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Mesenchymal stem cells traced by fluorescence and MRI double-image functional microspheres and application

A stem cell, dual-image technology, applied in general/multifunctional contrast agents, medical preparations containing active ingredients, preparations for in vivo experiments, etc., can solve the problem of undiscovered molecular probes and reduce the background Effects of fluorescence, minimal tissue damage, good biodegradability and biocompatibility

Active Publication Date: 2017-11-24
上海星绸生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no molecular probes with optical properties based on iridium compounds have been reported to track MSCs and perform dual-modal imaging

Method used

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  • Mesenchymal stem cells traced by fluorescence and MRI double-image functional microspheres and application
  • Mesenchymal stem cells traced by fluorescence and MRI double-image functional microspheres and application
  • Mesenchymal stem cells traced by fluorescence and MRI double-image functional microspheres and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The preparation method of the fluorescent and MRI double image functional microspheres of the present invention comprises the following steps:

[0037] (1) Weigh 2mg of ring metal iridium compound (Tris(2-(benzo[b]thiophen-2-yl)pyridineiridium(III) or Bis(2-benzo[b]thiophen-2-yl-pyridine)(acetylacetonate) iridium (III)) and 25mg PLA-PEG were mixed and dissolved with 1mL chloroform to obtain an oil phase. Prepare 100 μL of 10mg / mL Gd-DTPA contrast agent as the inner water phase.

[0038](2) The oil phase obtained in step (1) and the internal water phase were ultrasonically mixed 5 times, each time for 2 s (100 W, BILON92-II), to obtain colostrum.

[0039] (3) Add 4mL PVA aqueous solution (5wt%) to the product of step (2), and ultrasonically mix 5 times (2s, 100W) to form a double emulsion.

[0040] (4) Dilute the product of step (3) into PAA aqueous solution (5wt%, 40mL), stir overnight at room temperature, and avoid light to evaporate.

[0041] (5) Use a 50mL high-spe...

Embodiment 2

[0044] The preparation method of MSCs traced by fluorescent and MRI double image functional microspheres of the present invention comprises the following steps:

[0045] (1) HUCMSCs were normally subcultured, and the corresponding specifications were spread on the bottom of ordinary cell culture dishes.

[0046] (2) Mix the microsphere precipitation prepared in Example 1 with the MSCs medium, make a microsphere culture solution (0.2-1mg / mL) and place it in an incubator (37°C, 5% CO 2 ) and incubated for 24h.

[0047] (3) After 24 hours, HUCMSCs adhered to the wall, sucked off the culture medium, added the same volume of microsphere culture medium (0.2-1 mg / mL) incubated in step (2), and placed in an incubator (37 ° C, 5% CO 2 ), continue culturing for 2-4 days, preferably 2 days.

[0048] (4) After 2-4 days, remove the culture medium, wash twice with PBS, immerse in 5% paraformaldehyde for 30 minutes, wash twice with PBS, slide the bottom of the dish to make slide specimens....

Embodiment 3

[0055] The preparation method of the fluorescent and MRI double image functional microspheres of the present invention comprises the following steps:

[0056] (1) Weigh 2 mg of cyclometal iridium compound, mix with 25 mg of PLGA, PLA-PEG, and PLGA-PEG respectively, and dissolve with 1 mL of chloroform respectively to obtain an oil phase. Prepare 100 μL of Gd-DTPA contrast agent as the inner aqueous phase.

[0057] (2) The oil phase obtained in step (1) and the internal water phase were ultrasonically mixed 5 times (2s, 100W, BILON92-II) to obtain a colostrum.

[0058] (3) Add 4mL PVA aqueous solution (5wt%) to the product of step (2), and ultrasonically mix 5 times (2s, 100W) to form a double emulsion.

[0059] (4) Dilute the product of step (3) into PAA aqueous solution (5wt%, 40mL), stir overnight at room temperature, and avoid light to evaporate.

[0060] (5) Use a 50mL high-speed centrifuge tube to collect the solution containing microspheres in step (4), wash it once wi...

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Abstract

The invention relates to mesenchymal stem cells traced by fluorescence and MRI double-image functional microspheres. The mesenchymal stem cells endocytose the fluorescence and MRI double-image functional microspheres. The fluorescence and MRI double-image functional microspheres comprise the following components, by mass, of 70%-99.9% of polymers, 0.05%-20% of cyclic metal iridium compounds and 0.05%-10% of gadolinium agents, wherein the polymers are degradable. The invention further discloses an application of the mesenchymal stem cells in preparation of tracers and / or therapeutic agents for bone defect repairing and / or an osteoporosis disease. According to the prepared mesenchymal stem cells marked by the fluorescence and MRI double-image functional microspheres, stable single photon and double photon fluorescence intensity image signals and stable MRI image signals can be provided for observing the migration process of the stem cells in bone defect repairing and osteoporotic treatment, meanwhile, bone defects can be repaired, and osteoporotic bone defects can be treated.

Description

technical field [0001] The invention relates to the technical field of molecular probes, in particular to a mesenchymal stem cell traced by fluorescent and MRI double image function microspheres and its application. Background technique [0002] At present, osteoporosis (osteoporosis, OP), osteoporotic fractures and other diseases have received widespread attention as a global public health problem. Early and accurate diagnosis, reasonable treatment and monitoring of treatment effects are important for preventing or reducing the fragility caused by OP. Fractures are critical. The pathogenesis of OP is still unclear. It is generally believed that the bone homeostasis is broken, the number and function of osteoclasts and osteoblasts are out of balance, and bone resorption is greater than bone formation, which eventually causes OP. OP is often accompanied by a decrease in the proliferation and osteogenic differentiation of mesenchymal stem cells, and a decrease in the osteogen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K49/00A61K49/10A61K49/12A61K49/18A61P19/10A61P19/08
CPCA61K35/28A61K49/0002A61K49/0019A61K49/0054A61K49/0091A61K49/0097A61K49/105A61K49/126A61K49/1818A61K49/1896
Inventor 汤光宇徐虹邱裕友胡剑唐翠松熊祚钢张琦
Owner 上海星绸生物科技有限公司
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