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Application of engineered exosome in skin regeneration and repair of diabetic foot ulcer and preparation method of engineered exosome

A diabetic foot ulcer and skin regeneration technology, applied in the fields of biomedicine and tissue engineering, can solve the problems of patients' psychological trauma, affect the patient's living standard and quality, and limited treatment methods, so as to promote wound healing, good therapeutic effect, and easy to spread Effect

Inactive Publication Date: 2020-08-25
中怡(深圳)医疗科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The skin ulcers caused by vascular lesions are different from ordinary skin wounds. At present, the clinical treatment methods for diabetic foot ulcers are limited, generally anti-infective drugs, and even face the risk of amputation. However, long-term use of antibiotics will cause certain side effects Drug resistance, amputation will seriously affect the living standard and quality of patients, and make patients suffer from physical and psychological trauma for a long time

Method used

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  • Application of engineered exosome in skin regeneration and repair of diabetic foot ulcer and preparation method of engineered exosome
  • Application of engineered exosome in skin regeneration and repair of diabetic foot ulcer and preparation method of engineered exosome
  • Application of engineered exosome in skin regeneration and repair of diabetic foot ulcer and preparation method of engineered exosome

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Embodiment 1

[0043] The invention provides an application of an engineered exosome in skin regeneration and repair of diabetic foot ulcers and a preparation method thereof.

[0044] 1. Extraction and identification of exosomes from bone marrow mesenchymal stem cells

[0045] 1. Extraction of exosomes from bone marrow mesenchymal stem cells

[0046] The exosomes were extracted by ultra-high-speed centrifugation, and the steps were as follows:

[0047] Step (1): BMSCs (purchased from Cyagen) were subcultured to passage 3-5 with BMSCs complete medium (purchased from Cyagen), and the culture was further expanded until the BMSCs were grown to confluence At about 80%, discard the medium and replace it with serum-free DMEM-L medium to starve for 48 hours, and collect the starved medium;

[0048] Step (2): The culture obtained in step (1) was centrifuged at 2000×g for 10 min at 4°C, the precipitate was discarded, and the supernatant was collected;

[0049]Step (3): Centrifuge the supernatant ob...

Embodiment 2

[0070] The invention provides an application of engineered exosomes in skin regeneration and repair of diabetic foot ulcers and a preparation method thereof.

[0071] 1. Extraction and identification of hematopoietic stem cell exosomes

[0072] 1. Extraction of exosomes from hematopoietic stem cells

[0073] Exosomes were extracted by ultra-high-speed centrifugation, and the steps were as follows:

[0074] Step (1): Hematopoietic stem cells (purchased from ATCC) are subcultured with hematopoietic stem cell medium (purchased from Stemcell) to 3-5 generations, and further expanded for culture. When the hematopoietic stem cells grow to a confluence of about 80%, the medium is discarded and replaced. Starve the serum-free DMEM-L medium for 48 hours, and collect the starved medium;

[0075] Step (2): Centrifuge the culture obtained in step (1) at 2000×g for 10 min at 4°C, discard the precipitate, and collect the supernatant;

[0076] Step (3): Centrifuge the supernatant obtained...

Embodiment 3

[0094] The invention provides an application of engineered exosomes in skin regeneration and repair of diabetic foot ulcers and a preparation method thereof.

[0095] 1. Extraction and identification of exosomes from adipose-derived stem cells

[0096] 1. Extraction of exosomes from adipose-derived stem cells

[0097] Exosomes were extracted by ultra-high-speed centrifugation, and the steps were as follows:

[0098]Step (1): The adipose stem cells (purchased from Cyagen) were subcultured to 3-5 generations with adipose stem cell medium (purchased from Cyagen), and the culture was further expanded. When the adipose stem cells grew to about 80% confluence, the medium was discarded and replaced Starve the serum-free DMEM-L medium for 48h, and collect the starved medium;

[0099] Step (2): The culture obtained in step (1) was centrifuged at 2000×g for 10 min at 4°C, the precipitate was discarded, and the supernatant was collected;

[0100] Step (3): centrifuge the supernatant o...

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Abstract

The invention discloses an application of an engineered exosome in skin regeneration and repair of diabetic foot ulcer and a preparation method of the engineered exosome. The engineered exosome is characterized in that the exosome is used as a gene vector to load a gene for promoting vascular regeneration, and meanwhile, the exosome derived from stem cells contains various active factors to promote anti-infection of wound surface and repair of the blood vessel network and nerve. The exosome derived from adult stem cells is used for replacing stem cells which are strictly used clinically due toimmunogenicity and tumorigenicity, and the gene cargos of the synergistically loaded active molecules have the characteristics of multiple functions and good treatment effect, so that the exosome canbe widely applied to clinical repair of skin wound defects, particularly the regeneration and the repair of diabetic foot ulcerated skin.

Description

technical field [0001] The invention belongs to the fields of biomedicine and tissue engineering, in particular to the application of a novel engineered exosome in skin regeneration and repair of diabetic foot ulcers and a preparation method thereof. Background technique [0002] Exosomes are extracellular vesicles with a diameter of about 40-200nm secreted by various types of cells. They contain different kinds of microRNAs, proteins, lipids and other biologically active substances, and carry out information transmission and regulation between different cells. Signal transduction between cells plays an important role in many physiological and pathological processes such as wound repair, inflammatory response and tumor development. Stem cells are stem cells with high self-renewal ability and multi-directional differentiation potential, which have been used in clinical treatment of many diseases. However, direct use of stem cells for clinical treatment has a greater risk of ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K35/28A61K38/18A61K38/19A61P17/02A61P3/10A61P9/14C12N5/10
CPCA61K35/28A61K38/18A61K38/1825A61K38/1841A61K38/1866A61K38/191A61K48/0008A61K48/005A61P3/10A61P9/14A61P17/02C07K14/475C07K14/495C07K14/50C07K14/525C12N5/0647C12N5/0663C12N5/0667
Inventor 王江林查瑶
Owner 中怡(深圳)医疗科技集团有限公司
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