CN1970094A - Process for preparing hyaluronic acid-chitosan biomembrane - Google Patents
Process for preparing hyaluronic acid-chitosan biomembrane Download PDFInfo
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- CN1970094A CN1970094A CN 200610170264 CN200610170264A CN1970094A CN 1970094 A CN1970094 A CN 1970094A CN 200610170264 CN200610170264 CN 200610170264 CN 200610170264 A CN200610170264 A CN 200610170264A CN 1970094 A CN1970094 A CN 1970094A
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- hyaluronic acid
- chitosan
- biomembrane
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- butanediol bisglycidyl
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Abstract
The invention discloses a making method of hyaluronic acid-chitosan biomembrane, which adopts hyaluronic acid, carboxymethyl chitose, polyvinyl alcohol and butanediol diepihydrin as raw material.
Description
Technical field:
The application relates to a kind of biomembranous preparation method, is specifically related to a kind of preparation method that is used for preventing behind the glaucoma operation filtering the hyaluronic acid-chitosan biomembrane of passage scarring.
Background technology:
Glaucoma is the irreversibility diseases causing blindness of China first place, and operation is still the first-selected strategy of China's glaucoma treatment.But because biological inherent wound healing mechanism, the scarring that filters passage behind the glaucoma operation is the successful biggest obstacle of operation for glaucoma treatment.In recent years at the designed new operation of primary open angle glaucoma--the non-trabeculectomy associating SK glue that penetrates is implanted in and has obtained clinically using widely, wherein the effect of mechanical support filtration passage has been played in the implantation of SK glue, SK glue composition is the cross linkage hyaluronate sodium, early stage behind filtration surgery, kept the filtration passage, because continue flowing of aqueous humor, also delay to a certain extent and suppressed growth of fibroblasts, improved success rate of operation.But since at present the material of implanting in the Non Penetrating Trabecular Surgery (NPTS) art all from abroad, the expense cost height.We have carried out cost effectiveness analysis for this reason, penetrate operation and reduce a routine adverse events and need spend more 110,000 yuan of expense RMB by non-, reduce an example seriously adverse events need spend more 190,000 yuan of expense RMB.And by analyzing discovery, embedded material is the main source that this surgical selection expense increases.The cost that has only further reduction embedded material, this modus operandi could further be promoted in China.Hyaluronic acid be by glucuronic acid and N-acetylglucosamine with a kind of linear polysaccharide that glycosidic linkage alternately is formed by connecting, extensively be present in higher mammal and the zootic extracellular matrix.Studies show that in a large number hyaluronic acid has excellent biocompatibility and non-immunogenicity, has high viscoelasticity simultaneously.Chitosan is that biosphere is only second to cellulosic second largest class polysaccharide, research has both at home and abroad confirmed that chitosan has bioadhesive and multiple biological activity, comprise that chitosan can promote epithelial growth, suppress fibroblasts proliferation, the propagation that suppresses tumor cell, suppress microbial growths such as antibacterial, promote to organize the multiple effects such as reparation of wound surface.Therefore, chitosan and derivant thereof are applied in the clinical position widely, are used to prevent tissue adhesion, are made into artificial skin, promote the aspects such as skin repair of wound.
The present invention has utilized the biological nature of hyaluronic acid and chitosan, successfully synthesize hyaluronic acid-chitosan biomembrane, this biomembrane had both had the mechanical support effect of external product SK glue at the filtration surgery passage, has simultaneously the effect that suppresses fibroblast proliferation again, and the prices of raw and semifnished materials are cheap, product cost is significantly less than external imported product, will help the raising of glaucoma filtration surgery success rate and the extensive popularization of Non Penetrating Trabecular Surgery (NPTS) and carry out, and can reduce patient's medical expense.
Summary of the invention:
The present invention is that raw material prepares hyaluronic acid-chitosan biomembrane as follows with hyaluronic acid, carboxymethyl chitosan, polyvinyl alcohol, butanediol bisglycidyl ether etc.
1 gets purification good aseptic pyrogen-free hyaluronic acid, carboxymethyl chitosan, polyvinyl alcohol, and three's mass ratio is 8: 1: 1, and is soluble in water.
2 by calculating good amount adding cross-linking agent butanediol bisglycidyl ethereal solution, stirs.
Solid content is 40: 1 with the molal weight ratio of butanediol bisglycidyl ether.Solid content herein is meant formed gel-like substance in the solution of butanediol bisglycidyl ether adding step 1.
3 salivation method films.
Dry in 4 vacuum desiccators.
After 5 films were done, sterile razor blade was cut into small pieces.
Soak standby among the PBS of 6 aseptic pH7.4.
This product advantage: the biological membrane biological compatibility with this method development is good, has the effect of fibroblast proliferation under certain inhibition conjunctiva, and degradation process is slow, can not cause tangible inflammatory reaction and immunological rejection, and is with low cost.For suppressing to filter the scarring of passage behind the ophthalmology glaucoma filtration surgery, improving success rate of operation provides a kind of new material.
Description of drawings:
Fig. 1: experimental eye field of operation scanning electron microscope is figure as a result.
Fig. 2: contrast ocular operation domain scanning Electronic Speculum is figure as a result.
Fig. 3: experimental eye field of operation scanning electron microscope is figure as a result.
Fig. 4: contrast ocular operation domain scanning Electronic Speculum is figure as a result.
Fig. 5: experimental eye field of operation scanning electron microscope is figure as a result, amplifies 60,000 times.
Fig. 6: contrast ocular operation domain scanning Electronic Speculum is figure as a result, amplifies 30,000 times.
Fig. 7: experimental eye field of operation scanning electron microscope is figure as a result, amplifies 20,000 times.
Fig. 8: experimental eye field of operation scanning electron microscope is figure as a result, amplifies 100,000 times.
The specific embodiment
Embodiment 1: the preparation of hyaluronic acid-chitosan biomembrane
1 gets the good aseptic apyrogeneity hyaluronic acid 0.4g of purification, carboxymethyl chitosan 0.05g, and polyvinyl alcohol 0.05g is dissolved in the 20ml sterilized water, stirs 2 hours.
2 add cross-linking agent 1% butanediol bisglycidyl ether 1.3ml, stir 20min.
3 sialorrhea methods spread the colloid solution that stirs to 8 * 8cm
2Flat board on.
4 put into 40 ℃ of vacuum desiccators dry 1 day.
After 5 films were done, sterile razor blade was cut into 5 * 5cm
2Size.
The PBS washing of 6 aseptic pH7.4 was soaked 2 days.
7 bottle packing.
Embodiment 2: hyaluronic acid-chitosan biomembrane is to the inhibitory action of fibroblast in-vitro multiplication.
Fibroblast under the In vitro culture lagophthalmos conjunctiva uses diameter to make circular embodiment 1 biological diaphragm as the 6mm trepan.Place ready-formed ring film in 8 culture hole of first row of 96 orifice plates as experimental group, 8 culture hole of secondary series are not placed circular membrane and are organized in contrast, will cultivate the cell in the 4th generation with 5 * 10
3Individual/ml concentration is seeded in 96 orifice plates, selects the capable MTT colorimetric of cell of inoculation back 24h, 48h, 72h, repeats altogether 3 times.MTT colorimetric method result shows that the optical density value of experimental group 24h, 48h and 72h is respectively 0.2071 ± 8.264 * 10
-2, 0.1736 ± 7.963 * 10
-2With 0.1814 ± 4.749 * 10
-2, and matched group is 0.2837 ± 0.1095
-2, 0.2721 ± 8.332 * 10
-2With 0.3070 ± 5.556 * 10
-2, both gaps all have statistical significance (P＜0.05), prove at external synthetic hyaluronic acid-chitosan biomembrane to have the effect that suppresses fibroblast proliferation.
Embodiment 3: hyaluronic acid-chitosan biomembrane filters the effect of passage to the lagophthalmos filtration surgery:
The biomembrane of embodiment 1 preparation is implanted under the scleral flap of lagophthalmos filtration surgery, scanning electron microscope (seeing Fig. 1-8) is found chitosan blend film implantation region sclera bed smooth surface, as seen the sclera flaggy collagen of marshalling, there are a spot of propagation collagen fiber on the surface; The sclera bed rough surface of the contrast eye of implantable bioartificial film not, there are a large amount of propagation collagen fiber on the surface.Transmission electron microscope shows chitosan blend film implantation eye visible inflammatory cell and cell debris around the implantation film, while is dilatation of rough endoplasmic reticula in implanting the other fibroblast of film as seen, part is taken off granule, the synthetic proteins of showed cell and the function of secretory protein are affected, there are a large amount of fat to drip in the part fibroblast, the impaired transformation of showed cell function; As seen contrast eye contains the fibroblast of rough endoplasmic reticulum in a large number at two-layer scleral flap intersection, and sophisticated collagen fiber are arranged on every side.Illustrate that chitosan biomembrane has the effect that is suppressed to the synthetic collagen of fibrocyte.
Claims (1)
1, a kind of method for preparing hyaluronic acid-chitosan biomembrane, this method comprises the steps:
(1) get purification good aseptic pyrogen-free hyaluronic acid, carboxymethyl chitosan, polyvinyl alcohol, three's mass ratio is 8: 1: 1, and is soluble in water,
(2) add cross-linking agent butanediol bisglycidyl ethereal solution by calculating good amount, stir,
Solid content is 40: 1 with the molal weight ratio of butanediol bisglycidyl ether.Solid content herein is meant the butanediol bisglycidyl ether added formed gel-like substance in the solution of step (1),
(3) salivation method film,
It is (4) dry in the vacuum desiccator,
(5) after film was done, sterile razor blade was cut into small pieces,
(6) immersion is standby among the PBS of aseptic pH7.4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610170264 CN1970094A (en) | 2006-12-26 | 2006-12-26 | Process for preparing hyaluronic acid-chitosan biomembrane |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610170264 CN1970094A (en) | 2006-12-26 | 2006-12-26 | Process for preparing hyaluronic acid-chitosan biomembrane |
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| Publication Number | Publication Date |
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| CN1970094A true CN1970094A (en) | 2007-05-30 |
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| CN 200610170264 Pending CN1970094A (en) | 2006-12-26 | 2006-12-26 | Process for preparing hyaluronic acid-chitosan biomembrane |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105266952A (en) * | 2014-07-09 | 2016-01-27 | 首都医科大学附属北京同仁医院 | A device used in anti-glaucoma surgeries for preventing postoperative conjunctiva adhesion |
-
2006
- 2006-12-26 CN CN 200610170264 patent/CN1970094A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105266952A (en) * | 2014-07-09 | 2016-01-27 | 首都医科大学附属北京同仁医院 | A device used in anti-glaucoma surgeries for preventing postoperative conjunctiva adhesion |
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Open date: 20070530 |