Tissue-engineered dental pulp-dentin complex

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

Active Publication Date: 2014-08-27
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Description
  • Claims
  • Application Information

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

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  • Tissue-engineered dental pulp-dentin complex

Examples

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Effect test

Embodiment 1

[0024] Example 1. The effect of PDGF-BB on the 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 impacted teeth to extract teeth, rinse the blood on the surface of the isolated teeth with sterile saline; use a dental split 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 with gentle operation to avoid damage to the tissue as much as possible; rinse the removed pulp tissue with PBS twice, use sterile ophthalmic scissors to cut it to a tissue block of about 1 × 1 mm, and use a sterile cover glass The sheet covers the tissue block to prevent the tissue block from floating. Then, DMEM medium containing 20% ​​fetal bovine serum was added for culture. Change the medium every three days. When hDPSCs crawled out from t...

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 at a concentration of 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 1 ml of DMEM medium, resuspend the cells, add 5 μl of CM-DiI staining solution, incubate at 37 °C for 5 min, and then at 4 °C Incubate for 15min; ③Finally, centrifuge again and wash twice with PBS to remove the dye solution.

[0057] (2) Take the P3 generation hDPSCs, transfect Lenti-PDGF and Lenti-LacZ virus, when the cell confluence reaches more than 90%, trypsin digestion, collect the cells of each group, and prepare a cell suspension with a final concentration of 2 × 10 7cells / ml, i...

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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 the construction of a tissue-engineered dental pulp-dentin complex by a PDGF-BB gene-modified dental pulp stem cell. Background technique [0002] Irreversible pulpitis is a common and frequently-occurring disease in clinical practice. At present, root canal therapy is the conventional treatment method for this disease. However, the complete removal of pulp tissue will seriously affect the nutritional supply and normal metabolic activities of the teeth, resulting in tooth fragility. Enlarged and easily broken. Fortunately, stem cell-based pulp-dentin regeneration technology came into being. 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 ...

Claims

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Application Information

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