Application of mesenchymal stem cell apoptotic body to preparation of medicine for treating bone defects

A technology of mesenchymal stem cells and apoptotic bodies, applied in medicine, in the field of mesenchymal stem cell apoptotic bodies, can solve the problems that cannot fundamentally solve the problems of damaged bone cell repair and bone tissue regeneration, and achieve quality Stabilizes, promotes bone regeneration, and promotes the effect of new bone formation

Active Publication Date: 2021-10-29
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] One of the purposes of the present invention is to provide the application of mesenchymal stem cell apoptotic bodies in the preparation of drugs for the treatment of bone defects, and solve the problems in the prior art that cannot fundamentally solve the problem of damaged bone cell repair and bone tissue regeneration

Method used

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  • Application of mesenchymal stem cell apoptotic body to preparation of medicine for treating bone defects
  • Application of mesenchymal stem cell apoptotic body to preparation of medicine for treating bone defects
  • Application of mesenchymal stem cell apoptotic body to preparation of medicine for treating bone defects

Examples

Experimental program
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Effect test

Embodiment 1

[0043] Example 1: Induction of bone marrow mesenchymal stem cell apoptosis in vitro

[0044] Bone marrow mesenchymal stem cells cultured to the third generation 6 Inoculate to a 10cm culture dish at a density of one / dish and place at 37°C, 5% CO 2 The culture was continued under the conditions, and when the cell confluency reached 80%-85%, a dimethyl sulfoxide solution of staurosporine was added for induction, and the induction time was 4 hours.

[0045] Among them, the culture medium for the subculture of mesenchymal stem cells includes 500 parts by volume of α-MEM basal medium, 55.5 parts by volume of fetal bovine serum, penicillin with a concentration of 100 U / mL, and streptaline with a concentration of 100 U / mL. Mycin.

[0046] The solvent of staurosporine solution is dimethyl sulfoxide, 5mg staurosporine is dissolved in the dimethyl sulfoxide solution of 2.1435mL, the concentration that prepares is 5mm stock solution, staurosporine is in culture medium The working conc...

Embodiment 2

[0047] Example 2: Preparation of apoptotic bodies of mesenchymal stem cells

[0048] The supernatant from the induced apoptotic cells of Example 1 was collected in a sterile centrifuge tube (about 10ml in a 50ml tube), and the supernatant was centrifuged at 4°C and 800×g for 10min, and transferred to the Supernatant, remove cell debris; then centrifuge at 4°C, 16,000×g for 30 min, discard supernatant, collect precipitate, and resuspend in PBS; in order to remove the effect of reagents or serum in the medium, resuspend at 4 Centrifuge again at 16000×g for 30 min to obtain apoptotic bodies with higher purity as precipitates.

[0049] The diameter of the precipitated apoptotic body is 1-5 μm. The precipitated apoptotic bodies were then resuspended in 200 μL-400 μL PBS, aliquoted into sterile centrifuge tubes, and stored at -80°C for later use. At the same time, the basic morphology and markers of the apoptotic bodies were identified. The results are shown in Figure 1, Figure 1...

Embodiment 3

[0050] Example 3: Establishment, treatment and microCT detection of SD rat skull defect model

[0051] Select SD female rats about 4 weeks old, carry out intraperitoneal injection anesthesia with 1% sodium pentobarbital (40mg / kg), fix on the experimental table in prone position, connect the animal artificial respirator, the respiratory rate is about 86 times / min, The tidal volume was 15ml, and the respiratory ratio was set at 1:1. The head was depilated and skinned, and a drape was spread with disinfection. The skin was incised at the center of the skull at about the cranial suture, and the subcutaneous tissue was bluntly separated to visualize the skull. Connect the grinding machine, and cut the skull with a 5mm trephine at a speed of 1500r.p.m. or lower, so that the circular skull piece with a diameter of 5mm can be completely removed. During the operation, 1 drop of sterile saline was instilled every 2 seconds to flush the trephine and the skull (slow flushing speed is the...

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Abstract

The invention discloses an application of a mesenchymal stem cell apoptotic body to preparation of a medicine for treating bone defects, belongs to the technical field of biological medicines, and solves the problem that damaged bone cell repair and bone tissue regeneration cannot be fundamentally solved in the prior art. The invention provides the application of the mesenchymal stem cell apoptotic body to preparation of the medicine for treating bone defects. The invention also provides a medicinal preparation which comprises the mesenchymal stem cell apoptotic body and a pharmaceutically acceptable carrier. The inventor finds that the mesenchymal stem cell apoptotic body has the effects of promoting new bone formation and promoting bone regeneration for the first time, and has a good effect when being used for treating bone defects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of mesenchymal stem cell apoptotic bodies in the preparation of medicines for treating bone defects. Background technique [0002] Stem cells are a type of cells with self-replication and multi-directional differentiation capabilities. They can continuously renew themselves and transform into one or more cells that constitute human tissues or organs under specific conditions. Stem cells are the origin cells of the body and the progenitor cells that form various tissues and organs of the human body. Mesenchymal stem cells are a research branch of stem cells. Mesenchymal stem cells are clinically used to solve a variety of blood system diseases, cardiovascular diseases, liver cirrhosis, nervous system diseases, partial knee meniscectomy injury repair, autoimmune diseases, etc., but the mesenchymal stem cells are facing Difficulty in obtaining mater...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P19/08C12N5/073C12N5/0775
CPCA61K35/28C12N5/0663C12N5/0665C12N5/0667C12N5/0668C12N5/0605C12N5/0664A61P19/08C12N2509/00Y02A50/30
Inventor 田卫东李茂娇廖立
Owner SICHUAN UNIV
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