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Osteogenic differentiation inducer for mesenchymal stem cells and a preparation method of the osteogenic differentiation inducer

An osteogenic differentiation and stem cell technology, applied in the field of biomedical engineering, can solve the problems of low transduction efficiency of inducers, easy to induce immune response, large particles, etc., and achieve simple structure, no toxic side effects, and good biocompatibility. Effect

Active Publication Date: 2020-07-28
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, in order to achieve the delivery of inducers containing inducible siRNA into cells, traditional inducers often use non-viruses as carriers, resulting in low transduction efficiency of inducers, the target gene can only be expressed transiently, and the particles of the delivery vector are relatively small. Large, prone to immune response and clearance by the body

Method used

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  • Osteogenic differentiation inducer for mesenchymal stem cells and a preparation method of the osteogenic differentiation inducer
  • Osteogenic differentiation inducer for mesenchymal stem cells and a preparation method of the osteogenic differentiation inducer
  • Osteogenic differentiation inducer for mesenchymal stem cells and a preparation method of the osteogenic differentiation inducer

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Embodiment 1

[0040] In the present invention, by studying the biological mechanism characteristics of osteogenic differentiation of mesenchymal stem cells, a gold nano-core with small inner core volume and low biotoxicity is selected as a carrier of siRNA for inducing osteogenic differentiation of mesenchymal stem cells.

[0041] Wherein, the gold nano core is derived from Au-PEI (polyethyleneimine-gold nanoparticle composite). Au-PEI is obtained by mixing and reacting chloroauric acid solution and polyethyleneimine (PEI), heating and stirring, centrifuging and washing.

[0042] In the present invention, the chloroauric acid solution with a concentration of 2 to 3 mmol / L and the PEI solution with a mass fraction of 3 to 8% are mixed at a volume ratio of 10 to 15:1, and heated at 50 to 70°C for 15 to 25 minutes ; centrifuge at a speed of 6000-10000r / min for 3-8min, remove the supernatant and wash with deionized water to obtain Au-PEI particles.

[0043] The sequence of the siRNA for induci...

Embodiment 2

[0048] The present invention further studies loading display substances on the osteogenic differentiation inducer of mesenchymal stem cells described in Example 1, so that it can monitor the osteogenic differentiation of mesenchymal stem cells in real time. The present invention specifically studies a MMP-13 enzyme-responsive peptide segment linked to a fluorescent molecule—a FITC-containing polypeptide segment, which is used as a substance for real-time monitoring and display of inducing osteogenic differentiation of mesenchymal stem cells. The MMP-13 enzyme produced by osteoblasts (i.e., metalloproteinase 13 enzyme) cuts the FITC-containing polypeptide segment on the surface of the inducer, realizes specific cleavage of the polypeptide, and releases FITC fluorescent molecules; FITC fluorescent molecules are far away from gold After the nano-core, it emits fluorescence, so as to achieve the purpose of promoting and monitoring the osteogenic differentiation of mesenchymal stem ...

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Abstract

The invention provides an osteogenic differentiation inducer for mesenchymal stem cells and a preparation method of the osteogenic differentiation inducer. The osteogenic differentiation inducer for the mesenchymal stem cells comprises a gold nano core and siRNA which is modified on a surface of the gold nano core and is used for inducing osteogenic differentiation of the mesenchymal stem cells. The inducer has good biocompatibility and no toxic or side effect, is simple in structure and preparation process, can conveniently, efficiently and accurately carry the siRNA for inducing the osteogenic differentiation of the mesenchymal stem cells, safely induces human mesenchymal stem cells to be directionally differentiated into osteoblasts, and meanwhile monitors the cell differentiation condition in real time.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and in particular relates to a mesenchymal stem cell osteogenic differentiation inducer, a carrier carrying siRNA for inducing mesenchymal stem cell osteogenic differentiation, and a preparation method and application thereof. Background technique [0002] Osteoblasts and adipocytes are a common progenitor cell; bone marrow mesenchymal stem cells maintain the balance of differentiation, quantity and activity of the two in different conditions, precise humoral regulation and cell regulation in the body. The balance of adipocyte differentiation maintains the balance between intramedullary bone tissue and adipose tissue. Once the balance is broken, it can cause osteoblasts to decrease, decrease their activity, or increase their differentiation into adipocytes or increase their activity, leading to osteoporosis. occur. Therefore, how to regulate and induce the differentiation direction and bala...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N5/077
CPCC12N15/113C12N5/0654C12N2500/30C12N2506/1353
Inventor 黎锦明吴倩王锴鹏梁国海
Owner SOUTH CHINA NORMAL UNIVERSITY