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Applications of mesenchymal stem cell and genetically modified mesenchymal stem cell

A technology of stromal stem cells and uses, applied in the field of use of mesenchymal stem cells and genetically modified mesenchymal stem cells, can solve problems such as gaps in periodontal conditions

Inactive Publication Date: 2014-02-19
BEIJING STOMATOLOGY HOSPITAL CAPITAL MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, many scholars have tried to use various cells to treat periodontal bone tissue and soft tissue defects caused by periodontal disease and to promote periodontal tissue regeneration. However, rats, mice and dogs are mostly used as experimental animals. The teeth of these animals There is still a gap between the weekly situation and human beings

Method used

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  • Applications of mesenchymal stem cell and genetically modified mesenchymal stem cell
  • Applications of mesenchymal stem cell and genetically modified mesenchymal stem cell
  • Applications of mesenchymal stem cell and genetically modified mesenchymal stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Isolation, cultivation, expansion and gene modification of mesenchymal stem cells

[0070] (1) Isolation and culture of MSCs

[0071] MSCs from periodontal ligament, umbilical cord, fat or bone marrow were isolated by tissue block culture method or enzyme digestion method, and cultured to the third passage. The obtained MSCs were tested for cell surface markers and induced for osteogenesis and adipogenic differentiation, which proved to be MSCs. The specific isolation and cultivation methods are as follows:

[0072] Isolation and culture method of periodontal ligament MSC:

[0073] Aseptic extraction of human impacted third molars or orthodontics under anesthesia requires extraction of teeth. Freshly extracted teeth are immediately placed in a centrifuge tube filled with sterile PBS and antibiotics, and periodontal ligament stem cells are isolated and cultured within 12 hours. Gently peel off the periodontal ligament tissue in the middle 1 / 3 of the tooth ro...

Embodiment 2

[0084] The MSC biological property detection of embodiment 2HGF modification

[0085] According to the method of embodiment 1, after Ad-HGF infects MSC 48h, the expression level of HGF is detected by ELISA; The change of cell surface molecule is detected by flow cytometry; The change of cell proliferation ability is detected by Dy670 dye method; Anti-apoptotic effect. Taking bone marrow MSCs as an example, the biological properties of HGF-modified MSCs were detected.

[0086] (1) Detection of HGF expression level

[0087] After Ad-HGF infected MSCs, the expression of HGF was detected by ELISA. The results showed that after Ad-HGF infection, the expression level of HGF gradually increased until the 7th day (such as figure 1 shown).

[0088] (2) Effect of HGF modification on MSC cell surface molecules

[0089] Flow cytometry was used to detect the expression of various surface molecules CD29, CD73, CD166, HLA-ABC, HLA-DR, CD31, CD34 and CD45, and the results showed that infect...

Embodiment 3

[0095] Example 3 MSC Repair and Treatment Effects on Periodontal Bone Tissue and Soft Tissue Defects

[0096] Firstly, the experimental periodontitis model of Wuzhishan minipigs was established, and then, using cell membrane implantation and cell suspension injection technology, after CT images and clinical examination confirmed that the periodontitis model was established, the cell membrane implantation or cell suspension injection To the bone defect site of periodontitis, observe the repair of periodontal bone tissue and soft tissue defect, periodontal tissue pathological changes, blood test indicators (blood routine, blood biochemistry, Immunoglobulin) changes, changes in the expression of inflammatory factors, etc. Taking periodontal ligament MSC as an example, the therapeutic effect of MSC on periodontal bone tissue and soft tissue defects was tested.

[0097] (1) Establishment of minipig experimental periodontitis model

[0098] The minipig experimental periodontitis m...

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Abstract

The invention relates to applications of a mesenchymal stem cell and a genetically modified mesenchymal stem cell, especially relates to applications of a mesenchymal stem cell and a genetically modified mesenchymal stem cell in preparation of products for treating periodontal disease, repairing periodontal bone tissue or soft tissue defect and / or promoting regeneration of periodontal tissue. The invention further relates to a composition containing the mesenchymal stem cell and / or the genetically modified mesenchymal stem cell, and a cell patch. According to the invention, prosthodontic treatment effect of MSC (Mesenchymal Stem Cells) from different sources on the periodontal bone defect and periodontitis soft tissue is proved successfully, and a strong evidence for expanding seed cell source is provided; meanwhile, the application also proves that the prosthodontic treatment effect of MSC modified by an HGF (Hepatocyte Growth Factor) gene on periodontal bone defect and periodontitis soft tissue is superior to that of simple MSC, and shows that HGF and MSC can play a role in cooperation in prosthodontic treatment of periodontal bone tissue or soft tissue defect and / or promoting regeneration.

Description

technical field [0001] The present invention relates to the use of mesenchymal stem cells and genetically modified mesenchymal stem cells, especially mesenchymal stem cells and genetically modified mesenchymal stem cells are used in the preparation of treatment of periodontal disease and repair of various causes (such as periodontal disease ) caused periodontal bone tissue and soft tissue defects and / or use in products for periodontal tissue regeneration, the present invention also relates to genetically modified mesenchymal stem cells, and mesenchymal stem cells and / or genetically modified mesenchymal stem cells Compositions and cell sheets of mesenchymal stem cells. Background technique [0002] Periodontal disease is a common clinical disease in the oral cavity. It is an infectious disease characterized by the destruction of periodontal tissue (including periodontal ligament, cementum, alveolar bone and gums). The main clinical manifestations are gingival inflammation and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/12A61K35/28A61K35/44A61K35/50A61K38/18A61P1/02A61L27/38A61L27/54A61L31/16A61L31/00
Inventor 王松灵吴祖泽曹钰王华杜娟靳继德范志朋汪劲松张群伟
Owner BEIJING STOMATOLOGY HOSPITAL CAPITAL MEDICAL UNIV
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