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Method for evaluating osteoclastic bone absorption function

A technology of osteoclast and function, applied in the biological field, can solve the problems of restricting the study of osteoclast function factors, many factors affecting the results, poor accuracy, etc., and achieve the effects of avoiding result bias, convenient preparation and stable results

Active Publication Date: 2017-01-04
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Among the above methods, the first and second methods are indirect reflections of the function of osteoclasts, which can be used to infer the function of osteoclasts, but there are many factors that affect the results, and the accuracy is relatively poor
The third method can directly reflect the osteolytic function of osteoclasts, and can compare the strength of osteolytic function through the analysis of the number, area and depth of resorption lacuna, which is currently widely used, and the results are also A more accurate method, but this method has great limitations in the process of use
[0005] Based on the above reasons, so far, there is no better method to detect the osteolytic activity of osteoclasts, which greatly limits the research on the factors affecting the function of osteoclasts.

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  • Method for evaluating osteoclastic bone absorption function
  • Method for evaluating osteoclastic bone absorption function
  • Method for evaluating osteoclastic bone absorption function

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 The method for evaluating osteoclast bone resorption function of the present invention (1)

[0038] 1. Method

[0039] 1) Use a low-speed motor (3r / min) to drive an emery wheel, grind bovine femur cortex into powder, and store it in 75% alcohol. Specific use such as figure 1 The shown machine is milled, and other mills in the prior art can also be used.

[0040] 2) Use a 30 μm cell sieve to screen the bone powder smaller than 30 μm, centrifuge, pour off the alcohol, dry at 70°C, weigh a certain amount of bone powder into a pressure-resistant bottle, add a small amount of distilled water, and then perform high-pressure steam sterilization. After sterilization, add 75% alcohol to make a 2.5 mg / ml suspension.

[0041] 3) Place the bone powder suspension in an ultrasonic cleaner and mix it for 1 min, then add it dropwise to a 24-well cell culture plate in an ultra-clean bench, 1ml per well, and blow and suck several times before each drop , to mix well. Plac...

Embodiment 2

[0053] Embodiment 2 The method for evaluating osteoclast bone resorption function of the present invention (two)

[0054] 1) Use a low-speed motor (3r / min) to drive an emery wheel, grind bovine femur cortex into powder, and store it in 75% alcohol. Specific use such as figure 1 The shown machine is milled, and other mills in the prior art can also be used.

[0055] 2) Use a 20 μm cell sieve to screen the bone powder smaller than 20 μm, centrifuge, pour off the alcohol, dry at 70°C, weigh a certain amount of bone powder into a pressure-resistant bottle, add a small amount of distilled water, and then perform high-pressure steam sterilization. After sterilization, add 90% alcohol to make a 4mg / ml suspension.

[0056] 3) Place the bone powder suspension in an ultrasonic cleaner and mix it for 1 min, then add it dropwise to a 24-well cell culture plate in an ultra-clean bench, 1ml per well, and blow and suck several times before each drop , to mix well. Place the added culture...

Embodiment 3

[0062] Embodiment 3 The method for evaluating osteoclast bone resorption function of the present invention (3)

[0063] 1) Use a low-speed motor (3r / min) to drive an emery wheel, grind bovine femur cortex into powder, and store it in 75% alcohol. Specific use such as figure 1 The shown machine is milled, and other mills in the prior art can also be used.

[0064] 2) Use a 30 μm cell sieve to screen the bone powder smaller than 30 μm, centrifuge, pour off the alcohol, dry at 70°C, weigh a certain amount of bone powder into a pressure-resistant bottle, add a small amount of distilled water, and then perform high-pressure steam sterilization. After sterilization, add 70% alcohol to make a 5 mg / ml suspension.

[0065] 3) Place the bone powder suspension in an ultrasonic cleaner and mix it for 1 min, then add it dropwise to a 24-well cell culture plate in an ultra-clean bench, 1ml per well, and blow and suck several times before each drop , to mix well. Place the added culture ...

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Abstract

The invention provides a method for evaluating an osteoclastic bone absorption function. The method includes the steps of fixing bone meal with long diameter smaller than 30 micrometers to the bottom face of a cell culture plate, and comparing the before-culture light transmittance and the after-culture light transmittance of the cell culture plate by analyzing the before-culture and after-culture light density of a fixed point picture so as to reflect the bone meal absorption degree in the cell culture plate and evaluate the osteoclastic bone absorption function. The method is easier and more convenient to implement, solves the selective osteoclastic absorption problem, is more stable and reliable in result, has good operability and high sensitivity, and has important application value in research related to osteoclastic osteolysis activity detection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for evaluating the bone resorption function of osteoclasts and its application. Background technique [0002] Osteoporosis is a common and frequently-occurring disease in the aging society. Bone is constantly rebuilt through synthesis and degradation in the process of metabolism, and this process is completed by the joint action of osteoblasts and osteoclasts. Osteoclasts play a very important role in the process of bone metabolism, and their overactivation has a relatively direct effect on the occurrence and development of osteoporosis. The in vitro experiments of osteoclasts play an extremely important role in the study of the pathogenesis and treatment of osteoporosis. Among them, the evaluation of osteolytic activity of osteoclasts is often used as an observation index after treatment of various intervention factors, and plays an indispensable role in this ...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
CPCG01N33/5005
Inventor 漆祎鸣胡予任卫英陆逸沈志云
Owner ZHONGSHAN HOSPITAL FUDAN UNIV