CN108079383A - A kind of antitumor chitosan-nanometer hydroxyapatite of photo-thermal-carbon quantum dot stent, its preparation method and application - Google Patents
A kind of antitumor chitosan-nanometer hydroxyapatite of photo-thermal-carbon quantum dot stent, its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of preparation methods of the antitumor chitosan nano hydroxyapatite carbon quantum dot stent of photo-thermal, comprise the following steps：(1) carbon quantum dot is added in into glacial acetic acid solution, is configured to carbon quantum dot solution；(2) nanometer hydroxyapatite is added in carbon quantum dot solution made from step (1), abundant sonic oscillation makes nanometer hydroxyapatite be uniformly dispersed, and adds chitosan and is uniformly mixed, obtains mixed solution；(3) mixed solution that step (2) obtains is freezed, obtains lyophilized products；(4) added in into the lyophilized products that step (3) obtains in alkaline solution and glacial acetic acid, cleaning are freezed to get the antitumor chitosan nano hydroxyapatite carbon quantum dot stent of the photo-thermal again.The antitumor chitosan nano hydroxyapatite carbon quantum dot stent of photo-thermal that the preparation method of the present invention is prepared has porous structure, under near infrared light, there is good photo-thermal effect, has good antitumous effect.
Description
Technical field
It can be used as photo-thermal the present invention relates to chitosan-nanometer hydroxyapatite-carbon quantum dot material more particularly to one kind to resist
The preparation method and application of chitosan-nanometer hydroxyapatite-carbon quantum dot stent of tumour bone renovating material.
Background technology
The incidence of bone tumour accounts for the 2-3% of all tumor incidences, although overall incidence is not high, bone tumour is often led
Limbs of patient deformity or even threat to life are caused, is the major disease for seriously endangering human health.With the basic research of bone tumour
With deepening continuously for clinical technology, the particularly combined treatment of the raising of surgical technic and chemicotherapy, make the treatment of bone tumour
Effect is obviously improved, and particularly with malignant bone tumor, survival rate is improved by past 10-20% to present within 5 years
50%.However, bone tumour is postoperative to be often accompanied by huge bone defect and higher local relapse, the rate of transform, it is clinical treatment
Problem.Therefore, there is an urgent need in combination with antitumor and Bone Defect Repari new therapeutic strategy.
Bone tissue engineer is the research hotspot of Bone Defect Repari, however has antitumor and Bone Defect Repari effect material simultaneously at present
It is rarely reported.Photo-thermal therapy is a kind of new method for treating tumour, has huge clinical development potentiality.Photo-thermal therapy passes through profit
Entered with the material with photothermal conversion efficiency near tumor tissues, it will under the irradiation of external light source (being usually near infrared light)
Luminous energy is converted into thermal energy, further kills a kind of therapy of tumour cell.Carbon quantum dot is a kind of preferable photo-thermal material
Material has higher photothermal conversion efficiency and good biocompatibility.Meanwhile carbon quantum dot manufacturing cost is low, preparation method
Simply, take short.However, the photo-thermal bone repairing support for the carbon containing quantum dot reported at present almost without.
The content of the invention
On the one hand, the present invention provides a kind of antitumor chitosan of photo-thermal-nanometer hydroxyl for overcome the deficiencies in the prior art
The preparation method of base apatite-carbon quantum dot stent, the antitumor chitosan-nanometer hydroxyapatite-carbon of photo-thermal being prepared
Quantum dot stent has porous structure, under near infrared light, there is good photo-thermal effect, while has good antitumor
Effect.
The technical solution adopted by the present invention is：A kind of antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot branch of photo-thermal
The preparation method of frame, comprises the following steps：
(1) carbon quantum dot is added in into glacial acetic acid solution, is configured to carbon quantum dot solution；
(2) nanometer hydroxyapatite is added in carbon quantum dot solution made from step (1), makes nanometer hydroxyapatite point
It dissipates uniformly, adds chitosan and be uniformly mixed, obtain mixed solution；
(3) mixed solution that step (2) obtains is freezed, obtains lyophilized products；
(4) into the lyophilized products that step (3) obtains add in alkaline solution in and glacial acetic acid, cleaning, again freeze to get
Antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot the stent of photo-thermal.
In the above-mentioned technical solutions, chitosan and hydroxyapatite have good biocompatibility, bioactivity and rush
Into the ability of Bone Defect Repari, and the bone reparing biological material that chitosan and hydroxyapatite are formed is respectively provided with well in vivo, outside
Bone Defect Repari effect；Further, due to the addition of carbon quantum dot so that chitosan-nanometer hydroxyapatite-carbon of the invention
Quantum dot stent possesses the performance that photo-thermal effect is generated under near infrared light, so that stent has antitumor activity.
The selection of chitosan and nanometer hydroxyapatite is that inventor is tested by substantial amounts of creativeness from numerous materials
What screening obtained, inventor, which has found that the two is used in combination, can effectively make up respective deficiency.Inventor has found under study for action
The simple stent brittleness that nanometer hydroxyapatite is used to be prepared is big, and the simple support intensity being prepared using chitosan is not
Foot, and chitosan and being used in combination for nanometer hydroxyapatite play an important role of synergy, can effectively reduce the brittleness of stent,
Support intensity is improved, so that the performance of antitumor bone repairing support is more excellent.
It is further improved as to above-mentioned technical proposal, in the step (1), the concentration of carbon quantum dot is 1-3mg/
mL.When carbon quantum dot concentration is higher than 3mg/mL, material preparation is difficult, and when it is less than 1mg/mL, then photo-thermal effect is bad.
It is further improved as to above-mentioned technical proposal, the step (2) is specially：Nanometer hydroxyapatite is added in
In carbon quantum dot solution made from step (1), abundant sonic oscillation makes nanometer hydroxyapatite be uniformly dispersed, and adds chitosan
It is uniformly mixed, obtains mixed solution；
It is further improved as to above-mentioned technical proposal, in the step (2), in mixed solution, the concentration of chitosan
For 1-5%w/v, the content of nanometer hydroxyapatite is 30-70wt%, and when the two concentration is too low, the stent of preparation cannot be into
Type when the concentration is too high, then exists and prepares the problem of difficult.Inventor is by largely testing discovery when the concentration of chitosan is
2-4%w/v, when the content of nanometer hydroxyapatite is 50-70wt%, antitumor bone repairing support obtained can preferably expire
The requirement of sufficient antibacterial bone repairing support.
It should be noted that if without special instruction, the calculation formula for the concentration unit %w/v being referred to herein is：(substance
Quality/solution volume) × 100%, wherein the unit of the volume of quality/solution of substance be g/mL；For example, shell herein
The concentration of glycan is obtained by (volume of the quality of chitosan/component A) × 100%.
It is further improved as to above-mentioned technical proposal, in the step (2), chitosan, nano hydroxyapatite lime stone
Weight ratio with carbon quantum dot is 10:15:3.At this point, the antitumor activity of antitumor bone repairing support obtained can be optimal.
Be further improved as to above-mentioned technical proposal, in the step (2), in mixed solution, carboxylic chitosan it is dense
It spends for 2%w/v, nanometer hydroxyapatite content is 60%.Brace aperture size prepared by the concentration is more suitable, branch obtained
Frame performance is more preferable.
It is further improved as to above-mentioned technical proposal, the step (3) is implemented by following steps：By mixed solution
It moves into container, after -80 DEG C of freeze overnights, moves in freeze dryer and freeze；Preferably, in the step (3), freeze-drying time 12
~30h, more preferably for 24 hours.Both it can ensure that mixed solution thoroughly freezed when for 24 hours, while manufactured porous structure is more stable, suitable
Preferably.
The container can be culture plate, such as 48 well culture plates.
It is further improved as to above-mentioned technical proposal, in the step (5), freeze-drying time is 12~30h, is preferably
24h.It can both ensure that stent thoroughly freezed for 24 hours, while save the time, the performance of obtained stent is more preferable.
It is further improved as to above-mentioned technical proposal, the alkaline solution is sodium hydroxide solution；Preferably, hydrogen-oxygen
Change the concentration of sodium solution as 0.5~1M, more preferably 0.5M.When concentration is 0.5M, glacial acetic acid can be preferably neutralized.
On the other hand, the present invention also provides the antitumor shells of photo-thermal that preparation method in accordance with the above is prepared to gather
Sugar-nanometer hydroxyapatite-carbon quantum dot stent.
Another aspect, the present invention also provides the antitumor chitosan-nanometer hydroxyapatite of photo-thermal in accordance with the above-
Application of the carbon quantum dot stent as antitumor bone renovating material.
Compared with the prior art, beneficial effects of the present invention are：
(1) antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent prepared by the present invention has porous structure,
It is distributed between main 20~80 μm of pore size.
(2) preparation method of antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of the invention adds in carbon
Quantum point process is simple, and stent is made to possess photo-thermal effect under near infrared light.
(3) antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent prepared by the present invention is trained altogether with tumour cell
It supports, after ten minutes, tumour cell relative activity about can be down to 35% or so, and material has good for 808nm near infrared lights
Antitumor activity.
Description of the drawings
Fig. 1 is the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot of photo-thermal that the embodiment of the present invention 1 is prepared
Stent outside drawing.
Fig. 2 is the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot of photo-thermal that the embodiment of the present invention 1 is prepared
The stereoscan photograph of stent.
Fig. 3 is the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot of photo-thermal that the embodiment of the present invention 1 is prepared
Photo-thermal effect figure of the stent under 808nm near infrared lights.
Fig. 4 is the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot of photo-thermal that the embodiment of the present invention 1 is prepared
The antitumor activity testing result figure of stent.
Specific embodiment
To better illustrate the object, technical solutions and advantages of the present invention, below in conjunction with specific embodiment to the present invention
It is described further.
Embodiment 1
With the glacial acetic acid solution that ultra-pure water configuration concentration is 2%, 30mg carbon quantum dots are added in, magnetic stirring apparatus is in room temperature item
It is stirred until homogeneous under part.It is gradually added into 300mg nanometer hydroxyapatites and 200mg chitosans is added to carbon quantum dot solution, it is fixed
Hold to 10mL, magnetic stirring apparatus is stirred under normal temperature condition to substantially uniformity.Above-mentioned mixed solution is added in 48 well culture plates,
It is put into freeze overnight in -80 DEG C of refrigerators.48 orifice plates are moved in freeze dryer and are freezed for 24 hours.Add in 0.5M NaOH solutions in and vinegar
Acid, ultra-pure water are cleaned repeatedly to neutrality, are put into -80 DEG C of refrigerators and are freezed, and are freezed using freeze dryer and are obtained stent afterwards for 24 hours, are
Antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot the stent of photo-thermal.
Obtained stent is immersed in 500uL PBS solutions, gives 808nm near infrared lights 1W/cm210min is irradiated, is surveyed
Warm pin measures PBS solution temperature change.With 5 × 104A osteosarcoma cell UMR-106 is co-cultured, and 808nm near-infrareds were given in 1 day
Light 1W/cm210min is irradiated, discards culture medium, adds in 10% CCK-8 solution, 2h measures absorbance, detection branch after 450nm
Frame is to the inhibitory action of osteosarcoma cell.Antitumor chitosan-the nanometer hydroxyapatite of photo-thermal that the present embodiment 1 is prepared-
Carbon quantum dot stent appearance is as shown in Figure 1.
Fig. 2 is the scanning electron microscope of antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent manufactured in the present embodiment
Figure, as shown in Figure 2, the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of photo-thermal prepared by the present embodiment 1 is more
Pore structure.
Fig. 3 is the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of photo-thermal that the present embodiment 1 is prepared
Photo-thermal effect under 808nm near infrared lights, from the figure 3, it may be seen that photo-thermal prepared by the present embodiment 1 is immersed in 500uL PBS
In, after near infrared light 10min, PBS can be warming up to 52.6 DEG C.
Fig. 4 is the cell toxicant of antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent prepared by the present embodiment 1
Property testing result, as shown in Figure 4, the present embodiment 1 prepare the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot of photo-thermal
Stent is compared with chitosan alone-nanometer hydroxyapatite stent, and after near infrared light, versus cell activity is
34.05%, material prepared has good antitumor activity.
Embodiment 2
With the glacial acetic acid solution that ultra-pure water configuration concentration is 2%, 20mg carbon quantum dots are added in, magnetic stirring apparatus is in room temperature item
It is stirred until homogeneous under part.It is gradually added into 300mg nanometer hydroxyapatites and 200mg chitosans is added to carbon quantum dot solution, it is fixed
Hold to 10mL, magnetic stirring apparatus is stirred under normal temperature condition to substantially uniformity.Above-mentioned mixed solution is added in 48 well culture plates,
It is put into freeze overnight in -80 DEG C of refrigerators.48 orifice plates are moved in freeze dryer and are freezed for 24 hours.Add in 0.5M NaOH solutions in and vinegar
Acid, ultra-pure water are cleaned repeatedly to neutrality, are put into -80 DEG C of refrigerators and are freezed, and are freezed using freeze dryer and are obtained stent afterwards for 24 hours, are
Antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot the stent of photo-thermal.
Photo-thermal manufactured in the present embodiment is immersed in 500uL PBS, and after near infrared light 10min, PBS can be warming up to
47.5℃.Antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot the stent of photo-thermal manufactured in the present embodiment gathers with simple shell
Sugar-nanometer hydroxyapatite stent is compared, and after near infrared light, versus cell activity is 40.17%, material prepared tool
There is good antitumor activity.
Embodiment 3
With the glacial acetic acid solution that ultra-pure water configuration concentration is 2%, 30mg carbon quantum dots are added in, magnetic stirring apparatus is in room temperature item
It is stirred until homogeneous under part.It is gradually added into 150mg nanometer hydroxyapatites and 100mg chitosans is added to carbon quantum dot solution, it is fixed
Hold to 10mL, magnetic stirring apparatus is stirred under normal temperature condition to substantially uniformity.Above-mentioned mixed solution is added in 48 well culture plates,
It is put into freeze overnight in -80 DEG C of refrigerators.48 orifice plates are moved in freeze dryer and are freezed for 24 hours.Add in 0.5M NaOH solutions in and vinegar
Acid, ultra-pure water are cleaned repeatedly to neutrality, are put into -80 DEG C of refrigerators and are freezed, and are freezed using freeze dryer and are obtained stent afterwards for 24 hours, are
Antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot the stent of photo-thermal.
Photo-thermal manufactured in the present embodiment is immersed in 500uL PBS, and after near infrared light 10min, PBS can be warming up to
53.1℃.Antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot the stent of photo-thermal manufactured in the present embodiment gathers with simple shell
Sugar-nanometer hydroxyapatite stent is compared, and after near infrared light, versus cell activity is 31.84%, material prepared tool
There is good antitumor activity.
Embodiment 4
With the glacial acetic acid solution that ultra-pure water configuration concentration is 2%, 30mg carbon quantum dots are added in, magnetic stirring apparatus is in room temperature item
It is stirred until homogeneous under part.It is gradually added into 450mg nanometer hydroxyapatites and 300mg chitosans is added to carbon quantum dot solution, it is fixed
Hold to 10mL, magnetic stirring apparatus is stirred under normal temperature condition to substantially uniformity.Above-mentioned mixed solution is added in 48 well culture plates,
It is put into freeze overnight in -80 DEG C of refrigerators.48 orifice plates are moved in freeze dryer and are freezed for 24 hours.Add in 0.5M NaOH solutions in and vinegar
Acid, ultra-pure water are cleaned repeatedly to neutrality, are put into -80 DEG C of refrigerators and are freezed, and are freezed using freeze dryer and are obtained stent afterwards for 24 hours, are
Antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot the stent of photo-thermal.
Photo-thermal manufactured in the present embodiment is immersed in 500uL PBS, and after near infrared light 10min, PBS can be warming up to
50.5℃.Antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot the stent of photo-thermal manufactured in the present embodiment gathers with simple shell
Sugar-nanometer hydroxyapatite stent is compared, and after near infrared light, versus cell activity is 37.29%, material prepared tool
There is good antitumor activity.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention and from the embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (10)
1. a kind of preparation method of the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of photo-thermal, which is characterized in that
Comprise the following steps：
(1) carbon quantum dot is added in into glacial acetic acid solution, is configured to carbon quantum dot solution；
(2) nanometer hydroxyapatite is added in carbon quantum dot solution made from step (1), nanometer hydroxyapatite is made to disperse
It is even, it adds chitosan and is uniformly mixed, obtain mixed solution；
(3) mixed solution that step (2) obtains is freezed, obtains lyophilized products；
(4) added in into the lyophilized products that step (3) obtains in alkaline solution and glacial acetic acid, cleaning are freezed to get described again
Antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot the stent of photo-thermal.
2. the preparation side of the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of photo-thermal according to claim 1
Method, which is characterized in that in the step (1), the concentration of carbon quantum dot solution is 1-3%w/v, is preferably 3%w/v.
3. the preparation side of the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of photo-thermal according to claim 1
Method, which is characterized in that in the step (2), in mixed solution, the concentration of chitosan is 1-5%w/v, nanometer hydroxyapatite
Content be 30-70wt%.
4. the preparation side of the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of photo-thermal according to claim 1
Method, which is characterized in that in the step (2), in mixed solution, the weight of chitosan, nano hydroxyapatite lime stone and carbon quantum dot
Amount is than being 10:15:3.
5. the preparation side of the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of photo-thermal according to claim 1
Method, which is characterized in that the step (3) is implemented by following steps：Mixed solution is moved into container, -80 DEG C of freeze overnights
Afterwards, move in freeze dryer and freeze.
6. the preparation side of the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of photo-thermal according to claim 1
Method, which is characterized in that in the step (3), freeze-drying time is 12~30h, preferably for 24 hours.
7. the preparation side of the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of photo-thermal according to claim 1
Method, which is characterized in that in the step (4), freeze-drying time is 12~30h, preferably for 24 hours.
8. the preparation side of the antitumor chitosan-nanometer hydroxyapatite-carbon quantum dot stent of photo-thermal according to claim 1
Method, which is characterized in that the alkaline solution is sodium hydroxide solution；
Preferably, the concentration of sodium hydroxide solution is 0.5M.
9. according to the antitumor chitosan of photo-thermal-nanometer hydroxyl that preparation method according to any one of claims 1 to 8 is prepared
Base apatite-carbon quantum dot stent.
10. the antitumor chitosan-nanometer hydroxyapatite of photo-thermal according to claim 9-carbon quantum dot stent is as anti-
The application of tumour bone renovating material.
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| CN111228490A (en) * | 2020-02-25 | 2020-06-05 | 南通大学 | Preparation method of calcium-phosphorus composite porous carbon nanofiber photothermal reagent |
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| CN114129777A (en) * | 2020-09-03 | 2022-03-04 | 天津大学 | Preparation and application of photoresponse composite nano coating |
| CN114129777B (en) * | 2020-09-03 | 2022-11-29 | 天津大学 | Preparation and application of photoresponse composite nano coating |
| CN114275751A (en) * | 2022-02-16 | 2022-04-05 | 湖南大学 | Preparation method of hexagonal macroporous hydroxyapatite, product and application thereof |
| CN115779082A (en) * | 2022-12-28 | 2023-03-14 | 石河子大学 | Multi-linked drug-loaded phospholene thermosensitive hydrogel osteosarcoma-resistant drug delivery system and preparation method thereof |
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