Tissue engineering conduction bundle and construction method and application thereof

Abstract

The invention relates to the technical field of bioengineering. At present, many problems and difficulties exist in treating atrioventricular block through gene transfection and cell transplantation, and the current situation of researchment of biological pacing therapy is considered, so that a method that a tissue engineering conduction bundle is constructed in vitro by utilizing conduction bundle cells and scaffold materials so as to be transplanted to heart for repairing or reconstructing heart conduction path is an ideal solution of atrioventricular block therapy subject. However, no researchment of construction of the tissue engineering conduction bundle is reported at home and abroad. The invention aims to provide the tissue engineering conduction bundle and a construction method thereof. The construction method is characterized in that: heart conduction cells obtained by inducing collagen sponge and marrow mesenchymal stem cells are utilized to construct the tissue engineering conduction bundle. The tissue engineering conduction bundle provided by the invention has the characteristic of electrocardio conduction, can be used for treatment of transplant, and brings hope to treatment of arrhythmia diseases caused by atrioventricular block.

Description

technical field

[0001] The invention relates to the technical field of biomedical engineering, in particular to a tissue-engineered conduction bundle for treating heart atrioventricular block disease, a preparation method and application 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, so that patients can avoid the implantation of electronic cardiac pacemakers, and achieve the ideal therapeutic state advocated by biological cardiac pacing. However, exogenous gene introduction therapy faces many difficulties, such as lack of long-term stable expression of genes in vivo and difficulty in effective regulation; high-efficiency, safe, and well...

Examples

Embodiment 1

[0028] Example 1: Induced canine cardiac conduction cells were implanted into collagen sponges to construct tissue-engineered conduction bundles

[0029] 1. Preparation of cardiac conduction cells by inducing canine bone marrow mesenchymal stem cells

[0030] (1) Isolation and culture of canine bone marrow mesenchymal stem cells and canine atrial appendage myocytes

[0031] A. Isolation and culture of 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 (G...

Embodiment 2

[0053] Example 2: The rest are the same as in Example 1, and the concentration of paracrine factors endothelin-1 and neuregulin-1 is selected to be 1.5×10 in the induction of bone marrow mesenchymal stem cells to the direction of cardiac conduction cells. -8 M, the induction culture time is selected as 6 days, and the cell planting density is selected as 4×10 in the tissue engineering conduction bundle construction. 5 piece / cm 3 .

Embodiment 3

[0054] Example 3: The rest is the same as in Example 1, and the bone marrow mesenchymal stem cells and atrial appendage muscle cells are selected from rats.

technical field

[0001] The invention relates to the technical field of biomedical engineering, in particular to a tissue-engineered conduction bundle for treating heart atrioventricular block disease, a preparation method and application 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, so that patients can avoid the implantation of electronic cardiac pacemakers, and achieve the ideal therapeutic state advocated by biological cardiac pacing. However, exogenous gene introduction therapy faces many difficulties, such as lack of long-term stable expression of genes in vivo and difficulty in effective regulation; high-efficiency, safe, and well...

Examples

Embodiment 1

[0028] Example 1: Induced canine cardiac conduction cells were implanted into collagen sponges to construct tissue-engineered conduction bundles

[0029] 1. Preparation of cardiac conduction cells by inducing canine bone marrow mesenchymal stem cells

[0030] (1) Isolation and culture of canine bone marrow mesenchymal stem cells and canine atrial appendage myocytes

[0031] A. Isolation and culture of 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 (G...

Embodiment 2

[0053] Example 2: The rest are the same as in Example 1, and the concentration of paracrine factors endothelin-1 and neuregulin-1 is selected to be 1.5×10 in the induction of bone marrow mesenchymal stem cells to the direction of cardiac conduction cells. -8 M, the induction culture time is selected as 6 days, and the cell planting density is selected as 4×10 in the tissue engineering conduction bundle construction. 5 piece / cm 3 .

Embodiment 3

[0054] Example 3: The rest is the same as in Example 1, and the bone marrow mesenchymal stem cells and atrial appendage muscle cells are selected from rats.

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