CN101244274A - Ferrite/bacille calmette-guerin vaccine/temperature sensing gel rubber integration material and preparation thereof - Google Patents
Ferrite/bacille calmette-guerin vaccine/temperature sensing gel rubber integration material and preparation thereof Download PDFInfo
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- CN101244274A CN101244274A CNA2008100154555A CN200810015455A CN101244274A CN 101244274 A CN101244274 A CN 101244274A CN A2008100154555 A CNA2008100154555 A CN A2008100154555A CN 200810015455 A CN200810015455 A CN 200810015455A CN 101244274 A CN101244274 A CN 101244274A
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- guerin vaccine
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Abstract
The invention discloses a ferrite/bcg (bacillus calmette-guerin)/ temperature sensitive gel composite material, comprising weight percentage 5% to 15% bcg, 5% to 15% tri-iron tetroxide, 70% to 90% chitosan/Beta-sodium glycerophosphate xerogel; which adopts the technical proposal that: firstly, nanometer tri-iron tetroxide is prepared by the coprecipitation method; secondly, chitosan is dissolved into a dilute hydrochloric acid; quantitative prepared tri-iron tetroxide and bcg are evenly dispersed; Beta-sodium glycerophosphate solution is added; and then the mixed liquid is turned into gel by water bath heating; thirdly, the gel is freeze dried or vacuum dried to get chitosan xerogel; fourthly, the xerogel is grinded and then sieved to get the corresponding composite material. The composite material has the advantages that with the attaching-wall targeting and sustained releasing function, as well as the action of outside magnetic field, the power of the composite material can attach to the bladder inner wall, rather than being discharged with urine, so that the stay in the bladder is prolonged, and the efficacy of bcg is reinforced.
Description
Technical field
The present invention relates to chitosan/sodium gel is that carrier, ferroso-ferric oxide are the Biocomposite material that targeting material and lyophilized bacillus calmette-guerin vaccine are formed, and belongs to material science and biomedical Biocomposite material technical field of intersecting.
Background technology
At present, 90%～95% bladder cancer is the transitional epithelial cell cancer clinically, and the ill initial stage shows as the shallow cancer of table more.The treatment of the shallow bladder cancer of his-and-hers watches mainly relies on transurethral resection or electricity to iron.Hospital carries out transurethral resection of bladder tumor (Transurethral resection of bladder tunor) TURBT operation, and this operation often has residual cancer cell, and tumor recurs easily, do not deal with as postoperative, relapse rate can reach 60%～90%.Intravesical perfusion bacillus calmette-guerin vaccine (BCG) is acknowledged as the best approach of prevention tumor of bladder postoperative recurrence at present.But in this method, the time of staying of BCG in bladder is 1-2 hour only, promptly is excreted with urine, had a strong impact on its curative effect, and repeatedly perfusion of postoperative, brings very big misery to patient.Therefore, prolong bacillus calmette-guerin vaccine in the intravesical time of staying, it is significant to strengthen its drug effect.At biomedical sector, magnetic carrier can be used for the transmission of medicine, and load has the magnetic-particle of slow releasing pharmaceutical medicine to be transported to the predetermined diseased region of human body to carry out sustained release by means of the guide effect of externally-applied magnetic field.Ferroso-ferric oxide has excellent biological compatibility and magnetic property, especially when ferriferrous oxide particles size during less than 25nm, has superparamagnetism, and this makes it become a kind of magnetic targeting locator material of extensive use.Chitosan-sodium glycerophosphate thermosensitive hydrogel system is the novel drug-supplying system that just is suggested in 2000, exists with collosol state under preservation condition, but just can realize the irreversible transformation of colloidal sol to gel in 37 ℃ of following short periods.Because this gel rubber system has good biocompatibility and degradable characteristic, therefore be subjected to extensive concern as a kind of pharmaceutical carrier gradually.
Summary of the invention
The objective of the invention is to overcome the deficiency that bacillus calmette-guerin vaccine fails to give full play to its curative effect, thereby provide a kind of can targeting adherent prolongation bacillus calmette-guerin vaccine in the intravesical time of staying, slowly discharge the ferrite/bacillus calmette-guerin vaccine/thermosensitive hydrogel composite material of bacillus calmette-guerin vaccine, a kind of preparation method of this composite material is provided simultaneously.
Ferrite/bacillus calmette-guerin vaccine of the present invention/thermosensitive hydrogel composite material comprises lyophilized bacillus calmette-guerin vaccine, ferroso-ferric oxide and chitosan/sodium xerogel, the percentage by weight of each component is: lyophilized bacillus calmette-guerin vaccine 5%-15%, ferroso-ferric oxide 5%-15%, chitosan/sodium xerogel 70%-90%.
The preparation method of above-mentioned ferrite/bacillus calmette-guerin vaccine/thermosensitive hydrogel composite material may further comprise the steps:
(1) utilize coprecipitation to prepare nano ferriferrous oxide:
The dose volume ratio is that 1.75: 1 and concentration are 0.1molL respectively
-1Fe
3+And Fe
2+Solution is at mechanical agitation and logical N
2Under the condition,, calculate according to the stereometer of mixed solution and to prepare gained Fe in theory two kinds of solution mix homogeneously
3O
4Gravimetric value, add this Fe
3O
4The surfactant polyethylene-6000 (PEG-6000) of theoretical weight value 0.1%-1%; Stir after 15 minutes to mixed solution and dripping 1molL
-1NaOH solution, until pH value of solution=9, obtain black precipitate, under 70 ℃ of water-baths, make the black precipitate ripening, left standstill 5 hours, and removed the supernatant, the particle that generates is washed repeatedly with distilled water and dehydrated alcohol, the surfactant of flush away particle surface and foreign ion, at 50 ℃ of following vacuum dryings, grinding is sieved, and obtains nano ferriferrous oxide granule;
(2) chitosan is dissolved in 0.1molL
-1In the hydrochloric acid, fully stir, be mixed with the chitosan settled solution of 15.5-22.5mg/ml, successively add the ferroso-ferric oxide and the lyophilized bacillus calmette-guerin vaccine of step (1) preparation, stirring or ultra-sonic dispersion make its mix homogeneously; With deionized water sodium is mixed with the solution of 0.7-1.0g/ml, then with sodium solution and chitosan solution according to 1: 9 mixed of volume ratio, mix the back and continue to stir 10-30 minute;
(3) step (2) gained mixed liquor is placed in the 30-40 ℃ of thermostat water bath, make it change gel into;
(4) gel with step (3) gained carries out lyophilization or vacuum drying, gets rid of moisture, obtains containing the chitosan xerogel of bacillus calmette-guerin vaccine and ferroso-ferric oxide;
(5) xerogel with step (4) gained grinds, crosses 100 orders and obtains ferrite/bacillus calmette-guerin vaccine/thermosensitive hydrogel composite material with top sieve.
The composite material of the present invention preparation has the adherent and slow-release function of targeting, because the adding of ferroso-ferric oxide, do not excrete with urine thereby make this composite material powder body be affixed on the bladder inwall under can the outside magnetic field effect, has prolonged it in time that intravesical stops.In addition,, make bacillus calmette-guerin vaccine in the slow degradation process of gel, constantly discharge, strengthened the drug effect of bacillus calmette-guerin vaccine owing to adopt chitosan/sodium gel systems as carrier.
The specific embodiment
Embodiment 1
1) the dose volume ratio is that 1.75: 1 and concentration are 0.1molL respectively
-1FeCl
3And FeSO
4Solution is at mechanical agitation and logical N
2Under the condition, with two kinds of solution mix homogeneously.Add 0.1% (the volume gained Fe after theory is calculated that is joined two kinds of solution with step 1
3O
4Quality be benchmark) surfactant polyethylene-6000 (PEG-6000).Stir after 15 minutes to mixed solution and dripping 1molL
-1NaOH solution, until pH value of solution=9, obtain black precipitate.Under 70 ℃ of water-baths, make its ripening.Left standstill 5 hours, and removed the supernatant, the particle that generates is washed the surfactant of flush away particle surface and foreign ion repeatedly with distilled water and dehydrated alcohol.At 50 ℃ of following vacuum dryings, ground 100 mesh sieves, obtain Fe
3O
4Granule.
2) be that 95% chitosan is dissolved in 4.5ml 0.1molL with the 90mg deacetylation
-1In the hydrochloric acid, fully stirred 4 hours, be mixed with the chitosan solution of 20mg/ml.Add the ferroso-ferric oxide that makes in 70mg (percentage by weight the is 10.3%) step 1), the method that adopts mechanical agitation to add ultra-sonic dispersion is uniformly dispersed it.Add 70mg (percentage by weight is 10.3%) bacillus calmette-guerin vaccine then, stirred 1 hour.450mg sodium (percentage by weight of chitosan and sodium is 79.4%) is dissolved in the 0.5ml deionized water, be mixed with the solution of 0.9g/ml, drop in the chitosan solution according to 1: 9 ratio of volume ratio then, stir while dripping, solution mixes the back to be continued to stir 20 minutes.
3) with step 2) the gained mixed liquor places in 30 ℃ of thermostat water baths, makes it change gel into.
4) gel with the step 3) gained carries out vacuum drying, gets rid of moisture, obtains containing the chitosan xerogel of bacillus calmette-guerin vaccine and ferroso-ferric oxide.
5) xerogel with the step 4) gained grinds, crosses 200 mesh sieves and obtains corresponding composite material.
After measured, the main performance of the ferroso-ferric oxide/bacillus calmette-guerin vaccine that makes/integrated powder body of thermosensitive hydrogel: 48 hours bacillus calmette-guerin vaccine release rates are about 80%, and ferriferrous oxide particles is uniformly dispersed in gel, and biocompatibility is better.
Embodiment 2
1) the dose volume ratio is that 1.75: 1 and concentration are 0.1molL respectively
-1FeCl
3And FeSO
4Solution is at mechanical agitation and logical N
2Under the condition, with two kinds of solution mix homogeneously.Add 0.5wt% (the volume gained Fe after theory is calculated that is joined two kinds of solution with step 1
3O
4Quality be benchmark) surfactant polyethylene-6000 (PEG-6000).Stir after 15 minutes to mixed solution and dripping 1molL
-1NaOH solution, until pH value of solution=9, obtain black precipitate.Under 70 ℃ of water-baths, make its ripening.Left standstill 5 hours, and removed the supernatant, the particle that generates is washed the surfactant of flush away particle surface and foreign ion repeatedly with distilled water and dehydrated alcohol.At 50 ℃ of following vacuum dryings, ground 100 mesh sieves, obtain Fe
3O
4Granule.
2) be that 95% chitosan is dissolved in 4.5ml 0.1molL with the 80mg deacetylation
-1In the hydrochloric acid, fully stirred 3 hours, be mixed with the chitosan solution of 17.8mg/ml.Add the ferroso-ferric oxide that makes in 60mg (percentage by weight the is 8.1%) step 1), the method that adopts mechanical agitation to add ultra-sonic dispersion is uniformly dispersed it.Add 100mg (percentage by weight is 13.5%) bacillus calmette-guerin vaccine then, stirred 1 hour.500mg sodium (percentage by weight of chitosan and sodium is 78.4%) is dissolved in the 0.5ml deionized water, be mixed with the solution of 1g/ml, drop in the chitosan solution according to 1: 9 ratio of volume ratio then, stir while dripping, solution mixes the back to be continued to stir 30 minutes.
3) with step 2) the gained mixed liquor places in 35 ℃ of thermostat water baths, makes it change gel into.
4) gel with the step 3) gained carries out lyophilization, gets rid of moisture, obtains containing the chitosan xerogel of bacillus calmette-guerin vaccine and ferroso-ferric oxide.
5) xerogel with the step 4) gained grinds, crosses 300 mesh sieves and obtains corresponding composite material.
After measured, the main performance of the ferroso-ferric oxide/bacillus calmette-guerin vaccine that makes/integrated powder body of thermosensitive hydrogel: 36 hours bacillus calmette-guerin vaccine release rates are about 80%, and ferriferrous oxide particles is uniformly dispersed in gel, and biocompatibility is better.
Embodiment 3
1) the dose volume ratio is that 1.75: 1 and concentration are 0.1molL respectively
-1FeCl
3And FeSO
4Solution is at mechanical agitation and logical N
2Under the condition, with two kinds of solution mix homogeneously.Add 0.7wt% (the volume gained Fe after theory is calculated that is joined two kinds of solution with step 1
3O
4Quality be benchmark) surfactant polyethylene-6000 (PEG-6000).Stir after 15 minutes to mixed solution and dripping 1molL
-1NaOH solution, until pH value of solution=9, obtain black precipitate.Under 70 ℃ of water-baths, make its ripening.Left standstill 5 hours, and removed the supernatant, the particle that generates is washed the surfactant of flush away particle surface and foreign ion repeatedly with distilled water and dehydrated alcohol.At 50 ℃ of following vacuum dryings, ground 100 mesh sieves, obtain Fe
3O
4Granule.
2) be that 95% chitosan is dissolved in 4.5ml 0.1molL with the 70mg deacetylation
-1In the hydrochloric acid, fully stirred 3 hours, be mixed with the chitosan solution of 15.5mg/ml.Add the ferroso-ferric oxide that makes in 90mg (percentage by weight the is 15%) step 1), the method that adopts mechanical agitation to add ultra-sonic dispersion is uniformly dispersed it.Add 90mg (percentage by weight is 15%) bacillus calmette-guerin vaccine then, stirred 1 hour.350mg sodium (percentage by weight of chitosan and sodium is 70%) is dissolved in the 0.5ml deionized water, be mixed with the solution of 0.7g/ml, drop in the chitosan solution according to 1: 9 ratio of volume ratio then, stir while dripping, solution mixes the back to be continued to stir 10 minutes.
3) with step 2) the gained mixed liquor places in 40 ℃ of thermostat water baths, makes it change gel into.
4) gel with the step 3) gained carries out vacuum drying, gets rid of moisture, obtains containing the chitosan xerogel of bacillus calmette-guerin vaccine and ferroso-ferric oxide.
5) xerogel with the step 4) gained grinds, crosses 400 mesh sieves and obtains corresponding composite material.
After measured, the main performance of the ferroso-ferric oxide/bacillus calmette-guerin vaccine that makes/integrated powder body of thermosensitive hydrogel: 30 hours bacillus calmette-guerin vaccine release rates are about 80%, and ferriferrous oxide particles is uniformly dispersed in gel, and biocompatibility is better.
Embodiment 4
1) the dose volume ratio is that 1.75: 1 and concentration are 0.1molL respectively
-1FeCl
3And FeS0
4Solution is at mechanical agitation and logical N
2Under the condition, with two kinds of solution mix homogeneously.Add 1wt% (the volume gained Fe after theory is calculated that is joined two kinds of solution with step 1
3O
4Quality be benchmark) surfactant polyethylene-6000 (PEG-6000).Stir after 15 minutes to mixed solution and dripping 1molL
-1NaOH solution, until pH value of solution=9, obtain black precipitate.Under 70 ℃ of water-baths, make its ripening.Left standstill 5 hours, and removed the supernatant, the particle that generates is washed the surfactant of flush away particle surface and foreign ion repeatedly with distilled water and dehydrated alcohol.At 50 ℃ of following vacuum dryings, ground 100 mesh sieves, obtain Fe
3O
4Granule.
2) be that 95% chitosan is dissolved in 4.5ml 0.1molL with the 100mg deacetylation
-1In the hydrochloric acid, fully stirred 5 hours, be mixed with the chitosan solution of 22.2mg/ml.Add the ferroso-ferric oxide that makes in 30mg (percentage by weight the is 5.35%) step 1), the method that adopts mechanical agitation to add ultra-sonic dispersion is uniformly dispersed it.Add 30mg (percentage by weight is 5.35%) bacillus calmette-guerin vaccine then, stirred 1 hour.400mg sodium (percentage by weight of chitosan and sodium is 89.3%) is dissolved in the 0.5ml deionized water, be mixed with the solution of 0.8g/ml, drop in the chitosan solution according to 1: 9 ratio of volume ratio then, stir while dripping, solution mixes the back to be continued to stir 15 minutes.
3) with step 2) the gained mixed liquor places in 37 ℃ of thermostat water baths, makes it change gel into.
4) gel with the step 3) gained carries out vacuum drying, gets rid of moisture, obtains containing the chitosan xerogel of bacillus calmette-guerin vaccine and ferroso-ferric oxide.
5) xerogel with the step 4) gained grinds, crosses 200 mesh sieves and obtains corresponding composite material.
After measured, the main performance of the ferroso-ferric oxide/bacillus calmette-guerin vaccine that makes/integrated powder body of thermosensitive hydrogel: 72 hours bacillus calmette-guerin vaccine release rates are about 80%, and ferriferrous oxide particles is uniformly dispersed in gel, and biocompatibility is better.
Claims (2)
1. ferrite/bacillus calmette-guerin vaccine/thermosensitive hydrogel composite material, it is characterized in that: comprise lyophilized bacillus calmette-guerin vaccine, ferroso-ferric oxide and chitosan/sodium xerogel, the percentage by weight of each component is lyophilized bacillus calmette-guerin vaccine 5%-15%, ferroso-ferric oxide 5%-15%, chitosan/sodium xerogel 70%-90%.
2. the preparation method of the described ferrite/bacillus calmette-guerin vaccine of claim 1/thermosensitive hydrogel composite material, this ferrite/bacillus calmette-guerin vaccine/thermosensitive hydrogel composite material comprises lyophilized bacillus calmette-guerin vaccine, ferroso-ferric oxide and chitosan/sodium xerogel, and the percentage by weight of each component is lyophilized bacillus calmette-guerin vaccine 5%-15%, ferroso-ferric oxide 5%-15%, chitosan/sodium xerogel 70%-90%; It is characterized in that: preparation method may further comprise the steps:
(1) utilize coprecipitation to prepare nano ferriferrous oxide:
The dose volume ratio is that 1.75: 1 and concentration are 0.1molL respectively
-1Fe
3+And Fe
2+Solution is at mechanical agitation and logical N
2Under the condition,, calculate according to the stereometer of mixed solution and to prepare gained Fe in theory two kinds of solution mix homogeneously
3O
4Gravimetric value, add this Fe
3O
4The surfactant polyethylene-6000 of theoretical weight value 0.1%-1%; Stir after 15 minutes to mixed solution and dripping 1molL
-1NaOH solution, until pH value of solution=9, obtain black precipitate, under 70 ℃ of water-baths, make the black precipitate ripening, left standstill 5 hours, and removed the supernatant, the particle that generates is washed repeatedly with distilled water and dehydrated alcohol, the surfactant of flush away particle surface and foreign ion, at 50 ℃ of following vacuum dryings, grinding is sieved, and obtains nano ferriferrous oxide granule;
(2) chitosan is dissolved in 0.1molL
-1In the hydrochloric acid, fully stir, be mixed with the chitosan settled solution of 15.5-22.5mg/ml, successively add the ferroso-ferric oxide and the lyophilized bacillus calmette-guerin vaccine of step (1) preparation, stirring or ultra-sonic dispersion make its mix homogeneously; With deionized water sodium is mixed with the solution of 0.7-1.0g/ml, then with sodium solution and chitosan solution according to 1: 9 mixed of volume ratio, mix the back and continue to stir 10-30 minute;
(3) step (2) gained mixed liquor is placed in the 30-40 ℃ of thermostat water bath, make it change gel into;
(4) gel with step (3) gained carries out lyophilization or vacuum drying, gets rid of moisture, obtains containing the chitosan xerogel of bacillus calmette-guerin vaccine and ferroso-ferric oxide;
(5) xerogel with step (4) gained grinds, crosses 100 orders and obtains ferrite/bacillus calmette-guerin vaccine/thermosensitive hydrogel composite material with top sieve.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105477647A (en) * | 2015-12-02 | 2016-04-13 | 常州大学 | Graphene quantum dot/chitosan xerogel preparation and application of graphene quantum dot/chitosan xerogel to fluorescent imaging and drug sustained release |
| CN109806239A (en) * | 2019-04-11 | 2019-05-28 | 重庆市肿瘤研究所 | A kind of microcapsules of the treatment of the controllable in drug release in tumour cell breast cancer |
| CN110575566A (en) * | 2019-09-25 | 2019-12-17 | 重庆理工大学 | Magnetic-response natural vascular matrix gel scaffold material and preparation method thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1593385A (en) * | 2004-06-22 | 2005-03-16 | 天津大学 | Gel capable of injecting temperature sensitive complex and its preparation method |
| CN100478032C (en) * | 2005-10-28 | 2009-04-15 | 中国科学院过程工程研究所 | Preparation method of injection type pH and glucose sensitive hydrogel |
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2008
- 2008-03-27 CN CNB2008100154555A patent/CN100560136C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105477647A (en) * | 2015-12-02 | 2016-04-13 | 常州大学 | Graphene quantum dot/chitosan xerogel preparation and application of graphene quantum dot/chitosan xerogel to fluorescent imaging and drug sustained release |
| CN109806239A (en) * | 2019-04-11 | 2019-05-28 | 重庆市肿瘤研究所 | A kind of microcapsules of the treatment of the controllable in drug release in tumour cell breast cancer |
| CN110575566A (en) * | 2019-09-25 | 2019-12-17 | 重庆理工大学 | Magnetic-response natural vascular matrix gel scaffold material and preparation method thereof |
| CN110575566B (en) * | 2019-09-25 | 2021-09-14 | 重庆理工大学 | Magnetic-response natural vascular matrix gel scaffold material and preparation method thereof |
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