Medical appliance containing recombinant hCREG protein and preparation method of medical appliance
A medical device and protein technology, which is applied in the field of medical devices containing recombinant hCREG protein and its preparation, can solve the problem of low expression level, achieve the effects of inhibiting cell proliferation, avoiding allergic and inflammatory reactions, and promoting re-endothelialization
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Embodiment 1
[0051] Example 1 Preparation of recombinant hCREG glycoprotein and its coronary stent
[0052] 1. Preparation of Microporous Recombinant hCREG Glycoprotein Elution Scaffold
[0053] 1.1 Preparation of recombinant hCREG glycoprotein
[0054] 1.1.1 Construction of recombinant gene vector Since 1999, the mRNA differential display technology was used to successfully clone the human CREG gene from the human internal thoracic artery VSMCs cultured in vitro for the first time, and the hCREG open reading frame with a stop codon mutation was amplified by RT-PCR technology Frame, the hCREG RT-PCR primers of the stop codon mutation are as follows: the upstream primer is 5'-aa ggatcc atggccgggctatcccgc-3' (SEQ ID NO.1), and the downstream primer is: 5'-gc gaattc Gcactgaactgtgacatttaatattcttctgg-3' (SEQ ID NO .2), where the capital letter represents the stop codon of the C / G mutation.
[0055] The recombinant human CREG protein expression vector pcDNA3.1-his / myc-hCREG with his tag protei...
Embodiment 2
[0066] Embodiment 2 Preparation of drug-protein combined scaffold
[0067] The surface of the coronary stent in this embodiment is uniformly distributed with recombinant hCREG glycoprotein and rapamycin. Rapamycin is distributed on the outer layer of the stent, and the recombinant hCREG glycoprotein of the present application is distributed in the lumen of the stent.
[0068] Stent with micro-pores: the stent is made of 316L medical stainless steel substrate, and the surface is uniformly equipped with micro-pores of about 500 nm.
[0069] Preparation of three-sided rapamycin drug stent: prepare rapamycin drug with a concentration of 1% in acetone solution. The inner surface of the stent is protected, and rapamycin is sprayed on the outer surface of the stent.
[0070] Preparation of recombinant hCREG protein-eluting scaffolds: the recombinant hCREG glycoprotein is immobilized to the inner lumen of the scaffold by using the principle of physical adsorption of micron holes. T...
Embodiment 3
[0072] Example 3 Preparation of double-drug protein combined scaffold
[0073] The surface of the coronary stent in this embodiment is uniformly distributed with recombinant hCREG glycoprotein, rapamycin and paclitaxel. Rapamycin and paclitaxel were distributed on the outer surface of the stent, and recombinant hCREG glycoprotein was distributed in the lumen of the stent.
[0074] Stent with micro-pores: the stent is made of 316L medical stainless steel substrate, and the surface is uniformly equipped with micro-pores of about 500 nm.
[0075] Prepare a three-sided rapamycin drug stent: protect the inner surface of the stent, spray rapamycin and paclitaxel on the outer surface of the stent, the drug concentration is 1%, the drug solvent adopts tetrahydrofuran, and the space between rapamycin and paclitaxel is The mass ratio is 1:1.
[0076] Preparation of recombinant hCREG protein-eluting stent (drug on the outer surface, protein on the inner surface): the recombinant hCREG ...
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