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Bioactive material promoted stem cell secreted exosome in treatment of corneal injuries

A biomaterial and bioactive technology, applied in the field of tissue engineering and new medicine, can solve the problems of increased drug usage, poor patient compliance, inflammatory response, etc., and achieve the effect of enhancing retention rate, improving stability and promoting recovery

Active Publication Date: 2018-11-06
BEIJING HEALTH & BIOTECH (H&B) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the current method of ocular drug administration is multiple administrations, but repeated drug stimulation may produce inflammatory reactions, and the amount of drugs used increases, and patient compliance is poor

Method used

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  • Bioactive material promoted stem cell secreted exosome in treatment of corneal injuries
  • Bioactive material promoted stem cell secreted exosome in treatment of corneal injuries
  • Bioactive material promoted stem cell secreted exosome in treatment of corneal injuries

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Experimental program
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Effect test

Embodiment 1

[0017] Embodiment 1, the present invention provides a method for extraction and identification of stem cell-derived exosomes.

[0018] Cell culture medium, double antibody, trypsin and other reagents were purchased from Gibco Company; cell culture consumables were purchased from Corning Company.

[0019] The serum required for cell culture is fetal bovine serum with exosomes removed, purchased from BI Company, and the processing steps are as follows: put the fetal bovine serum in an ultracentrifuge tube, centrifuge at 120,000 g at 4 °C for 2 h, and take it in an ultra-clean bench. and filtered with a 0.22 μm syringe filter, and stored in a -80°C refrigerator for later use.

[0020] For cell biology experiments such as cell subculture, cryopreservation and recovery, please refer to "Animal Cell Culture (Sixth Edition)".

[0021] Collect hP-MSCs conditioned medium containing exosomes: when cultured at 75 cm 2 The hP-MSCs in the cell culture flasks were in the logarithmic growt...

Embodiment 2

[0031] Embodiment 2, the present invention provides the preparation method of chitosan hydrogel.

[0032] Preparation method of temperature-responsive chitosan hydrogel: Dissolve chitosan (Mn = 50,000) powder in 0.1 M acetic acid, filter sterilize with 0.22 μm filter, and prepare a solution of 2% chitosan in an ice bath .

Embodiment 3

[0033] Embodiment 3, the present invention provides a method for delivering exosomes using hydrogel wrapping.

[0034] Dissolve 0.2% hydrogel solution and stem cell-derived exosomes in equal volumes to prepare a 1× solution containing exosomes, and keep it at 4°C all the time. Keep the syringe at 4°C pre-cooling during use. .

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Abstract

The invention relates to biomaterial hydrogel eye drops containing stem cell secreted exosome, good in cellular biocompatibility and biological activity. Active hydrogel can serve as the eye drops, and the exosome is slowly released into damaged tissue; the hydrogel can also enhance retention and local concentration of the exosome at injured sites to promote repair of eye tissue injuries, promotecorneal cells to survive and proliferate, reduce apoptosis, promote functional recovery of the injured sites, inhibit pathological angiogenesis and inhibit eye inflammation; meanwhile, after the active hydrogel is combined with the bioactive material, residence time of the exosome can be prolonged, drug treating times can be reduced, and accurate control of drug dosage is facilitated.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering and new medicine. Specifically, it relates to the preparation of a bioactive material hydrogel eye drop combined with stem cell-derived exosomes, and also relates to the therapeutic effect of using such a material as an eye drop in the repair of corneal tissue damage. Background technique [0002] Stem cells are widely used in a variety of diseases because of their totipotency and multi-directional differentiation potential. Stem cells can be divided into adult stem cells and embryonic stem cells. Among them, mesenchymal stem cells are a variety of adult stem cells that only express low levels of HLA / MHC. Class I molecules with low immunogenicity and certain immunosuppressive ability. It exhibits pluripotency and is capable of differentiating in vitro and in vivo into the phenotype of many connective tissue cell types such as osteoblasts, chondrocytes and adipocytes. become an importa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K35/22A61K35/34A61K35/44A61K9/08A61K47/36A61K47/38A61P27/02A61P17/02
CPCA61K9/0019A61K9/0048A61K9/08A61K35/22A61K35/28A61K35/34A61K35/44A61K47/36A61K47/38A61P17/02A61P27/02
Inventor 李宗金陶红燕
Owner BEIJING HEALTH & BIOTECH (H&B) CO LTD
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