A tissue engineered myocardial patch and its preparation and application
A technique of tissue engineering and cardiac muscle, applied in the field of regenerative medicine, can solve the problems of lack of biological activity, large difference in extracellular matrix, etc., to improve mechanical strength and toughness, facilitate implantation operation, and improve biological activity Effect
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Embodiment 1
[0027] Preparation of 1% (w / v, g / ml) chitosan (5000 kDa), 1% (w / v, g / ml) hyaluronic acid (4000000 kDa) and 3% (w / v, g / ml) acidic The gelatin (120bloom g) of the gel solution 10 g, after which the crosslinking agent glutaraldehyde was added, the crosslinking agent final concentration was 0.01% (w / v, g / ml), and the crosslinking conditions were temperature 23 ° C. The humidity was 60%, obtained a gel film, and then neutralized with a 0.1 M glycine solution and (soaked time of 1 h), washed and dried (18 ° C, moisture 20%.) After obtaining the imitation extracellular matrix base film. Thereafter, rat myocardial cells and vascular endothelial cells were inoculated on this base membrane (two types of cells were 2: 1) and 2 days were cultured to form a first layer of cell layer. The polylatine solution of 4% (w / v, g / ml) is added above the first layer of cell layer (200 microliters / cm) 2 The same cell suspension is then cultured, and the second layer of cell layer is ...
Embodiment 2
[0029] Preparation of 10% (w / v, g / ml) chitosan (300000 kDa), 0.1% (w / v, g / ml) hyaluronic acid (1000000 kDa) and 15% (w / v, g / ml) base The gel (150bloom g) of the gel solution 10 g, after which the crosslinking agent formaldehyde was added, the crosslinking agent final concentration was 3% (w / v, g / ml), and the crosslinking conditions were temperature 18 ° C, humidity. 80%, obtained a gel film, and then subjected to a 0.1M glycine solution and (a soaking time of 2.5 h), washed and dried (25 ° C, humidity 20%.) After obtaining an extracellular matrix base film. Thereafter, rats were inoculated on this base membrane (rat formation of rats) and endothelial cells (two types of cells were 3: 2) and cultured for 2 days to form a first layer of cell layer. Cell adhesives 3% (w / v, g / ml) fibrin solution (200 microliters / cm) above the first layer of cell layer 2The same cell suspension is then cultured, and the second layer of cell layer is formed. Add cell adhesive 1% (w...
Embodiment 3
[0031] Preparation of 5% (w / v, g / ml) chitosan (100000 kDa), 0.4% (w / v, g / ml) hyaluronic acid (2000000 kDa) and 8% (w / v, g / ml) base The gel (200bloom g) of the gel solution 10g, after which the crosslinking agent is added to the crosslinking reaction, the final concentration of the crosslinking agent is 0.5% (w / v, g / ml), and the crosslinking conditions are temperature 25 ° C. The humidity is 50%, and the gel film is obtained, and then the lysate and dried (20 ° C, 20% by weight of 20%.) Are obtained, and then the extracellular matrix base film is obtained. Thereafter, the cardiomyocytes, vascular endothelial cells and fibroblasts of rats with murderous stem cells were inoculated on this base film (three types of cells were 3: 2: 1) and cultured for 2 days to form the first A layer of cell layer. Cell adhesives 6% (w / v, g / ml) Matrigel (200 microliters / cm) above the first layer of cell layer 2 The same cell suspension is then cultured, and the second layer of ce...
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