CN104017769A - Application of ruthenium complex modified nano selenium in promotion of mesenchymal stem cell in-vitro osteoblast differentiation - Google Patents
Application of ruthenium complex modified nano selenium in promotion of mesenchymal stem cell in-vitro osteoblast differentiation Download PDFInfo
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Abstract
The invention belongs to the technical field of promotion of mesenchymal stem cell in-vitro osteoblast differentiation, and discloses an application of ruthenium complex modified nano selenium in promotion of mesenchymal stem cell in-vitro osteoblast differentiation. The invention provides the application of the ruthenium complex modified nano selenium in promotion of mesenchymal stem cell in-vitro osteoblast differentiation. The nano selenium can promote the mesenchymal stem cell in-vitro osteoblast differentiation of human umbilical cord, the differentiation balance is exsited between the mesenchymal stem cell ossification differentiation and esterification differentiation, so that the nano selenium has the effect for inhibiting esterification differentiation of mesenchymal stem cells, and can effectively labeling the mesenchymal stem cell. According to the invention, a ruthenium complex is used for modifying nano selenium, other auxiliary reagents are required for adding during a preparation process, the products system is simple, and the product can be directly preserved and used. The employed modification agent can greatly reduce the cytotoxicity of the ruthenium complex modified nano selenium, usage amount of the medicine for cells can be increased, external discharge is reduced, and the medicine in the cell can keep as a high level.
Description
Technical field
The invention belongs to and promote mescenchymal stem cell Osteoblast Differentiation technical field, particularly a kind of ruthenium complexe decorated nanometer selenium is in the application promoting in mescenchymal stem cell Osteoblast Differentiation.
Background technology
Along with human longevity prolongation and automobile are ridden instead of walk, the osteopathias such as osteoporosis, osteoporotic fracture have become serious major global public health problem.The generation of osteopathia is bone forming and the unbalanced performance of bone resorption in bone metabolism.The roughly process that bone is rebuild is: bone resorption → bone forming → bone is static → and bone resorption, such one moves in circles.Scleroblast (Osteoblast, OB) and osteoclast (Osteoclast, OC) are two kinds of main cells that maintain bone form and size during bone is rebuild.OB derives from mescenchymal stem cell, and energy promoting bone growing starts bone forming thereby secrete the cytokines such as a large amount of osso-albumins, ground substance of bone, and these factors get up to control propagation, maturation and the activation of OC by OC coupling simultaneously.Promoting mescenchymal stem cell osteoblast differentiation, realize bone forming regeneration, is prevent or treat a treatment machine meeting that reduces the osteopathia causing because of bone content, has attracted special concern in bone tissue engineer.
Umbilical cord mesenchymal stem cells separates and obtains from neonatal umbilical cord, and source is abundant, and the convenience of drawing materials, simple is without any side effects to newborn infant and puerpera.Umbilical cord immunity system is in the primitive stage, and immunogenicity is low, has reduced the graft-rejection after cell therapy.In addition, umbilical cord derived stem cell is more original, and proliferation and differentiation ability is strong, can be used for the treatment of bone tissue engineer or hemopoietic system, in the clinical application of mescenchymal stem cell, has vast potential for future development.The bone forming agent that is applied to clinically at present promoting bone growing mainly comprises two classes, fluorochemical and Rat parathyroid hormone 1-34.But though the toxicity of fluorochemical is little, long-term a large amount of uses can cause osteosclerosis, bone strength declines; The low dose of intermittently administration of Rat parathyroid hormone 1-34 is can promoting bone growing, but heavy dose continues medication and suppresses osteoblast activity.These problems are restricted its clinical application.
In recent years, seeking efficient, nontoxicly, have no side effect, have the medicine of the promoting bone growing of biocompatibility, is that stem cell is applied to the large crucial of bone tissue engineer treatment osteopathia.Wherein Nano medication is based on its high-performance: small-size effect, can realize efficient Cell uptake; Bigger serface, can the multiple functional groups of carrier band or active centre; Be convenient to biological degradation or absorption etc., Nano medication is paid close attention to widely and is paid much attention at field of medicaments.
Selenium (Se) is one of trace element of needed by human, it have anti-oxidant, delay senility and anti-cancer, anticancer effect, selenium nano particles (SeNPs) toxicity is low, and aspect biological activity, has more advantage.So far find no and close nanometer selenium for promoting the report of mescenchymal stem cell Osteoblast Differentiation.
Summary of the invention
In order to overcome the shortcoming and deficiency of above-mentioned prior art, primary and foremost purpose of the present invention is to provide a kind of ruthenium complexe decorated nanometer selenium in the application promoting in mescenchymal stem cell Osteoblast Differentiation.
Object of the present invention realizes by following proposal:
Ruthenium complexe decorated nanometer selenium is in the application promoting in mescenchymal stem cell Osteoblast Differentiation.
Described ruthenium complexe decorated nanometer selenium is activeconstituents, has the effect that promotes mescenchymal stem cell Osteoblast Differentiation.
Described ruthenium complexe decorated nanometer selenium refers to the nanometer selenium that ruthenium complexe-X modifies, and wherein, X is citric acid, aspartic acid, L-glutamic acid, vitamin A acid or vitamins C, and ruthenium complexe is bipyridyl ruthenium title complex or phenanthroline ruthenium complexe.
Described ruthenium complexe decorated nanometer selenium (X-Ru@Se) preferably prepares by the following method:
Get Na
2seO
3solution, bipyridyl ruthenium title complex or phenanthroline ruthenium complexe solution and water mix, and after the lower X of dropping of stirring solution, continue to stir, and obtain red nano colloidal sol, room temperature placement, and centrifugal, washing, dry, obtains ruthenium complexe decorated nanometer selenium.
Described Na
2seO
3strength of solution is 0.1～1.0mol/L.
Described bipyridyl ruthenium title complex or phenanthroline ruthenium complexe strength of solution are 0.02～0.2mmol/L.
Described X strength of solution is 0.1～1.0mol/L.
Preferably, Na used
2seO
3the mol ratio of solution, bipyridyl ruthenium title complex or phenanthroline ruthenium complexe solution and water is 1:0.2:10.
X solution used and Na
2seO
3the mol ratio of solution is 0.1:1～2:1.
Preferably, described continuation is stirred and is referred to stir 2～5h.
Preferably, while dripping X solution, keeping system pH is 9～11, preferably regulates by hydrochloric acid, more preferably regulates by 0.1mol/L hydrochloric acid.
Preferably, the time that described room temperature is placed is 24h.
Preferably, the red nano colloidal sol preparing can be in 4 DEG C of storages.
Preferably in preparation process, water used is pure water or the ultrapure water of being prepared by Milli-Q system.
In above-mentioned preparation process, by regulating X liquor capacity to regulate the particle diameter of made nanometer selenium.
The present invention, with respect to prior art, has following advantage and beneficial effect:
(1) the invention provides ruthenium complexe decorated nanometer selenium in the application promoting in mescenchymal stem cell Osteoblast Differentiation.Nanometer selenium of the present invention can not only promote the effect of human umbilical cord mesenchymal stem cells Osteoblast Differentiation, because the Osteoblast Differentiation of mescenchymal stem cell exists differentiation balance with becoming ester differentiation, therefore nanometer selenium of the present invention has the effect that mescenchymal stem cell becomes fat to break up that suppresses simultaneously, and can significant notation mescenchymal stem cell.
(2) the ruthenium complexe decorated nanometer selenium that prepared by the present invention, preparation process is simple without other auxiliary reagent of interpolation, product system, and product can directly be preserved and use.
(3) modifier that the present invention adopts has greatly reduced the cytotoxicity of ruthenium complexe decorated nanometer selenium, and can increase the ingestion of medicines amount of cell, reduces outer row, thereby ensures that in cell, medicine maintains higher level.
Brief description of the drawings
Fig. 1 is the perspective Electronic Speculum figure of the nanometer selenium of bipyridyl ruthenium title complex-citric acid modification.
Fig. 2 is the perspective Electronic Speculum figure of the nanometer selenium of citric acid modification.
Fig. 3 is that ruthenium complexe decorated nanometer selenium promotes stem cell Osteoblast Differentiation colored graph.
Fig. 4 is that ruthenium complexe decorated nanometer selenium inhibition stem cell becomes fat differential stain figure.
Fig. 5 is that ruthenium complexe decorated nanometer selenium activates BMP signal path, promotes the immunofluorescence image that p-Smad1/5 expresses.
Fig. 6 is the laser co-focusing image of ruthenium complexe decorated nanometer selenium significant notation mescenchymal stem cell.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited to this.
As long as the raw materials used purity of the present invention reaches analytical pure above, source all can be buied from market.Selected cell be the 4th generation human umbilical cord mesenchymal stem cells (cell separation from ewborn infant umbilical cord obtains, and ewborn infant umbilical cord is provided by No.1 Hospital Attached to Jinan Univ., takes from the pregnant baby of healthy puerpera.)
Embodiment 1: the preparation of the nanometer selenium (Cit-Ru@Se) of bipyridyl ruthenium title complex-citric acid modification
[Ru (bpy)
2(dhipH
3)] (ClO
4)
2preparation: cis-[Ru (bpy)
2cl
2] 2H
2o (Guangzhou eastward fight Hua Bo Instrument Ltd.) (0.156g, 0.3mmol) and 3,4-dihydroxyl-imidazo [4,5-f] [1,10]-phenanthroline (dhipH
3) (Guangzhou eastward fight Hua Bo Instrument Ltd.) (0.098g, 0.3mmol) be dissolved in 10mL ethylene glycol, the lower 120 DEG C of heating 6h of argon shield.Solution gradually becomes shiny red.Be chilled to after room temperature the saturated NaClO of limit agitation and dropping
4, produce immediately a large amount of red precipitates.Stir leave standstill after by solution suction filtration, crude product through wash dry after, separate with neutral alumina (200 order) post.Elutriant is the mixed solvent of V (toluene): V (acetonitrile)=1:4, collects main red stripes component, and pressure reducing and steaming solvent obtains shiny red [Ru (bpy)
2(dhipH
3)] (ClO
4)
2crystal.
[Ru (phen)
2(dhipH
3)] (ClO
4)
2preparation method the same, with [Ru (phen)
2(dhipH
3)] (ClO
4)
2(Guangzhou eastward fight Hua Bo Instrument Ltd.) (0.17g, 0.3mmol) replaces cis-[Ru (bpy)
2cl
2] 2H
2o.
(1) by [Ru (bpy)
2(dhipH
3)] (ClO
4)
2[Ru (phen)
2(dhipH
3)] (ClO
4)
2be dissolved in respectively the storing solution that is mixed with 0.2mmol/L in pure water, and be stored in 4 DEG C for subsequent use;
(2) get 2.9410g Trisodium Citrate and be dissolved in 10mL pure water and be mixed with the solution of 1mol/L, and be stored in 4 DEG C for subsequent use;
(3) by 1.7294g Na
2seO
3be dissolved in the solution that 10mL pure water is mixed with 1mol/L, and be stored in 4 DEG C for subsequent use;
(4) get respectively 1mL Na
2seO
3solution, 2mL[Ru (bpy)
2(dhipH
3)] (ClO
4)
2storing solution and 10mL ultrapure water mix, and obtain solution A; 1mL1mol/L sodium citrate solution is added drop-wise in solution A under constantly stirring, and in process, keeping pH value is 9～11; Continue to stir 2～5h, obtain burgundy Nano sol; Observe by TEANAI-10 type transmission electron microscope (TEM), as shown in Figure 1.Obtain the dispersion system of nanoparticle, its particle diameter is at 100～120nm.At room temperature stable existence of this nanoparticle, easily preserves; Centrifugal, washing, dry, obtain the nanometer selenium (Cit-Ru@Se) of bipyridyl ruthenium title complex-citric acid modification.
(5) get 1mL Na
2seO
3solution, drips 2mL0.1mol/L sodium citrate solution, and regulating its pH value is 9～11.Under constantly stirring, the sodium borohydride solution of 0.5mL0.1mol/L is added drop-wise in above-mentioned solution, continue to stir 4～6h, obtain bolarious citric acid modification nanometer selenium (Cit-Ru@Se) colloidal sol.Its pattern is observed by TEANAI-10 type transmission electron microscope (TEM), as shown in Figure 2.Obtain the dispersion system of nanoparticle, its particle diameter is at 70～95nm.At room temperature stable existence of this nanoparticle, easily preserves.
(6) nanometer selenium of the nanometer selenium that bipyridyl ruthenium title complex-aspartic acid is modified, the nanometer selenium that bipyridyl ruthenium title complex-L-glutamic acid is modified, the modification of bipyridyl ruthenium title complex-vitamin A acid and the same step of nanometer selenium preparation method (4) that bipyridyl ruthenium title complex-vitamins C is modified, just change aspartic acid solution, glutamic acid solution, vitamin A acid solution or the vitamin c solution for 1mol/L into 1mol/L sodium citrate solution.
(7) the same step of the preparation method of the nanometer selenium of phenanthroline ruthenium complexe-citric acid modification (4) is only [the Ru (bpy) 0.2mmol/L
2(dhipH
3)] (ClO
4)
2storing solution changes 0.2mmol/L[Ru (phen) into
2(dhipH
3)] (ClO
4)
2storing solution.
Embodiment 2: the experiment of the external promotion stem cell Osteoblast Differentiation of nanometer selenium (Cit-Ru@Se) of bipyridyl ruthenium title complex-citric acid modification
The separation and Culture of human umbilical cord mesenchymal stem cells (HUMSCs): according to hospital's regulation and through family members' agreement, get normal foot monthly output healthy babies umbilical cord tissue from operating table, be soaked in DMEM.In Bechtop, take out umbilical cord, fully wash blood residual on umbilical cord with D-Hanks ' balance liquid, remove after umbilical vein, Umbilical artery and umbilical cord adventitia, peel off Wharton glue, and shredded as about 1mm
3tissue block.Tissue block is moved in collagenase II (1g/L), add a little DMEM, in 37 DEG C of isothermal vibration instrument, digest, until Wharton glue all digests.Celliferous Digestive system is sucked to aseptic centrifuge tube, add the DMEM of 30 times of volumes repeatedly to blow and beat dilution, after the centrifugal 10min of 1500r/min, abandon supernatant liquor, with the DMEM/F12 substratum piping and druming dilution containing 15%FBS, cell suspension is uniformly distributed, be transferred in common culturing bottle, at 37 DEG C, 5%CO
2in incubator, cultivate.After cell attachment, within the 3rd day, carry out half amount and change liquid, discard not attached cell, within later every 3～4 days, change liquid 1 time.Under inverted microscope, observe, cell reaches after 90% fusion, with the trysinization of 1.25g/L, goes down to posterity in the ratio of 1:2～1:3, continuous amplification cultivation.
Human umbilical cord mesenchymal stem cells (HUMSCs, 5 × 10
5the every hole of cell) be inoculated in respectively in 24 hole tissue culturing plates, be placed in 5%CO
2, 37 DEG C of incubators are cultivated 3 days.Basic medium is removed, change into the induced liquid that contains osteogenic induction composition (osteoinductive agent OS, DMEM (H)+10%FBS+ is dual anti-+ 10mmol/L sodium β-glycerophosphate+0.1 μ mol/L dexamethasone+50mg/L Vc), and the bipyridyl ruthenium complex decorating nanometer selenium (ultimate density is 5 μ g/mL) or the NaF (ultimate density is 1mmol/L) that add respectively embodiment 1 to prepare, cultivate 7 and 14 days.The formation of mineralising joint knot is dyeed by sodium alizarinsulfonate (Alizarin Red S).Absorb substratum, rinse after cell 2 times with the PBS of precooling, fix 10～15min with 100% ethanol.PBS rinses after 2 drying at room temperature, adds Alizarin red staining reagent, and after the colour developing of 30min staining reagent, flowing water is taken pictures after rinsing and being dried.And dissolve, extract the mineralising tubercle of intracellular dyeing with 20% methyl alcohol and 10% acetic acid, detect its OD value at 450nm place use NanoDrop spectrophotometry.
Mineralising promotion rate=[OD
sample-OD
blank]/[OD
control-OD
blank] × 100%.
The HUMSCs that osteogenic induction (OS) is processed is used as Normal group, and 1mmol/mL NaF is as positive control.Through the osteogenic induction of 14 days, the mineralization degree of HUMSCs was assessed by Alizarin red staining.With respect to Normal group, the nanometer selenium (Cit@Se) of the nanometer selenium of bipyridyl ruthenium complex decorating of the present invention (Cit-Ru@Se), citric acid modification, the mineralization degree of NaF all have larger reinforced effects.Wherein, as shown in Figure 3, Cit-Ru@Se of the present invention has strengthened approximately 28.34%, and 15.68% of 21.9% and the NaF enhancing strengthening with respect to Cit@Se has more obvious effect.It is good fat-soluble and water-soluble that experimental result shows that the nanometer selenium of bipyridyl ruthenium complex decorating of the present invention has, and easily absorbed internalization by mescenchymal stem cell.In test, mescenchymal stem cell Osteoblast Differentiation is had to good promoter action in vitro, better than the curative effect of the nanometer selenium of NaF and citric acid modification, show good using value, be applicable to being prepared into promoting bone growing medicine.
Embodiment 3: nanometer selenium (Cit-Ru@Se) the vitro inhibition stem cell of bipyridyl ruthenium title complex-citric acid modification becomes the experiment of fat differentiation
Human umbilical cord mesenchymal stem cells (every hole 5 × 10
5cell) be inoculated in 24 hole tissue culturing plates, be placed in 5%CO
2, 37 DEG C of incubators are cultivated 3 days.Basic medium is removed, change into the induced liquid that contains into fat induction composition (fat inductor AS, DMEM (L)+10%FBS+10 μ mol/L dexamethasone+200 μ mol/L indomethacin+0.5mmol/L3-isobutyl--1-methyl xanthines (IBMX)+10mg/L Regular Insulin), and the bipyridyl ruthenium complex decorating nanometer selenium (ultimate density is 5 μ g/mL) or the NaF (ultimate density is 1mmol/L) that add respectively embodiment 1 to prepare, induce 9 and 18 days.In substratum, adding ruthenium complexe decorated nanometer selenium of the present invention is experimental group, the positive control group of NaF.Fat granule in the adipocyte of differentiation of stem cells is evaluated by oil red O stain method.Absorb nutrient solution, the PBS of precooling rinses after cell 2 times, fixes 10～15min with 4% poly formic acid.PBS rinses after 2 drying at room temperature, adds oil red staining reagent, and after the colour developing of 15min staining reagent, after soft flushing is dry, Microscopic observation is taken pictures.
Become fat differentiation inhibiting rate=[OD
sample-OD
blank]/[OD
control-OD
blank] × 100%.
The HUMSCs that becomes fat induction (AS) to process is used as Normal group, and 1mmol/mL NaF is as positive control.Through the osteogenic induction of 18 days, the one-tenth fat situation of HUMSCs was assessed by oil red O stain.With respect to normal experimental group, the nanometer selenium inhibiting rate that ruthenium complexe of the present invention is modified obviously improves.Wherein, as shown in Figure 4, the one-tenth fat of the nanometer selenium (Cit-Ru@Se) of bipyridyl ruthenium title complex-citric acid modification, the nanometer selenium (Cit@Se) of citric acid modification, NaF breaks up the inhibiting rate approximately 35.81%, 27.53%, 14.90% being subject to.Experimental result shows that the nanometer selenium (X-Ru@Se) of ruthenium complexe modification of the present invention becomes fat differentiation to have very strong restraining effect to mescenchymal stem cell in testing in vitro.Derived from Mesenchymal Stem Cells is between scleroblast and adipocyte, to exist plasticity-significantly, and both conversions are mutual.The nanometer selenium (X-Ru@Se) of bipyridyl ruthenium complex decorating of the present invention is broken up and increases its Osteoblast Differentiation by suppressing mescenchymal stem cell lipoblast, is to prevent or treat a potential treatment machine meeting that reduces the osteopathia causing because of bone content.
Embodiment 4: the impact that the nanometer selenium (Cit-Ru@Se) of bipyridyl ruthenium title complex-citric acid modification is expressed BMP signal path key protein p-Smad1/5
Smad1/5 albumen is the key protein that BMP signal path relates to.In the time that BMP signal path is activated, Smad1/5 albumen carries out phosphorylation, generate p-Smad1/5 albumen, p-Smad1/5 albumen and Smad4 protein binding form the expression that community enters karyon regulatory Osteoblast Differentiation genes involved, thereby promote stem cell Osteoblast Differentiation.
Human umbilical cord mesenchymal stem cells is taking density as 1 × 10
7individual cell is inoculated into focus in culture dish and cultivates 3 days at 37 DEG C.Then substratum is changed, and add respectively 2mL bipyridyl ruthenium complex decorating of the present invention nanometer selenium, Noggin (final concentration is 10 μ g/mL) (Noggin, noggin,---BMP signal pathway inhibitor.Purchased from Sigma) osteogenic induction substratum.Hatch after 4 hours for 37 DEG C, PBS cleans 3 times for cell, with 4% paraformaldehyde fixed cell 30min, then contains penetrating 10 minutes of the PBS of 0.25%Triton X-100.Add subsequently pSmad1/5 4 DEG C of overnight incubation, then at room temperature hatch 1 hour with two antibody of FITC mark.Finally, cell is dyeed with DAPI.Laser co-focusing picture is taken and is obtained with Zeiss Laser Scanning Confocal Microscope.
As can be seen from Figure 5, the green fluorescence intensity maximum of p-Smad1/5 albumen in the lower mescenchymal stem cell of the nanometer selenium of bipyridyl ruthenium complex decorating of the present invention (Cit-Ru@Se) effect.The raising greatly that in the lower mescenchymal stem cell of nanometer selenium (X-Ru@Se) effect that ruthenium complexe is modified, the expression level of p-Smad1/5 albumen is processed than positive controls NaF.This illustrates that bipyridyl ruthenium complex decorating nanometer selenium provided by the invention can serve as the medicine of promoting bone growing.In addition, the result of Fig. 5 also shows, in mescenchymal stem cell Osteoblast Differentiation process, thereby bipyridyl ruthenium complex decorating nanometer selenium promotes stem cell Osteoblast Differentiation by activating BMP signal path.
Embodiment 5: the cell imaging of nanometer selenium (Cit-Ru@Se) the significant notation mescenchymal stem cell of bipyridyl ruthenium title complex-citric acid modification
Human umbilical cord mesenchymal stem cells is with 1 × 10
4be inoculated in laser co-focusing ware, be placed in 37 DEG C, 5%CO
2incubator is cultivated after 24h, adds the nanometer selenium (X-Ru@Se) of bipyridyl ruthenium complex decorating in the burnt ware of each copolymerization, and ultimate density is 10 μ g/mL.Add after the ruthenium complexe decorated nanometer selenium that embodiment 1 prepares, cell is put back to incubator and is continued to cultivate after 12h, with PBS washed cell 3 times, removes the nanoparticle that is not absorbed internalization.Again cell is put back to incubator and continued to cultivate 12h, 48h.Substratum in burnt copolymerization ware is changed into the basic medium of serum-free, take the cell image of the mescenchymal stem cell that obtains ruthenium complexe decorated nanometer selenium mark with Zeiss Laser Scanning Confocal Microscope.
Fig. 6 demonstration adds after mescenchymal stem cell 12h, the 48h of Cit-Ru@Se mark, and the variation of its fluorescence intensity is little, illustrates that the nanometer selenium (X-Ru@Se) of bipyridyl ruthenium complex decorating of the present invention can stable labelling mescenchymal stem cell.By in conjunction with examples of implementation noted earlier, illustrate that ruthenium complexe decorated nanometer selenium is that a biocompatibility is good, effectively the cell marking material of promoting bone growing.
Above-described embodiment is preferably embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and principle, substitutes, combination, simplify; all should be equivalent substitute mode, within being included in protection scope of the present invention.
Claims (6)
1. ruthenium complexe decorated nanometer selenium is in the application promoting in mescenchymal stem cell Osteoblast Differentiation.
2. ruthenium complexe decorated nanometer selenium according to claim 1 is in the application promoting in mescenchymal stem cell Osteoblast Differentiation, it is characterized in that: described ruthenium complexe decorated nanometer selenium refers to the nanometer selenium that ruthenium complexe-X modifies, wherein, X is citric acid, aspartic acid, L-glutamic acid, vitamin A acid or vitamins C, and ruthenium complexe is bipyridyl ruthenium title complex or phenanthroline ruthenium complexe.
3. ruthenium complexe decorated nanometer selenium according to claim 1, in the application promoting in mescenchymal stem cell Osteoblast Differentiation, is characterized in that: described ruthenium complexe decorated nanometer selenium prepares by the following method: get Na
2seO
3solution, bipyridyl ruthenium title complex or phenanthroline ruthenium complexe solution and water mix, and after the lower X of dropping of stirring solution, continue to stir, and obtain red nano colloidal sol, room temperature placement, and centrifugal, washing, dry, obtains ruthenium complexe decorated nanometer selenium.
4. ruthenium complexe decorated nanometer selenium according to claim 3, in the application promoting in mescenchymal stem cell Osteoblast Differentiation, is characterized in that: described Na
2seO
3strength of solution is 0.1～1.0mol/L; Described bipyridyl ruthenium title complex or phenanthroline ruthenium complexe strength of solution are 0.02～0.2mmol/L; Described X strength of solution is 0.1～1.0mol/L.
5. ruthenium complexe decorated nanometer selenium according to claim 3, in the application promoting in mescenchymal stem cell Osteoblast Differentiation, is characterized in that: Na used
2seO
3the mol ratio of solution, bipyridyl ruthenium title complex or phenanthroline ruthenium complexe solution and water is 1:0.2:10; X solution used and Na
2seO
3the mol ratio of solution is 0.1:1～2:1.
6. ruthenium complexe decorated nanometer selenium according to claim 3, in the application promoting in mescenchymal stem cell Osteoblast Differentiation, is characterized in that: described continuation is stirred and referred to stir 2～5h; The time that described room temperature is placed is 24h.
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| CN104383543A (en) * | 2014-11-24 | 2015-03-04 | 暨南大学 | Application of chiral nano-selenium material supported siRNA in preparation of antitumor drug |
| CN104383543B (en) * | 2014-11-24 | 2017-11-21 | 暨南大学 | Chiral nanometer selenium material load siRNA is preparing the application of antineoplastic |
| CN111560243A (en) * | 2020-04-06 | 2020-08-21 | 华南师范大学 | Near-infrared controlled UCNPs carrier and preparation method and application thereof |
| CN111560243B (en) * | 2020-04-06 | 2023-01-03 | 华南师范大学 | Near-infrared controlled UCNPs carrier and preparation method and application thereof |
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