CN104940982B - The preparation method of dressing skin - Google Patents
The preparation method of dressing skin Download PDFInfo
- Publication number
- CN104940982B CN104940982B CN201510025968.4A CN201510025968A CN104940982B CN 104940982 B CN104940982 B CN 104940982B CN 201510025968 A CN201510025968 A CN 201510025968A CN 104940982 B CN104940982 B CN 104940982B
- Authority
- CN
- China
- Prior art keywords
- skin
- xenogenesis
- dressing
- preparation
- virus
- 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.)
- Expired - Fee Related
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Materials For Medical Uses (AREA)
Abstract
The invention discloses a kind of preparation method of dressing skin, comprise the following steps：By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.3~0.5mm is obtained；Xenogenesis cladding is soaked with NaCl solution successively, sterile water wash, with 37 DEG C of mass fractions for 0.2~0.5% neutral protein enzyme solutions soak, sterile water wash obtain xenogenesis skin；And xenogenesis skin progress inactivation treatment is obtained into dressing skin, inactivation treatment includes：Xenogenesis skin is heated 60~120 minutes at 50~60 DEG C, and by xenogenesis skin gamma-ray irradiation.The preparation method of above-mentioned dressing skin is simple to operate, with low cost, can tetra- kinds of viruses of complete inactivation PRV, VSV, PV1 and PPV, inactivation of virus efficiency is significantly improved, and solves and the problem of complete inactivation PV1 is viral is individually unable to gamma-ray irradiation, improve the security of dressing skin.
Description
Technical field
The present invention relates to animal derived biological dressing Viral inactivation techniques field, more particularly to a kind of preparation of dressing skin
Method.
Background technology
It is existing be used to burning, heating power wound or the xenogenesis skin that uses of chemical injury be often for pigskin.Pigskin and the skin texture of people
Similar, it preferably can be sticked on the surface of a wound, played reduction surface of a wound moisture evaporation, reduced electrolyte loss, barrier bacterium and control
The effect of infection is made, clinical effectiveness is good, be a kind of preferably alternative allograft skin, obtain wider in burn, scald field
General application.Pigskin must be strictly controlled as a kind of animal derived Wound dressing skin, its viral carrying amount.Therefore,
The selection of virus inactivation technology is most important.At present, burn and scald mainly used with pigskin gamma-rays (gamma rays) irradiate into
Row inactivation of virus.
According to the regulation of national Bureau of Drugs Supervision associated documents, in every step inactivation step, viral reduction amount reaches 4Logs (i.e.
LgTCID50/ ml) more than, it is believed that inactivation of viruses method is effective；And deactivation of each step to virus can be superimposed.Virus drop
Low amounts is bigger, the virus-free detection of blind passage three generations, illustrates that inactivation technology effect is better, the security of dressing skin is higher.Research discovery,
Although using gamma-ray irradiation the viral reduction amount of pigskin can be made to reach more than 4Logs, it is believed that this method is effectively, grey containing spinal cord
Cell blind passage three generations is added after matter inflammation virus sabinI types virus (PV1) sample irradiation and cytopathy occurs, illustrates PV1 viruses
Inactivation is not thorough enough.To improve the security performance of dressing skin, it should try further to improve inactivation of virus efficiency.
The content of the invention
Based on this, it is necessary to for inactivation of virus it is inefficient the problem of there is provided a kind of inactivation of virus efficiency and security
The preparation method of high dressing skin.
A kind of preparation method of dressing skin, comprises the following steps：
By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.3~0.5mm is obtained；
The xenogenesis cladding is soaked with NaCl solution successively, sterile water wash, with 37 DEG C of mass fractions be 0.2~
0.5% neutral protein enzyme solutions soak, sterile water wash obtains xenogenesis skin；And
Xenogenesis skin progress inactivation treatment is obtained into dressing skin, the inactivation treatment includes：By the xenogenesis skin 50
Heated 60~120 minutes at~60 DEG C, and by the xenogenesis skin gamma-ray irradiation.
In one of the embodiments, the concentration of the NaCl solution is 0.5~2mol/L, the NaCl solution immersion
Time is 12~24h.
In one of the embodiments, the time of the neutral protein enzyme solutions immersion is 2~6h.
In one of the embodiments, the concrete operations that xenogenesis skin is heated at 50~60 DEG C to 60~120 minutes
For：Xenogenesis skin is placed in oscillator, temperature control is heated 60~120 minutes at 50~60 DEG C in water bath with thermostatic control.
In one of the embodiments, the frequency of oscillation of the oscillator is 100~110rpm.
In one of the embodiments, the time of the heating is 90~120min.
In one of the embodiments, the gamma-rays by radio isotope cobalt -60 produce, irradiation dose be 15~
25kGy。
In one of the embodiments, the irradiation dose is 25kGy.
In one of the embodiments, it is described xenogenesis skin progress inactivation treatment is obtained into dressing skin before, will be described
Xenogenesis skin is washed with sterile saline.
In one of the embodiments, the concentration of the NaCl solution is 1mol/L, the time of the NaCl solution immersion
For 20h, the mass fraction of the neutral protein enzyme solutions is 0.35%, and the time of the neutral protein enzyme solutions immersion is 4h.
The preparation method of above-mentioned dressing skin is simple to operate, with low cost, can tetra- kinds of complete inactivation PRV, VSV, PV1 and 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 shows
Write and improve, tetra- kinds of Viral diagnosis of obtained dressing skin PRV, VSV, PV1 and PPV are qualified, solve and individually use gamma-ray irradiation
The problem of complete inactivation PV1 is viral is unable to, the security of dressing skin is improved.
Embodiment
For the ease of understanding the present invention, the present invention is described more fully below.And give the preferable of the present invention
Experiment.But, the present invention can be realized in many different forms, however it is not limited to experiment described herein.On the contrary,
The purpose for providing these experiments is to make the understanding to the disclosure more thorough comprehensive.
Unless otherwise defined, all of technologies and scientific terms used here by the article is with belonging to technical field of the invention
The implication that technical staff is generally understood that is identical.Term used in the description of the invention herein is intended merely to description tool
The purpose of the experiment of body, it is not intended that in the limitation present invention.Term as used herein " and/or " include one or more correlations
Listed Items arbitrary and all combination.
The preparation method of the dressing skin of one embodiment, comprises the following steps：
S100：By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis skin that thickness is 0.3~0.5mm is obtained
Material.
Preferably, the thickness of xenogenesis cladding is 0.3mm.
Preferably, xenogenesis skin raw material is pigskin raw material, and pigskin is similar to the skin texture of people, and it can preferably stick to wound
On face, play a part of reducing surface of a wound moisture evaporation, reduce electrolyte loss, barrier bacterium and infection control, clinical effectiveness is good,
It is a kind of preferably alternative allograft skin, wide application is obtained in burn, scald field.
Pigskin must be strictly controlled as a kind of animal derived Wound dressing skin, its viral carrying amount.Therefore, it is sick
The selection of malicious inactivation technology is most important.According to " traditional Chinese medicines prison note [2002] 160 ", " state's food medicine prison note [2008] 7 " file
Regulation, selects following four indicator virus.
Pseudorabies virus (PRV), belongs to herpetoviridae, is that double-stranded DNA has after birth virus.To fatsolvents such as ether, chloroforms,
The sensitivity such as formalin and ultraviolet irradiation, is the stronger herpesviral of a species resistance.
Vesicular stomatitis virus (VSV), belongs to Rhabdoviridae, is that strand RNA has after birth virus.It is not acidproof, it is seen that light, purple
Outside line and fatsolvent can inactivate it, and the resistance to the Physicochemical factor is similar to foot and mouth disease virus.
Poliovirus sabinI types (PV1), category enterovirus section is Microrna without after birth virus.Due to without born of the same parents
Film, to physical and chemical factor better resistance.
Pig parvoviral (PPV), belongs to Parvoviridae, is small DNA viral without after birth.Due to without after birth, to physics and chemistry because
Sub- resistance is extremely strong.
Viral reduction amount is represented that R=inactivates provirus titre-inactivation restrovirus titre by R.Viral reduction amount R >=
4Logs, judges that the step is effective.Viral complete inactivation, refers to after inactivation of virus, the reduction amount of viral load reach 4Logs with
On, and the virus-free detection of blind passage three generations.
S200：Xenogenesis cladding is soaked with NaCl solution successively, sterile water wash, with 37 DEG C of mass fractions be 0.2~
0.5% neutral protein enzyme solutions soak, sterile water wash obtains xenogenesis skin.
Especially, the concentration of NaCl solution is 0.5~2mol/L, and the time of NaCl solution immersion is 12~24h.
Especially, neutral egg enzyme solutions are the aqueous solution.Especially, the time of neutral protein enzyme solutions immersion is 2~6h.
Preferably, the concentration of NaCl solution is 1mol/L, and the time of NaCl solution immersion is 20h, neutral protein enzyme solutions
Mass fraction be 0.35%, neutral protein enzyme solutions immersion time be 4h.
Sterilized water be by high-temperature steam method, chemical method, ozone approach or physical filtering method by water microorganism kill or
Water obtained from filtering out.Specifically, in the present embodiment, sterilized water is using at hollow fiber ultrafiltration membrane physical filtering method
Manage obtained water.
Especially, the number of times of sterile water wash is 2~4 times.Using sterile water wash, the chips such as pig hair are removed clean.
Especially, sterile water wash can be using ultrasonic wave cleaning.Preferably, the time cleaned every time is 10 minutes.
Xenogenesis skin raw material by taking skin, is taken off cell processing, intactly saves epidermis by step S100 and step S200
Structure.
S300：Xenogenesis skin is heated 60~120 minutes at 50~60 DEG C, the xenogenesis skin that virus is tentatively inactivated is obtained.
Before xenogenesis skin progress inactivation treatment obtained into dressing skin, xenogenesis skin is washed with sterile saline.Sterile life
Reason salt solution is that the physiological saline configured is made through high pressure steam sterilization.The mass fraction of sterile saline be 0.85~
0.9%.
The concrete operations that xenogenesis skin is heated 60~120 minutes at 50~60 DEG C are：Xenogenesis skin is placed in oscillator,
Temperature control is heated 60~120 minutes at 50~60 DEG C in water bath with thermostatic control.
More particularly, the frequency of oscillation of oscillator is 100~110rpm (rev/min).
Xenogenesis skin after being handled using step S300, the reduction amount of four kinds of indicator virus quantity is reached more than 4Logs, said
Bright use step S300 can effectively inactivate above-mentioned four kinds of indicator viruses.
It is noted that the time of heating can make tri- kinds of viral loads of PRV, VSV and PV1 of addition more than 5min
Reduction amount reaches that more than 4Logs, the i.e. step are effective for the inactivation of PRV, VSV and PV1 virus.The time of heating exceedes
15min can complete inactivation add PRV and two kinds of VSV have after birth virus.
When the time of heating is 60~120min, you can tri- kinds of viruses of PRV, VSV and PV1 that complete inactivation is added.It is more excellent
Choosing, the time of heating is 90~120min.PPV viruses are after one step S300 inactivations, although the drop of PPV viral loads
Low amounts reaches more than 4Logs, it is believed that step S300 is effective for PPV inactivation of virus；But blind passage three generations virus is still active.Cause
This single step S300 on the one hand can be by tri- kinds of viral complete inactivations of PRV, VSV and PV1, and on the other hand exist to go out completely
The shortcoming of PPV viruses living.
S400：The xenogenesis skin that virus is tentatively inactivated, with gamma-ray irradiation, obtains dressing skin.
It is preferred that, the xenogenesis skin that virus is tentatively inactivated is placed on aluminum foil sack using sterile saline as liquid is preserved
Middle sealing preserve is to carry out gamma-ray irradiation.
Gamma-rays is produced by radio isotope cobalt -60.Especially, irradiation dose is 15~25kGy.Irradiation dose mistake
Greatly, other materials denaturation such as protein of xenogenesis skin is caused, therefore control can make in the zone of reasonableness of inactivation of virus.It is preferred that
Ground, irradiation dose is 25kGy.
After bacterium in xenogenesis skin is irradiated by gamma-rays, physiological dysfunction, it is impossible to grow, in addition it is dead.
Tri- kinds of viral complete inactivations of PRV, VSV and PPV can be compensate for step S300 and are unable to complete inactivation by step S400
The shortcoming of PPV viruses.In addition, PV1 viruses are after one step S400 inactivations, although the reduction amount of PV1 viral loads reaches
More than 4Logs, it is believed that step S400 is also effective for PV1 inactivation of virus；But blind passage three generations virus is still active.Therefore it is single
Step S400 on the one hand on the other hand the viral complete inactivations of tri- kinds of PRV, VSV and PPV can be existed and be unable to complete inactivation PV1
The shortcoming of virus.
Therefore, the preparation method of above-mentioned dressing skin, can tetra- kinds of viruses of complete inactivation PRV, VSV, PV1 and PPV.It can manage
Solution, step S300 and step S400 order can be exchanged, i.e., by xenogenesis skin gamma-ray irradiation, obtain what virus was tentatively inactivated
Xenogenesis skin；The xenogenesis skin that virus is tentatively inactivated is heated to 50~60 DEG C, and the time of heating is 60~120 minutes, obtains dressing
Skin.
Above-mentioned dressing skin is soaked 20 minutes before use, being installed and being put into 35~45 DEG C of water with Aluminium Foil Package, carries out rewarming；Will
After surface of a wound routine debridement, the surface of a wound is covered in being taken out under aseptic condition, conventional wrapping and dressing.
The preparation method of above-mentioned dressing skin is simple to operate, with low cost, can tetra- kinds of complete inactivation PRV, VSV, PV1 and PPV
Virus, four kinds of viral reduction amounts are respectively：Reduction amount R >=13.76Logs of PRV viruses；The reduction amount R of VSV viruses >=
12.25Logs；Reduction amount R >=12.01Logs of PV1 viruses；Reduction amount R >=10.75Logs of PPV viruses.With single γ
The virus inactivation technology of x ray irradiation x is compared, and the preparation method inactivation of virus efficiency of above-mentioned dressing skin is significantly improved, and what is obtained applies
Expect that skin tetra- kinds of Viral diagnosis of PRV, VSV, PV1 and PPV are qualified, solve and complete inactivation PV1 is individually unable to gamma-ray irradiation
Viral the problem of, improve the security of dressing skin.
It is specific embodiment below.
The xenogenesis skin raw material that embodiment 1~8 and comparative example 1 are used is pigskin raw material.
Embodiment 1
By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.3mm is obtained, by xenogenesis skin
Material soaks 20h with 1mol/L NaCl solution at room temperature successively, is with sterile water wash, with 37 DEG C of mass fraction
0.35% neutral protein enzyme solutions immersion 4h, then obtains xenogenesis skin with sterile water wash.
Xenogenesis skin is placed in oscillator, the frequency of oscillation of oscillator is 100rpm.Temperature control is to 60 DEG C in water bath with thermostatic control,
After heating 120 minutes, the xenogenesis skin that virus is tentatively inactivated is obtained.The xenogenesis skin that virus is tentatively inactivated, be with irradiation dose
25kGy gamma-ray irradiation, obtains dressing skin.
Embodiment 2
By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.5mm is obtained, by xenogenesis skin
Material soaks 12h with 2mol/L NaCl solution at room temperature successively, is 0.5% with sterile water wash, with 37 DEG C of mass fractions
Neutral protein enzyme solutions immersion 6h, xenogenesis skin is then obtained with sterile water wash.
Xenogenesis skin is placed in oscillator, the frequency of oscillation of oscillator is 110rpm.Temperature control is to 55 DEG C in water bath with thermostatic control,
After heating 90 minutes, the xenogenesis skin that virus is tentatively inactivated is obtained.The xenogenesis skin that virus is tentatively inactivated, be with irradiation dose
15kGy gamma-ray irradiation, obtains dressing skin.
Embodiment 3
By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.4mm is obtained, by xenogenesis skin
Material soaks 24h with 0.5mol/L NaCl solution at room temperature successively, is with sterile water wash, with 37 DEG C of mass fraction
0.2% neutral protein enzyme solutions immersion 2h, then obtains xenogenesis skin with sterile water wash.
Xenogenesis skin is placed in oscillator, the frequency of oscillation of oscillator is 105rpm.Temperature control is to 50 DEG C in water bath with thermostatic control,
After heating 120 minutes, the xenogenesis skin that virus is tentatively inactivated is obtained.The xenogenesis skin that virus is tentatively inactivated, be with irradiation dose
20kGy gamma-ray irradiation, obtains dressing skin.
Embodiment 4
By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.3mm is obtained, by xenogenesis skin
Material soaks 20h with 1mol/L NaCl solution at room temperature successively, is 0.35% with sterile water wash, with 37 DEG C of mass fractions
Neutral protein enzyme solutions immersion 4h, xenogenesis skin is then obtained with sterile water wash.
Xenogenesis skin is placed in oscillator, the frequency of oscillation of oscillator is 110rpm.Temperature control is to 60 DEG C in water bath with thermostatic control,
The time of heating is 60 minutes, obtains the xenogenesis skin that virus is tentatively inactivated.The xenogenesis skin that virus is tentatively inactivated, uses irradiation dose
For 25kGy gamma-ray irradiation, dressing skin is obtained.
Embodiment 5
By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.3mm is obtained, by xenogenesis skin
Material soaks 20h with 1mol/L NaCl solution at room temperature successively, is 0.35% with sterile water wash, with 37 DEG C of mass fractions
Neutral protein enzyme solutions immersion 4h, xenogenesis skin is then obtained with sterile water wash.
The gamma-ray irradiation for being 25kGy with irradiation dose by xenogenesis skin, obtains the xenogenesis skin that virus is tentatively inactivated.By virus
The xenogenesis skin tentatively inactivated is placed in oscillator, and the frequency of oscillation of oscillator is 100rpm.Temperature control is to 60 DEG C in water bath with thermostatic control,
The time of heating is 120 minutes, obtains dressing skin.
Embodiment 6
By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.5mm is obtained, by xenogenesis skin
Material soaks 12h with 2mol/L NaCl solution at room temperature successively, is 0.5% with sterile water wash, with 37 DEG C of mass fractions
Neutral protein enzyme solutions immersion 6h, xenogenesis skin is then obtained with sterile water wash.
The gamma-ray irradiation for being 15kGy with irradiation dose by xenogenesis skin, obtains the xenogenesis skin that virus is tentatively inactivated.By virus
The xenogenesis skin tentatively inactivated is placed in oscillator, and the frequency of oscillation of oscillator is 100rpm.Temperature control is to 55 DEG C in water bath with thermostatic control,
The time of heating is 90 minutes, obtains dressing skin.
Embodiment 7
By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.4mm is obtained, by xenogenesis skin
Material soaks 24h with 0.5mol/L NaCl solution at room temperature successively, is with sterile water wash, with 37 DEG C of mass fraction
0.2% neutral protein enzyme solutions immersion 2h, then obtains xenogenesis skin with sterile water wash.
The gamma-ray irradiation for being 20kGy with irradiation dose by xenogenesis skin, obtains the xenogenesis skin that virus is tentatively inactivated.By virus
The xenogenesis skin tentatively inactivated is placed in oscillator, and the frequency of oscillation of oscillator is 105rpm.Temperature control is to 50 in water bath with thermostatic control
DEG C, the time of heating is 120 minutes, obtains dressing skin.
Embodiment 8
By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.3mm is obtained, by xenogenesis skin
Material soaks 20h with 1mol/L NaCl solution at room temperature successively, is 0.35% with sterile water wash, with 37 DEG C of mass fractions
Neutral protein enzyme solutions immersion 4h, xenogenesis skin is then obtained with sterile water wash.
The gamma-ray irradiation for being 25kGy with irradiation dose by xenogenesis skin, obtains the xenogenesis skin that virus is tentatively inactivated.By virus
The xenogenesis skin tentatively inactivated is placed in oscillator, and the frequency of oscillation of oscillator is 110rpm.Temperature control is to 60 DEG C in water bath with thermostatic control,
The time of heating is 60 minutes, obtains dressing skin.
Comparative example 1
By the depilation of xenogenesis skin raw material, handled by conventional method, obtain xenogenesis skin.It is with irradiation dose by xenogenesis skin
25kGy gamma-ray irradiation, obtains dressing skin.
The dressing skin that embodiment 1~8 and comparative example 1 are obtained, is immersed in 20mL sterile salines after 1h,
0.2ml physiological saline is taken out, 1.8ml sterile salines are added, then through 10 times of dilutions, obtains sample solution, virus drop is determined
Degree, virus titer is used to represent viral load.Method of testing is as follows, virus titer test result such as table 1.
Make detection cell to tetra- kinds of sensitive Vero and PK-15 cell lines of virus of PRV, VSV, PV1 and PPV, use
Microdose cytopathic effect assay.By sample solution, add and be inoculated with 96 orifice plates of detection cell, each dilution factor does the repetition of 8 holes, puts
37 DEG C, 5%CO2It is incubated in incubator.
It need to be incubated 72 hours with Vero cell detection PRV, VSV and PV1 viruses, light Microscopic observation simultaneously records cytopathy
(CPE) situation.It need to be incubated 120 hours with PK-15 cell detection PPV viruses, elder generation is in light Microscopic observation and records cytopathy feelings
Condition.Because PPV cytopathogenic effect is weaker (especially in the reduction of PPV activity and higher dilution factor), so need to be in cytopathy
On the basis of judgement, (Tissue Culture Plate is washed the specific anti-PPV of increase fluorescent antibody staining through PBS, 4% paraformaldehyde is consolidated
The step process such as fixed, washing, 2%TritonX-100 ruptures of membranes, washing, fluorescent antibody staining, washing), and under fluorescence microscope
Observation.The cell infected by PPV visible beautiful emerald green fluorescence spot in the case where occurring without lesion, this method can be bright
The aobvious specificity for improving PPV detections and sensitivity.The limiting value of this method detection is -0.5Logs.Virus titer presses KarberShi
Method 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 |
The dressing skin that embodiment 1~8 and comparative example 1 are obtained, carries out virus living by the experiment of the generation of cell blind passage 3
Property test.In the overall process in the generation of cell blind passage 3, it is positive (+) that cytopathy, which occurs, in any time, illustrates that virus is not complete
Full inactivation；It is negative (-) not occur cytopathy, illustrates virus by complete inactivation.Cell blind passage three generations's experimental result is shown 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 | (-) | (-) | (+) | (-) |
From table 1 it follows that comparative example 1, tri- kinds of virus titers of PRV, VSV and PPV are no more than -0.5Logs, and
PV1 virus titers are 0.13Logs.Tetra- kinds of Viral diagnosis results of PRV, VSV, PV1 and PPV of embodiment 1~8 are qualified, PRV,
Tetra- kinds of virus titers of VSV, PV1 and PPV are no more than -0.5Logs.
From Table 2, it can be seen that after the sample cell blind passage three generations of comparative example 1, the cytopathy of detection PV1 viruses
It is the positive, illustrates the PPV viruses added without complete inactivation.After the sample cell blind passage three generations of embodiment 1~8, detection
Tetra- kinds of viral cytopathies of PRV, VSV, PV1 and PPV are feminine gender, illustrate the tetra- kinds of viruses of PRV, VSV, PV1 and PPV added
Complete inactivation.
Therefore, the preparation method of above-mentioned dressing skin can tetra- kinds of viruses of complete inactivation PRV, VSV, PV1 and PPV, using above-mentioned
The dressing skin that the preparation method of dressing skin is obtained, tetra- kinds of Viral diagnosis results of PRV, VSV, PV1 and PPV are qualified.
Described above to test the several embodiments for only expressing the present invention, it describes more specific and detailed, but not
Therefore the limitation to the scope of the claims of the present invention can be interpreted as.It should be pointed out that coming for one of ordinary skill in the art
Say, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection of the present invention
Scope.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (9)
1. a kind of preparation method of dressing skin, it is characterised in that comprise the following steps：
By the depilation of xenogenesis skin raw material, fat and cuticula are scraped off, the xenogenesis cladding that thickness is 0.3~0.5mm is obtained；
The xenogenesis cladding is soaked with NaCl solution successively, sterile water wash, with 37 DEG C of mass fractions be 0.2~0.5%
Neutral protein enzyme solutions immersion, sterile water wash obtain xenogenesis skin, wherein the neutral proteinase solution immersion time be 2
~6h；And
Xenogenesis skin progress inactivation treatment is obtained into dressing skin, the inactivation treatment includes：By the xenogenesis skin 50~60
Heated 60~120 minutes at DEG C, and by the xenogenesis skin gamma-ray irradiation.
2. the preparation method of dressing skin according to claim 1, it is characterised in that the concentration of the NaCl solution is 0.5
~2mol/L, the time of the NaCl solution immersion is 12~24h.
3. the preparation method of dressing skin according to claim 1, it is characterised in that it is described by xenogenesis skin at 50~60 DEG C
The heating concrete operations of 60~120 minutes are：Xenogenesis skin is placed in oscillator, temperature control is at 50~60 DEG C in water bath with thermostatic control
Heating 60~120 minutes.
4. the preparation method of dressing skin according to claim 3, it is characterised in that the frequency of oscillation of the oscillator is
100~110rpm.
5. the preparation method of dressing skin according to claim 1, it is characterised in that the time of the heating is 90~
120min。
6. the preparation method of dressing skin according to claim 1, it is characterised in that the gamma-rays is by radio isotope
Cobalt -60 is produced, and irradiation dose is 15~25kGy.
7. the preparation method of dressing skin according to claim 6, it is characterised in that the irradiation dose is 25kGy.
8. the preparation method of dressing skin according to claim 1, it is characterised in that described to be inactivated the xenogenesis skin
Processing is obtained before dressing skin, and the xenogenesis skin is washed with sterile saline.
9. the preparation method of dressing skin according to claim 1, it is characterised in that the concentration of the NaCl solution is
1mol/L, the time of the NaCl solution immersion is 20h, and the mass fraction of the neutral protein enzyme solutions is 0.35%, described
The time of neutral protein enzyme solutions immersion is 4h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510025968.4A CN104940982B (en) | 2015-01-19 | 2015-01-19 | The preparation method of dressing skin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510025968.4A CN104940982B (en) | 2015-01-19 | 2015-01-19 | The preparation method of dressing skin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104940982A CN104940982A (en) | 2015-09-30 |
| CN104940982B true CN104940982B (en) | 2017-09-22 |
Family
ID=54156395
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510025968.4A Expired - Fee Related CN104940982B (en) | 2015-01-19 | 2015-01-19 | The preparation method of dressing skin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104940982B (en) |
Families Citing this family (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 |
Family Cites Families (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 |
| KR100947553B1 (en) * | 2008-02-14 | 2010-03-12 | 주식회사 바이오랜드 | 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 |
-
2015
- 2015-01-19 CN CN201510025968.4A patent/CN104940982B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN104940982A (en) | 2015-09-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104940982B (en) | The preparation method of dressing skin | |
| AT392905B (en) | METHOD FOR THE TREATMENT OF A BLOOD coagulation factor VIII CONCENTRATE | |
| Chaufour et al. | Evaluation of disinfection and sterilization of reusable angioscopes with the duck hepatitis B model | |
| CN113621584B (en) | Staphylococcus aureus bacteriophage and antibacterial application thereof | |
| Ab Hamid et al. | Scanning electron microscopic assessment on surface morphology of preserved human amniotic membrane after gamma sterilisation | |
| JPWO2010067873A1 (en) | Antiviral sanitary textile products | |
| Gugerell et al. | Viral safety of APOSECTM: a novel peripheral blood mononuclear cell derived-biological for regenerative medicine | |
| Osumi et al. | Acceleration of wound healing by ultrasound activation of TiO2 in Escherichia coli‐infected wounds in mice | |
| CN104027347B (en) | The application of first group-beta-cyclodextrin in preparation control BmNPV infected silkworm cell drug | |
| Paggiaro et al. | Effect of different human tissue processing techniques on SARS-CoV-2 inactivation-review | |
| Spaulding et al. | Chemical disinfection | |
| CN108079363A (en) | A kind of kit and its application that cell processing is taken off for animal tissue | |
| Straube et al. | In vitro influence of d/l-lactic acid, sodium chloride and sodium nitrite on the infectivity of feline calicivirus and of ECHO virus as potential surrogates for foodborne viruses | |
| CN101757616B (en) | Safe freeze-dried mammal thrombin preparation and preparation method thereof | |
| CN101012455A (en) | Deactivation method of biochemistry substance, preparing method of cardiomyopeptidin and use of cardiomyopeptidin | |
| CN104450630A (en) | Method for culturing goose parvovirus by using goose embryo fibroblast line | |
| CN109337868A (en) | Utilize the method for VAK technology immune cell activated in vitro | |
| CN101461817A (en) | Use of 1,2,3,4,6-five-O-gallnut acyl radical-b-D-glucose | |
| CN108969804A (en) | A kind of preparation method of allogeneic dressing skin | |
| CN104546895B (en) | Application of lambda-carrageenan oligosaccharide to preparation of anti-rabies virus drug | |
| CN105028501A (en) | Disinfectant for pharmacy | |
| CN111068086A (en) | Method for disinfecting environment-friendly medical material | |
| JP4132151B2 (en) | How to deactivate prions | |
| CN116676278A (en) | Platelet virus inactivation method and application of troxerutin in platelet virus inactivation | |
| Bae et al. | Virus inactivation during the manufacture of a collagen type I from bovine hides |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170922 Termination date: 20180119 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |