Biological mixed artificial blood vessel and preparation method thereof
An artificial blood vessel and hybrid technology, which is applied in the field of bioengineering to achieve the effects of facilitating angiogenesis, promoting endothelial repair, and regulating angiogenesis
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Embodiment 1
[0039] The preparation of embodiment 1 biohybrid artificial blood vessel
[0040] 1. Preparation of Inner Main Body Vascular Stent
[0041] 1) Preparation of natural polymer artificial blood vessels by electrospinning: prepare a PCL electrospinning solution with a concentration of 10 g / ml with hexafluoroisopropanol, and stir until completely dissolved; the instrument used in the experiment is a YFSP-T high-voltage electrospinning machine . Put the electrospinning solution in a 10ml syringe, select the No. 7 needle, set the voltage to 15KV, the advancing speed of the syringe pump is 1ml / h, the receiving distance is 15cm, the drum receiving device is connected to a 3mm steel pipe for collection, and the drum speed is 150r / h min, the obtained polymer tubular fiber polymer is used as the inner main body vascular stent;
[0042] Prepared by the acellular matrix method:
[0043] 2) The prepared allogeneic acellular matrix stent is derived from the pig's cardiovascular system. Aft...
Embodiment 2
[0049] Example 2 Artificial blood vessels promote the growth and activity of vascular endothelial cells
[0050] ① Preparation of HUVECs culture and extract
[0051] HUVECs were cultured in RPMI1640 with 1% double antibody (penicillin / streptomycin) and 10% FBS serum at 37°C and 5% CO2, and the state of the cells was observed regularly for medium replacement or passage.
[0052] Peracetic acid was diluted with sterile PBS to a final concentration of 0.1% (v / v). It is best to use it now. Cut the prepared Gelatin, GelMA, and PCL materials into 1cm×1cm size, soak in peracetic acid for 1h. Pour off the peracetic acid, carefully rinse the material with sterile PBS, shake to wash off the peracetic acid, wash 3 times, 1 min each time. Soak in sterile PBS for 3 hours, discard the PBS, and finally soak in sterile PBS and store at 4°C. According to ISO10993-12:2012(2012), the ratio of the surface area of the material to the extraction medium is 6cm 2 / ml, the material was soaked i...
Embodiment 3
[0055] Example 3 Artificial blood vessels promote the migration of vascular endothelial cells
[0056] The effects of Glatin, GelMA and PCL materials on cell migration were observed by scratch method. Digest cells in logarithmic growth phase to make 1x10 5 cells / ml cell suspension, inoculate 12-well plate with 1ml / well; when the HUVECs in the well grow to a single layer, starve the cells with serum-free medium for 4 hours; then use a 10ml pipette tip to mark "ten" on the well plate After adding PBS to wash for 3 times, add medium and extract, randomly select 6 fields of view to take pictures under the microscope, and record it as the scratch width of 0h; cultivate in 37℃, 50% COZ incubator for 12h and 36h respectively Take pictures to observe the migration of cells. Such as Figure 4 As shown, compared with the control group, the GelMA and PCL groups significantly promoted the migration of vascular endothelial cells after culturing for 12h and 36h. Therefore, it is enough t...
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