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Effect of LncRNA-TUG1 on regulating and controlling PDLSCs osteogenic differentiation and tissue regeneration

A technology for osteogenic differentiation and tissue regeneration, applied in medical preparations containing active ingredients, bone/connective tissue cells, digestive system, etc.

Active Publication Date: 2017-11-17
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the role of LncRNA-TUG1 in periodontal ligament stem cells and its correlation with osteogenic differentiation and tissue regeneration of periodontal ligament stem cells

Method used

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  • Effect of LncRNA-TUG1 on regulating and controlling PDLSCs osteogenic differentiation and tissue regeneration
  • Effect of LncRNA-TUG1 on regulating and controlling PDLSCs osteogenic differentiation and tissue regeneration
  • Effect of LncRNA-TUG1 on regulating and controlling PDLSCs osteogenic differentiation and tissue regeneration

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Biological characteristics of periodontal ligament stem cells

[0034] 1. Isolation and culture of human periodontal ligament stem cells

[0035] Under anesthesia, aseptically extract the impacted third molars or the first and second premolars used for orthodontic treatment, gently peel off the periodontal tissue around it, take the periodontal tissue in the middle, wash it repeatedly with PBS, cut it into pieces, and put it in the Digest in the digestion solution containing type I collagenase (3g / L) and Dispase II enzyme (4g / L), digest at 37°C for 40 minutes, collect the cells through a 70um cell sieve, centrifuge at 1000 rpm for 5 minutes, and remove the upper liquid , resuspended into a single-cell suspension with fresh culture medium. Cells were seeded at 25cm 2 In a cell culture flask, in α-MEM medium (containing 20% ​​fetal bovine serum, 2 mmol / L glutamine, 100 U / ml penicillin and 100 μg / ml streptomycin) at 37°C, 5% CO 2 For culture, replace the cultu...

Embodiment 2

[0041] Example 2 Screening of LncRNA-TUG1, and its cell localization and expression changes in osteogenic induction 1. Periodontal ligament stem cells were induced to osteogenic on 0d and 7d respectively, RNA was extracted, and the screening was confirmed by RT-qPCR The target non-coding RNA was obviously expressed, and the trend of osteogenesis induction was carried out at different times. .

[0042] Experimental results such as figure 2 As shown, the significantly expressed LncRNA-TUG1 was screened out, and the expression trend of TUG1 in the osteogenic differentiation of PDLSCs was preliminarily confirmed.

[0043] 2. Lentiviral vector construction

[0044] Primers were designed according to the human LncRNA-TUG1 sequence (ID: 55000), and the primers were synthesized and purified by Shanghai Lingke Biotechnology Co., Ltd. After annealing, it is connected to the linearized pGenesil-1 plasmid to construct the expression vector lentivirus of LncRNA-TUG1 (silent), and inhib...

Embodiment 3

[0073] Example 3 LncRNA-TUG1 regulates tissue regeneration of periodontal ligament stem cells in vivo

[0074] The periodontal ligament stem cell complex was implanted subcutaneously in the back of nude mice, and its tissue formation ability was observed.

[0075] 1. Transfect periodontal ligament stem cells with lentiviral sh-NC empty vector and sh-TUG1-2# respectively

[0076] Human periodontal ligament stem cells were cultured by tissue block method and enzyme digestion method, and the vigorously growing third-generation periodontal ligament stem cells were inoculated on a 60mm culture dish, and the culture components were α-MEM medium (containing 15% fetal bovine serum, 2 mmol / L glutamine, 100U / ml penicillin, 100μg / ml streptomycin). The periodontal ligament stem cells were transfected with lentivirus sh-NC empty vector and sh-TUG1-2# respectively, and the experiment was divided into three groups: (1) normal cultured periodontal ligament stem cell group (2) sh-NC empty ve...

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Abstract

The invention discloses an effect of LncRNA-TUG1 on regulating and controlling PDLSCs osteogenic differentiation and tissue regeneration. Based upon in vivo and in vitro experiments of researching the effect of the LncRNA-TUG1 on regulating and controlling PDLSCs osteogenic differentiation and tissue regeneration, the LncRNA-TUG1 plays an important role in regulating and controlling the osteogenic differentiation and tissue regeneration of periodontal ligament stem cells; and the LncRNA-TUG1, as a potential target in tissue regeneration therapy, can promote the development of novel LncRNA-TUG1 related medicines or factors, and the LncRNA-TUG1 can be finally applied to the field of regenerative medicine, so that real precision medicine in the clinical field can be achieved.

Description

technical field [0001] The invention relates to the technical field of oral tissue regeneration, and in particular to the role of LncRNA-TUG1 in regulating PDLSCs osteogenic differentiation and tissue regeneration. Background technique [0002] Oral stem cell research provides the possibility for the biological regeneration and biological function restoration of human teeth and periodontal tissue. In recent years, the research progress on dental stem cells and tissue engineering technology has brought new research directions to biological regeneration. Dental stem cells have the advantages of strong vitality, convenient material collection, and low immune rejection. They can be used for the regeneration of soft and hard tissues in many parts of the human body, such as the repair and regeneration of teeth and periodontal tissue, post-traumatic eye repair, skin healing and regeneration, Tissue reconstruction of fat cells, self-repair of the immune system, etc. Periodontal li...

Claims

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Application Information

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IPC IPC(8): C12N5/077A61K45/00A61P1/02
CPCA61K45/00C12N5/0654C12N2506/1392
Inventor 魏福兰王春玲何琴谷秀格
Owner SHANDONG UNIV
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