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Polypeptide co-modified naringin nanoliposomes for promoting proliferation and osteogenic differentiation of dental pulp stem cells and preparation method and application thereof

A technology of naringin nano- and dental pulp stem cells, which is applied in the field of polypeptide co-modified naringin nano-liposomes and its preparation, can solve the problems of increasing cytotoxicity, limiting human application, and low cell uptake rate, etc. Effects of drug concentration, increased uptake, and good proliferation of dental pulp stem cells

Active Publication Date: 2021-06-04
BEIJING STOMATOLOGY HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Studies have found that naringin has antibacterial, anti-inflammatory, anti-viral, anti-cancer, anti-mutation, anti-allergic, anti-ulcer, analgesic, lowering blood pressure, lowering blood sugar and other activities, and can lower blood cholesterol and reduce the formation of thrombus , improve local microcirculation and nutrient supply, often used to prevent and treat food accumulation, abdominal distension, cough with phlegm, dysentery, diarrhea and cardiovascular and cerebrovascular diseases. At present, some scholars have conducted research on the role of naringin in promoting osteogenesis However, researchers have found that due to the poor water solubility of naringin, the cell uptake rate is low. In order to increase cell uptake and enhance drug efficacy, it is usually necessary to use a large amount of organic solvents and increase the dosage concentration in the process of cell and animal experiments, so that Greatly increased its cytotoxicity, which in turn caused great difficulties and risks to the development of related preparations and actual clinical applications, limiting its human application

Method used

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  • Polypeptide co-modified naringin nanoliposomes for promoting proliferation and osteogenic differentiation of dental pulp stem cells and preparation method and application thereof
  • Polypeptide co-modified naringin nanoliposomes for promoting proliferation and osteogenic differentiation of dental pulp stem cells and preparation method and application thereof
  • Polypeptide co-modified naringin nanoliposomes for promoting proliferation and osteogenic differentiation of dental pulp stem cells and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: A preparation method of polypeptide co-modified naringin nanoliposomes for promoting the proliferation and osteogenic differentiation of dental pulp stem cells

[0039]1. Reagent materials and equipment

[0040] 1. Reagent materials

[0041] Naringin, acetonitrile (chromatographically pure) (Sinopharm Chemical Reagent Co., Ltd.);

[0042] Phosphatidylcholine, lecithin EPC, cholesterol, distearoylphosphatidylethanolamine-polyethylene glycol-carboxyl (Shanghai Shunna Biological Co., Ltd.);

[0043] 1-Ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC, SIGMA, USA);

[0044] Tetrahydrofuran, disodium hydrogen phosphate, anhydrous methanol, petroleum ether (Shanghai Chemical Reagent Factory).

[0045] 2. Instruments and equipment

[0046] SB20001 precision electronic balance (Shanghai Huyueming Scientific Instrument Co., Ltd.);

[0047] RV-5A rotary evaporator (Shanghai Yiheng Scientific Instrument Co., Ltd.);

[0048] LC-16 high performance liqui...

Embodiment 2

[0066] Example 2: Effects of polypeptide co-modified naringin nanoliposomes on the proliferation of dental pulp stem cells

[0067] After the successful preparation of peptide co-modified naringin nanoliposomes, we conducted cell in vitro experiments to study the effect of the nanoliposomes on the osteogenic differentiation of dental pulp stem cells. First, use the MTT experiment to determine the effective concentration of naringin raw materials on the osteogenic differentiation of dental pulp stem cells, and then compare and observe the effects of polypeptide co-modified naringin nanoliposomes on the proliferation of dental pulp stem cells, including observing each time period with an inverted microscope Morphological changes of dots, CCK-8 detection of stem cell activity changes, life and death staining to observe the survival of stem cells, flow cytometry to detect cell cycle and apoptosis, and q-PCR to detect the expression of vascular growth factor VEGF.

[0068] The expe...

Embodiment 3

[0085] Example 3: Effect of polypeptide co-modified naringin nanoliposomes on osteogenic differentiation of dental pulp stem cells

[0086] In order to observe the effect of RGD&TAT polypeptide co-modified naringin nanoliposomes on the osteogenic differentiation of dental pulp stem cells, related experiments on osteogenesis were carried out, including alizarin red staining, ALP staining, detection of ALP, OCN, RUNX2 by q-PCR , BSP, OSX and other osteogenic gene expression. These experimental results showed that RGD&TAT polypeptide co-modified naringin nanoliposomes had a better effect on promoting osteogenic differentiation of dental pulp stem cells than other experimental groups.

[0087] (1) Alizarin red staining to evaluate osteogenesis

[0088] In the alizarin red staining experiment, it can be observed that on the 14th and 21st days of administration, the RGD&TAT polypeptide co-modified naringin nanoliposome group showed the best osteogenic effect ( Figure 11 ).

[00...

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Abstract

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to polypeptide co-modified naringin nanoliposomes for promoting proliferation and osteogenic differentiation of dental pulp stem cells and a preparation method and application thereof . The naringin nanoliposomes disclosed by the invention are prepared by co-modification of RGD peptide and TAT peptide. The polypeptide co-modified naringin nanoliposomes prepared by the method of the invention can obviously reduce the administration concentration, increase the uptake of drugs by cells and promote the exertion of drug effects of the naringin, and cell experiments prove that the polypeptide co-modified naringin nanoliposomes show no obvious cytotoxicity, on the premise that the safety of the preparation is fully guaranteed, the naringin nanoliposomes show a good effect of promoting proliferation of the dental pulp stem cells and have an excellent capability of promoting osteogenic differentiation of the dental pulp stem cells, so that the naringin nanoliposomes have wide related pharmaceutical preparation research and development and clinical application prospects.

Description

technical field [0001] The invention belongs to the technical field of pharmaceutical preparations, and in particular relates to a polypeptide co-modified naringin nanoliposome for promoting the proliferation and osteogenic differentiation of dental pulp stem cells, its preparation method and application. Background technique [0002] As a national treasure of our country, traditional Chinese medicine has obvious and definite curative effects in the treatment of various diseases. The traditional Chinese medicine Rhizoma Rhizoma Rhizoma Drynariae belongs to the kidney and liver meridian. It has the functions of nourishing the kidney and strengthening the bones, and relieving pain from injuries. It is one of the commonly used traditional Chinese medicines in the Department of Orthopedics and Traumatology. It is rich in active ingredients such as flavonoids, alkaloids, and phenols, among which naringin is one of its main active ingredients. Studies have found that naringin has...

Claims

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Application Information

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IPC IPC(8): A61K47/69A61K31/7048A61K47/62A61K47/65A61P19/02A61P19/08
CPCA61K31/7048A61K47/65A61K47/62A61K47/6911A61P19/08A61P19/02
Inventor 张栋梁初晓阳杨锴
Owner BEIJING STOMATOLOGY HOSPITAL CAPITAL MEDICAL UNIV
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