Construction method of tissue engineering blood vessel

A tissue engineering and construction method technology, applied in the construction of tissue engineered blood vessels, in the field of in vitro construction of vascular grafts with autologous living tissue materials, can solve the problems of mechanical properties, compliance restricting wide application, infectious diseases, antigen residues, etc. Effects of immune rejection and inflammatory response, perfect cytocompatibility

Inactive Publication Date: 2008-09-10
XUANWU HOSPITAL OF CAPITAL UNIV OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recombinant extracellular matrix materials represented by collagen matrix have incomparable advantages in biocompatibility with traditional materials. However, problems in mechanical properties and compliance have greatly restricted the use of such materials

Method used

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  • Construction method of tissue engineering blood vessel
  • Construction method of tissue engineering blood vessel
  • Construction method of tissue engineering blood vessel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1. Reagents and materials

[0050] 1. Phosphate buffer saline (PBS solution): a small packet of commercially available PBS powder (Gibco), dissolved in 1000ml of distilled water, adjusted to pH 7.2, filtered and sterilized, aliquoted, and placed at 4°C for later use.

[0051] 2. M199 culture medium: a small bag of commercially available M199 powder (Gibco), add 1.2g of sodium bicarbonate, 1.5mol / LHEPES 10ml, three distilled water to 1L, adjust the pH to 7.2, filter and sterilize, and pack in 250ml, Store at 4°C for later use.

[0052] 3. DMEM culture medium: a small bag of commercially available DMEM powder (Gibco), add 1.2g of sodium bicarbonate, 1.5mol / LHEPES 10ml, three distilled water to 1L, adjust the pH to 7.2, filter and sterilize, pack in 250ml, Store at 4°C for later use.

[0053] 4. Endothelial cells and internal circulation culture medium: M199 culture medium containing 15-20% FBS and 0.1 mg / ml endothelial cell growth supplement (ECGS, purchased from Instit...

Embodiment 2

[0069] Example 2. Results evaluation

[0070] Specimens were collected from the fresh autologous living tissue biological tubes and tissue engineered blood vessels involved in Example 1, and mechanical measurements and biological tests were carried out according to the following methods.

[0071] 1. Method

[0072] (1) Mechanical testing

[0073] 1. Determination of tensile strength at break: cut the specimen into a plate shape, and break it on a tensile machine. The measured maximum tensile force is divided by the cross-sectional area of ​​the experimental material, which is the material tensile strength of the specimen, indicating the self-activity. Stretch resistance of tissue materials.

[0074] 2. Determination of elongation at break: Cut the specimen into a plate shape, break it on a tensile machine, record the change in length of the sample from being stretched to being broken, and divide it by the original length of the sample, the resulting percentage is The elonga...

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Abstract

The invention relates to a construction method of a tissue engineering blood vessel which belongs to the tissue engineering field. The construction method of the tissue engineering blood vessel includes the following steps: (1) tubular autologous living tissue bed frame material is connected to a continuous perfusion bioreactor under the vitro aseptic condition, and the outside of a vessel lumen is filled with culture medium; (2) endothelial cells obtained from the vitro culture medium is suspended at M199 or vascular endothelium-like cells generating from the induction of bone marrow mono nucleus cells are suspended at endothelial progenitor cell culture medium and are inoculated on the inner surface of the vessel lumen according to a high density planting method; (3) after the endothelial cells or the vascular endothelium-like cells are adhered, the inner cycle and the external cycle of the vessel lumen are communicated and the blood circulation is simulated to provide proper pressure stimulation and shear stress stimulation and the endothelial cells or the vascular endothelium-like cells are cultured for 5-7 days in a carbon dioxide incubator. The construction method has the advantages that perfect cell compatibility and enough mechanical strength are provided, the defects of immunological rejection and inflammation reaction, etc. caused by heterogenetic material are overcome, and the preparation time is short.

Description

technical field [0001] The invention relates to a method for constructing tissue engineering blood vessels, more specifically a method for constructing blood vessel grafts in vitro with autologous living tissue materials, belonging to the field of tissue engineering. Background technique [0002] At present, cardiovascular disease has become one of the diseases that seriously endanger human health, especially vascular occlusive disease, which ultimately requires surgery to replace diseased blood vessels. Currently commonly used vascular graft materials include artificial blood vessels made of non-degradable synthetic materials, homogeneous or heterogeneous blood vessels and autologous blood vessels that have been processed in different ways. The main problems of artificial blood vessel materials are thrombosis and coagulation problems; the same or heterogeneous blood vessels mainly have problems such as rejection, calcification, and poor durability. Autologous blood vessels...

Claims

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Application Information

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IPC IPC(8): A61L27/38
Inventor 张建陈晓松李建新谷涌泉俞衡锡陈兵齐力行李学锋郭连瑞罗涛崔世军徐东吴应锋
Owner XUANWU HOSPITAL OF CAPITAL UNIV OF MEDICAL SCI
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