CN104940982A - Preparation method of dressing skin - Google Patents
Preparation method of dressing skin Download PDFInfo
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Abstract
The invention discloses a preparation method of a dressing skin. The preparation method comprises the following steps of: removing hairs of xenoskin raw materials, and scraping to remove fat and a cuticle so as to obtain a xenoskin material with the thickness of 0.3-0.5 mm; sequentially immersing by using NaCl solution, cleaning by using sterile water, immersing by using neutral protease with the mass fraction of 0.2-0.5% at 37 DEG C, and cleaning by using sterile water so as to obtain a xenoskin; and inactivating the xenoskin so as to obtain the dressing skin, wherein the inactivating step comprises the steps of heating the xenoskin at 50-60 DEG C for 60-120 MIN and irradiating the xenoskin by using gamma rays. The preparation method of the dressing skin is simple to operate and low in cost; four kinds of viruses including PRV (Pseudorabies Virus), VSV (Vesicular Stomatitis Virus), PV1 (Poliovirus 1) and PPV (Porcine Parvovirus) can be completely inactivated; the virus inactivating efficiency is obviously increased; the problem that PV1 cannot be completely inactivated only by irradiating gamma rays can be solved; and therefore, the safety of the dressing skin is increased.
Description
Technical field
The present invention relates to animal derived biological dressing Viral inactivation techniques field, particularly relate to a kind of preparation method of dressing skin.
Background technology
Existing for burning, the xenogenesis skin that adopts of heating power wound or chemical injury is often for Corii Sus domestica.Corii Sus domestica is similar to the skin texture of people, it can stick on wound surface preferably, play the effect reducing the evaporation of wound surface moisture, reduce electrolyte loss, intercept antibacterial and infection control, clinical effectiveness is good, be a kind of good alternative allograft skin, obtain applying more widely in burn, scald field.Corii Sus domestica is as a kind of animal derived Wound dressing skin, and its viral carrying amount must strictly be controlled.Therefore, the selection of virus inactivation technology is most important.At present, burn and scald Corii Sus domestica mainly utilizes gamma-rays (gamma ray) irradiation to carry out inactivation of virus.
According to the regulation of national Bureau of Drugs Supervision associated documents, often walking in inactivation step, viral reducing amount reaches 4Logs (i.e. LgTCID
50/ ml) more than, think that inactivation of viruses method is effective; And each step can superpose the deactivation of virus.Virus reducing amount is larger, and blind passage three generations is virus-free to be detected, and illustrate that inactivation technology effect is better, the safety of dressing skin is higher.Research finds, although utilize gamma-ray irradiation that Corii Sus domestica virus reducing amount can be made to reach more than 4Logs, think that the method is effective, but all there is cytopathy containing adding cell blind passage three generations after poliovirus sabinI type virus (PV1) sample irradiation, illustrating that PV1 inactivation of virus is thorough not.For improving the security performance of dressing skin, should manage to improve inactivation of virus efficiency further.
Summary of the invention
Based on this, be necessary for the not high problem of inactivation of virus efficiency, the preparation method of a kind of inactivation of virus efficiency and the high dressing skin of safety is provided.
A preparation method for dressing skin, comprises the following steps:
Xenogenesis skin raw material is lost hair or feathers, scrapes degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.3 ~ 0.5mm;
By described xenogenesis skin material successively with NaCl solution immersion, sterile water wash, with the mass fractions of 37 DEG C be 0.2 ~ 0.5% neutral protease solution soaking, sterile water wash obtain xenogenesis skin; And
Described xenogenesis skin is carried out inactivation treatment and obtains dressing skin, described inactivation treatment comprises: heated 60 ~ 120 minutes at 50 ~ 60 DEG C by described xenogenesis skin, and by described xenogenesis skin gamma-ray irradiation.
Wherein in an embodiment, the concentration of described NaCl solution is 0.5 ~ 2mol/L, and the time that described NaCl solution is soaked is 12 ~ 24h.
Wherein in an embodiment, the time of described neutral protease solution soaking is 2 ~ 6h.
Wherein in an embodiment, describedly by the concrete operations that xenogenesis skin heats 60 ~ 120 minutes at 50 ~ 60 DEG C be: xenogenesis skin is placed in agitator, and in water bath with thermostatic control, temperature control heats 60 ~ 120 minutes at 50 ~ 60 DEG C.
Wherein in an embodiment, the frequency of oscillation of described agitator is 100 ~ 110rpm.
Wherein in an embodiment, the time of described heating is 90 ~ 120min.
Wherein in an embodiment, described gamma-rays is produced by radiosiotope cobalt-60, and irradiation dose is 15 ~ 25kGy.
Wherein in an embodiment, described irradiation dose is 25kGy.
Wherein in an embodiment, described xenogenesis skin physiological saline solution before inactivation treatment obtains dressing skin, washs by described being carried out by described xenogenesis skin.
Wherein in an embodiment, the concentration of described NaCl solution is 1mol/L, and the time that described NaCl solution is soaked is 20h, and the mass fraction of described neutral protein enzymatic solution is 0.35%, and the time of described neutral protease solution soaking is 4h.
The preparation method of above-mentioned dressing skin is simple to operate, with low cost, can complete inactivation PRV, VSV, PV1 and PPV tetra-kinds of viruses, compared with the virus inactivation technology of single gamma-ray irradiation, the preparation method inactivation of virus efficiency of above-mentioned dressing skin significantly improves, the dressing skin PRV obtained, VSV, PV1 and PPV tetra-kinds of Viral diagnosis are all qualified, solving independent gamma-ray irradiation can not the problem of complete inactivation PV1 virus, improves the safety of dressing skin.
Detailed description of the invention
For the ease of understanding the present invention, below the present invention is described more fully.And give and of the present inventionly preferably to test.But the present invention can realize in many different forms, is not limited to test described herein.On the contrary, the object providing these to test be make the understanding of disclosure of the present invention more comprehensively thorough.
Unless otherwise defined, all technology used herein and scientific terminology are identical with belonging to the implication that those skilled in the art of the present invention understand usually.The object of term used in the description of the invention herein just in order to describe concrete test, is not intended to be restriction the present invention.Term as used herein " and/or " comprise arbitrary and all combinations of one or more relevant Listed Items.
The preparation method of the dressing skin of one embodiment, comprises the following steps:
S100: xenogenesis skin raw material is lost hair or feathers, scrapes degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.3 ~ 0.5mm.
Preferably, the thickness of xenogenesis skin material is 0.3mm.
Preferably, xenogenesis skin raw material is Corii Sus domestica raw material, Corii Sus domestica is similar to the skin texture of people, it can stick on wound surface preferably, play the effect reducing the evaporation of wound surface moisture, reduce electrolyte loss, intercept antibacterial and infection control, clinical effectiveness is good, is a kind of good alternative allograft skin, obtains applying more widely in burn, scald field.
Corii Sus domestica is as a kind of animal derived Wound dressing skin, and its viral carrying amount must strictly be controlled.Therefore, the selection of virus inactivation technology is most important.According to " No. [2002] 160, traditional Chinese medicines prison note ", " No. [2008] 7, state's food medicine prison note " file regulation, select following four kinds of indicator viruses.
Pseudorabies virus (PRV), belongs to herpetoviridae, for double-stranded DNA has after birth virus.To the fatsolvent such as ether, chloroform, the sensitivity such as formalin and ultraviolet radiation is the herpesvirus that a species resistance is stronger.
Vesicular stomatitis virus (VSV), belongs to Rhabdoviridae, for strand RNA has after birth virus.Not acidproof, visible ray, ultraviolet and fatsolvent can make its deactivation, similar to foot and mouth disease virus to the resistance of the Physicochemical factor.
Poliovirus sabinI type (PV1), belongs to enterovirus section, for Microrna is without after birth virus.Due to without after birth, to physical and chemical factor better resistance.
Pig parvoviral (PPV), belongs to Parvoviridae, for small DNA is without after birth virus.Due to without after birth, extremely strong to physical and chemical factor resistance.
Virus reducing amount is represented by R, virus titer after the titre-deactivation of R=deactivation provirus.Virus reducing amount R >=4Logs, judges that this step is effective.Virus complete inactivation, after referring to inactivation of virus, the reducing amount of viral load reaches more than 4Logs, and blind passage three generations is virus-free detects.
S200: by xenogenesis skin material successively with NaCl solution immersion, sterile water wash, with the mass fractions of 37 DEG C be 0.2 ~ 0.5% neutral protease solution soaking, sterile water wash obtain xenogenesis skin.
Especially, the concentration of NaCl solution is 0.5 ~ 2mol/L, and the time that NaCl solution is soaked is 12 ~ 24h.
Especially, neutral egg enzymatic solution is aqueous solution.Especially, the time of neutral protease solution soaking is 2 ~ 6h.
Preferably, the concentration of NaCl solution is 1mol/L, and the time that NaCl solution is soaked is 20h, and the mass fraction of neutral protein enzymatic solution is 0.35%, and the time of neutral protease solution soaking is 4h.
Sterilized water is microorganism in water is killed or filtered out by high-temperature steam method, chemical method, ozone approach or physical filtering method the water obtained.Particularly, in the present embodiment, sterilized water is the water adopting the process of hollow fiber ultrafiltration membrane physical filtering method to obtain.
Especially, the number of times of sterile water wash is 2 ~ 4 times.Adopt sterile water wash, the chips such as Pilus Sus domestica are removed clean.Especially, sterile water wash can adopt ultrasonic waves for cleaning.Preferably, the time of each cleaning is 10 minutes.
Step S100 and step S200 is by xenogenesis skin raw material through bark fetching, and de-cell process, intactly saves the structure of epidermis.
S300: heated 60 ~ 120 minutes at 50 ~ 60 DEG C by xenogenesis skin, obtains the xenogenesis skin of the preliminary deactivation of virus.
Xenogenesis skin is carried out before inactivation treatment obtains dressing skin, xenogenesis skin physiological saline solution is washed.Physiological saline solution is obtained through high pressure steam sterilization by the normal saline configured.The mass fraction of physiological saline solution is 0.85 ~ 0.9%.
Be: xenogenesis skin is placed in agitator that temperature control heats 60 ~ 120 minutes at 50 ~ 60 DEG C in water bath with thermostatic control by the concrete operations that xenogenesis skin heats 60 ~ 120 minutes at 50 ~ 60 DEG C.
More particularly, the frequency of oscillation of agitator is 100 ~ 110rpm (rev/min).
Adopt the xenogenesis skin after step S300 process, the reducing amount of four kinds of indicator virus quantity all reaches more than 4Logs, illustrates that employing step S300 can effective above-mentioned four kinds of indicator viruses of deactivation.
It is worth mentioning that, the time of heating can make the reducing amount of PRV, VSV and PV1 tri-kinds of viral loads added reach more than 4Logs more than 5min, namely this step is effective for the deactivation of PRV, VSV and PV1 virus.Can there be after birth virus the time of heating by PRV and VSV two kinds that add of complete inactivation more than 15min.
The time of heating, when being 60 ~ 120min, gets final product PRV, VSV and PV1 tri-kinds of viruses that complete inactivation adds.Preferred, the time of heating is 90 ~ 120min.PPV virus, after one step S300 deactivation, although the reducing amount of PPV viral load reaches more than 4Logs, thinks that step S300 is effective for PPV inactivation of virus; But blind passage three generations virus still has activity.Therefore single step S300 mono-aspect can by PRV, VSV and PV1 tri-kinds of viral complete inactivations, and existing on the other hand can not the shortcoming of complete inactivation PPV virus.
S400: by the xenogenesis skin of preliminary for virus deactivation, with gamma-ray irradiation, obtain dressing skin.
Preferably, by the xenogenesis skin of preliminary for virus deactivation using physiological saline solution as conserving liquid, be placed on sealing in aluminum foil sack and preserve to carry out gamma-ray irradiation.
Gamma-rays is produced by radiosiotope cobalt-60.Especially, irradiation dose is 15 ~ 25kGy.Irradiation dose is excessive, causes other material degeneration such as the protein of xenogenesis skin, therefore controls can make in the zone of reasonableness of inactivation of virus.Preferably, irradiation dose is 25kGy.
After antibacterial in xenogenesis skin is subject to gamma-rays irradiation, physiological dysfunction, failure to thrive, even dead.
Step S400 can by PRV, VSV and PPV tri-kinds of viral complete inactivations, and compensate for step S300 can not the shortcoming of complete inactivation PPV virus.In addition, PV1 virus, after one step S400 deactivation, although the reducing amount of PV1 viral load reaches more than 4Logs, thinks that step S400 is also effective for PV1 inactivation of virus; But blind passage three generations virus still has activity.Therefore single step S400 mono-aspect can by PRV, VSV and PPV tri-kinds of viral complete inactivations, and existing on the other hand can not the shortcoming of complete inactivation PV1 virus.
Therefore, the preparation method of above-mentioned dressing skin, can complete inactivation PRV, VSV, PV1 and PPV tetra-kinds of viruses.Be appreciated that the order of step S300 and step S400 can exchange, by xenogenesis skin gamma-ray irradiation, obtain the xenogenesis skin of the preliminary deactivation of virus; The xenogenesis skin of preliminary for virus deactivation is heated to 50 ~ 60 DEG C, and the time of heating is 60 ~ 120 minutes, obtains dressing skin.
Before above-mentioned dressing skin uses, install with Aluminium Foil Package the water putting into 35 ~ 45 DEG C and soak 20 minutes, carry out rewarming; After conventional for wound surface debridement, take out under aseptic condition and be covered in wound surface, routine is wrapped up and is changed dressings.
The preparation method of above-mentioned dressing skin is simple to operate, with low cost, can complete inactivation PRV, VSV, PV1 and PPV tetra-kinds of viruses, and four kinds of viral reducing amounts are respectively: the reducing amount R >=13.76Logs of PRV virus; Reducing amount R >=the 12.25Logs of VSV virus; Reducing amount R >=the 12.01Logs of PV1 virus; Reducing amount R >=the 10.75Logs of PPV virus.Compared with the virus inactivation technology of single gamma-ray irradiation, the preparation method inactivation of virus efficiency of above-mentioned dressing skin significantly improves, the dressing skin PRV obtained, VSV, PV1 and PPV tetra-kinds of Viral diagnosis are all qualified, solving independent gamma-ray irradiation can not the problem of complete inactivation PV1 virus, improves the safety of dressing skin.
It is below specific embodiment.
The xenogenesis skin raw material that embodiment 1 ~ 8 and comparative example 1 adopt is Corii Sus domestica raw material.
Embodiment 1
Xenogenesis skin raw material is lost hair or feathers, scrape degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.3mm, by xenogenesis skin material successively at room temperature with the NaCl solution of 1mol/L soak 20h, with sterile water wash, with the mass fractions of 37 DEG C be 0.35% neutral protease solution soaking 4h, then obtain xenogenesis skin by sterile water wash.
Xenogenesis skin is placed in agitator, and the frequency of oscillation of agitator is 100rpm.Temperature control to 60 DEG C in water bath with thermostatic control, heated after 120 minutes, obtained the xenogenesis skin of the preliminary deactivation of virus.By the xenogenesis skin of preliminary for virus deactivation, be the gamma-ray irradiation of 25kGy with irradiation dose, obtain dressing skin.
Embodiment 2
Xenogenesis skin raw material is lost hair or feathers, scrape degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.5mm, by xenogenesis skin material successively at room temperature with the NaCl solution of 2mol/L soak 12h, with sterile water wash, with the mass fractions of 37 DEG C be 0.5% neutral protease solution soaking 6h, then obtain xenogenesis skin by sterile water wash.
Xenogenesis skin is placed in agitator, and the frequency of oscillation of agitator is 110rpm.Temperature control to 55 DEG C in water bath with thermostatic control, heated after 90 minutes, obtained the xenogenesis skin of the preliminary deactivation of virus.By the xenogenesis skin of preliminary for virus deactivation, be the gamma-ray irradiation of 15kGy with irradiation dose, obtain dressing skin.
Embodiment 3
Xenogenesis skin raw material is lost hair or feathers, scrape degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.4mm, by xenogenesis skin material successively at room temperature with the NaCl solution of 0.5mol/L soak 24h, with sterile water wash, with the mass fractions of 37 DEG C be 0.2% neutral protease solution soaking 2h, then obtain xenogenesis skin by sterile water wash.
Xenogenesis skin is placed in agitator, and the frequency of oscillation of agitator is 105rpm.Temperature control to 50 DEG C in water bath with thermostatic control, heated after 120 minutes, obtained the xenogenesis skin of the preliminary deactivation of virus.By the xenogenesis skin of preliminary for virus deactivation, be the gamma-ray irradiation of 20kGy with irradiation dose, obtain dressing skin.
Embodiment 4
Xenogenesis skin raw material is lost hair or feathers, scrape degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.3mm, by xenogenesis skin material successively at room temperature with the NaCl solution of 1mol/L soak 20h, with sterile water wash, with the mass fractions of 37 DEG C be 0.35% neutral protease solution soaking 4h, then obtain xenogenesis skin by sterile water wash.
Xenogenesis skin is placed in agitator, and the frequency of oscillation of agitator is 110rpm.Temperature control to 60 DEG C in water bath with thermostatic control, the time of heating is 60 minutes, obtains the xenogenesis skin of the preliminary deactivation of virus.By the xenogenesis skin of preliminary for virus deactivation, be the gamma-ray irradiation of 25kGy with irradiation dose, obtain dressing skin.
Embodiment 5
Xenogenesis skin raw material is lost hair or feathers, scrape degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.3mm, by xenogenesis skin material successively at room temperature with the NaCl solution of 1mol/L soak 20h, with sterile water wash, with the mass fractions of 37 DEG C be 0.35% neutral protease solution soaking 4h, then obtain xenogenesis skin by sterile water wash.
Be the gamma-ray irradiation of 25kGy by xenogenesis skin irradiation dose, obtain the xenogenesis skin of the preliminary deactivation of virus.The xenogenesis skin of preliminary for virus deactivation is placed in agitator, and the frequency of oscillation of agitator is 100rpm.Temperature control to 60 DEG C in water bath with thermostatic control, the time of heating is 120 minutes, obtains dressing skin.
Embodiment 6
Xenogenesis skin raw material is lost hair or feathers, scrape degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.5mm, by xenogenesis skin material successively at room temperature with the NaCl solution of 2mol/L soak 12h, with sterile water wash, with the mass fractions of 37 DEG C be 0.5% neutral protease solution soaking 6h, then obtain xenogenesis skin by sterile water wash.
Be the gamma-ray irradiation of 15kGy by xenogenesis skin irradiation dose, obtain the xenogenesis skin of the preliminary deactivation of virus.The xenogenesis skin of preliminary for virus deactivation is placed in agitator, and the frequency of oscillation of agitator is 100rpm.Temperature control to 55 DEG C in water bath with thermostatic control, the time of heating is 90 minutes, obtains dressing skin.
Embodiment 7
Xenogenesis skin raw material is lost hair or feathers, scrape degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.4mm, by xenogenesis skin material successively at room temperature with the NaCl solution of 0.5mol/L soak 24h, with sterile water wash, with the mass fractions of 37 DEG C be 0.2% neutral protease solution soaking 2h, then obtain xenogenesis skin by sterile water wash.
Be the gamma-ray irradiation of 20kGy by xenogenesis skin irradiation dose, obtain the xenogenesis skin of the preliminary deactivation of virus.The xenogenesis skin of preliminary for virus deactivation is placed in agitator, and the frequency of oscillation of agitator is 105rpm.Temperature control to 50 DEG C in water bath with thermostatic control, the time of heating is 120 minutes, obtains dressing skin.
Embodiment 8
Xenogenesis skin raw material is lost hair or feathers, scrape degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.3mm, by xenogenesis skin material successively at room temperature with the NaCl solution of 1mol/L soak 20h, with sterile water wash, with the mass fractions of 37 DEG C be 0.35% neutral protease solution soaking 4h, then obtain xenogenesis skin by sterile water wash.
Be the gamma-ray irradiation of 25kGy by xenogenesis skin irradiation dose, obtain the xenogenesis skin of the preliminary deactivation of virus.The xenogenesis skin of preliminary for virus deactivation is placed in agitator, and the frequency of oscillation of agitator is 110rpm.Temperature control to 60 DEG C in water bath with thermostatic control, the time of heating is 60 minutes, obtains dressing skin.
Comparative example 1
Xenogenesis skin raw material is lost hair or feathers, through conventional method process, obtains xenogenesis skin.Be the gamma-ray irradiation of 25kGy by xenogenesis skin irradiation dose, obtain dressing skin.
By the dressing skin that embodiment 1 ~ 8 and comparative example 1 obtain, be immersed in 20mL physiological saline solution after 1h, take out 0.2ml normal saline, add 1.8ml physiological saline solution, then through 10 times of dilutions, obtain sample solution, measure virus titer, virus titer is for representing viral load.Method of testing is as follows, and virus titer test result is as table 1.
Select Vero and the PK-15 cell strain responsive to PRV, VSV, PV1 and PPV tetra-kinds virus to do to detect cell, adopt Microdose cytopathic effect assay.By sample solution, add in 96 orifice plates inoculated and detect cell, each dilution factor does 8 holes and repeats, and puts 37 DEG C, 5%CO
2hatch in incubator.
Need hatch 72 hours by Vero cell detection PRV, VSV and PV1 virus, light Microscopic observation also records cytopathy (CPE) situation.120 hours need be hatched by PK-15 cell detection PPV virus, first record cytopathy situation at light Microscopic observation.Due to the cytopathogenic effect of PPV more weak (especially when PPV activity reduces and dilution factor is higher), so on the basis that need judge in cytopathy, increase fluorescent antibody staining step process such as () Tissue Culture Plate is fixed through PBS washing, 4% paraformaldehyde, wash, 2%TritonX-100 rupture of membranes, washing, fluorescent antibody staining, washings of specificity anti-PPV, and at fluorescence microscopy Microscopic observation.The emerald green fluorescence speckle that the cell infected by PPV is beautiful as seen when there is not pathological changes, the method can significantly improve the specificity and sensitivity that PPV detects.The ultimate value that the method detects is-0.5Logs.Virus titer is pressed KarberShi method and is calculated.
Table 1
| PRV | VSV | PV1 | PPV | |
| Embodiment 1 | ≤-0.5 | ≤-0.5 | ≤-0.5 | ≤-0.5 |
| Embodiment 2 | ≤-0.5 | ≤-0.5 | ≤-0.5 | ≤-0.5 |
| Embodiment 3 | ≤-0.5 | ≤-0.5 | ≤-0.5 | ≤-0.5 |
| Embodiment 4 | ≤-0.5 | ≤-0.5 | ≤-0.5 | ≤-0.5 |
| Embodiment 5 | ≤-0.5 | ≤-0.5 | ≤-0.5 | ≤-0.5 |
| Embodiment 6 | ≤-0.5 | ≤-0.5 | ≤-0.5 | ≤-0.5 |
| Embodiment 7 | ≤-0.5 | ≤-0.5 | ≤-0.5 | ≤-0.5 |
| Embodiment 8 | ≤-0.5 | ≤-0.5 | ≤-0.5 | ≤-0.5 |
| Comparative example 1 | ≤-0.5 | ≤-0.5 | 0.13 | ≤-0.5 |
By the dressing skin that embodiment 1 ~ 8 and comparative example 1 obtain, carry out virus activity test by the experiment of cell blind passage 3 generation.In the overall process in cell blind passage 3 generation, any period occurs that cytopathy is the positive (+), virus is described not by complete inactivation; All do not occur that cytopathy is negative (-), virus is described by complete inactivation.Cell blind passage three generations experimental result is in table 2.
Table 2
| PRV | VSV | PV1 | PPV | |
| Embodiment 1 | (-) | (-) | (-) | (-) |
| Embodiment 2 | (-) | (-) | (-) | (-) |
| Embodiment 3 | (-) | (-) | (-) | (-) |
| Embodiment 4 | (-) | (-) | (-) | (-) |
| Embodiment 5 | (-) | (-) | (-) | (-) |
| Embodiment 6 | (-) | (-) | (-) | (-) |
| Embodiment 7 | (-) | (-) | (-) | (-) |
| Embodiment 8 | (-) | (-) | (-) | (-) |
| Comparative example 1 | (-) | (-) | (+) | (-) |
As can be seen from Table 1, comparative example 1, PRV, VSV and PPV tri-kinds of virus titers are no more than-0.5Logs, and PV1 virus titer is 0.13Logs.PRV, VSV, PV1 and PPV tetra-kinds of Viral diagnosis results of embodiment 1 ~ 8 are all qualified, and PRV, VSV, PV1 and PPV tetra-kinds of virus titers are no more than-0.5Logs.
As can be seen from Table 2, after the sample cell blind passage three generations of comparative example 1, the cytopathy detecting PV1 virus is the positive, illustrates that the PPV virus added does not have complete inactivation.After the sample cell blind passage three generations of embodiment 1 ~ 8, the cytopathy detecting PRV, VSV, PV1 and PPV tetra-kinds viral is feminine gender, and PRV, VSV, PV1 and PPV tetra-kinds of viruses complete inactivation added is described.
Therefore, the preparation method of above-mentioned dressing skin can complete inactivation PRV, VSV, PV1 and PPV tetra-kinds of viruses, and adopt the dressing skin that the preparation method of above-mentioned dressing skin obtains, PRV, VSV, PV1 and PPV tetra-kinds of Viral diagnosis results are qualified.
The above test only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but therefore can not be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (10)
1. a preparation method for dressing skin, is characterized in that, comprises the following steps:
Xenogenesis skin raw material is lost hair or feathers, scrapes degrease and horny layer, obtain the xenogenesis skin material that thickness is 0.3 ~ 0.5mm;
By described xenogenesis skin material successively with NaCl solution immersion, sterile water wash, with the mass fractions of 37 DEG C be 0.2 ~ 0.5% neutral protease solution soaking, sterile water wash obtain xenogenesis skin; And
Described xenogenesis skin is carried out inactivation treatment and obtains dressing skin, described inactivation treatment comprises: heated 60 ~ 120 minutes at 50 ~ 60 DEG C by described xenogenesis skin, and by described xenogenesis skin gamma-ray irradiation.
2. the preparation method of dressing skin according to claim 1, is characterized in that, the concentration of described NaCl solution is 0.5 ~ 2mol/L, and the time that described NaCl solution is soaked is 12 ~ 24h.
3. the preparation method of dressing skin according to claim 1, is characterized in that, the time of described neutral protease solution soaking is 2 ~ 6h.
4. the preparation method of dressing skin according to claim 1, it is characterized in that, describedly by the concrete operations that xenogenesis skin heats 60 ~ 120 minutes at 50 ~ 60 DEG C be: xenogenesis skin is placed in agitator, and in water bath with thermostatic control, temperature control heats 60 ~ 120 minutes at 50 ~ 60 DEG C.
5. the preparation method of dressing skin according to claim 4, is characterized in that, the frequency of oscillation of described agitator is 100 ~ 110rpm.
6. the preparation method of dressing skin according to claim 1, is characterized in that, the time of described heating is 90 ~ 120min.
7. the preparation method of dressing skin according to claim 1, is characterized in that, described gamma-rays is produced by radiosiotope cobalt-60, and irradiation dose is 15 ~ 25kGy.
8. the preparation method of dressing skin according to claim 7, is characterized in that, described irradiation dose is 25kGy.
9. the preparation method of dressing skin according to claim 1, is characterized in that, described xenogenesis skin physiological saline solution before inactivation treatment obtains dressing skin, washs by described being carried out by described xenogenesis skin.
10. the preparation method of dressing skin according to claim 1, it is characterized in that, the concentration of described NaCl solution is 1mol/L, and the time that described NaCl solution is soaked is 20h, the mass fraction of described neutral protein enzymatic solution is 0.35%, and the time of described neutral protease solution soaking is 4h.
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| CN106963956A (en) * | 2017-03-01 | 2017-07-21 | 广州格雷特生物科技有限公司 | The inactivation technology of duck tembusu virus viruses in yolk antibody |
| CN108969804A (en) * | 2018-08-24 | 2018-12-11 | 江西省科星生物工程有限公司 | A kind of preparation method of allogeneic dressing skin |
| CN110833630A (en) * | 2019-12-17 | 2020-02-25 | 温州施乐康医疗器械有限公司 | Dressing for promoting wound healing and application thereof |
| CN113648436A (en) * | 2021-09-03 | 2021-11-16 | 江西省科星生物工程有限公司 | Inactivation treatment device for preparing allogeneic dressing skin |
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| CN101954116A (en) * | 2010-08-31 | 2011-01-26 | 长沙达瑞奇实业有限公司 | Manufacturing method of xenoskin used for burns and scalds |
| CN102218162A (en) * | 2011-05-24 | 2011-10-19 | 山西奥瑞生物材料有限公司 | Preparation method of homologous acellular dermal matrix |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101468213A (en) * | 2007-12-28 | 2009-07-01 | 郭翔 | Method for preparing irradiation crosslinking heterogeneous skin acellular matrix and products produced thereby |
| US20090208551A1 (en) * | 2008-02-14 | 2009-08-20 | Bioland Ltd. | Biological implantation material and method for preparing same |
| CN101954116A (en) * | 2010-08-31 | 2011-01-26 | 长沙达瑞奇实业有限公司 | Manufacturing method of xenoskin used for burns and scalds |
| CN102218162A (en) * | 2011-05-24 | 2011-10-19 | 山西奥瑞生物材料有限公司 | Preparation method of homologous acellular dermal matrix |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106963956A (en) * | 2017-03-01 | 2017-07-21 | 广州格雷特生物科技有限公司 | The inactivation technology of duck tembusu virus viruses in yolk antibody |
| CN108969804A (en) * | 2018-08-24 | 2018-12-11 | 江西省科星生物工程有限公司 | A kind of preparation method of allogeneic dressing skin |
| CN110833630A (en) * | 2019-12-17 | 2020-02-25 | 温州施乐康医疗器械有限公司 | Dressing for promoting wound healing and application thereof |
| CN113648436A (en) * | 2021-09-03 | 2021-11-16 | 江西省科星生物工程有限公司 | Inactivation treatment device for preparing allogeneic dressing skin |
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