CN103004752A - Skin preserving fluid with mycose as raw material, preparation method of skin preserving fluid and skin preservation method - Google Patents
Skin preserving fluid with mycose as raw material, preparation method of skin preserving fluid and skin preservation method Download PDFInfo
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- CN103004752A CN103004752A CN2013100188177A CN201310018817A CN103004752A CN 103004752 A CN103004752 A CN 103004752A CN 2013100188177 A CN2013100188177 A CN 2013100188177A CN 201310018817 A CN201310018817 A CN 201310018817A CN 103004752 A CN103004752 A CN 103004752A
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- skin
- mycose
- preserving fluid
- trehalose
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Abstract
The invention discloses a skin preserving fluid with mycose as a raw material. The skin preserving fluid with mycose as the raw material contains 45-165 g/L of mycose and 4.5-16.5 g/L of calf serum. The preparation method of the skin preserving fluid comprises the following steps of: taking mycose dry powder, mixing the mycose dry powder with distilled water to dissolve the mycose dry powder so as to prepare a mycose solution with the concentration of 45-165 g/L; adding the calf serum and mixing uniformly; and adjusting the pH value of the uniformly mixed solution to 7.0-7.2. The skin preserving method disclosed by the invention comprises the steps of: cleaning fresh allogeneic skin with sterile normal saline for 2-5 times, spin-drying the skin and putting the skin in mycose preservation solution prepared in advance to be soaked for more than 8 hours. The skin preserving fluid with mycose as the raw material, disclosed by the invention, has a easy material source and a relatively simple operation process, is suitable for long-distance transportation, and has a relatively long preservation time; and various damages to isolated skin caused by ischemia are reduced, and effective vitality of the isolated skin is kept, so that transportation and clinical operations can be achieved. The allogeneic skin after transplantation can recover function per se rapidly.
Description
Technical field
The present invention relates to a kind of human body skin take trehalose as raw material and preserve liquid and preparation method thereof.
Background technology
The at present domestic cryogenic freezing that adopts is preserved skin more, and its major defect is that the recrystallization of producing in the rewarming process produces harmful ice crystal damage to skin.Studies show that when only having rewarming speed to reach 3.3 ~ 17 ℃/s that just can avoid the ice crystal damage, so fast rewarming speed is difficult to reach when clinical practice.Often can add dimethyl sulfoxide (DMSO) as the protectant of cell at Cryopreservation skin.Because of its freeze proof effect, be used for the protection of cell cryopreservation, dimethyl sulfoxide (DMSO) can reduce the cell freezing point, reduces the formation of ice crystal, alleviates the radical pair primary cellular defect, changes biomembrane to the permeability of electrolyte, medicine, poisonous substance and metabolism.But there are some researches show that there is certain toxic action in dimethyl sulfoxide (DMSO), hydrophobic group has an effect with protein, causes protein denaturation, has blood vessel toxicity and liver renal toxicity.When freezing skin is used for clinical rewarming, may pollute skin.
The patent No. is: the patent disclosure of 89105183.X a kind of method of preserving skin with honey, heterogenous skin is preserved with soaking in Mel.Utilize honey as Protector.Can the long period preserve skin.Method is that heterogenous skin is carried out disinfection, puts into the container that fills honey after soaking and preserve.Contain a large amount of sugar in the honey, more than 20 seed amino acids, more than ten plant enzyme, vitamin, hormone, antibiotic, efficient alcohol, resin mineral matter etc., have sterilization, antibacterial characteristics.The processing procedure of preserving the skin method with honey is fairly simple, but honey is because of the difference of honeybee kind, honeybee source, environment, and its chemical composition has very big-difference, difficult quality guarantee, in practice difficult assurance.
Summary of the invention
One of purpose of the present invention is: provide a kind of isolated skin vigor of preserving apparently higher than the preserving fluid for skin take trehalose as raw material of traditional vitrification.
Another object of the present invention is to: provide that a kind of preparation technology is simple, method of operating is easy, convenient transportation, the simple described preserving fluid for skin take trehalose as raw material of drawing materials is preserved liquid and preparation method thereof.
Another object of the present invention is to: a kind of long, safe and reliable vitrification method of isolated skin vigor, time of preserving is provided.
Preserving fluid for skin take trehalose as raw material of the present invention contains trehalose 45 ~ 165g/L, contains calf serum 4.5 ~ 16.5g/L.
The preparation method of the preserving fluid for skin take trehalose as raw material of the present invention may further comprise the steps: (1) gets trehalose dry powder, and with the distilled water mix and blend and make it dissolving, to become concentration be the 45-165g/L aqueous trehalose in preparation;
(2) add calf serum, stir;
(3) solution that stirs is regulated pH value to 7.0-7.2.
The distilled water temperature is in the time of 25 ℃ ± 2 ℃ among the preparation method of the preserving fluid for skin take trehalose as raw material of the present invention, and adding trehalose dry powder also stirs, and it is fully dissolved.
Vitrification method of the present invention is: fresh Skin allograft is cleaned 2-5 time through stroke-physiological saline solution, put into after the drying and prepared the immersion of trehalose preservation liquid more than 8 hours, consumption is as the criterion with the submergence preservation object.
The present invention draws materials easily, and operating process is relatively simple, can adapt to long-distance transport, and the holding time is longer; Reduce ischemic to the various damages that isolated skin causes, make the isolated skin vigor of remaining valid, in order to finish transportation, clinical operation.Make heterogenous skin rapid restore funcitons after transfer operation.
Embodiment
Trehalose is a kind of safety, reliable natural carbohydrate, and self property is highly stable, and various bioactivators is had magical protective effect.Scientists is found, but desert plant leaf roll cypress can miracle be brought back to life behind the nearly withered chance water of arid; Alpine plant is brought back to life grass can anti-ly cross the ice and snow severe cold; Some insects are not freezed under the conditions such as high temperature and dry dehydration high and cold, do not thirst, and are exactly the life miracle that their body intracellular trehalose is created.Trehalose has magical protective effect to organism; because trehalose can form the diaphragm of uniqueness at cell surface under the mal-conditions such as high temperature, high and cold, hyperosmosis and dry dehydration; protected protein matter molecule consistency inactivation effectively, thereby the life process of the body that sustains life and production feature.And other carbohydrates such as occurring in nature such as sucrose, glucose etc. all do not have this function.Contain protein, fat, sugar, mineral salt, vitamins and other nutritious components in the calf serum, the Skin Cell of the ischemic that exsomatizes is played certain protective effect.Summary: trehalose has magical protective effect to organism, is because trehalose forms unique diaphragm, effectively protected protein matter molecule consistency inactivation at cell surface under the adverse circumstances such as high temperature, high and cold, hyperosmosis and dry dehydration.Thereby keep the biological characteristic protection to skin.The purpose of vitrification is to reduce the various damages that ischemic causes isolated skin, in order to finish storage, and the In transit vigor of remaining valid.The invention provides a kind of storage procedures of different conventional arts, utilize aqueous trehalose as Protector, can preserve skin in general refrigerator (4 ~ 10 ℃) long period, method be Skin allograft after stroke-physiological saline solution is cleaned, put into aqueous trehalose and soak, get final product through the cobalt-60γray irradiation sterilization.
The present invention is described in detail below in conjunction with embodiment.
Embodiment 1:
The inventive method may further comprise the steps: the configuration of aqueous trehalose: get clean container, import 1000ml distilled water (25 ℃), slowly add 45g trehalose dry powder, the limit edged stirs with glass bar, dry powder is dissolved fully, add an amount of NaHCO3, regulate pH value to 7.0 filtration sterilization.
(2) calf serum of newly buying is stored in-20 ℃ of refrigerators, as for 4 ℃ of refrigerator defrostings about 10 hours, then as for deactivation complement component 3 in 56 ℃ of thermostat water baths 0 minute, constantly shakes serum bottle in the water-bath and makes and be heated evenly filtration sterilization before using.
(3) at normal temperatures put into preservation object (skin) after (25 ℃) adding calf serum 4.5g makes and seal, namely join namely with allowing it freely cool off about 2 hours.Put into 4 ℃ of Refrigerator stores
(4) the preservation liquid in the container is as the criterion with submergence skin.
Embodiment 2:
Get clean container, import 1000ml distilled water (26 ℃), slowly add 100g trehalose dry powder, the limit adds just stirs with glass bar, and dry powder is dissolved fully, adds an amount of NaHCO3, regulates pH value to 7.1 filtration sterilization.
(2) calf serum of newly buying is stored in-20 ℃ of refrigerators, as for 4 ℃ of refrigerator defrostings about 10 hours, then as for deactivation complement component 3 in 56 ℃ of thermostat water baths 0 minute, constantly shakes serum bottle in the water-bath and makes and be heated evenly filtration sterilization before using.
(3) at normal temperatures put into preservation object (skin) after (24 ℃) adding calf serum 10g makes and seal, namely join namely with allowing it freely cool off about 2 hours.Put into 4 ℃ of Refrigerator stores
(4) the preservation liquid in the container is as the criterion with submergence skin.
Embodiment 3:
Get clean container, import 1000ml distilled water (25 ℃), slowly add 165g trehalose dry powder, the limit adds just stirs with glass bar, and dry powder is dissolved fully, adds an amount of NaHCO3, regulates pH value to 7.2 filtration sterilization.
(2) calf serum of newly buying is stored in-20 ~-30 ℃ of refrigerators, as for 4 ± 2 ℃ of refrigerator defrostings 10 ~ 11 hours, then as for deactivation complement component 3 in 58 ± 2 ℃ of thermostat water baths 0 minute, constantly shakes serum bottle in the water-bath and makes and be heated evenly filtration sterilization before using.
(3) at normal temperatures put into preservation object (skin) after (25 ℃) adding calf serum 16.5g prepares and seal, namely join namely with allowing it freely cool off about 2 ~ 3 hours.Put into 4 ± 2 ℃ of Refrigerator stores
(4) the preservation liquid in the container is as the criterion with submergence skin.
The Skin allograft selection criteria
(1) gets skin: get healthy donor's fresh cadaver skin, get the split-thickness skin graft that thickness is 0.4mm with manual dermatome
(2) age: 18-50 one full year of life
(3) draw materials the time: be no more than 8 hours under the normal temperature, be no more than 24 hours below 4 ℃.
(4) without tuberculosis, hepatitis, syphilis, acquired immune deficiency syndrome (AIDS) or rheumatism medical history.
(5) without severe trauma, mainly comprise large-area burns, MSW.
(6) serum detects: it is negative that HIV, HbsAg, HCV and syphilis serum detect reflection.
Below provide and adopt the detection method of MTT Skin Cell activity to adopt method to compare to conventional method and the present invention.
MTT arranges positive control (fresh skin) and negative control when measuring.
Use mtt assay detection of skin active (the fresh skin value is 100)
Experiment shows, the skin vitality of preserving with aqueous trehalose is apparently higher than traditional salt solution store method.For clinical practice provides the better quality Skin allograft.
Claims (4)
1. the preserving fluid for skin take trehalose as raw material is characterized in that: contain trehalose 45 ~ 165g/L, contain calf serum 4.5 ~ 16.5g/L.
2. the preparation method of preserving fluid for skin as claimed in claim 1, it is characterized in that: (1) gets trehalose dry powder, and with the distilled water mix and blend and make it dissolving, it is the 45-165g/L aqueous trehalose that preparation becomes concentration;
(2) add calf serum, stir;
(3) solution that stirs is regulated pH value to 7.0-7.2.
3. the preparation method of preserving fluid for skin as claimed in claim 2 is characterized in that: the distilled water temperature adds trehalose dry powder and also stirs, and it is fully dissolved in the time of 25 ℃ ± 2 ℃.
4. vitrification method is characterized in that: fresh Skin allograft is cleaned 2-5 time through stroke-physiological saline solution, put into after the drying and prepared trehalose and preserve the liquid immersion more than 8 hours, consumption is as the criterion with the submergence preservation object.
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| CN2013100188177A CN103004752A (en) | 2013-01-18 | 2013-01-18 | Skin preserving fluid with mycose as raw material, preparation method of skin preserving fluid and skin preservation method |
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| CN2013100188177A CN103004752A (en) | 2013-01-18 | 2013-01-18 | Skin preserving fluid with mycose as raw material, preparation method of skin preserving fluid and skin preservation method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108338160A (en) * | 2018-04-27 | 2018-07-31 | 张长水 | A kind of hair follicle preserves liquid and preparation method thereof |
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| CN1044575A (en) * | 1989-10-11 | 1990-08-15 | 中国人民解放军第二○四医院 | Preserve the method for bone, cartilage, skin with honey |
| CN1220824A (en) * | 1998-09-10 | 1999-06-30 | 河南省新乡市第二人民医院 | Preserving fluid for skin |
| CN101366976A (en) * | 2008-09-03 | 2009-02-18 | 陕西瑞盛生物科技有限公司 | Humanized heterogenous cell epimatrix material and preparation method thereof |
| RU2371916C1 (en) * | 2008-06-04 | 2009-11-10 | Государственное научное учреждение ордена "Знак Почета" Научно-исследовательский институт сельского хозяйства Крайнего Севера Сибирского отделения Российской академии сельскохозяйственных наук (ГНУ НИИСХ КС СО Россельхозакадемии) | Preservation method of reindeer unossified antlers using infrared radiation |
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| CN101870962A (en) * | 2010-06-12 | 2010-10-27 | 中国科学院西北高原生物研究所 | Method for constructing Tibetan antelope skin fibroblast line |
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2013
- 2013-01-18 CN CN2013100188177A patent/CN103004752A/en active Pending
Patent Citations (6)
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| CN1044575A (en) * | 1989-10-11 | 1990-08-15 | 中国人民解放军第二○四医院 | Preserve the method for bone, cartilage, skin with honey |
| CN1220824A (en) * | 1998-09-10 | 1999-06-30 | 河南省新乡市第二人民医院 | Preserving fluid for skin |
| RU2371916C1 (en) * | 2008-06-04 | 2009-11-10 | Государственное научное учреждение ордена "Знак Почета" Научно-исследовательский институт сельского хозяйства Крайнего Севера Сибирского отделения Российской академии сельскохозяйственных наук (ГНУ НИИСХ КС СО Россельхозакадемии) | Preservation method of reindeer unossified antlers using infrared radiation |
| CN101366976A (en) * | 2008-09-03 | 2009-02-18 | 陕西瑞盛生物科技有限公司 | Humanized heterogenous cell epimatrix material and preparation method thereof |
| CN101720753A (en) * | 2009-12-09 | 2010-06-09 | 中国人民解放军第四军医大学 | Cryopreservation solution of tissue engineering products and application method thereof |
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| 徐洋 等: "皮肤及皮肤相关组织细胞的深低温冻存研究进展", 《内蒙古医学杂志》 * |
Cited By (1)
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| CN108338160A (en) * | 2018-04-27 | 2018-07-31 | 张长水 | A kind of hair follicle preserves liquid and preparation method thereof |
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Application publication date: 20130403 |