Biological agent for promoting proliferation and migration of dental pulp stem cells

A technology of dental pulp stem cells and biological preparations, applied in the field of biological preparations that promote the proliferation and migration of dental pulp stem cells, can solve problems such as crown damage and dental caries, and achieve the effect of promoting proliferation and migration

Active Publication Date: 2020-09-22
青岛赛尔斯干细胞库有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If left untreated, it can lead to cavities in the tooth and eventually the complete destruction of the crown

Method used

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  • Biological agent for promoting proliferation and migration of dental pulp stem cells
  • Biological agent for promoting proliferation and migration of dental pulp stem cells
  • Biological agent for promoting proliferation and migration of dental pulp stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] (1) Wash 200g of dried capers with clean water and dry them;

[0044] (2) Crush the dried capers into powder, add 1L of 95% ethanol to soak for 24 hours, filter and dry;

[0045] (3) Add 2L of deionized water to the dried caper powder, extract at 90°C for 3 hours, and centrifuge to obtain crude caper polysaccharide extract A;

[0046] (4) Concentrate the crude caper polysaccharide extract A to 400ml under reduced pressure;

[0047] (4) Use the Sevage method to remove protein to obtain crude caper polysaccharide extract B, repeat the Sevage method to remove protein twice, and obtain crude caper polysaccharide extract C;

[0048] (5) Add 400 ml of 85% ethanol to the crude caper polysaccharide extract C for 12 hours to obtain the caper polysaccharide extract;

[0049] (6) Caper polysaccharide extract was freeze-dried to obtain caper polysaccharide.

Embodiment 2

[0051] Primary culture of human dental pulp stem cells

[0052] (1) Immerse the caries-free teeth with complete tooth surfaces collected in our hospital into sterile PBS with double antibiotics, wash them repeatedly, and remove the soft tissues such as periodontal ligament attached to the root and neck of the tooth;

[0053] (2) Use a chisel to split the tooth longitudinally along the long axis of the tooth, use sterile forceps to take out the pulp tissue, cut off the 2mm part of the tip, and place it in a sterile EP tube;

[0054] (3) After rinsing with PBS for 3 times, add 500 μl of PBS and cut the pulp tissue with scissors, put the EP tube in a centrifuge, centrifuge at 1000rpm / min for 5 minutes, retain the precipitate, and remove the supernatant;

[0055] (4) Add type I collagenase, place in a cell constant temperature incubator, and digest at 37°C for 45 minutes;

[0056] (5) Add an appropriate amount of α-MEM medium containing 10% FBS to stop the digestion, centrifuge a...

Embodiment 3

[0060] Effects of caper polysaccharides on the proliferation of dental pulp stem cells

[0061] (1) Inoculate 100 μl of third-generation dental pulp stem cells in logarithmic growth phase in a 96-well plate, with 2000 cells per well;

[0062] (2) After adhering to the wall, discard the medium, add caper polysaccharides of 0 μg / ml, 50 μg / ml, 100 μg / ml, 200 μg / ml and 400 μg / ml respectively, 100 μl per well, and set 3 replicates in each group, The experiment was repeated three times;

[0063] (3) At 24h, 48h and 72h respectively, use CCK-8 to detect the absorbance at a wavelength of 450nm.

[0064] Table 1 Absorbance of dental pulp stem cells treated with different concentrations of caper polysaccharide

[0065]

[0066] The experimental results are shown in Table 1. It can be seen from the table that the caper polysaccharide can effectively promote the proliferation of dental pulp stem cells, and the 200 μg / ml caper polysaccharide has the best effect on promoting the prolifer...

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Abstract

The invention provides a biological agent for promoting proliferation and migration of dental pulp stem cells, and belongs to the technical field of stem cells. CCK-8, fluorescent quantitative PCR, Transwell experiments and Western Blot experiments prove that the capparis spinosa polysaccharide can effectively promote proliferation and cell migration of dental pulp stem cells, and possibility is provided for further developing application of the dental pulp stem cells in the aspect of dental restoration.

Description

technical field [0001] The invention belongs to the technical field of dental pulp stem cells, in particular to a biological preparation for promoting proliferation and migration of dental pulp stem cells. Background technique [0002] Tooth decay is commonly known as tooth decay, and tooth decay is a common oral bacterial disease. When caries develops to the dentin, bacteria and the toxic products they secrete will invade the pulp tissue through the dentin tubules, resulting in varying degrees of damage to the dentin. Dental caries can be secondary to pulpitis and periodontitis, and can even cause alveolar bone and jaw inflammation. If left untreated, it can lead to cavities in the tooth and eventually the complete destruction of the crown. [0003] Dental pulp stem cells are a kind of mesenchymal stem cells with self-renewal and multilineage differentiation existing in dental pulp tissue. Dental pulp cells can differentiate into different cell types under the action of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C08B37/00
CPCC08B37/0003C12N5/0664C12N2501/90
Inventor 刘金凤杨桂梅
Owner 青岛赛尔斯干细胞库有限公司
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