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A tissue-engineered pulp-dentin complex

A technique of tissue engineering and dentin, applied in dental prosthesis, dentistry, dental preparations, etc., to achieve the effects of promoting tooth regeneration, enhancing tooth forming ability, and promoting proliferation

Active Publication Date: 2017-01-18
SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Research in this area has not yet been reported in the literature

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  • A tissue-engineered pulp-dentin complex
  • A tissue-engineered pulp-dentin complex
  • A tissue-engineered pulp-dentin complex

Examples

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Effect test

Embodiment 1

[0024] Example 1. Effect of PDGF-BB on biological activity of human dental pulp stem cells (hDPSCs) in vitro

[0025] Step 1. Isolation, culture and identification of human dental pulp stem cells

[0026] Clinical collection of orthodontic needs or extraction of impacted teeth, rinse the blood on the surface of the isolated teeth with sterile saline; use a dental drill to cut a circle around the neck of the tooth to facilitate the exposure of the pulp cavity; use a pulp extraction needle Gently remove the pulp tissue, operate gently, and try to avoid damage to the tissue; rinse the removed pulp tissue twice with PBS, cut it into a tissue piece with a size of about 1×1mm with sterile ophthalmic scissors, and cover it with a sterile cover glass Cover the tissue block with a sheet to prevent the tissue block from floating. Then add DMEM medium containing 20% ​​fetal bovine serum for cultivation. The medium was changed every three and a half volumes. When hDPSCs climbed out fro...

Embodiment 2

[0054] Example 2. In vivo stem cell recruitment effect of PDGF-BB gene modified human dental pulp stem cells (hDPSCs) and ectopic regeneration ability of dental pulp-dentin complex

[0055] Step 1. In vivo recruitment experiment of human dental pulp stem cells in nude mice

[0056] (1) The cell membrane of hDPSCs was labeled with CM-DiI dye, and the concentration of the dye solution was 1 μg / mL. The specific steps are as follows: ① Trypsinize the cells, centrifuge at 1000 rpm for 5 min; ② Discard the supernatant, add 1ml DMEM culture medium, resuspend the cells, add 5μl CM-DiI staining solution, incubate at 37°C for 5min, then 4°C Incubate for 15 minutes; ③ Finally, centrifuge again and wash twice with PBS to remove the staining solution.

[0057] (2) Take the P3 generation hDPSCs, transfect them with Lenti-PDGF and Lenti-LacZ viruses, and when the cell confluency reaches over 90%, digest with trypsin, collect cells from each group, and prepare a cell suspension with a final ...

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Abstract

The invention discloses a tissue-engineered dental pulp-dentin complex. The tissue-engineered dental pulp-dentin complex is constructed by dental pulp stem cells modified by PDGF-BB (Platelet-Derived Growth Factor-BB) genes, and is prepared by the following steps: (1) transfecting dental pulp stem cells which are cultured separately in vitro by using the PDGF-BB genes; (2) loading a product obtained in the step (1) onto a biological scaffold material to construct the engineered dental pulp-dentin complex. The tissue-engineered dental pulp-dentin complex has the advantages that multiplication of the dental pulp stem cells, differentiation of grown dents and secretion of grown blood vessel inducible factors can be facilitated by PDGF-BB, and the recruitment of stem cells can be facilitated to accelerate the growth of dents; in a nude mouse subcutaneous ectopic grown dent model, the mass of regenerated dentins is increased remarkably in an experimental group of dental pulp stem cells modified by PDGF-BB. The application advantages of the dental pulp steam cells modified by PDGF-BB in the regeneration of the dental pulp-dentin complex are verified, and a novel treatment method is provided for the problem of tooth regeneration.

Description

technical field [0001] The invention relates to the field of tooth regeneration, in particular to a PDGF-BB gene-modified dental pulp stem cell for constructing a tissue-engineered dental pulp-dentine complex. Background technique [0002] Irreducible pulpitis is a clinically common and frequently-occurring disease. At present, root canal therapy is the routine treatment for this disease, but the complete removal of pulp tissue will seriously affect the nutrition supply and normal metabolic activities of teeth, resulting in tooth brittleness. enlarged and easily broken. Fortunately, stem cell-based pulp-dentin regeneration technology has emerged. Dental pulp stem cells isolated from deciduous teeth, orthodontic extractions, and impacted wisdom teeth provide ideal seed cells for pulp-dentin regeneration. However, the pulp cavity is mainly connected with the autologous tissue through the narrow root canal-apical foramen, which is also the main channel for the nutrition suppl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K6/02
Inventor 张文杰张茂林蒋欣泉张志愿
Owner SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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