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Periodontal defect repair system containing Let-7a

A technology of let-7a and repair system, applied in genetically modified cells, cells modified by introducing foreign genetic material, bone/connective tissue cells, etc., can solve the problem of limited repair effect of periodontal defect repair system

Active Publication Date: 2021-09-21
STOMATOLOGICAL HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention intends to provide a periodontal defect repair system containing Let-7a to solve the technical problem that the repair effect of the periodontal defect repair system based on periodontal ligament stem cells is limited

Method used

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  • Periodontal defect repair system containing Let-7a
  • Periodontal defect repair system containing Let-7a
  • Periodontal defect repair system containing Let-7a

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Isolation and identification of periodontal ligament stem cells (PDLSCs)

[0034] The process of isolation, culture and passage of PDLSCs is as follows:

[0035]Premolars extracted due to orthodontic needs were collected, and the inclusion and exclusion criteria for sample collection were as follows:

[0036] Inclusion criteria: 12-18 years old; healthy without systemic diseases; healthy without caries; tooth root development completed.

[0037] Exclusion criteria: severe systemic diseases; dental caries or periapical lesions; root canal treatment; severe root resorption.

[0038] Soak the teeth in a PBS solution containing 2% bis-antibody, and store them in an ice box at low temperature. The storage time should not exceed 2 hours, and the next step is required. Transfer the isolated teeth to the ultra-clean bench, add new PBS solution containing 5% double-antibody to the isolated tooth, tighten the cap of the centrifuge tube, shake it up and down manually ...

Embodiment 2

[0043] Example 2: Preparation of PDLSCs cell aggregates and let-7a transfection

[0044] The P3 generation PDLSCs cells were routinely digested, neutralized, and counted in 3×10 5 seeded in six-well plates. The cells were cultured with complete medium, and the induction solution was added when the cells grew confluent to a cell density of 80%, and the induction solution was replaced every 3 days. Use the induction medium to induce the cultured cells for 7 days (37°C conventional culture in the incubator) to obtain PDLSCs cell aggregates. For the obtained PDLSCs cell aggregates, see Figure 4 .

[0045] The mature sequence of mmu-let-7a (MIMAT0000521) was obtained through the mirbase database. Let-7a mimics, inhibitor, and negative control were designed by RiboBio based on the mature sequence of let-7a. The sequence is as follows:

[0046] mmu-let-7a mature sequence: 5'-UGAGGUAGUAGGUUGUAUAGUU-3' (SEQ ID NO.1)

[0047] let-7a mimic sense strand: 5'-UGAGGUAGUAGGUUGUAUAGUU-3' ...

Embodiment 3

[0052] Example 3: In vivo repair of jaw defects with PDLSCs cell aggregates

[0053] The specific process for the preparation of PDLSCs cell aggregates transfected with let-7a mimic is as follows:

[0054] The media used include the following:

[0055] Complete medium (first medium): use α-MEM to prepare a medium containing 10% FBS and 1% double antibody (penicillin double antibody); induction solution (second medium): 50 μg / mL Complete medium of vitamin C; medium without antibiotics (the third medium): a medium containing 10% FBS was prepared with α-MEM.

[0056] Aggregate induction (to obtain the first cell aggregate): P3 generation PDLSCs cells were routinely digested, neutralized, and counted, and 3×10 5 seeded in six-well plates. Use the complete medium (the first medium) to culture the cells, and when the cells grow confluent to a cell density of 80%, add the second medium (replace the first medium with the second medium), and replace the second culture every 3 days ...

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Abstract

The invention relates to the technical field of bone defect repair, in particular to a periodontal defect repair system containing Let-7a. The system comprises a third cell polymer and hydroxyapatite-beta tricalcium phosphate wrapped in the third cell polymer; the third cell polymer is transfected with let-7a mimic; the let-7a mimic is a double-stranded RNA, and the sequence of a sense strand of the let-7a mimic is as shown in SEQ ID NO. 2. According to the scheme, the technical problem that the effect of repairing bone defects by using periodontal ligament stem cells is poor can be solved. According to the scheme, the osteogenic differentiation capacity of the periodontal ligament stem cells is regulated and controlled by using Let-7a, and a feasible way is provided for rapid and effective repair of periodontal defects in combination with a periodontal ligament stem cell polymer form cost scheme.

Description

technical field [0001] The invention relates to the technical field of bone defect repair, in particular to a periodontal defect repair system containing Let-7a. Background technique [0002] Periodontitis is a common inflammatory disease that affects the tooth-supporting tissues. The main treatment methods currently used clinically are plaque control (including supragingival scaling and subgingival scaling) and periodontal flap surgery, which can prevent the further loss of periodontal supporting tissue, but cannot make the missing tissue get a good recovery. Regeneration; the rise of regenerative surgery has brought new hope for periodontal treatment, including guided tissue regeneration, periodontal bone grafting, and direct delivery of bioactive polypeptide growth factors to the root surface, including platelet-derived growth factor (PDGF ) and insulin-like growth factor-i (IGF-I), bone morphogenetic proteins (BMPs), etc., but these regenerative operations have narrow a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/12A61L27/50A61L27/54C12N5/0775C12N5/077C12N5/10C12N15/113
CPCA61L27/3834A61L27/3865A61L27/3895A61L27/12A61L27/50A61L27/54C12N5/0654C12N15/113A61L2430/12A61L2430/40A61L2300/412A61L2300/258C12N2509/00C12N2509/10C12N2500/38C12N2506/1392C12N2310/141C12N2510/00
Inventor 杨德琴于洋杨诗瑶
Owner STOMATOLOGICAL HOSPITAL OF CHONGQING MEDICAL UNIV
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