CN101690716A - Calcium alginate-chitosan sustained-release microsphere carrying growth hormone and application thereof - Google Patents
Calcium alginate-chitosan sustained-release microsphere carrying growth hormone and application thereof Download PDFInfo
- Publication number
- CN101690716A CN101690716A CN200910019437A CN200910019437A CN101690716A CN 101690716 A CN101690716 A CN 101690716A CN 200910019437 A CN200910019437 A CN 200910019437A CN 200910019437 A CN200910019437 A CN 200910019437A CN 101690716 A CN101690716 A CN 101690716A
- Authority
- CN
- China
- Prior art keywords
- microsphere
- growth hormone
- chitosan
- solution
- syringe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a calcium alginate-chitosan sustained-release microsphere carrying growth hormone. The sustained-release microsphere comprises the principal components including the growth hormone, the calcium alginate and the chitosan. The sustained-release microsphere can be used for preparing the drugs for fracture healing. The sustained-release microsphere has good sphericity and narrow particle size distribution range and can further sustain the release and absorption of the drugs. The microsphere features simple and efficient shaping process and mild shaping conditions. The growth hormone has high entrapment rate and activity, and organic solvents are unnecessary, etc.
Description
Technical field
The present invention relates to a kind of sustained-release micro-spheres, especially a kind ofly be loaded with the calcium alginate of growth hormone, the gel micro-ball of chitosan, and relate to the application of this microsphere.Belong to biological chemical field.
Background technology
Studies show that growth hormone has certain facilitation to general fracture; Other there are some researches show that body inner growth hormone secretion level reduces with increasing the minimizing of age and estrogen secretion, and the growth hormone reduction can cause abnormal bone metabolism and fracture repair ability to reduce.Fracture healing process still is subjected to adjustings such as endocrine hormone and cell growth factor and influences except that closely related with blood supply and mechanics factor, and wherein, growth hormone plays a part certain.Growth hormone can act on osseous tissue by different approach and mode, and IGFs plays important transmission and mediation therein, IGFs both can indirectly be produced and be released in the blood circulation by acting on liver by growth hormone, also can directly act on osteoblast and the chondrocyte film growth hormone receptor (GH-R) and induces its generation IGFs and act on the part.The IGFs effect is comparatively complicated, and IGFs can improve the reaction of osteoblast to growth hormone on the one hand; " coupling " between formation of IGFs adjustable bone and bone resorption on the other hand.IGF discharges in osseous tissue during bone resorption, promotes osteoblastic bone formation effect as the paracrine factor that postpones, and fills the bone resorption chamber to produce new bone; The IGF product of osteoclast is regulated the osteoblastic quantity in bone resorption position as paracrine factor simultaneously; And the IGF that osteoblast produces has determined the filling operation in bone resorption chamber as the autocrine factor.IGFs comprises IGF-I, IGF-II, IGF-I receptor, IGF-II receptor, and IGF connects albumen and IGFBP protease, thereby its effect also must be subjected to many-side, multilevel adjusting.And growth hormone has promoted a large amount of myeloid elements to invade external callus and the mineralising that influences the callus tissue also plays a role, growth hormone is mainly by promoting the bone conversion that osseous tissue is worked, because the bone conversion is positive balance the bone amount is increased at trophophase, and after menopause, be negative balance owing to the bone conversion, then cause the bone amount to reduce.More with report about growth hormone to the union of fracture experimental study of effect, think significant to the union of fracture effect of additional exogenous growth hormone dosage, Mechanical test results shows that it is the most obvious that the maximal dose group increases the mechanical property effect.Except that dosage factor, administration frequency more should be paid attention to, studies show that, the growth hormone base concentration is below the 10ng/ml in the female rats blood plasma, and peak value can reach 200～800ng/ml, and its secretion cycle is 30～60 minutes, and after exogenous supplementation with growth hormones, growth hormone concentration is higher than foundation level and can keeping 6～8 hours in the blood plasma, and this is quite useful to union of fracture.Thereby, science and reasonably administration frequency should follow the secretion frequency and the rule of the individual endogenous growth hormone of administration.And in the different times administration of union of fracture, the result also there are differences, and there are some researches show, growth hormone (was fractured back 20 days in) mainly in early days to the facilitation of union of fracture.
The fluctuation of growth hormone (GH) level is in the range of normal value at 0.68-9.01ng/ml, and GH begins to increase 10 of nights, and 12 of nights are the highest to last till 2:00 AM, and this moment, the lasting high level of GH helped the synthetic and growth promoter of albumen of body.At present, people use growth hormone can only adopt injection system, have inconvenience and cause suffering to patient, how to make its medication more convenient and can promote better that the healing of fracturing is noticeable day by day.
CN961157143 discloses a kind of microsphere supported method of chitin for preparing, it is to add alkali liquor and Reducing agent in chitin powder, generate chitosan, generate soluble-salt with acid reaction again, aqueous solution with this product disperses in organic solvent, curing molding, and the chitin of producing sphere diameter scope 0.001～10mm is microsphere supported.Has bioaffinity, porous, stability.Be used for cell embedding, microbial immobilized, antibacterial immobilization fermentation, aspects such as zooblast adhere-wall culture, miniature slow release and chromatography.CN2003101247037 discloses a kind of chitin-alginic acid calcium gel micro-ball soft tissue enhancement material, coats chitosan film by calcium alginate gel bead, and mixes the formation gel with sodium alginate, and wherein the particle diameter of calcium alginate gel bead is 1-2000 μ m.CN2004100160670 discloses a kind of triple complex microsphere preparation, preparation mainly consisted of model drug, calcium alginate, chitosan, Vicryl Rapide.This complex microsphere can make Partial Protein class and polypeptide drug be protected in hydrophilic sodium alginate-chitosan microcapsules environment and obviously reduce the prominent phenomenon of releasing, and reduces not exclusively to discharge, and can form the release mode of regulating medicine by changing PLGA.Existing many sustained-release micro-spheres with bioaffinity; though other drug is had good embedding, slow release and protective effect; but all can not be used for embedding, slow release and protection to growth hormone; so that after the growth hormone oral administration in gastrointestinal tract activity be not destroyed and can slow release, make growth hormone bring into play more dauer effect.
Summary of the invention
Deficiency at the prior art existence, technical problem to be solved by this invention is, a kind of calcium alginate-chitosan sustained-release microsphere carrying growth hormone is provided so that after the growth hormone oral administration in gastrointestinal tract activity be not destroyed and can be by slow release, make the more persistent effect of growth hormone performance.
For solving the problems of the technologies described above, the technical scheme that the present invention takes is, a kind of calcium alginate-chitosan sustained-release microsphere carrying growth hormone, and its main component is growth hormone, calcium alginate, chitosan, is prepared from through the following step:
At first precision is measured 10IU/mL growth hormone injection and sodium alginate soln, with in 1: 4 syringe of packing into behind the volume ratio mix homogeneously; The positive pole of impulse electric field preparing instrument is connected with syringe metal orifice, negative pole is connected with placing coagulation bath solution metal conductive ring, between orifice and coagulation bath, form pulse static field, pulse electric field device instructed voltage 380V, pulse frequency 120Hz, pulsewidth 2ms; Syringe is released liquid, overcomes inherent viscous force of sodium alginate soln and surface tension under the electric field force effect, makes the mixed solution drop splash into coagulation bath and solidifies, and forms microsphere;
Then microsphere is leached, and mix the back vibration with 1: 10 volume ratio, form even semi permeability thin film, obtain composite gel microsphere at microsphere surface with the film forming chitosan solution; Again composite gel microsphere vacuum lyophilization is got product;
Wherein: used sodium alginate soln concentration is 5～20mg/ml, and coagulation bath solution is 5～30mg/ml CaCl
2Solution, film forming chitosan solution concentration is 5～20mg/ml.
Above-mentioned calcium alginate-chitosan sustained-release microsphere carrying growth hormone, its preparation process is as follows:
At first precision is measured 10IU/mL growth hormone injection and sodium alginate soln, with in 1: 4 syringe of packing into behind the volume ratio mix homogeneously; The positive pole of impulse electric field preparing instrument is connected with syringe metal orifice, the orifice aperture is 450um～600um, negative pole is connected with placing coagulation bath solution metal conductive ring, between orifice and coagulation bath, form pulse static field, pulse electric field device instructed voltage 380V, pulse frequency 120Hz, pulsewidth 2ms; Syringe is released liquid, the material liquid flow velocity is 8mL/h～50mL/h, overcomes inherent viscous force of sodium alginate soln and surface tension under the electric field force effect, makes the mixed solution drop splash into coagulation bath and solidifies, form microsphere, orifice and liquid level distance are 1cm～5cm;
Then microsphere is leached, and mix 170rpm vibration 15min in the adding film forming chitosan solution with 1: 10 volume ratio, form even semi permeability thin film, obtain composite gel microsphere at microsphere surface with the film forming chitosan solution; Again with composite gel microsphere at-75 ℃ of pre-freeze 2h, in vacuum 3 * 10
3Mbar, temperature-45 a ℃ lyophilization 24h get product;
Wherein: used sodium alginate soln concentration is 15mg/ml, and coagulation bath solution is 20mg/mlCaCl
2Solution, film forming chitosan solution concentration is 10mg/ml.
Above-mentioned calcium alginate-chitosan sustained-release microsphere carrying growth hormone, in the preparation, its syringe metal orifice preferred 450um in aperture, syringe is released the preferred 8mL/h of feed liquid flow velocity, orifice and the preferred 2cm of liquid level distance.
Above-mentioned calcium alginate-chitosan sustained-release microsphere carrying growth hormone is used to prepare the medicine for the treatment of union of fracture.
The present invention relates to have the chitosan of physiologically active, it has positive charge cation group rare in the universe behind deacetylation, and is alkalescence, and this is for the health of organism, and especially human beings'health is significant.After chitosan is absorbed by the body, can bundle, adsorb undesirable substances such as fat electronegative in the human body, toxin, can blood fat reducing, activating cell, enhancing immune function of human body, healing time is shortened, thus alleviate the misery of fracture patient.In addition, chitosan has biological degradability, macromolecule carrier as control drug release in the slow release formulation has obvious superiority, and having avoided more present macromolecular materials (as silicone rubber etc.) is that the patient brings unnecessary operation trouble and painful as pharmaceutical carrier.
Microsphere good sphericity of the present invention, particle size distribution range is narrow, can make the release of medicine absorb more lasting.The microsphere forming process is easy, efficient, the condition of molding gentleness, and growth hormone envelop rate and active high need not organic solvent etc.
In the raw material involved in the present invention, the growth hormone injection is the product of Changchun Jinsai Medicine Co.,Ltd, sodium alginate, calcium chloride, chitosan are provided by Chinese Marine University, are industry common agents, all the other unspecified industry common agents that are.
Instrument and equipment involved in the present invention is common instrument except that having particularly pointed out.
The specific embodiment
The present invention is described in more detail below in conjunction with embodiment.
Embodiment 1
The calcium alginate-chitosan sustained-release microsphere carrying growth hormone of present embodiment, its main component are growth hormone, calcium alginate, chitosan, adopt the following step preparation:
At first precision is measured 10IU/mL growth hormone injection and 15mg/ml sodium alginate soln, with in 1: 4 syringe of packing into behind the volume ratio mix homogeneously; The positive pole of impulse electric field preparing instrument is connected with syringe metal orifice, the orifice aperture is 450um, negative pole is connected with placing coagulation bath solution metal conductive ring, between orifice and coagulation bath, form pulse static field, pulse electric field device instructed voltage 380V, pulse frequency 120Hz, pulsewidth 2ms; Syringe is released liquid, and the material liquid flow velocity is 8mL/h, overcomes inherent viscous force of sodium alginate soln and surface tension under the electric field force effect, makes the mixed solution drop splash into 20mg/ml CaCl
2Coagulation bath solidifies in the solution, forms microsphere, and orifice and liquid level distance are 2cm.Then microsphere is leached, and mix with 1: 10 volume ratio with 10mg/ml film forming chitosan solution, the 170rpm 15min that vibrates forms even semi permeability thin film at microsphere surface, obtains composite gel microsphere; Again with composite gel microsphere at-75 ℃ of pre-freeze 2h, in vacuum 3 * 10
3Mbar, temperature-45 a ℃ lyophilization 24h get product.
Embodiment 2
The calcium alginate-chitosan sustained-release microsphere carrying growth hormone of present embodiment, its main component are growth hormone, calcium alginate, chitosan, adopt the following step preparation:
At first precision is measured 10IU/mL growth hormone injection and 20mg/ml sodium alginate soln, with in 1: 4 syringe of packing into behind the volume ratio mix homogeneously; The positive pole of impulse electric field preparing instrument is connected with syringe metal orifice, the orifice aperture is 600um, negative pole is connected with placing coagulation bath solution metal conductive ring, between orifice and coagulation bath, form pulse static field, pulse electric field device instructed voltage 380V, pulse frequency 120Hz, pulsewidth 2ms; Syringe is released liquid, and the material liquid flow velocity is 25mL/h, overcomes inherent viscous force of sodium alginate soln and surface tension under the electric field force effect, makes the mixed solution drop splash into 30mg/ml CaCl
2Coagulation bath solidifies in the solution, forms microsphere, and orifice and liquid level distance are 5cm.Then microsphere is leached, and mix with 1: 10 volume ratio with 20mg/ml film forming chitosan solution, the 170rpm 15min that vibrates forms even semi permeability thin film at microsphere surface, obtains composite gel microsphere; Again with composite gel microsphere at-75 ℃ of pre-freeze 2h, in vacuum 3 * 10
3Mbar, temperature-45 a ℃ lyophilization 24h get product.
Embodiment 3
The calcium alginate-chitosan sustained-release microsphere carrying growth hormone of present embodiment, its main component are growth hormone, calcium alginate, chitosan, adopt the following step preparation:
At first precision is measured 10IU/mL growth hormone injection and 5mg/ml sodium alginate soln, with in 1: 4 syringe of packing into behind the volume ratio mix homogeneously; The positive pole of impulse electric field preparing instrument is connected with syringe metal orifice, the orifice aperture is 500um, negative pole is connected with placing coagulation bath solution metal conductive ring, between orifice and coagulation bath, form pulse static field, pulse electric field device instructed voltage 380V, pulse frequency 120Hz, pulsewidth 2ms; Syringe is released liquid, and the material liquid flow velocity is 50mL/h, overcomes inherent viscous force of sodium alginate soln and surface tension under the electric field force effect, makes the mixed solution drop splash into 5mg/ml CaCl
2Coagulation bath solidifies in the solution, forms microsphere, and orifice and liquid level distance are 1cm.Then microsphere is leached, and mix with 1: 10 volume ratio with 5mg/ml film forming chitosan solution, the 170rpm 15min that vibrates forms even semi permeability thin film at microsphere surface, obtains composite gel microsphere; Again with composite gel microsphere at-75 ℃ of pre-freeze 2h, in vacuum 3 * 10
3Mbar, temperature-45 a ℃ lyophilization 24h get product.
Embodiment 4
Present embodiment is to the detection of specific sustained-release micro-spheres of the present invention and analysis.
1, the microsphere sphere diameter is measured
Utilize biological inverted microscope to read 100 medicine carrying microballoons sphere diameters at random, with arithmetic mean of instantaneous value as microsphere average grain diameter d.
2, growth hormone entrapment efficiency determination in the microsphere
Application fetches-Hydrolyze method detects growth hormone microsphere carrying drug ratio and envelop rate.Precision takes by weighing cryodesiccated microsphere 20mg, behind 5ml simulated intestinal fluid dissolving microsphere, 1 * 10
4Centrifugal 10min under the rpm condition keeps supernatant.It is that 7.4 PBS1.0ml disperses again that precipitation adds pH value, and centrifugal back keeps supernatant; Precipitation is dissolved in 10ml 0.1mol/L sodium hydroxide, and is centrifugal after fully mixing; Isolate supernatant, merge with preceding step supernatant; With in the 0.1mol/L hydrochloric acid and blended supernatant.Growth hormone content in the mixing supernatant is measured according to Pharmacopoeia of the People's Republic of China III portion (version in 2005) appendix VIB albuminometry the 2nd method (Lowry method).The envelop rate of microsphere calculates by following formula:
Envelop rate=(actual measurement carrying growth hormone amount/theoretical carrying growth hormone amount) * 100%.
3, carrying growth hormone microsphere form and Structure Mechanism
The configuration of carrying growth hormone microsphere before the lyophilizing, form rounding, sphericity are good.The particle size distribution of carrying growth hormone microsphere is comparatively even after the lyophilizing, good dispersion.Microsphere is not slick spheroid, and there are many holes on the surface, and this helps drug molecule and slowly discharges from microsphere inside and prevent that digestive enzyme the gastrointestinal tract from entering in the microsphere destruction to growth hormone.
4, Ca
2+Concentration is to the influence of the carrying growth hormone microsphere envelop rate of preparation
What table 1 was listed is at Ca
2+Concentration was respectively 0.5%, 1%, 2%, 3% o'clock, under 1.5% Algin, the 1% chitosan condition to the influence of carrying growth hormone microsphere envelop rate of preparation.Result in the table 1 shows, Ca
2+During concentration 2%, the envelop rate of growth hormone is the highest in the microsphere, and microspherulite diameter does not have significant change.
Table 1Ca
2+Concentration is to the influence of the carrying growth hormone microsphere envelop rate of preparation
5, sodium alginate concentration is to the influence of medicine carrying microballoons particle diameter, envelop rate
What table 2 was listed is that sodium alginate concentration was respectively Ca at 0.5%, 1%, 1.5%, 2% o'clock
2+Under concentration 2%, the 1% chitosan condition to the preparation the carrying growth hormone microspherulite diameter and the influence of envelop rate.The result shows that sodium alginate concentration is big more, and the envelop rate of growth hormone is high more in the microsphere, and microspherulite diameter is the carrying growth hormone microspherulite diameter minimum that is in the preparation of 1.5% sodium alginate, and envelop rate is all greater than 90%.
Table 2 sodium alginate concentration is to the influence of medicine carrying microballoons particle diameter, envelop rate
Algin concentration
0.5%
1%
1.5%
2%
Envelop rate
75%
84%
93%
96%
Particle diameter
130um
145um
150um
190um
6, chitosan concentration is to the influence of medicine carrying microballoons particle diameter, envelop rate
Table 3 chitosan concentration is to the influence of medicine carrying microballoons particle diameter, envelop rate
Chitosan concentration
0.5%
1%
2%
Envelop rate
95%
96%
96%
Particle diameter
150um
155um
155um
What table 3 was listed is that chitosan concentration was respectively Ca at 0.5%, 1%, 2% o'clock
2+Under concentration 2%, the 1.5% sodium alginate condition to the preparation carrying growth hormone microsphere envelop rate and the influence of particle diameter.Result in the table 3 shows that chitosan concentration is to envelop rate and grain diameter influence's minimum of growth hormone in the microsphere.Envelop rate all greater than 90% situation under, adopt 0.5% chitosan concentration microspheres prepared particle diameter minimum.
7, the orifice aperture is to the influence of medicine carrying microballoons particle diameter, envelop rate
Table 4 is listed is orifice aperture when being respectively 450um, 600um, 700um, 1.5% Algin, Ca
2+Under concentration 2%, the 1% chitosan condition to the particle diameter of microsphere of preparation and the influence of envelop rate.The result shows that the aperture is big more, and microspherulite diameter is big more, and envelop rate is more little.When the aperture was respectively 450um and 600um, envelop rate had actual production and is worth greater than 80%.
Table 4 orifice aperture is to the influence of medicine carrying microballoons particle diameter, envelop rate
The orifice aperture
450um
600um
700um
Envelop rate
94%
87%
70%
Particle diameter
150um
175um
220um
8, the material liquid flow velocity is to the influence of medicine carrying microballoons particle diameter, envelop rate
Table 5 is listed is material liquid flow velocity when being respectively 8mL/h, 25mL/h, 50mL/h, 1.5% Algin, Ca
2+Under concentration 2%, the 1% chitosan condition to the particle diameter of microsphere of preparation and the influence of envelop rate.The result shows that the material liquid flow velocity is not remarkable to the influence of envelop rate, and envelop rate is all about 80%.And to the influence of particle diameter clearly, particle diameter increased rapidly after the material liquid flow velocity surpassed 50mL/h, when the material liquid flow velocity more hour, the microspherulite diameter of preparation is also more little.Find microsphere preparation process inventor, too fast when the material liquid flow velocity, when reaching 50mL/h, electric discharge phenomena usually appear, and under the effect of high-voltage arc, liquid disperses, and produces a large amount of magnetic tape trailer minimicrospheres, and big microsphere wherein breaks out-of-shape easily; Electric arc also can destroy the activity of growth hormone.Thereby adopt less material liquid flow velocity to help reducing microspherulite diameter.
Table 5 material liquid flow velocity is to the influence of medicine carrying microballoons particle diameter, envelop rate
The material liquid flow velocity
8mL/h
25mL/h
50mL/h
Envelop rate
90%
92%
95%
Particle diameter
150um
200um
260um
9, liquid level is apart from the influence to medicine carrying microballoons particle diameter, envelop rate
Table 6 is listed is liquid level when being respectively 1cm, 2cm, 5cm, 1.5% Algin, Ca
2+Under concentration 2%, the 1% chitosan condition to the particle diameter of insulin microsphere of preparation and the influence of envelop rate.Envelop rate and particle diameter are subjected to the influence of liquid level distance all to embody the existence of extreme value.When the liquid level distance was respectively 1cm, 2cm, 5cm, envelop rate was all more than 90%; And when the liquid level distance was 2cm, particle diameter had all reached minimum.
Table 6 liquid level is apart from the influence to medicine carrying microballoons particle diameter, envelop rate
The liquid level distance
1cm
2cm
5cm
Envelop rate
95%
90%
96%
Particle diameter
175um
135um
220um
Embodiment 5
Present embodiment is the application test to medicine carrying microballoons of the present invention.
Agents useful for same: 1, simulated gastric fluid: precision is measured the 2.25ml concentrated hydrochloric acid, is settled to 250ml with distilled water, records pH=1.30.2, simulated intestinal fluid: accurate weighing sodium hydroxide 0.4g and potassium dihydrogen phosphate 1.7g, after the distilled water dissolving, standardize solution records pH=7.5 to 250ml.3, the microsphere of embodiment 1 preparation.
Test method:
Adopt culture method, precision takes by weighing microsphere 10mg, places tool plug test tube, adds the 20ml release medium.Sample is put into 37 ℃ of constant temperature table shakes, is the 50rpm vibration with the rotating speed.Regularly take out sample cell, 1 * 10
4Behind the rpm frozen centrifugation 10min, take out supernatant 2ml, add the fresh release medium of equivalent again.Albuminous content as blank, relatively discharges the content of growth hormone in the liquid with the release supernatant of blank microsphere indirectly in the different time sections supernatant by the absorption photometric value.
Experimental result is: in the in-vitro simulated gastric juice, microsphere not swelling and institute's carrying growth hormone is not released substantially; In simulated intestinal fluid, microspheres swell up and institute's carrying growth hormone are released.The PH sensitivity and the medicine that show microspheres swell up thus discharge with flooding mechanism, and freeze dried microsphere exists many fold to reach than small holes at not swollen situation lower surface, and it keeps apart the medicine in the microsphere and extraneous digestive enzyme; After entering intestinal juice, microsphere generation swelling, the fold of microsphere surface disappears and open in the cavity, and this provides passage for the infiltration of dissolution medium and the diffusion of medicine.Can reach a conclusion thus: medicine carrying microballoons of the present invention is fit to be applied to prepare the medicine that is loaded with growth hormone for the treatment of union of fracture very much.
Claims (4)
1. calcium alginate-chitosan sustained-release microsphere carrying growth hormone, its main component is growth hormone, calcium alginate, chitosan, is prepared from through the following step:
At first precision is measured 10IU/mL growth hormone injection and sodium alginate soln, with in 1: 4 syringe of packing into behind the volume ratio mix homogeneously; The positive pole of impulse electric field preparing instrument is connected with syringe metal orifice, negative pole is connected with placing coagulation bath solution metal conductive ring, between orifice and coagulation bath, form pulse static field, pulse electric field device instructed voltage 380V, pulse frequency 120Hz, pulsewidth 2ms; Syringe is released liquid, overcomes inherent viscous force of sodium alginate soln and surface tension under the electric field force effect, makes the mixed solution drop splash into coagulation bath and solidifies, and forms microsphere;
Then microsphere is leached, and mix the back vibration with 1: 10 volume ratio, form even semi permeability thin film, obtain composite gel microsphere at microsphere surface with the film forming chitosan solution; Again composite gel microsphere vacuum lyophilization is got product;
Wherein: used sodium alginate soln concentration is 5～20mg/ml, and coagulation bath solution is 5～30mg/ml CaCl
2Solution, film forming chitosan solution concentration is 5～20mg/ml.
2. calcium alginate-chitosan sustained-release microsphere carrying growth hormone according to claim 1, its preparation process is as follows:
At first precision is measured 10IU/mL growth hormone injection and sodium alginate soln, with in 1: 4 syringe of packing into behind the volume ratio mix homogeneously; The positive pole of impulse electric field preparing instrument is connected with syringe metal orifice, the orifice aperture is 450um～600um, negative pole is connected with placing coagulation bath solution metal conductive ring, between orifice and coagulation bath, form pulse static field, pulse electric field device instructed voltage 380V, pulse frequency 120Hz, pulsewidth 2ms; Syringe is released liquid, the material liquid flow velocity is 8mL/h～50mL/h, overcomes inherent viscous force of sodium alginate soln and surface tension under the electric field force effect, makes the mixed solution drop splash into coagulation bath and solidifies, form microsphere, orifice and liquid level distance are 1cm～5cm;
Then microsphere is leached, and mix 170rpm vibration 15min in the adding film forming chitosan solution with 1: 10 volume ratio, form even semi permeability thin film, obtain composite gel microsphere at microsphere surface with the film forming chitosan solution; Again with composite gel microsphere at-75 ℃ of pre-freeze 2h, in vacuum 3 * 10
3Mbar, temperature-45 a ℃ lyophilization 24h get product;
Wherein: used sodium alginate soln concentration is 15mg/ml, and coagulation bath solution is 20mg/mlCaCl
2Solution, film forming chitosan solution concentration is 10mg/ml.
3. calcium alginate-chitosan sustained-release microsphere carrying growth hormone according to claim 2 is characterized in that: when this sustained-release micro-spheres of preparation, syringe metal orifice aperture is 450um, and it is 8mL/h that syringe is released the feed liquid flow velocity, and orifice and liquid level distance are 2cm.
4. claim 1,2 or 3 each described calcium alginate-chitosan sustained-release microsphere carrying growth hormones are used to prepare the medicine for the treatment of union of fracture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910019437A CN101690716A (en) | 2009-10-13 | 2009-10-13 | Calcium alginate-chitosan sustained-release microsphere carrying growth hormone and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910019437A CN101690716A (en) | 2009-10-13 | 2009-10-13 | Calcium alginate-chitosan sustained-release microsphere carrying growth hormone and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101690716A true CN101690716A (en) | 2010-04-07 |
Family
ID=42079467
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910019437A Pending CN101690716A (en) | 2009-10-13 | 2009-10-13 | Calcium alginate-chitosan sustained-release microsphere carrying growth hormone and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101690716A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102258481A (en) * | 2011-08-19 | 2011-11-30 | 薛巍 | Method for preparing self-assembled medicine-carried microspheres by combining high-voltage electrostatic liquid droplet method and layer-by-layer self-assembly method |
| CN102428961A (en) * | 2011-11-15 | 2012-05-02 | 农业部环境保护科研监测所 | Method for preparing nano MgO sustained-release sterilizing granules |
| CN103495210A (en) * | 2013-09-26 | 2014-01-08 | 福州大学 | Chitosan-hydroxylapatite in-situ loaded icariin composite microspheres |
| CN103751791A (en) * | 2014-01-20 | 2014-04-30 | 中国海洋大学 | Method for preparing composite chitosan nanogel immobilized multi-layer capsules |
| CN103757917A (en) * | 2014-01-26 | 2014-04-30 | 江汉大学 | Method for preparing chitosan-based composite material with electroactivity |
| CN108414393A (en) * | 2018-06-13 | 2018-08-17 | 安徽中医药大学 | A method of measuring entrapment efficiency of liposome |
| CN114762732A (en) * | 2021-01-13 | 2022-07-19 | 梦芊科技知识产权有限公司 | Controlled release microspheres and anti-aging application thereof |
| CN115057741A (en) * | 2022-07-04 | 2022-09-16 | 西南交通大学 | Preparation method of calcium alginate-chitosan microbead slow-release fertilizer difficult to swell |
| CN115282127A (en) * | 2022-04-21 | 2022-11-04 | 广州医科大学附属第二医院 | Chitosan calcium alginate nano-particle microsphere and preparation method thereof |
| CN115590826A (en) * | 2022-10-21 | 2023-01-13 | 北京安奇生物医药科技有限公司（Cn） | Active protein mucosal drug delivery preparation and preparation method and application thereof |
-
2009
- 2009-10-13 CN CN200910019437A patent/CN101690716A/en active Pending
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102258481A (en) * | 2011-08-19 | 2011-11-30 | 薛巍 | Method for preparing self-assembled medicine-carried microspheres by combining high-voltage electrostatic liquid droplet method and layer-by-layer self-assembly method |
| CN102428961A (en) * | 2011-11-15 | 2012-05-02 | 农业部环境保护科研监测所 | Method for preparing nano MgO sustained-release sterilizing granules |
| CN102428961B (en) * | 2011-11-15 | 2013-07-17 | 农业部环境保护科研监测所 | Method for preparing nano MgO sustained-release sterilizing granules |
| CN103495210A (en) * | 2013-09-26 | 2014-01-08 | 福州大学 | Chitosan-hydroxylapatite in-situ loaded icariin composite microspheres |
| CN103495210B (en) * | 2013-09-26 | 2015-03-04 | 福州大学 | Chitosan-hydroxylapatite in-situ loaded icariin composite microspheres |
| CN103751791B (en) * | 2014-01-20 | 2016-01-06 | 中国海洋大学 | The preparation method of the gel immobilized multilamellar capsule of a kind of chitosan composite Nano |
| CN103751791A (en) * | 2014-01-20 | 2014-04-30 | 中国海洋大学 | Method for preparing composite chitosan nanogel immobilized multi-layer capsules |
| CN103757917B (en) * | 2014-01-26 | 2016-02-17 | 江汉大学 | The preparation method of the electroactive chitosan-based composite of a kind of tool |
| CN103757917A (en) * | 2014-01-26 | 2014-04-30 | 江汉大学 | Method for preparing chitosan-based composite material with electroactivity |
| CN108414393A (en) * | 2018-06-13 | 2018-08-17 | 安徽中医药大学 | A method of measuring entrapment efficiency of liposome |
| CN114762732A (en) * | 2021-01-13 | 2022-07-19 | 梦芊科技知识产权有限公司 | Controlled release microspheres and anti-aging application thereof |
| CN115282127A (en) * | 2022-04-21 | 2022-11-04 | 广州医科大学附属第二医院 | Chitosan calcium alginate nano-particle microsphere and preparation method thereof |
| CN115282127B (en) * | 2022-04-21 | 2023-08-29 | 广州医科大学附属第二医院 | Chitosan calcium alginate nanoparticle microsphere and preparation method thereof |
| CN115057741A (en) * | 2022-07-04 | 2022-09-16 | 西南交通大学 | Preparation method of calcium alginate-chitosan microbead slow-release fertilizer difficult to swell |
| CN115057741B (en) * | 2022-07-04 | 2023-06-20 | 西南交通大学 | Preparation method of calcium alginate-chitosan microsphere slow-release fertilizer not easy to swell |
| CN115590826A (en) * | 2022-10-21 | 2023-01-13 | 北京安奇生物医药科技有限公司（Cn） | Active protein mucosal drug delivery preparation and preparation method and application thereof |
| CN115590826B (en) * | 2022-10-21 | 2023-12-19 | 北京安必奇生物科技有限公司 | Active protein mucous membrane administration preparation and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101690716A (en) | Calcium alginate-chitosan sustained-release microsphere carrying growth hormone and application thereof | |
| CN103040727A (en) | Preparation method of drug and protein sustained-release alginate hybrid gel | |
| KR100535319B1 (en) | Drug composition with controlled drug release rate | |
| EP1722759A2 (en) | Chitosan containing solution | |
| CN101249077A (en) | Preparation of degradable pollutant polyalcohol stephanoporate microballoons and uses thereof | |
| TW200529869A (en) | Delivery system for drug and cell therapy | |
| CN1180569A (en) | Preparation for medicine feeding through nose | |
| CN107007875B (en) | Enzyme and temperature dual-responsiveness drug-loaded hydrogel and preparation method and application thereof | |
| CN101756908A (en) | Hydroxyapatite micro-sphere with polyester coating and preparation method thereof | |
| CN113318224B (en) | Double-wheel-shaped nano-particles and preparation method thereof | |
| CN105617362A (en) | Novel insulin-phospholipid-chitosan self-assembled microparticle carrier and preparation thereof | |
| CN108096225B (en) | Nanofiber membrane containing calcitonin liposome and preparation method and application thereof | |
| US20110201680A1 (en) | formulation of silymarin with high efficacy and prolonged action and the preparation method thereof | |
| CN101511170B (en) | Sublimable sustained release delivery system and method of making same | |
| JP2009523791A (en) | Drug containing dextran having a molecular weight of 1.0 to 100 kDa and preparation method thereof | |
| CN102657871A (en) | Oral slow release preparation, entrapment material and preparation method | |
| CN115337272B (en) | Natural polysaccharide-based chemical-physical double-crosslinked hydrogel particles and preparation and application thereof | |
| CN1198100A (en) | Chitosan drug delivery system | |
| CN113197842B (en) | Cannabidiol injectable hydrogel, preparation method and application thereof | |
| CN101708175A (en) | Sustained release microsphere medicinal preparation using PHBHHx as carrier and method for preparing same | |
| CN101502490A (en) | Ketoprofen bioadhesive gel microsphere and preparation method thereof | |
| CN108553424B (en) | Preparation method of oral amoxicillin pH response type nano carrier and product thereof | |
| CN1315530C (en) | Method of preparing system for controlled release of medication in protein category | |
| Arulanandraj et al. | NANOTECHNOLOGICAL APPROACH TO ENHANCE THE STABILITY AND BIOAVAILABILITY OF THE HERBAL DRUG" MURVA" | |
| CN102274485B (en) | Antibacterial peptide sustained-release microsphere preparation and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20100407 |