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Application of a Nanocomposite of Functionalized Polyamide-Amine Dendrimers

A technology of amine dendrimers and nanocomposites, applied in the field of targeted gene transfection with polymer nanocarriers

Inactive Publication Date: 2016-01-20
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Retrieval of related literature and patents at home and abroad shows that there is no report on the method of using functionalized dendrimers and their nanoparticles as carriers for gene transfection of bone marrow mesenchymal stem cells

Method used

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  • Application of a Nanocomposite of Functionalized Polyamide-Amine Dendrimers
  • Application of a Nanocomposite of Functionalized Polyamide-Amine Dendrimers
  • Application of a Nanocomposite of Functionalized Polyamide-Amine Dendrimers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Weigh NH 2 - PEG-COOH 11.58 mg, dissolved in 5 mL DMSO. While stirring, 6-MAL with a dry weight of 1.785 mg dissolved in 1 mL of DMSO solution was added dropwise, and reacted for 8 hours to obtain a MAL-PEG-COOH solution. Add 1mL RGD-SH DMSO solution (4mg / mL) dropwise to the above MAL-PEG-COOH solution, and react for 12h to obtain RGD-PEG-COOH solution. Weigh 4.0 mg of EDC and 2.4 mg of NHS, dissolve them in 1 mL of DMSO, add the EDC solution and NHS solution drop by drop to the RGD-PEG-COOH solution, and stir for 3 hours. Weigh the fifth generation polyamidoamine dendrimer (G5.NH 2 ) 15.06 mg, dissolved in 5 mL DMSO. Add the activated RGD-PEG-COOH solution dropwise to G5.NH 2 solution, magnetically stirred at room temperature, and reacted for 3d to obtain G5.NH 2 -(PEG-RGD) 10 . Weigh mPEG-COOH11.58mg, dissolved in 5mLDMSO. Weigh 2.0 mg EDC and 1.2 mg NHS, dissolve them in 1 mL DMSO respectively, add the EDC solution and NHS solution dropwise to the mPEG-COOH s...

Embodiment 2

[0072] G5.NH 2 , Comparative Example 1, Comparative Example 2, the G5.NH prepared by the method of Comparative Example 3 and Example 1 2 -mPEG 20 , {(Au 0 ) 25 -G5.NH 2 -mPEG 20}, G5.NH 2 -(PEG-RGD) 10 -mPEG 10 , {(Au 0 ) 25 -G5.NH 2 -(PEG-RGD) 10 -mPEG 10}Under different N / P ratio conditions (1:1, 2.5:1, 5:1) respectively form vector / hBMP-2pDNA complexes with 5 μghBMP-2pDNA through electrostatic interaction, so that the final volume is fixed at 100 μL, and incubated at room temperature 20min, then add 1mL LPBS. Its hydrodynamic particle size and surface potential were characterized by Malvern laser particle size analyzer (Malvern, MK, 633nm laser), the results are as attached to the description figure 1 shown. The results showed that with the increase of N / P ratio, the hydrodynamic particle size of the composites decreased, almost all below 200nm; while the surface potential of most of the composites increased with the increase of N / P ratio, but Also around 20...

Embodiment 3

[0074] to have V　3 Bone marrow mesenchymal stem cells hMSCs expressing integrin as model cells to test G5.NH 2 , the cytotoxicity of the materials prepared in Comparative Example 1, Comparative Example 2, Comparative Example 3 and Example 1. Take 1.5×10 4 Density per well hMSCs were seeded in 96-well plates, cultured in 100 μL α-MEM medium supplemented with 100 U / mL penicillin, 100 U / mL streptomycin and 10% FBS, and cultured at 37 °C for 24 h at a concentration of 5% carbon dioxide. Then the medium was replaced with G5.NH containing G5. 2 , G5.NH 2 -mPEG 20 , {(Au 0 ) 25 -G5.NH 2 -mPEG 20}, G5.NH 2 -(PEG-RGD) 10 -mPEG 10 and {(Au 0 ) 25 -G5.NH 2 -(PEG-RGD) 10 -mPEG 10} cell culture medium and cells were co-cultured for 24 hours, then 20 μL of MTT (5.0 mg / mL) solution was added, and the culture was continued for 4 hours. Remove the culture medium in the wells, add 150 μL DMSO to each well, shake on the shaker for 30 minutes, and test the absorbance value with a...

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Abstract

The invention relates to application of a functionalized polyamide-amine dendrimer nanocomposite, and particularly relates to application of a functionalized polyamide-amine dendrimer nanocomposite which carries hBMP-2pDNA and induces hMSCs to be differentiated into a carrier of osteoblast. The preparation process of the functionalized polyamide-amine dendrimer nanocomposite is simple, the experiment conditions are mild, and the experiment is easy to operate. The functionalized polyamide-amine dendrimer nanocomposite carrier is a good gene carrier which can successfully carry hBMP-2pDNA and induce mesenchymal stem cells to be differentiated into osteoblast, and has good application prospect in repair and rebuilding of bone tissues.

Description

technical field [0001] The invention belongs to the field of targeted gene transfection of polymer nanocarriers, and particularly relates to a nanocomplex of functionalized polyamide-amine dendrimers as a loaded hBMP-2pDNA (human bone morphogenetic protein particle DNA) to induce hMSCs (bone marrow interstitial DNA) Mesenchymal stem cells) differentiated into osteoblasts. Background technique [0002] Gene therapy, as a new technology born from the combination of modern medicine and molecular biology, is a novice in the treatment of diseases by introducing exogenous normal genes into target cells to correct or compensate for diseases caused by gene deletions or abnormalities. As early as 1968, American scientist Michael Blazer first proposed the concept of gene therapy in the medical field. But it was not until 1990 that the first relevant clinical trials were conducted in humans. Over the past decade, many clinical trials have focused on gene therapy as a novel medicinal ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K47/34C08G83/00A61L27/18A61L27/54A61P19/08
Inventor 史向阳孔令丹侯文秀
Owner DONGHUA UNIV
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