CN105126084A - Collagen composition and preparation method thereof - Google Patents
Collagen composition and preparation method thereof Download PDFInfo
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- CN105126084A CN105126084A CN201510456175.8A CN201510456175A CN105126084A CN 105126084 A CN105126084 A CN 105126084A CN 201510456175 A CN201510456175 A CN 201510456175A CN 105126084 A CN105126084 A CN 105126084A
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Abstract
The invention provides a collagen composition. The collagen composition comprises an epidermal growth factor, collagen, blood platelet extract and water, wherein mass concentration of the blood platelet extract in the collagen composition is 0.5-1.5mg/mL. The collagen composition has the advantages that by extracting inner components of blood platelets and combining the collagen with the EGF (epidermal growth factor), the collagen composition is capable of achieving hemostasis quickly and promoting wound healing; the collagen composition can be used for hemostasis directly and quickly and is good in effect and capable of enhancing inflammation and immune responses by the aid of the extracted inner components of the blood platelets and is capable of accelerating wound healing by combining the EGF to promote epidermal cell growth; all the ingredients can be degraded in a human body, are free of any toxic or side effect and irritation and have no residue after being degraded.
Description
Technical field
The invention belongs to biomedicine technical field, particularly relate to a kind of collagen composition and preparation method thereof.
Background technology
Collagen protein is biopolymer, is the main component in animal connective tissue, is also the functional protein that mammal in-vivo content is maximum, distribution is the widest, accounts for 25% ~ 30% of total protein.With the transmission of the formation organized, maturation, cell-tocell, and joint lubrication, wound healing, calcification, blood coagulation and aging etc. have close relationship.Collagen protein is also one of critical raw material of biotech industry most, in all extensive application such as medical material, cosmetics, food industry.
In prior art, also exist and collagen protein is combined with some biomaterials the hemorrhage playing anastalsis, the Chinese patent being 200710115108.5 as application number discloses a kind of hemostatic material, its main raw material(s) is chitosan, collagen protein and sodium carboxymethyl cellulose, but, this hemostatic material only has the effect of hemostasis, hemorrhage wound not only be there is no to the effect of Promotive union, also may stimulate damaged hemorrhage wound, and, the chitosan that this hemorrhage is selected and sodium carboxymethyl cellulose are foreign substance, there is certain toxic and side effects.
Summary of the invention
The invention provides a kind of collagen composition and preparation method thereof, collagen composition provided by the invention has the function of hemostasis and bonding wound simultaneously, and has no side effect.
The invention provides a kind of collagen composition, comprising: epithelical cell growth factor, collagen protein, platelet extract and water;
The mass concentration of described platelet extract in described collagen composition is 0.5 ~ 1.5mg/mL.
Preferably, the mass concentration of described epithelical cell growth factor in described collagen composition is 80 ~ 120ng/mL.
Preferably, in described collagen composition, the mass fraction of collagen protein is 40 ~ 50%.
Preferably, described platelet extract obtains according to following steps:
A) umbilical blood is centrifugal, extract platelet;
B) adopt buffer by resuspended for described platelet, obtain thrombocyte suspension, by described thrombocyte suspension multigelation 3 ~ 8 times, obtain platelet freeze thawing liquid;
C) described platelet freeze thawing liquid adopts the rotating speed of 6000 ~ 10000g to carry out centrifugal, obtains platelet extract.
Preferably, described steps A) specifically comprise:
By umbilical blood with the centrifugal 5 ~ 20min of the rotating speed of 200 ~ 500g, get upper plasma;
By upper plasma with the centrifugal 1 ~ 10min of the rotating speed of 200 ~ 500g, get supernatant;
By supernatant with the centrifugal 1 ~ 10min of the rotating speed of 1000 ~ 1500g, obtain platelet.
Preferably, described step B) in freeze thawing comprise freezing and melt; Described freezing temperature is-85 ~-75 DEG C;
The temperature of described thawing is 30 ~ 40 DEG C.
Preferably, described step C) in centrifugal temperature be 1 ~ 10 DEG C;
Described step C) in the centrifugal time be 20 ~ 40min.
Preferably, the pH value of described collagen composition is 6.5 ~ 7.0.
The invention provides a kind of preparation method of collagen composition, comprise the following steps:
1) umbilical blood is centrifugal, extract platelet;
2) adopt buffer by resuspended for described platelet, obtain thrombocyte suspension, by described thrombocyte suspension multigelation 3 ~ 8 times, obtain platelet freeze thawing liquid;
3) described platelet freeze thawing liquid adopts the rotating speed of 6000 ~ 10000g to carry out centrifugal, obtains platelet extract;
4) epithelical cell growth factor, collagen solution and platelet extract are mixed, obtain collagen composition, in described collagen composition, the mass concentration of platelet extract is 0.5 ~ 1.5mg/mL.
Preferably, described step 4) also comprise before:
After collagen protein is mixed with glacial acetic acid, carry out degerming and deactivation successively, obtain collagen solution, and then epithelical cell growth factor, collagen solution and platelet extract are mixed, obtain collagen composition.
The invention provides a kind of collagen composition, comprise epithelical cell growth factor, collagen protein, platelet extract and water, the mass concentration of described platelet extract in described collagen composition is 0.5 ~ 1.5mg/mL.The present invention is by extracting platelet internal component, incorporating collagen albumen and epithelical cell growth factor (EGF), energy quick-acting haemostatic powder and bonding wound, because be extract the platelet internal component obtained, can directly act on hemostasis, speed is fast, effective, strengthening inflammation and immunoreation, promote epidermal growth in conjunction with EGF, accelerating wound.Further, above-mentioned each composition all can at people's vivo degradation, without any side effects, has no stimulation, without any residual after degraded.
Accompanying drawing explanation
In order to be illustrated more clearly in the embodiment of the present invention or technical scheme of the prior art, be briefly described to the accompanying drawing used required in embodiment or description of the prior art below, apparently, accompanying drawing in the following describes is only embodiments of the invention, for those of ordinary skill in the art, under the prerequisite not paying creative work, other accompanying drawing can also be obtained according to the accompanying drawing provided.
Fig. 1 is the Type B wound schematic diagram in the embodiment of the present invention;
Fig. 2 is the Type B wound healing figure that the collagem membrane using the embodiment of the present invention 4 to obtain pastes.
Detailed description of the invention
The invention provides a kind of collagen composition, comprising: epithelical cell growth factor, collagen protein, platelet extract and water,
The mass concentration of described platelet extract in described collagen composition is 0.5 ~ 1.5mg/mL.
Collagen composition provided by the invention comprises epithelical cell growth factor (EGF), described epithelical cell growth factor has another name called oligopeptide-1, it is a kind of active substance in human body, the mass concentration of described epithelical cell growth factor in described collagen composition is preferably 80 ~ 120ng/mL, be more preferably 85 ~ 115ng/mL, most preferably be 90 ~ 110ng/mL.The source of the present invention to described epithelical cell growth factor does not have special restriction, buys commercial goods.As, the recombinant human epidermal growth factor lyophilized powder of 01.ExCell (CB) the .CB031-0041 model that ExCell (CB) company can be adopted to provide.
Collagen composition provided by the invention comprises collagen protein, described collagen protein preferably adopts animal collagen, the mass fraction of described collagen protein in described collagen composition is preferably 40 ~ 50%, is more preferably 43 ~ 48%, most preferably is 45 ~ 46%.The source of the present invention to described collagen protein does not have special restriction, adopts commercial goods, e.g., and the animal collagen of the QB2732-2005 model that bio tech ltd of Xu Er U.S., Shijiazhuang can be adopted to provide.
Collagen composition provided by the invention comprises platelet extract, and the mass concentration of described platelet extract in described collagen composition is 0.5 ~ 1.5mg/mL, is preferably 0.8 ~ 1.3mg/mL, is more preferably 1 ~ 1.1mg/mL.
α granule and sensitization granule are contained, fibrinogen, platelet factor 4, cathepsin A, cathepsin D, acid hydrolase etc. in α granule in platelet inside; Sensitization granule contains 5-hydroxy tryptamine, ADP, ATP, calcium ion, epinephrine, antiplasmin, pyrophosphoric acid etc.These factor component produce thrombin by complicated change, are that the Fibrinogen in blood becomes fibrin, and the fibrin interweaved mutually makes platelet plug tangle into blood clot with hemocyte, i.e. thrombosis.Along with the formation of thrombosis, intra platelet free calcium TXA2.; Dense granule and α granule are by discharging the various active materials such as ADP, 5-hydroxy tryptamine, platelet factor 4, beta-thromboglobulin, thrombospondin, cell growth factor, blooc coagulation factor V, VII, Ⅻ and vascular permeability factor with surface connecting system (SCS), these active substances are by activating other platelet in blood, promote vasoconstriction, the various ways such as short fibrin formation strengthen hemostasis.In the present invention, the protein ingredient of the described platelet extract mainly all kinds of factor.
In the present invention, described platelet extract preferably obtains according to following steps:
A) umbilical blood is centrifugal, extract platelet;
B) adopt buffer by resuspended for described platelet, obtain thrombocyte suspension, by described thrombocyte suspension multigelation 3 ~ 8 times, obtain platelet freeze thawing liquid;
C) described platelet freeze thawing liquid adopts the rotating speed of 6000 ~ 10000g to carry out centrifugal, obtains platelet extract.
The present invention is centrifugal by umbilical blood, extract obtain platelet, the present invention preferably by umbilical blood with the centrifugal 5 ~ 20min of the rotating speed of 200 ~ 500g, get upper plasma; By upper plasma with the centrifugal 1 ~ 10min of the rotating speed of 200 ~ 500g, get supernatant; By supernatant with the centrifugal 1 ~ 10min of the rotating speed of 1000 ~ 1500g, obtain platelet; The present invention more preferably with the centrifugal 10 ~ 15min of the rotating speed of 300 ~ 400g, gets upper plasma; By upper plasma with the centrifugal 5 ~ 7min of the rotating speed of 300 ~ 400g, get supernatant; By supernatant with the centrifugal 5 ~ 7min of the rotating speed of 1200 ~ 1300g, obtain platelet.
After obtaining platelet, the present invention adopts buffer by resuspended for described platelet, obtains thrombocyte suspension, and in the present invention, described buffer solution is preferably phosphate buffered solution, and the pH value of described buffer solution is preferably 7 ~ 8, is more preferably 7.2 ~ 7.5; The consumption of the present invention to described buffer solution does not have special restriction.In the present invention, described resuspended be technological means well known to those skilled in the art.
The present invention is by the thrombocyte suspension multigelation that obtains 3 ~ 8 times, obtain platelet freeze thawing liquid, the present invention is preferably by described thrombocyte suspension freeze thawing 4 ~ 5 times, obtain platelet freeze thawing liquid, described freeze thawing comprises freezing and melts, described freezing temperature is preferably-85 ~-75 DEG C, is more preferably-80 DEG C; The temperature of described thawing is preferably 30 ~ 40 DEG C, is more preferably 35 ~ 37 DEG C.
The platelet freeze thawing liquid obtained carries out centrifugal with the rotating speed of 6000 ~ 10000g by the present invention, obtain platelet extract, the platelet freeze thawing liquid obtained preferably carries out centrifugal with the rotating speed of 7000 ~ 8000g by the present invention, to remove platelet membrane and other cell debriss, then get supernatant, described supernatant is platelet extract.In the present invention, the described time centrifugal to platelet freeze thawing liquid is preferably 20 ~ 40min, is more preferably 30 ~ 35min, and the described temperature centrifugal to platelet freeze thawing liquid is preferably 1 ~ 10 DEG C, is more preferably 4 ~ 8 DEG C.After obtaining platelet extract, the present invention is preferably frozen for subsequent use at-80 DEG C.
In the present invention, described platelet extract is the protein ingredient of all kinds of factor in platelet mainly, and described platelet extract can directly act on hemostasis, and anthemorrhagic speed is fast, effective.
Present invention also offers a kind of preparation method of collagen composition, comprise the following steps:
1) umbilical blood is centrifugal, extract platelet;
2) adopt buffer by resuspended for described platelet, obtain thrombocyte suspension, by described thrombocyte suspension multigelation 3 ~ 8 times, obtain platelet freeze thawing liquid;
3) described platelet freeze thawing liquid adopts the rotating speed of 6000 ~ 10000g to carry out centrifugal, obtains platelet extract;
4) epithelical cell growth factor, collagen solution and platelet extract are mixed, obtain collagen composition, in described collagen composition, the mass concentration of platelet extract is 0.5 ~ 1.5mg/mL.
In the present invention, described step 1), 2), 3) for extracting the preparation method of platelet extract from umbilical blood, the method is consistent with the preparation method of platelet extract in technique scheme, does not repeat them here.
After obtaining platelet extract, epithelical cell growth factor, collagen solution and platelet extract mix by the present invention, obtain collagen composition, collagen protein preferably mixes with glacial acetic acid by the present invention, carry out degerming and deactivation successively, obtain collagen solution, then epithelical cell growth factor, collagen solution and platelet extract are mixed, obtain collagen composition.In the present invention, the mass concentration of described glacial acetic acid is preferably 0.1 ~ 0.5%, and the consumption of the present invention to described glacial acetic acid is not particularly limited.The filter screen that the mixture of described collagen protein and glacial acetic acid is preferably crossed 0.22 μm by the present invention is degerming, and then constant temperature 1 ~ 2min at 39 ~ 41 DEG C, carries out deactivation, obtain collagen solution.
The present invention is preferably existing is dissolved in collagen solution by described platelet extract, and then is dissolved in above-mentioned mixed liquor by epithelical cell growth factor solution, obtains collagen composition.In the present invention, described epithelical cell growth factor solution is the mixed liquor of epithelical cell growth factor and phosphate buffered solution.The mass concentration of described phosphate buffered solution is preferably 0.1 ~ 0.5%, and consumption does not do particular restriction.
After obtaining collagen composition, in order to make it be more suitable for human body environment, the present invention preferably adds alkaline medium wherein and its pH value is adjusted to 6.5 ~ 7.0, and described collagen composition forms colloid simultaneously.In the present invention, described alkaline medium is preferably sodium hydroxide solution, and the molar concentration of described alkaline medium is preferably 1 ~ 2mol/L, and the process of above-mentioned adjust ph is preferably carried out under the constant temperature of 1 ~ 10 DEG C, is more preferably 4 ~ 5 DEG C.
The mass concentration of the collagen composition mesocuticle cell growth factor that the present invention finally obtains is 80 ~ 120ng/mL; In collagen composition, the mass fraction of collagen protein is 40 ~ 50%; The mass concentration of platelet extract in described collagen composition is 0.5 ~ 1.5mg/mL, and the mass fraction of water is 50 ~ 60%.
After obtaining collagen composition, the present invention is coated on medical pressure-sensing adhesive plaster, 10cm
2circular area in coating 2mL collagen composition, sterile sealing, obtains collagem membrane and pastes.
In order to ensure the result of use that described collagen composition and collagem membrane paste, the present invention preferably aseptically completes described collagen composition and collagem membrane pastes preparation, more preferably completes in the superclean bench of ten thousand grades of clean areas.
The invention provides a kind of collagen composition, comprise epithelical cell growth factor, collagen protein and platelet extract, the mass concentration of described platelet extract in described collagen composition is 0.5 ~ 1.5mg/mL.The present invention is by extracting platelet internal component, incorporating collagen albumen and epithelical cell growth factor (EGF), energy quick-acting haemostatic powder and bonding wound, because be extract the platelet internal component obtained, can directly act on hemostasis, speed is fast, effective, strengthening inflammation and immunoreation, promote epidermal growth in conjunction with EGF, accelerating wound.Further, above-mentioned each composition all can at people's vivo degradation, without any side effects, has no stimulation, without any residual after degraded.
In order to further illustrate the present invention, below in conjunction with embodiment, a kind of collagen composition provided by the invention and preparation method thereof being described in detail, but can not limiting the scope of the present invention being understood as.
In the examples below, epithelical cell growth factor is the recombinant human epidermal growth factor lyophilized powder of 01.ExCell (CB) the .CB031-0041 model that ExCell (CB) company provides; The animal collagen of the QB2732-2005 model that collagen protein provides for bio tech ltd of Xu Er U.S., Shijiazhuang.
Embodiment 1
100ml umbilical blood, the centrifugal 10min of 400g, extracts upper plasma;
Upper plasma, the centrifugal 5min of 400g, extracts supernatant;
The centrifugal 5min of supernatant 1200g, removes supernatant liquid, obtains platelet.
Resuspended with 10mL phosphate buffer (pH7.2), by the continuous multigelation of thrombocyte suspension (-80 DEG C/37 DEG C) 5 times obtained, then the centrifugal 30min of 8000g is to remove platelet membrane and other cell debris at 4 DEG C, and the supernatant-80 DEG C obtaining 2mL mass concentration 2g/mL is frozen for subsequent use.
100 μ g recombinant human epidermal growth factor lyophilized powders, mix with the PBS of 2ml ,-80 DEG C for subsequent use.
Animal collagen is dissolved in 0.1% glacial acetic acid, makes solution, 0.22 μm of strainer filtering is degerming, constant temperature 1min at 39 DEG C.
Thaw EGF solution and platelet supernatant fluid, mixes to collagen solution the EGF solution of 0.8mL and 0.25ml platelet extract phosphate buffer to 500mL;
It is 6.5 plastics that the NaOH solution slowly adding 1mol/L under 4 DEG C of constant temperatures is adjusted to PH, obtains collagen composition.In the collagen composition obtained, EGF is 80ng/ml, platelet extract 0.5mg/ml, and collagen protein quality percentage ratio is 40%, moisture 60%.
Get the medical pressure-sensing adhesive plaster of sterilization treatment, 10cm
2circular area in add 2mL finished gel, aseptic sealed packages obtains collagem membrane and pastes.
Embodiment 2
100ml umbilical blood, the centrifugal 10min of 400g, extracts upper plasma;
Upper plasma, the centrifugal 5min of 400g, extracts supernatant;
The centrifugal 5min of supernatant 1200g, removes supernatant liquid, obtains platelet.
Resuspended with 10mL phosphate buffer (pH7.2), by the continuous multigelation of thrombocyte suspension (-80 DEG C/37 DEG C) 5 times obtained, then the centrifugal 30min of 8000g is to remove platelet membrane and other cell debris at 4 DEG C, and the supernatant-80 DEG C obtaining 2mL mass concentration 2g/mL is frozen for subsequent use.
100 μ g recombinant human epidermal growth factor lyophilized powders, mix with the PBS of 2ml ,-80 DEG C for subsequent use.
Animal collagen is dissolved in 0.1% glacial acetic acid, makes solution, 0.22 μm of strainer filtering is degerming, constant temperature 1min at 39 DEG C.
Thaw EGF solution and platelet supernatant fluid, mixes to collagen solution the EGF solution of 1.2mL and 0.75mL platelet extract phosphate buffer to 500mL;
It is 6.5 plastics that the NaOH solution slowly adding 1mol/L under 4 DEG C of constant temperatures is adjusted to PH, obtains collagen composition.In the collagen composition obtained, EGF is 120ng/ml, platelet extract 1.5mg/ml, and collagen protein quality percentage ratio is 50%, moisture 50%.
Get the medical pressure-sensing adhesive plaster of sterilization treatment, 10cm
2circular area in add 2mL finished gel, aseptic sealed packages obtains collagem membrane and pastes.
Embodiment 3
100ml umbilical blood, the centrifugal 10min of 400g, extracts upper plasma;
Upper plasma, the centrifugal 5min of 400g, extracts supernatant;
The centrifugal 5min of supernatant 1200g, removes supernatant liquid, obtains platelet.
Resuspended with 10mL phosphate buffer (pH7.2), by the continuous multigelation of thrombocyte suspension (-80 DEG C/37 DEG C) 5 times obtained, then the centrifugal 30min of 8000g is to remove platelet membrane and other cell debris at 4 DEG C, and the supernatant-80 DEG C obtaining 2mL mass concentration 2g/mL is frozen for subsequent use.
100 μ g recombinant human epidermal growth factor lyophilized powders, mix with the PBS of 2ml ,-80 DEG C for subsequent use.
Animal collagen is dissolved in 0.1% glacial acetic acid, makes solution, 0.22 μm of strainer filtering is degerming, constant temperature 1min at 39 DEG C.
Thaw EGF solution and platelet supernatant fluid, mixes to collagen solution the EGF solution of 1mL and 0.5mL platelet extract phosphate buffer to 500mL;
It is 6.5 plastics that the NaOH solution slowly adding 1mol/L under 4 DEG C of constant temperatures is adjusted to PH, obtains collagen composition.In the collagen composition obtained, EGF is 100ng/ml, platelet extract 1mg/ml, and collagen protein quality percentage ratio is 45%, moisture 55%.
Get the medical pressure-sensing adhesive plaster of sterilization treatment, 10cm
2circular area in add 2mL finished gel, aseptic sealed packages obtains collagem membrane and pastes.
Embodiment 4
100ml umbilical blood, the centrifugal 10min of 400g, extracts upper plasma;
Upper plasma, the centrifugal 5min of 400g, extracts supernatant;
The centrifugal 5min of supernatant 1200g, removes supernatant liquid, obtains platelet.
Resuspended with 10mL phosphate buffer (pH7.2), by the continuous multigelation of thrombocyte suspension (-80 DEG C/37 DEG C) 5 times obtained, then the centrifugal 30min of 8000g is to remove platelet membrane and other cell debris at 4 DEG C, and the supernatant-80 DEG C obtaining 2mL mass concentration 2g/mL is frozen for subsequent use.
100 μ g recombinant human epidermal growth factor lyophilized powders, mix with the PBS of 2ml ,-80 DEG C for subsequent use.
Animal collagen is dissolved in 0.1% glacial acetic acid, makes solution, 0.22 μm of strainer filtering is degerming, constant temperature 1min at 39 DEG C.
Thaw EGF solution and platelet supernatant fluid, mixes to collagen solution the EGF solution of 1.1mL and 0.6mL platelet extract phosphate buffer to 500mL;
It is 6.5 plastics that the NaOH solution slowly adding 1mol/L under 4 DEG C of constant temperatures is adjusted to PH, obtains collagen composition.In the collagen composition obtained, EGF is 110ng/ml, platelet extract 1.2mg/ml, and collagen protein quality percentage ratio is 50%, moisture 50%.
Get the medical pressure-sensing adhesive plaster of sterilization treatment, 10cm
2circular area in add 2mL finished gel, aseptic sealed packages obtains collagem membrane and pastes.
Comparative example 1
The chitose haemostatic of acquisition is prepared according to the publication number Chinese patent that is CN104114178A.
Application examples
Get 2 weeks mices 10, carry out back unhairing, 5 to carry out 1cm dark, and the wound model process of 2cm long back, is designated as A type wound; 5 are carried out diameter 2cm, and the epidermis of degree of depth 2mm is removed, and be designated as Type B wound, as shown in Figure 1, Fig. 1 is the Type B wound schematic diagram in the embodiment of the present invention.
Collagem membrane in Example 1 ~ 4 pastes and hemorrhage in comparative example 1 and binding agent process wound respectively, within every 4 hours, observes once, calculating wound healing time and between the blood stream stopping time.Result as shown in table 1 and Fig. 2, table 1 be the product that the embodiment of the present invention 1 ~ 4 and comparative example 1 obtain wound healing time and between the blood stream stopping time, Fig. 2 is the Type B wound healing figure using the collagem membrane that obtains of the embodiment of the present invention 4 to paste.
The wound healing time of the product that table 1 embodiment of the present invention 1 ~ 4 and comparative example 1 obtain and between the blood stream stopping time
| Comparative example 1 | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 |
| A type wound healing time | 5 days 8 hours | 3 days 12 hours | 3 days 4 hours | 3 days 8 hours | 3 days 4 hours |
| The Type B wound hemostasis time | 65 seconds | 45 seconds | 38 seconds | 42 seconds | 38 seconds |
| Type B wound healing time | 10 days 12 hours | 6 days 12 hours | 5 days 20 hours | 6 days 4 hours | 5 days 16 hours |
As can be seen from Table 1, for category-A wound, the speed of wound healing that the collagem membrane that employing the embodiment of the present invention 1 ~ 4 provides pastes is obviously faster than comparative example 1, and the speed of wound healing of embodiment 2 and embodiment 4 is the fastest.
For category-B wound, the anthemorrhagic speed that the collagem membrane that employing the embodiment of the present invention 1 ~ 4 provides pastes and speed of wound healing are obviously faster than comparative example 1, and the anthemorrhagic speed of embodiment 2 and embodiment 4 is the fastest, and the wound healing of embodiment 4 is consuming time the shortest.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. a collagen composition, comprising: epithelical cell growth factor, collagen protein, platelet extract and water;
The mass concentration of described platelet extract in described collagen composition is 0.5 ~ 1.5mg/mL.
2. collagen composition according to claim 1, is characterized in that, the mass concentration of described epithelical cell growth factor in described collagen composition is 80 ~ 120ng/mL.
3. collagen composition according to claim 1, is characterized in that, in described collagen composition, the mass fraction of collagen protein is 40 ~ 50%.
4. collagen composition according to claim 1, is characterized in that, described platelet extract obtains according to following steps:
A) umbilical blood is centrifugal, extract platelet;
B) adopt buffer by resuspended for described platelet, obtain thrombocyte suspension, by described thrombocyte suspension multigelation 3 ~ 8 times, obtain platelet freeze thawing liquid;
C) described platelet freeze thawing liquid adopts the rotating speed of 6000 ~ 10000g to carry out centrifugal, obtains platelet extract.
5. collagen composition according to claim 4, is characterized in that, described steps A) specifically comprise:
By umbilical blood with the centrifugal 5 ~ 20min of the rotating speed of 200 ~ 500g, get upper plasma;
By upper plasma with the centrifugal 1 ~ 10min of the rotating speed of 200 ~ 500g, get supernatant;
By supernatant with the centrifugal 1 ~ 10min of the rotating speed of 1000 ~ 1500g, obtain platelet.
6. collagen composition according to claim 4, is characterized in that, described step B) in freeze thawing comprise freezing and melt; Described freezing temperature is-85 ~-75 DEG C;
The temperature of described thawing is 30 ~ 40 DEG C.
7. collagen composition according to claim 4, is characterized in that, described step C) in centrifugal temperature be 1 ~ 10 DEG C;
Described step C) in the centrifugal time be 20 ~ 40min.
8. collagen composition according to claim 1, is characterized in that, the pH value of described collagen composition is 6.5 ~ 7.0.
9. a preparation method for collagen composition, comprises the following steps:
1) umbilical blood is centrifugal, extract platelet;
2) adopt buffer by resuspended for described platelet, obtain thrombocyte suspension, by described thrombocyte suspension multigelation 3 ~ 8 times, obtain platelet freeze thawing liquid;
3) described platelet freeze thawing liquid adopts the rotating speed of 6000 ~ 10000g to carry out centrifugal, obtains platelet extract;
4) epithelical cell growth factor, collagen solution and platelet extract are mixed, obtain collagen composition, in described collagen composition, the mass concentration of platelet extract is 0.5 ~ 1.5mg/mL.
10. preparation method according to claim 9, is characterized in that, described step 4) also comprise before:
After collagen protein is mixed with glacial acetic acid, carry out degerming and deactivation successively, obtain collagen solution, and then epithelical cell growth factor, collagen solution and platelet extract are mixed, obtain collagen composition.
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| CN108272529A (en) * | 2018-01-29 | 2018-07-13 | 孟国路 | A kind of tack dura mater sticking patch and preparation method thereof |
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