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Injectable myocardial tissue engineering product applying OPF-based hydrogel as liquid scaffold

A tissue engineering and hydrogel technology, applied in the field of injectable myocardial tissue engineering products, can solve the problems that have not yet been reported, and achieve the effects of facilitating treatment operations, promoting myocardial tissue regeneration, and simple operation technology.

Inactive Publication Date: 2010-01-06
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, OPF hydrogel is mainly used in cartilage and bone tissue engineering, and it has not been reported for the research and development of injectable myocardial tissue engineering products.

Method used

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  • Injectable myocardial tissue engineering product applying OPF-based hydrogel as liquid scaffold
  • Injectable myocardial tissue engineering product applying OPF-based hydrogel as liquid scaffold
  • Injectable myocardial tissue engineering product applying OPF-based hydrogel as liquid scaffold

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: the preparation of OPF

[0032] Dissolve 30 g of polyethylene glycol with a molecular weight of 1000 in 250 ml of toluene, and after azeotropic distillation, dissolve the dry polyethylene glycol in 250 ml of anhydrous dichloromethane. Fumaryl chloride (0.3 mol) and triethylamine were then added dropwise to the polyethylene glycol solution over 5 hours while the reaction flask was maintained at 0°C and stirred at room temperature for 1-2 days to ensure maximum conversion. After the reaction, the solvent was removed by rotary evaporation, after which the product was dissolved in 500 ml ethyl acetate. The solution was filtered to remove the salt formed by chloride ions and triethylamine during the reaction, and the oligomer was recrystallized twice in ethyl acetate. Finally, pure OPF was precipitated in diethyl ether and dried in vacuo (figure 1 ).

[0033] Purified OPF was stored at -20°C and sterilized before use by exposure to ethylene oxide for 16 hours...

Embodiment 2

[0034] Embodiment 2: Histocompatibility detection of OPF hydrogel

[0035] 0.1ml of OPF hydrogel was injected and implanted into the myocardium of S-D rats, and they were sacrificed at 1, 2, and 4 weeks respectively. Two rats were taken each time. After the specimens were taken out, paraffin sections were made for HE staining to observe the inflammatory reaction. One week after surgery, the OPF hydrogel was widely distributed in the myocardial tissue space, and a little inflammatory cells were mixed in between ( figure 2A); 2 weeks after operation, the OPF hydrogel was partially degraded and surrounded by inflammatory cell infiltration ( figure 2 B); No obvious OPF hydrogel can be seen in the 4-week group after operation, and a small amount of inflammatory cell infiltration can be seen ( figure 2 C).

Embodiment 3

[0036] Embodiment 3 Preparation of cardiomyocytes derived from mouse embryonic stem cells (mES)

[0037] Preparation of mouse embryonic fibroblast feeder layer: kill BALB / C mice at 13-15 days pregnant, remove the head and internal organs of the embryos under aseptic conditions, wash them thoroughly with PBS, cut them into pieces and suspend them in Digest in stages in 0.25% trypsin solution. Stop the digestion at an appropriate time, aspirate the supernatant and centrifuge to collect the cells. Cell viability was identified by the trypan blue exclusion method, and counted at 5×10 8 cells / L were resuspended in H-DMEM medium containing 10% FBS, 37°C, 5% CO 2 and cultured overnight in a saturated humidity incubator. The next day, the cells were frozen for later use or directly prepared as feeder cells: MEFs were treated with a medium containing 10 mg / L mitomycin C for 2.5-3 hours, and washed sufficiently with PBS to remove residual components. Adding H-DMEM medium containing ...

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Abstract

The invention discloses an injectable myocardial tissue engineering product applying OPF hydrogel as a liquid scaffold, in particular, the product applies the OPF hydrogel as the scaffold and combines seed cells from different sources, such as embryonic stem cells, mesenchymal stem cells, myocardial cells and the like, and the repair effect of the product on myocardial infarction regions is observed after the product is injected and transplanted into the certain regions of an animal myocardial infarction model. The product built by the scaffold can improve the retention rates and the survival rates of the seed cells, promote regeneration of the myocardial tissues, increase the ventricular wall thickness of the infarction regions, reshape the original ventricle and improve the cardiac function. The product has the characteristic of injectability and is convenient for treatment and operation, thus avoiding the risks brought by the operations such as cardiac arrest and extracorporeal circulation, etc. The invention is simple in operating process and mild in implementation conditions, provides a new product for the myocardial tissue engineering and is of great significance in the development of clinically treating heart diseases by tissue-engineered myocardium.

Description

technical field [0001] The invention belongs to the field of tissue engineering and regenerative medicine, in particular to an injectable myocardial tissue engineering product made of oligoethylene glycol fumarate (OPF) hydrogel as a scaffold. Background of the invention [0002] The morbidity and mortality of ischemic heart disease and subsequent heart failure are increasing year by year globally. Cardiovascular system disease has become one of the main causes of death and disability in the population, especially the elderly, and is the main cause of death in the medical field today. One of the puzzles. Current treatment methods include drugs, coronary intervention, coronary artery bypass surgery, etc. Although these methods can alleviate the patient's symptoms to a certain extent, they are difficult to fundamentally restore the number of cardiomyocytes and improve the diastolic and systolic function of the heart. Heart disease patients often need organ transplantation at ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L31/06A61L31/16A61K38/18A61K9/00A61P9/10
Inventor 王常勇王海滨郝彤段翠密
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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