Preparation method of engineering periodontium

A periodontal tissue and engineering technology, applied in the field of engineering periodontal tissue preparation, can solve the problems of periodontal tissue regeneration that have not yet been established, materials cannot be completely degraded, immune rejection of the body, etc., and achieve rich extracellular matrix. , to avoid poor compatibility, the effect of repairing periodontal defects

Inactive Publication Date: 2017-06-27
XIAN TISSUE ENG & REGENERATIVE MEDICINE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Japanese scholar Iwata et al. studied periodontal tissue regeneration by applying a composite of multilayer periodontal ligament cell membrane and PGA membrane. The composite cell membrane was implanted into the alveolar bone defect of Beagle dogs, and the periodontal defect was detected after 6 weeks. , the transplanted periodontal ligament cell membrane can promote the regeneration of alveolar bone and periodontal ligament, but biomaterials are still needed as scaffolds for periodontal tissue regeneration, and there are problems such as biomaterials may cause immune rejection of the body and materials cannot be completely degraded
[0006] In summary, there is no report on the use of biomaterial-engineered periodontal tissue for periodontal tissue regeneration.

Method used

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  • Preparation method of engineering periodontium
  • Preparation method of engineering periodontium
  • Preparation method of engineering periodontium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0034] Embodiment 1, one of the preparation methods of engineered periodontal tissue

[0035] Its preparation steps are as follows:

[0036] Step 1. Periodontal ligament stem cell culture: Peel off the periodontal ligament tissue of the deciduous teeth of the minipig and cut it into pieces, digest it with 0.2g / L type Ⅰ collagenase and 0.1g / L Dispase enzyme for 1 hour until the tissue is loose, and use a pipette Blow and beat to fully disperse the periodontal ligament tissue. Sieve through a sieve to form a single cell suspension, discard the supernatant after centrifugation, then add periodontal ligament stem cell culture medium to resuspend the cells, transfer to a cell culture bottle, 37 ° C, 5% CO 2 Cultured in an incubator, the medium was changed every other day, and the cells were cultured to 90% confluency before being passaged to obtain the first generation of periodontal ligament stem cells.

[0037] The composition of the periodontal ligament stem cell culture mediu...

Embodiment ２

[0044] Embodiment 2, the second preparation method of engineered periodontal tissue

[0045] Step 1. Periodontal ligament stem cell culture: Peel and shred the periodontal ligament tissue of the permanent teeth of minipigs, digest with 0.5g / L type Ⅰ collagenase and 0.3g / L Dispase enzyme for 0.5 hours until the tissue is loose, and use The periodontal ligament tissue was fully dispersed by pipetting. Sieve through a sieve to form a single cell suspension, discard the supernatant after centrifugation, then add special culture medium to resuspend the cells, transfer to a cell culture bottle, 37 ° C, 5 % CO 2 Cultivate in an incubator, change the medium every other day, and culture the cells until they reach 90% confluence to obtain periodontal ligament stem cells.

[0046] The composition of the periodontal ligament stem cell culture medium is: 10% (v / v) fetal bovine serum, 0.3 mg / mL glutamine, 90 ng / mL basic fibroblast growth factor in commercial α-MEM culture medium .

[004...

Embodiment ３

[0053] Embodiment 3, the third preparation method of engineered periodontal tissue

[0054] Step 1. Periodontal ligament stem cell culture: Mince the periodontal ligament tissue of deciduous teeth of miniature pigs, digest with 0.1 g / L type Ⅰ collagenase and 0.4 g / L Dispase enzyme for 1.5 hours until the tissue is loose, and sieve to obtain For single cell suspension, discard the supernatant after centrifugation, then add dental pulp cell culture medium to resuspend the cells, and place at 37°C, 5% CO 2 Cultured under the same conditions, the medium was changed every other day, and the cells were cultured to 90% confluency and then passaged to obtain the first generation of periodontal ligament stem cells.

[0055] The composition of the periodontal ligament stem cell culture medium is: in commercial α-MEM culture medium, 10% (v / v) fetal bovine serum, 0.4mg / mL glutamine, 125ng / mL basic fibroblast growth factor.

[0056] Step 2. Screening of periodontal ligament stem cells: t...

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Abstract

The invention relates to a preparation method of engineering periodontium, which is used for reconstruction of periodontium. The periodontium comprises periodontal ligament stem cells and an extracellular matrix secreted and synthesized by the periodontal ligament stem cells. The periodontal ligament stem cells are subjected to amplification culture to form multi-layered periodontal ligament stem cell assemblies, and then the periodontal ligament stem cell assemblies are arranged layer by layer and molded to form the engineering periodontium. The periodontium can form a new dentinal structure in the human body, so that periodontal defects can be repaired. The periodontium is simple in preparation, good in plasticity, moderate in cell density, abundant in extracellular matrix content, free of support material, and short in in-vitro culture time.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering, and in particular relates to a preparation method of engineered periodontal tissue for reconstruction and regeneration of periodontal tissue. Background technique [0002] Periodontitis is a common oral disease, and its pathological features are loss of periodontal attachment and alveolar bone recession, and the lesion process involves the supporting tissues of teeth such as gingiva, alveolar bone, periodontal ligament, and cementum, eventually leading to The loss of teeth brings great pain to patients. Therefore, exploring the method of periodontal tissue regeneration has very important clinical significance for promoting the regeneration of alveolar bone and periodontal ligament. [0003] At present, there are two main methods to promote periodontal regeneration clinically, bone or bioactive material transplantation and guided tissue regeneration (GTR). However, although the use of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38
CPCA61L27/3834A61L27/3865A61L27/3895
Inventor 张勇杰
Owner XIAN TISSUE ENG & REGENERATIVE MEDICINE RES INST
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