Kit and method for detecting porous dental hydroxy apatite
A hydroxyapatite and kit technology, which can be used in the detection/measurement of irregular teeth, dental preparations, dentistry, etc., can solve the problems of difficult differential diagnosis and poor MIH effect.
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Embodiment 2
[0185] Example 2 - Preparation and testing of porous hydroxyapatite probes
[0186] Materials and Methods
[0187] SMCC (N-hydroxysuccinimidyl succinimidyl 4-[N-maleimidomethyl]cyclohexanecarboxylate; CAS#: 64987-85-5) is a non-cleavable isoform Bifunctional cross-linking agent, has been The spacer arm reactively separates the amine and sulfhydryl groups. Amino Black (CAS#: 1064-48-8) is a general purpose blue / black dye, used here as a chromogenic reporter containing primary amine groups. Hemoglobin from cow (CAS#: 9008-02-0) comprises a heterotetramer of 2 pairs of polypeptide chains (α and β; respectively: SEQ ID NO 20 and SEQ ID NO 21). The beta chain has a single solvent-exposed thiol-containing cysteine residue, while the alpha chain has no cysteine.
[0188] SMCC (75 mM in dimethyl sulfoxide) was added to 9 volumes of amino black (37.5 mM in phosphate buffered saline (PBS, 137 mM NaCl, 2.7 mM KCl, 10 mM disodium hydrogen phosphate, pH 7.4)) and Incubate at 21°C...
Embodiment 3
[0195] Example 3 - Probes that specifically bind to porous enamel ( Figure 30 )
[0196] method
[0197] To test whether the probe in Example 2 specifically binds to porous enamel, a complex carious lesion was covered with the probe and rinsed thoroughly.
[0198] Photograph of a human first molar with a large area of caries (porous enamel, white opaque areas) before and after application of the probe ( Figure 30 ). Use a brush to apply the probe to the entire crown area for 1 minute. After application of the probe, the tooth was rinsed under running water for 10 seconds, photographed, and then the tooth was rinsed again for 2 minutes, and photographed.
[0199] result
[0200] Normal enamel is not labeled.
[0201] Clearly carious areas were marked specifically and clearly, but the markings were patchy in some places. Unmarked carious areas show a bright surface that is resistant to scratching, while marked areas have a dark surface that is easily scratched. T...
Embodiment 4
[0204] Example 4 - The probe is able to specifically detect early demineralization of the enamel surface (incipient caries model) ( Figure 31 )
[0205] method
[0206] To test whether the probe in Example 2 could specifically detect early caries, artificial caries lesions were created (before probe application) on normal enamel surfaces using strong acid spots.
[0207] Human first molars, caries-free prior to acid treatment, were photographed before and after probe application ( Figure 31 ). Three enamel areas were then exposed to acid (0.5 μl 85% H 3 PO 4 ) for 1, 3 or 10 minutes to introduce artificial carious lesions, followed by a two-minute rinse in 100 ml TBS (25 mM Tris pH 7.2, 160 mM NaCl), followed by an additional 2-minute rinse under running water. The tooth was air dried and the probe was applied to the entire area with a brush for 3 minutes. After application, unbound probes were first removed by wiping with absorbent paper and then rinsed under runnin...
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