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Tissue-engineered atrioventricular node and construction method and application thereof

A technology for tissue engineering and construction methods, applied in the field of biomedical engineering

Inactive Publication Date: 2012-06-13
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are no research reports on the construction of tissue-engineered AV nodes at home and abroad, and there are only a few case reports on the use of engineered tissue constructs to establish AV conduction pathways

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Inducing canine bone marrow mesenchymal stem cells to prepare cardiac pacemaker cells and conduction cells

[0030] 1. Isolation and culture of canine bone marrow mesenchymal stem cells, canine sinoatrial node cells and canine atrial appendage myocytes

[0031] (1) Isolation and culture of canine bone marrow mesenchymal stem cells

[0032]Bone marrow mesenchymal stem cells cultured by direct adherence method of canine whole bone marrow: extract 10ml of iliac bone marrow from adult hybrid dogs (Experimental Animal Center, Second Military Medical University, 15kg), centrifuge at 900×g for 10min, discard supernatant and fat; add red blood cells to lyse Liquid TrisNH 4 Cl, remove red blood cells at 4°C for 5 min, add 1×10 6 / ml was inoculated in a culture dish coated with fibronectin (FN) (Sigma Company), and the culture medium was 60% DMEM-F12 (Gibco Company), 10% FCS (Gibco Company), 10ng / ml EGF (cytolab Company) , 1000U / ml LIF (Chemicon). After 24 hours, t...

Embodiment 2

[0043] Example 2: Induced canine cardiac pacemaker cells and conduction cells were implanted into collagen sponges to construct tissue-engineered atrioventricular nodes

[0044] 1. Construction of tissue engineered atrioventricular junction

[0045] (1) Collagen sponge preparation

[0046] Collagen is derived from type I collagen of bovine Achilles tendon, and the collagen sponge is provided by Wuxi Bedi Biological Co., Ltd. Cut the collagen sponge into a size of 1.5×1.0×0.2cm, use cobalt 60 Irradiate for more than 12 hours for disinfection.

[0047] (2) Canine cardiac pacemaker cells and conduction cell mixture inoculated with collagen sponge

[0048] Take the induced cardiac pacemaker cells and conduction cells respectively, add 1ml of 0.25% trypsin (Gibco Company), and store at 37°C, 5% CO 2 Digest in the incubator for 1-2 minutes, observe under an inverted phase-contrast microscope, the cell body retracts into a round shape, add 1ml of DMEM / F12 medium (Gibco company) c...

Embodiment 3

[0054] Example 3: In vivo observation of conduction effect of tissue engineered atrioventricular node transplantation

[0055] Transplant the successfully constructed tissue-engineered conduction bundle into the subepicardium of the atrioventricular junction in dogs (Experimental Animal Center, Second Military Medical University, 15 kg) through thoracotomy, that is, to connect the tissue-engineered conduction bundle to the coronary sulcus of the heart In both atrioventricular myocardium, microsurgical suture; 3 days before transplantation, cyclosporine A (Beijing Shuanglu Pharmaceutical Co., Ltd., dose 0.25mg / day) and prednisolone (Shanghai General Pharmaceutical Co., Ltd. Ltd, the dose is 0.1mg / day) to suppress immune rejection.

[0056] The results show that the tissue engineering atrioventricular node of the present invention can survive in the host myocardium, and the cells in the transplanted tissue and the host cardiomyocytes have formed an electromechanical coupling; af...

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Abstract

The invention relates to the technical field of biomedical engineering. At present, the research about the construction of a tissue-engineered atrioventricular node is not reported in China and foreign countries. The invention provides a method for constructing the tissue-engineered atrioventricular node by using cardiac pacing cells and conduction cells which are obtained by inducing collagen sponge and bone marrow mesenchymal stem cells. The bone marrow mesenchymal stem cells are induced into the pacing cells through co-culture with sinoatrial node cells, and are induced into the cardiac pacing cells by a method for co-culture with auricular appendage muscle cells; and the two kinds of cells are used as seed cells, the collagen sponge is taken as a bracket, and the tissue-engineered atrioventricular node with a pacing characteristic and an electrocardio-conduction characteristic is constructed in vitro. Through animal experiments, the successfully constructed tissue-engineered atrioventricular node is transplanted to treat atrioventricular conduction block, and a good treatment effect is achieved. The tissue-engineered atrioventricular node which has the electrocardio-conduction characteristic and can be transplanted for treatment is obtained, and hope is brought to treatment of arrhythmia caused by heart atrioventricular conduction block.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to a tissue-engineered atrioventricular node for treating heart atrioventricular block disease and a preparation method thereof. Background technique [0002] At present, the idea of ​​biological pacing for the treatment of atrioventricular block is to use gene transfection or cell transplantation to establish a new stable pacemaker below the site of conduction block (such as the interventricular septum or the left and right ventricular walls) to provide and improve ventricular function. Autonomic rhythm, or introducing pacing genes and pacing cells into the conduction block site, repairing the damaged conduction tissue, so that patients can avoid the implantation of electronic cardiac pacemakers, and achieve the ideal therapeutic state advocated by biological cardiac pacing . However, although the transfection of pacing genes improves and produces the self-disciplin...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/24
Inventor 李玉泉张传森张喜杨向群郭金萍张璐萍黄会龙刘镇姬瑞娟
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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