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Applications of pH-responsive nanometer material in preparation of anti-bone resorption drugs with effects of osteoporosis prevention and treatment

A nanomaterial, osteoporosis technology, applied in the field of biomedicine, to shorten the life cycle, prevent and treat osteoporosis, and promote the effect of new bone formation and vascularization

Inactive Publication Date: 2018-02-23
窦策
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its inhibition of osteoclasts is non-selective, and both POC and mature OC are non-selectively inhibited

Method used

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  • Applications of pH-responsive nanometer material in preparation of anti-bone resorption drugs with effects of osteoporosis prevention and treatment
  • Applications of pH-responsive nanometer material in preparation of anti-bone resorption drugs with effects of osteoporosis prevention and treatment
  • Applications of pH-responsive nanometer material in preparation of anti-bone resorption drugs with effects of osteoporosis prevention and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. High-concentration pH-responsive nanomaterials inhibit the activity of mature osteoclasts

[0034] BMMs cells were divided into 1×10 3 cells / well were inoculated in a 96-well plate, and the medium was discarded after the cells were overgrown. ), the pH-responsive nano The materials were added to each well plate containing fresh medium, while RANKL (100 ng / mL) and M-CSF (50 ng / mL) were used to induce BMMS cells to differentiate into osteoclasts. Cultured for 24h and 72h, respectively, to observe the effect of pH-responsive nanomaterials RNAKL-induced proliferation of BMMS cells. At the observation point, the cell activity was detected by the CCK-8 method, the number of living cells was detected by the flow cytometer, and the cell-nanoparticle interaction was observed by the transmission electron microscope. The result is as figure 1 As shown in B, C, D and E, the abscissa is the dose of pH-responsive nanomaterials at different concentrations, and the ordin...

Embodiment 2

[0035] Example 2, pH-responsive nanomaterials early (within 72h) promote RANKL-induced osteoclast cell generation

[0036] Grouping: Group 1, no RANKL induction, with pH-responsive nanomaterials group; Group 2, RANKL (100ng / mL) induction, 1μg / mL, 10μg / mL, 100μg / ml pH-responsive nanomaterials experimental group; Group 3, RANKL (100ng / mL) induction, 1μg / mL, 10μg / mL, 100μg / mL trivalent ion experimental group.

[0037] BMMS cells were divided into 1 × 10 3 cells / well were inoculated in a 96-well plate, and the medium was discarded after the cells were overgrown. ), the pH-responsive nanomaterials were added to each well plate containing fresh medium, while RANKL (100ng / mL) and M-CSF (50ng / mL) were used to induce BMMs cells to differentiate into osteoclasts. Tartrate-resistant acid phosphatase (TRAP) staining was performed 72 hours after induction. After discarding the medium, wash with PBS 3 times, 3 min each time. Fix with 4% paraformaldehyde for 20 minutes, wash with PBS 3 t...

Embodiment 3

[0038] Immunofluorescent staining of FAK (Actin Cytoskeleton and Focal Adhesion Staining) was performed after induction for 72 hours. After the cells were taken out, the medium was discarded, washed twice with 1×PBS, fixed with 4% paraformaldehyde at room temperature for 20 min, and washed twice with 1×PBS. Permeate the cells with 0.1% Triton X-100 for 5 minutes, and wash twice with 1×PBS. Blocking buffer (1%BSAin 1×PBS) fixed for 30min. The primary antibody (Anti-Vinculin) was diluted to the working concentration (1:300) in Blocking buffer, and the cells were incubated at room temperature for 1 h. Wash with 1×wash buffer (0.05% Tween-20in 1×PBS) three times, 5-10min each time. The secondary antibody (Alexa Fluor 488Goat Anti-Mouse IgG (H+L) Antibody, Invitrogen) was diluted to the working concentration (1:500), and TRITCconjugated Phalloidin (1:500) was added at the same time. Incubate together for 1 h at room temperature. Wash with 1×wash buffer three times, 5-10min each...

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Abstract

The invention relates to applications of a pH-responsive nanometer material in preparation of anti-bone resorption drugs with effects of osteoporosis prevention and treatment. According to the presentinvention, by using the pH-responsive nanometer material, mature osteoclasts are inhibited in a targeted manner while the osteoclast precursor is retained, wherein the retained osteoclast precursor cells can promote the neovascularization function, such that the effective anti-bone resorption drug with effects of osteoporosis prevention and treatment can be formed, has the effect superior to thefirst-line anti-bone resorption drug diphosphate, and can provide important significance for osteoporosis treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of pH-responsive nanometer materials in the preparation of osteoporosis anti-bone resorption drugs. Background technique [0002] Osteoporosis (osteoporosis, OP) is caused by a variety of reasons, the amount of bone tissue per unit volume is reduced, and the bone density is reduced as a systemic, progressive, metabolic bone disease. OP is divided into primary and secondary, and primary OP is further divided into type I postmenopausal OP and type II senile OP. In all OP patients, decreased bone tissue volume per unit volume (BV / TV), decreased bone mineral density (BMD), and destroyed bone microarchitecture. At present, there are about 200 million OP patients in the world, and there is one OP patient in every three women over the age of 50. In an aging society, OP has become an important factor that threatens the health of the elderly and reduces the quality...

Claims

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Application Information

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IPC IPC(8): A61K33/44A61K31/722A61P19/10
CPCA61K31/722A61K33/44
Inventor 窦策董世武许建中
Owner 窦策
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