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Temperature sensitive biogel preparation and application thereof

A technology of gel preparation and sex biology, applied in the field of stem cell application, can solve the problems of limited separation effect and retrograde infection, and achieve the effects of improving viscosity and bioadhesion, preventing infection and preventing uterine intrauterine adhesion.

Active Publication Date: 2019-05-31
江苏拓弘生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are disadvantages such as limited separation effect and easy to cause retrograde infection.

Method used

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  • Temperature sensitive biogel preparation and application thereof
  • Temperature sensitive biogel preparation and application thereof
  • Temperature sensitive biogel preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Preparation of MSCs exosomes

[0037] 1) Human placental mesenchymal stem cells were plated at 1×10 6 / cm 2 The densities were inoculated in multiple culture flasks. Place the cell culture flask at 37°C, 5% CO 2 Culture in a humidified incubator, and change the medium every three or four days. The culture medium is a commonly used mammalian cell culture medium. The cell growth and morphological changes were observed under an inverted microscope. When the cells grew to 60% of the bottom of the culture flask, fresh culture medium was replaced. After 48 hours, the supernatant of each culture flask was collected and combined. Take 30 mL of the supernatant, centrifuge at 1000 × g for 15 min to remove cell debris, filter it through a 0.45 μm sterile filter into a 20 mL ultrafiltration centrifuge tube, and centrifuge at 1000 × g for 70 min at 4 °C to obtain exosome-containing The concentrated solution was transferred to an ultracentrifuge tube with 200 μL of suc...

Embodiment 2

[0039] The preparation of embodiment 2 cold solution method in situ gel

[0040] Take the MSCs exosomes prepared in Example 1, and mix the following raw materials by weight: 20 parts of MSCs exosomes, 26 parts of Poloxamer 407 (P407), 5 parts of Poloxamer 188 (P188), 0.3 part of sodium alginate, 0.1 part of polyethylene glycol, 0.5 part of chitosan, 48 parts of phosphate buffer and pure water. Under stirring conditions, the above substances were slowly added in proportion to sterile pure water for injection, and placed in a 4°C refrigerator for 24 hours to fully swell to obtain a clear and uniform solution, which is a temperature-sensitive biogel preparation. It was placed in a 4°C refrigerator for later use.

[0041] The inversion method was used to determine the gelation temperature of temperature-sensitive biogel formulations. Take 5mL of the above-prepared solution into a 1mL test tube, put it in a 15°C water bath, and slowly heat up at a rate of 1°C / min. When the soluti...

Embodiment 3

[0042] Example 3 Preparation of cold solution method in situ gel

[0043] Take the MSCs exosomes prepared in Example 1, and mix the following raw materials by weight: 20 parts of MSCs exosomes, 25 parts of Poloxamer 407 (P407), 7 parts of Poloxamer 188 (P188), 0.3 part of hydroxypropyl methylcellulose, 0.1 part of polyethylene glycol, 0.5 part of chitosan, 48 parts of phosphate buffer and pure water. Under stirring conditions, the above substances were slowly added in proportion to sterile pure water for injection, and placed in a 4°C refrigerator for 24 hours to fully swell to obtain a clear and uniform solution, which is a temperature-sensitive biogel preparation. It was placed in a 4°C refrigerator for later use.

[0044] The gelling temperature was measured according to the method of Example 2, and the results showed that the gelling temperature was 30-35°C.

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Abstract

The invention relates to a temperature sensitive biogel preparation comprising, by mass, 20-30 parts of mesenchymal stem cell exosome, 20-28 parts poloxamer 407, 5-10 parts of poloxamer 188, 0.9-2.4 parts of polymer stabilizer for adjusting gelation temperature and drug release characteristics and 32-54 parts of water. The biocompatible degradable polymer is used as a drug-loading matrix of the mesenchymal stem cell exosome, and a polymer stabilizer is added to adjust the gelation temperature and the drug release characteristics, so that the loaded MSCs exosome can be released continuously toreduce the dosing frequency, prevent leakage of liquid medicine, improve the bioavailability and provide the safe effective preparation convenient to apply clinically for treatment of IUA.

Description

technical field [0001] The invention relates to the technical field of stem cell application, in particular to a temperature-sensitive biogel preparation and application thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are a class of adult stem cells derived from mesoderm with self-renewal, proliferation and multi-directional differentiation potential, which can differentiate into osteoblasts, chondroblasts and adipocytes. MSCs can be isolated from various tissues such as bone marrow, adipose tissue, lung, liver, spleen, placenta, amniotic fluid, umbilical blood, and umbilical cord. MSCs have been widely used in tissue engineering and immunotherapy due to their ability of easy isolation, multi-directional differentiation, unique immunophenotype and immune regulation. [0003] Impairment of repair after endometrial injury is an important cause of common clinical diseases such as oligomenorrhea, intrauterine adhesions, amenorrhea and secondary infertility....

Claims

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Application Information

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IPC IPC(8): A61K9/06A61K47/10A61K47/36A61K47/38A61K35/28A61P15/00
Inventor 廖联明黄静廉云飞于峰祥
Owner 江苏拓弘生物科技有限公司
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