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A Screening System for Drugs Affecting Bone Mineralization

A technology of mineralization and drugs, applied in the direction of measuring devices, instruments, fluorescence/phosphorescence, etc., can solve the problem that the degree of change cannot be fully defined, and achieve the effects of reducing experimental costs, improving screening efficiency, and improving accuracy

Active Publication Date: 2016-03-30
BIOLOGY INST OF SHANDONG ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Although the analytical method published by Angeleen Fleming et al. using zebrafish as an animal model can indeed be used to screen drugs that affect mineralization, the degree of change cannot be fully defined by using the depth of the dye as a numerical quantitative result

Method used

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  • A Screening System for Drugs Affecting Bone Mineralization
  • A Screening System for Drugs Affecting Bone Mineralization
  • A Screening System for Drugs Affecting Bone Mineralization

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1. The effect of Alendronate on the mineralization of zebrafish

[0083] For mating with wild-type zebrafish, a septum was used the day before pairing to ensure embryonic stage consistency. Use the same pair of embryos to conduct experiments, and the time of embryos born is controlled within half an hour to reduce the difference in development time between embryos. The embryos are stored in a petri dish with embryo culture medium, so that the embryos are not easy to mold, and also keep them in the water. The mineral content is the same, put them into the incubator and raise them at 28°C for three days, during which the dead embryos need to be sucked out.

[0084]On the third day, use a dropper to select 6 wild-type embryos and place them in a 24-well plate, blot the water as dry as possible with a micropipette, first add 1ml of embryo culture medium, then add 1mM Alendronate 1μl, dilute 1000 times to 10μM, and adjust according to the concentration Adjust, stir ...

Embodiment 2

[0088] Example 2. Effects of Dorsomorphin on Zebrafish Mineralization

[0089] Use wild-type zebrafish for mating, and use a spacer the day before mating to ensure consistency of embryonic stages. Use the same pair of embryos to conduct experiments, and the time of embryos born is controlled within half an hour to reduce the difference in development time between embryos. The embryos are stored in a petri dish with embryo culture medium, so that the embryos are not easy to mold, and also keep them in water. The mineral content is the same, and they are kept in an incubator at 28°C for three days, during which the dead embryos need to be sucked out.

[0090] On the third day, use a dropper to select 20 wild-type embryos and place them in a 12-well plate. First add 999 μl of embryo culture solution, then add 1 μl of 1 mM Dorsomorphin, dilute 1000 times to 10 μM, and adjust according to the concentration. Use a micropipette tip to stir slightly or Shake on a shaker, the total vo...

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Abstract

The invention relates to a screening system of drugs influencing bone mineralization. The screening system of the drugs influencing the bone mineralization comprises a zebra fish embryo in a test group, a zebra fish embryo in a positive control group, a zebra fish embryo in a negative control group, a calcein dyeing agent, an image acquisition system, and image analysis and statistics software. Compared with a traditional dyeing method using alizarin red, the screening system of the drugs influencing the bone mineralization solves the problem that the traditional dyeing which is judged by the depth of color is influenced easily by external factors such as dyeing conditions, dyeing time and operation, and overcomes the disadvantages that the bone mineralization degree cannot be reflected exactly as the existence of the dye instead of the depth of the dye is utilized to evaluate the variation of the bone formation.

Description

technical field [0001] The invention relates to a screening system for drugs affecting bone mineralization, in particular to a screening system for bone mineralization drugs using calcein as a dye, and belongs to the technical field of drug screening. Background technique [0002] Due to the advancement of scientific and technological civilization, the average age of men and women is constantly increasing, and the aging population is increasing, which makes the number of patients with osteoporosis gradually increase. Osteoporosis is a disease in which the bone structure is loosened due to osteopenia and decreased bone density, and patients' bones are fragile and prone to fractures. This type of disease is common in the elderly over 55 years old and postmenopausal women. The reason is that the secretion of estrogen in postmenopausal women stops due to bone aging, resulting in a large loss of bone mass. In order to replenish the lost bone mass, most people will take a large a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G01N21/00
Inventor 韩利文刘可春蔡志杰萧崇德
Owner BIOLOGY INST OF SHANDONG ACAD OF SCI
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