CN1120023C - Centrifugal method for preparing cornea collagen film - Google Patents
Centrifugal method for preparing cornea collagen film Download PDFInfo
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- CN1120023C CN1120023C CN 98113956 CN98113956A CN1120023C CN 1120023 C CN1120023 C CN 1120023C CN 98113956 CN98113956 CN 98113956 CN 98113956 A CN98113956 A CN 98113956A CN 1120023 C CN1120023 C CN 1120023C
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- cornea
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- collagen
- collagen film
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Abstract
The present invention relates to a centrifugal method for preparing a cornea collagen film prepared from a collagen sample and a mould. The present invention is characterized in that after the collagen sample and film forming materials, namely polyvinyl alcohol are mixed, the mixture is put into a concave spherical surface arranged on the upper end of the mould, the mould is vertically and fixedly connected with the upper part of a shaft of a rotary device of which the rotating speed is from 200 to 800 revolutions per minute, a shaft of the mould and the shaft of the rotary device are positioned on the same shaft line, and the device is arranged and is dried in a thermotank of which the temperature is 30DEG C. After the device is operated for 1.3 to 2 hours, the mixture is centrifugated and dried into a transparent cornea collagen film of which the center is thin, and the periphery of the film is thick. The anoxia opportunity and the stimulating symptoms of the cornea can be reduced by wearing the film, and the film is favorable for a patient to use.
Description
Technical field: the present invention belongs to processing technique that forms cornea collagen film and the cornea collagen film product made from this technology.
Background technology: the method for producing cornea collagen film at present is more, but the collagem membrane of producing has its deficiency, can not satisfy patient's needs, easily makes the weary oxygen of patient's cornea central authorities after wearing, and edema produces irritation.One, static seasoning: the collagem membrane that this method processes is thicker, the film central thickness is greater than peripheral thickness, inner surface is rough, collagem membrane is worn on the cornea, the weary oxygen chance of cornea central authorities is increased, and two, the encapsulating and stamping method: the collagem membrane that this method forms easily breaks, and film edge is rough, the cutting film formation time is long, and program is loaded down with trivial details.
Summary of the invention: the objective of the invention is to propose a kind of cornea collagen film and method for making thereof that adopts the centrifuging preparation, the collagen that extracts purification is mixed the back add the mould upper end with a certain proportion of filmogen polyvinyl alcohol, place the axle top of rotary apparatus, utilize the action of centrifugal force of rotating shaft, be processed into collagem membrane.The characteristics of the film made from this method are that central authorities are thin, and periphery is thick, and the patient wears the back and reduces weary oxygen chance of cornea and cornea irritation.The present invention adopts the method for centrifugal method for preparing cornea collagen film to be: at first prepare the collagen sample, the mould of preparation cornea collagen film is provided.Get sterile fresh pig sclera, make homogenate, after neutral salt solution extracts, centrifugal 20 minutes, precipitate with distilled water wash, desalination, is put in 0.3% acetic acid solution again, placement added pepsin after 48 hours in 4 ℃ temperature; Ultracentrifugation is 20 minutes after placing 24 hours under 4 ℃ of environment, getting its supernatant stirs with salt, treat fully to separate out collagen solution ultracentrifugation 20 minutes again, collect thick precipitate, place bag filter, in distill water dialysis, obtain the collagen of purification, ultraviolet spectrometry absorption peak 232nm, SDS-PAGE electrophoresis showed α, β, γ bands of a spectrum composition are a large amount of I types, a small amount of III Collagen Type VI; Getting the neutral collagen of PH, to give over to sample standby, the collagen sample added a small amount of filmogen polyvinyl alcohol process cornea collagen film; It is characterized in that: the mixed liquor of collagen sample and filmogen polyvinyl alcohol is inserted in the matrix sphere of mould upper end, mould and revolution are that the top vertical fixing of axle of 200～800 rev/mins rotary apparatus is connected, and the axle that makes mould shaft and rotary apparatus is on same axis, rotary apparatus is positioned at temperature at 30 ℃ thermostatic drying chamber, the axle of rotary apparatus drove mould rotation after 1.3～2 hours, and is centrifugal, be dried to the transparency cornea collagem membrane.The cornea collagen film that makes with this method is the shape that central authorities are thin, periphery is thick.
Operation principle of the present invention: the present invention utilizes action of centrifugal force, forms the cornea collagen film that central authorities are thin, periphery is thick, transparent on forming mould.Its collagen sample is that the raw material of using with other preparation collagem membrane is the same, passes through extraction, purification process, verifies as qualified collagen sample.When the preparation collagem membrane, add the filmogen polyvinyl alcohol again and be mixed into liquid.This mixed liquor is put into the mould upper end matrix sphere that rustless steel is made, mould is fixed on the top of rotary apparatus such as rotating shaft, and vertical the connection, the axis of the axle of assurance device and the axis of mould are on the same axis, could guarantee that like this mould and rotary apparatus rotate on same axis, make matrix sphere interior mixed liquor in mould upper end under the centrifugal action of the axle of rotary apparatus, its revolution is 200～800 rev/mins, in the drying baker of 30 ℃ of constant temperature, centrifugal drying becomes the cornea collagen film that central authorities are thin, periphery is thick, transparent.
The present invention because the method is to adopt action of centrifugal force, directly is connected mould compared with prior art with the axle fixed vertical of rotary apparatus, guarantee in same axis rotation, and select 200～800 rev/mins, prevents that mixed liquor from splashing; Except raw material equally, add the moulding material polyvinyl alcohol again, under centrifugal drying, form the cornea collagen film that central authorities are thin, periphery is thick, transparent.This film film formation time is short, the efficient height, and the film of making is even, smooth, transparent, can change the central authorities and the peripheral thickness of its film by changing the revolution of rotary apparatus, satisfies the demand.After wearing this film, can reduce weary oxygen chance of cornea and cornea irritation, help the patient and use.
The specific embodiment: at first produce collagen sample (raw material); The fresh pig sclera that the cancellation poison is crossed is made homogenate; After the neutral salt solution extraction, centrifugal 20 minutes, precipitate distilled water wash, desalination, put into 0.3% acetic acid solution again, placement added pepsin after 48 hours in 4 ℃ of temperature, and ultracentrifugation is 20 minutes after placing 24 hours under 4 ℃ of environment, gets its supernatant and stirs with salt, treat fully to separate out collagen solution ultracentrifugation 20 minutes again, collect thick precipitate, place bag filter, in distill water dialysis, obtain the collagen of purification, getting the neutral collagen of PH, to make sample standby.Take out collagen solution 150 μ l, add 150 μ l polyvinyl alcohol and mix.Get height that stainless steel material is processed into and be 30 millimeters cylinder, upper surface concave spherical surface radius of curvature is 9 millimeters, 12 millimeters of peripheral linking part chord lengths, and the string of circumferential edges is 14.5 millimeters a mould, mould is vertically fixedlyed connected with the axle upper end of SYB～E infusion pump, and pump revolution is 300 rev/mins.Pump and mould are inserted constant temperature in 30 ℃ drying baker, work after 1.5 hours, can form the cornea collagen film that central authorities are thin, periphery is thick, transparent.
The central authorities and the peripheral thickness of the collagem membrane that centrifuging of the present invention and static method are made are compared as follows table:
| Method | The film number | Central thickness (mm) X ± S | Peripheral thickness (mm) X ± S | Difference L checks t | P |
| Static method | 15 | 0.0391±0.0034 | 0.0244±0.0014 | 15.137 | ＜0.01 |
| ≤ 200 rev/mins of centrifuging≤600 rev/min | 15 15 | 0.0279±0.0005 0.0207±0.0089 | 0.0246±0.0016 0.0317±0.0178 | 9.20 4.31 | ＜0.01 ＜0.01 |
Embodiments of the invention:
At first produce collagen sample (raw material); The fresh pig sclera that the cancellation poison is crossed is made homogenate; After extracting before the neutral salt solution, centrifugal 20 minutes, precipitation distilled water wash, desalination, put into 0.3% acetic acid solution again, place after 48 hours for 4 ℃ and add pepsin, placed 24 hours ultracentrifugations 20 minutes, and got its supernatant and stir for 4 ℃ with salt, treat fully to separate out collagen solution ultracentrifugation 20 minutes again, collect thick precipitate, place bag filter, in distill water dialysis, obtain the collagen of purification, getting the neutral collagen of PH, to make sample standby.Take out collagen solution 150 μ l, add 150 μ l polyvinyl alcohol and mix.Get height that stainless steel material is processed into and be 30 millimeters cylinder, upper surface concave spherical surface radius of curvature is 9 millimeters, 12 millimeters of peripheral linking part chord lengths, the mould that the string of circumferential edges is 14.5 millimeters, axle upper end mould is vertical and the SYB-E infusion pump is fixedly connected, and pump revolution is 300 rev/mins.Pump and mould are inserted constant temperature in 30 ℃ drying baker, work after 1.5 hours, can form the cornea collagen film that central authorities are thin, periphery is thick, transparent.
Claims (3)
1, adopt the method for the cornea collagen film of centrifuging preparation, at first prepare the collagen sample, the mould of preparation cornea collagen film is provided, it is characterized in that the mixed liquor of collagen sample and filmogen polyvinyl alcohol is inserted in the matrix sphere of mould upper end, mould and revolution are that the top vertical fixing of axle of 200～800 rev/mins rotary apparatus is connected, and the axle that makes mould shaft and rotary apparatus is on same axis, rotary apparatus is positioned at temperature at 30 ℃ thermostatic drying chamber, the axle of rotary apparatus drove the mould rotation after 1.3～2 hours, and is centrifugal, be dried to the transparency cornea collagem membrane.
2, adopt the cornea collagen film of centrifuging preparation as claimed in claim 1, it is characterized in that cornea collagen film is the shape that central authorities are thin, periphery is thick.
3, the cornea collagen film of employing according to claim 2 centrifuging preparation as claimed in claim 1, the central thickness that it is characterized in that cornea collagen film is 0.0207 ± 0.0089mm, the peripheral thickness of cornea collagen film is 0.0317 ± 0.0178mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 98113956 CN1120023C (en) | 1998-04-29 | 1998-04-29 | Centrifugal method for preparing cornea collagen film |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 98113956 CN1120023C (en) | 1998-04-29 | 1998-04-29 | Centrifugal method for preparing cornea collagen film |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1234214A CN1234214A (en) | 1999-11-10 |
| CN1120023C true CN1120023C (en) | 2003-09-03 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 98113956 Expired - Fee Related CN1120023C (en) | 1998-04-29 | 1998-04-29 | Centrifugal method for preparing cornea collagen film |
Country Status (1)
| Country | Link |
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| CN (1) | CN1120023C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1314461C (en) * | 2004-02-27 | 2007-05-09 | 四川大学 | Artificial comea having biological activital activity and preparing method |
| CN105412974B (en) * | 2015-11-27 | 2019-03-05 | 广州市朴道联信生物科技有限公司 | A kind of preparation method of double-layer structure cornea repair material |
| CN107213510B (en) * | 2017-05-16 | 2020-05-12 | 暨南大学 | Preparation method of high-strength curvature collagen membrane |
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1998
- 1998-04-29 CN CN 98113956 patent/CN1120023C/en not_active Expired - Fee Related
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| Publication number | Publication date |
|---|---|
| CN1234214A (en) | 1999-11-10 |
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