Nano-drug based on stem cells as well as preparation method and application of nano-drug
A nano-drug and stem cell technology, applied in the field of medicine, can solve problems such as short residence time and difficulty penetrating the bladder permeability barrier, and achieve the effect of improving mechanical pain sensitivity and improving symptoms of urination function
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Embodiment 1
[0046] The embodiment of the present application provides a method for preparing stem cell conditioned medium, specifically including:
[0047] see figure 1 , figure 1 The flow chart for the preparation of the stem cell conditioned medium provided in the examples of this application, using low-glucose DMEM medium containing 10% fetal bovine serum (FBS) to cultivate hUC-MSCs, culture and passage to the fourth generation, and replace it with low-glucose DMEM medium without FBS. The hUC-MSCs were cultured in the glucose DMEM medium to obtain the stem cell conditioned medium, specifically as follows: when the growth confluence of the hUC-MSC cells reached 80%, the medium was replaced with a low-glucose DMEM medium without FBS. After 48 hours, conditioned media from hUC-MSCs were collected and centrifuged (1500 xg, 10 minutes) to remove cell debris. Afterwards, centrifugation was performed using a centrifugal ultrafilter (3kDa cut-off, Millipore, Billerica, MA) at 4°C (4000×g, 40...
Embodiment 2
[0052] The embodiment of the present application provides a stem cell-based nanomedicine, which is PLGA / CM-TMC NPs, and the specific preparation method refers to the comparative example.
[0053] see figure 2 , figure 2 The flow chart for the preparation of PLGA / CM-TMC NPs provided in the examples of this application. Specifically, 0.2 mL of concentrated stem cell-conditioned medium was added to 1 mL of DCM containing 20 mg of PLGA polymer, and sonicated in an ice bath for 60 seconds (100 W) to obtain a W / O primary emulsion. Then, the above W / O primary emulsion was poured into 4 mL of 1% (w / v) PVA aqueous solution (containing 1.6 mg TMC), and sonicated in an ice bath for 60 seconds (100 W) to obtain a W / O / W complex lotion. Subsequently, the W / O / W double emulsion was added dropwise into 30 mL of 1% (w / v) PVA aqueous solution, and then stirred at room temperature for at least 3 hours to remove DCM. Finally, PLGA / CM-TMC NPs were collected by centrifugation for 30 min (15000...
Embodiment 3
[0055] The embodiment of the present application provides a stem cell-based nanomedicine, which is PCL / CM-TMC NPs, and the specific preparation method is as follows:
[0056] PCL polymer (molecular weight 60 kDa) was purchased from Aladdin Biochemical Technology Co., Ltd. (China). Add 0.2 mL of concentrated stem cell-conditioned medium to 1 mL of DCM containing 20 mg of PCL polymer and sonicate for 60 s (100 W) in an ice bath to obtain a W / O primary emulsion. Then, the above W / O primary emulsion was poured into 4 mL of 1% (w / v) PVA aqueous solution (containing 1.6 mg TMC), and sonicated in an ice bath for 60 seconds (100 W) to obtain a W / O / W complex lotion. Subsequently, the W / O / W double emulsion was added dropwise into 30 mL of 1% (w / v) PVA aqueous solution, and then stirred at room temperature for at least 3 hours to remove DCM. Finally, the PCL / CM-TMC NPs were collected by centrifugation for 30 min (15000 × g, 4 °C) and resuspended in ultrapure water, and the centrifugati...
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