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Preparation method of trinary composite stent of plasmid DNA / fibrin gel / polymer

A fibrin gel and ternary composite technology is applied in the field of preparation of plasmid DNA/fibrin gel/polymer ternary composite scaffolds, which can solve the problems of adverse reactions of hosts, easy inactivation, expensive growth factors, etc. Achieve the effect of repairing application value, promoting growth, and strengthening application value

Inactive Publication Date: 2015-07-15
ZHEJIANG UNIV
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  • Description
  • Claims
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Problems solved by technology

However, the differentiation of stem cells is regulated by the surrounding environment and growth factors. At present, most scaffolds are designed to load growth factors into the scaffolds, hoping that the loaded growth factors can promote the directional differentiation of stem cells; however, growth factors are expensive and easy to obtain. Inactivation, a large number of repeated use of growth factors will also bring adverse reactions to the host

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  • Preparation method of trinary composite stent of plasmid DNA / fibrin gel / polymer
  • Preparation method of trinary composite stent of plasmid DNA / fibrin gel / polymer
  • Preparation method of trinary composite stent of plasmid DNA / fibrin gel / polymer

Examples

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example 1

[0030] 1) Dissolve quaternized chitosan in pH 7.4 phosphate buffer, prepare a 1 mg / ml solution, and filter and sterilize through a 0.22 μm filter membrane; prepare a 500 μg / mL plasmid DNA capable of expressing BMP Aqueous solution; Plasmid DNA and quaternized chitosan solution were mixed in equal volumes, vigorously oscillated and mixed evenly, and left to stand to prepare plasmid DNA composite particles; the morphology and particle size distribution of plasmid DNA composite particles were respectively as follows figure 1 and figure 2 .

[0031] 2) Dissolve fibrinogen powder in normal saline to prepare a fibrinogen solution with a concentration of 40mg / mL; dissolve thrombin in 40mM calcium chloride solution to prepare a thrombin chloride solution with a concentration of 10U / mL calcium solution;

[0032] 3) Immerse the polylactic acid porous scaffold into ethanol, adopt the method of vacuumizing first, and then restore the pressure, and introduce ethanol into the polylactic ...

example 2

[0035] 1) Dissolve quaternized chitosan in pH 7.4 phosphate buffer, prepare a 2.5 mg / mL solution, filter and sterilize through a 0.22 μm filter membrane; prepare 1000 μg of plasmid DNA capable of expressing TGF-β1 / mL aqueous solution; the plasmid DNA and the quaternized chitosan solution were mixed in equal proportions, vigorously oscillated and mixed evenly, left to stand, and the plasmid DNA composite particles were prepared;

[0036] 2) Dissolve fibrinogen powder in normal saline to prepare a fibrinogen solution with a concentration of 80mg / mL; dissolve thrombin in 40mM calcium chloride solution to prepare a thrombin chloride solution with a concentration of 20U / mL calcium solution;

[0037] 3) Immerse the poly(lactic-co-glycolic acid) porous scaffold into ethanol, and introduce the ethanol into the poly(lactic-co-glycolic acid) porous scaffold by first vacuuming and then restoring the pressure until the scaffold is completely immersed in the ethanol middle; immerse the s...

example 3

[0041] 1) Dissolve quaternized chitosan in pH 7.4 phosphate buffer, prepare a 10 mg / mL solution, filter and sterilize through a 0.22 μm filter membrane; prepare a 100 μg / mL plasmid DNA capable of expressing IGF-1 mL of aqueous solution; the plasmid DNA and the quaternized chitosan solution were mixed in equal proportions, vigorously oscillated and mixed evenly, and left to stand to prepare plasmid DNA composite particles;

[0042] 2) Dissolve fibrinogen powder in normal saline to prepare a fibrinogen solution with a concentration of 10mg / mL; dissolve thrombin in 40mM calcium chloride solution to prepare a thrombin chloride solution with a concentration of 15U / mL calcium solution;

[0043] 3) Immerse the polyurethane porous stent in ethanol, and introduce the ethanol into the polyurethane porous stent by first vacuuming and then restoring the pressure until the stent is completely immersed in ethanol; immerse the ethanol-introduced stent in triple distilled water , using the m...

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Abstract

The invention discloses a preparation method of a trinary composite stent of plasmid DNA / fibrin gel / polymer. The preparation method comprises the following steps of: geometrically and uniformly mixing an N-trimethyl chitosan solution and a plasmid DNA solution to prepare plasmid DNA composite particles; dissolving fibrinogen in physiological saline to prepare a fibrinogen solution; dissolving thrombin in a calcium chloride solution to prepare a thrombin calcium chloride solution; sterilizing a polymer porous stent with ethanol; simultaneously filling the plasmid DNA composite particles, the fibrinogen solution and the thrombin calcium chloride solution with the same volumes in the polymer porous stent by utilizing negative pressure; and arranging the stent filled with the plasmid DNA composite particles, the fibrinogen solution and the thrombin calcium chloride solution in a constant-temperature oven to fully gelatinize the fiber protein. The preparation method is simple and convenient, uses materials with wide resources and can simulate the composition and the structure of the cartilage of a human body. The composite stent of the invention has favorable mechanical property and biological activity, can accelerate the restoration of the cartilage and the osseous tissue and have strong application values of cartilage and bone restoration.

Description

technical field [0001] The invention relates to a preparation method of a cartilage tissue regeneration scaffold in vivo, in particular to a preparation method of a plasmid DNA / fibrin gel / polymer ternary composite scaffold. Background technique [0002] Arthritis is a clinically common disease, and the articular cartilage damage caused by arthritis or sports trauma brings pain to many patients. Cartilage has active metabolism and limited repair ability. It has no blood vessels, cannot form fibrous clots after injury, has no migration of inflammatory cells, and no vascular undifferentiated cells into the damaged site, so it is not easy to repair by itself. So far, clinically, there is still a lack of effective methods to repair damaged cartilage tissue. Applying the methods and principles of regenerative medicine to repair cartilage tissue is an important means at present, and has achieved good results. Among them, the cartilage repair scaffold plays a very important role i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/48A61L27/54A61L27/56
Inventor 高长有王玮李博赵海光毛峥伟
Owner ZHEJIANG UNIV
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