biomimetic polypeptide of dentin matrix protein-1 (DMP-1), which can bind to dentine collagen fibers and initiate mineralization.</p><p><b>METHODS</b>A novel polypeptide was developed by connecting the collagen binding domain of DMP-1 "DSESSEEDR" to the hydrophilic C-terminal of amelogenin "TKREEVD". The polypeptide was synthetically prepared by standard solid-phase peptide synthesis. Human dentine slices were completely demineralized by hydrochloric acid to expose the dentine collagen. Fluorescein isothiocyanate coupled polypeptide was applied to the exposed dentine collagen. Fluorescent microscopy was used to examine the polypeptide specially bond to the dentine collagen. Nucleation and growth of hydroxyapatite was initiated by immersing the polypeptide into calcium chloride and sodium hypophosphate solutions respectively. Scanning electron microscopy (SEM), transmission electron microscopy (TEM) and selected area diffraction (SAD) were used to examine the hydroxyapatites formed. RESULTS Fluorescent dentine collagen was identified in the demineralized dentine specimens. Nucleation and growth of crystals were detected after immersing the polypeptide into calcium chloride and sodium hypophosphate solutions by SEM and TEM. SAD confirmed the crystals were hydroxyapatites.</p><p><b>CONCLUSION</b>The polypeptide of "DSESSEEDRTKREEVD" can simulate DMP-1 binding collagen and initiate hydroxyapatite nucleation and growth. It may be a potential molecular tool for dentine remineralization.</p>