CN1081379A - Human placental collagen and preparation method thereof - Google Patents
Human placental collagen and preparation method thereof Download PDFInfo
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- CN1081379A CN1081379A CN92108780A CN92108780A CN1081379A CN 1081379 A CN1081379 A CN 1081379A CN 92108780 A CN92108780 A CN 92108780A CN 92108780 A CN92108780 A CN 92108780A CN 1081379 A CN1081379 A CN 1081379A
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- collagen
- human placental
- placental collagen
- preparing human
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Abstract
A kind of method for preparing human placental collagen, it is characterized in that utilizing healthy puerpera's Placenta Hominis not have the peripheral part placenta tissue that meconium pollutes, after cleaning, smash, add processing such as pepsin digestion, extraction, mixing, can make the aseptic human placental collagen that has no side effect.Human placental collagen does not cause the operation receiveing person and produces immunoreation, and retention time is long, and orthopedic effect is better than the bovine collagen like product.Human placental collagen is used for filling miniature pitting skin injury.
Description
The invention belongs to the biomaterial for medical purpose field.
Be the injectivity bovine collagen in order to the biomaterial of filling miniature pitting skin defect (as cicatrix, wrinkle, acne scar etc.) in the world at present.Clinical proof bovine collagen may cause that human body is proteic immunoreation to xenogenesis.
Is proteic immunoreation for fear of human body to xenogenesis, seeks the biomaterial that retention time is long, orthopedic effect is better than bovine collagen, is used for shaping and beauty, filling miniature pitting skin defect, and human placental collagen is studied.
Get healthy puerpera's Placenta Hominis, freezing preservation is spent the night, and the peripheral part placenta tissue that next day, selected no meconium polluted shreds tissue with the acetone immersion after cleaning, and cleans.The Placenta Hominis that shreds is placed 0.5mol/L acetic acid and smash, add pepsin (0.1-2.9g/100g wet tissue) digestion.Centrifugal, stay precipitate A.Supernatant adds NaOH solution, abandons precipitation, and supernatant stays precipitation (collagen A) through dialysis, centrifugal.
Precipitate A is through NaCl(5-15%) wash, use Na
2HPO
4Extract repeatedly, centrifugal removal cleanout fluid, (0.1-0.4mol/L PH2.5-4.0) extracts precipitation, and is centrifugal, stays precipitation (collagen B) with citrate buffer solution.
With collagen A and collagen B press different proportion (10: 1,9: 1 ..., 1: 1) and mixing, phosphate buffer is dialysed to balance.
(0.01-0.2 μ/ml) and lignocaine, decentralized making it by No. 6 syringe needles is used to fill small skin depressed lesion after safety verification is qualified to add growth hormone.
Unless otherwise noted, more than operation is all carried out under aseptic, 0-4 ℃ condition.
This biomaterial does not cause the operation receiveing person and produces immunoreation, and retention time is long, and orthopedic effect is better than the bovine collagen like product.
Most preferred embodiment: get no hepatitis, acquired immune deficiency syndrome (AIDS) and other infectious disease, the free of contamination peripheral part of the Placenta Hominis of no heritability collagen is removed blood contamination to placenta tissue with the phosphate buffer spray irrigation of enzyme-containing inhibitor and is creamy white.
Tissue is shredded the back use the acetone soaked overnight, change secondary next day, the reuse phosphate buffer cleans 5 times.
Place 0.5mol/L acetic acid (PH2.5) to smash on the Placenta Hominis that shreds, and add pepsin (0.1-2.0g/100g wet tissue), digest 1-3 days (concrete dosage and natural law are determined according to the emulsifying degree of Placenta Hominis homogenate).Centrifugal 30 minutes, 14000rpm stayed precipitation (precipitate A).
Dropwise add 10mol/L NaOH in the supernatant and transfer PH to 7.2-7.8, abandon precipitation, supernatant is to 0.02mol/LNa
2HPO
4Dialysed 4-6 days, centrifugal, abandon supernatant, stay precipitation (collagen A).
Precipitate A is washed 8 times with 5 times of volume 10%NaCL, the 0.067mol/LNa of 5 times of volumes of reuse
2HPO
4Extract repeatedly 3 times, centrifugal removal supernatant, precipitation is extracted 4 times with the citrate buffer solution (PH37,0.15mol/L) of 5 times of volumes, and the centrifugal supernatant that goes stays precipitation (collagen B).
Collagen A and collagen B are pressed different proportion (1: 0,4: 1,3: 1,2: 1) mixing, phosphate buffer is dialysed to balance (PH7.0-7.4), add growth hormone (0.01-0.2 μ/ml) and lignocaine (0.1-0.5%).More than each position different and select according to patient and injection site.
Decentralized making it by No. 6 syringe needles is by coming into operation after the safety test.
Above process does not have specified otherwise and all carry out whole sterile workings under 0-4 ℃ of condition.
Claims (6)
1, a kind of method for preparing human placental collagen is characterized in that: the peripheral part placenta tissue with the no meconium of healthy puerpera's Placenta Hominis pollutes, and clean, shred and be placed in the acetic acid and smash, add pepsin digestion, the centrifugal precipitate A of staying; Supernatant is through the centrifugal precipitation (collagen A) of staying of dialysing; Precipitate A is washed through NaCl, uses Na
2HPO
4Extract, centrifugal removal cleanout fluid, precipitation is extracted with citrate buffer solution, the centrifugal precipitation (collagen B) of staying; Collagen A and collagen B dialyse to balance to phosphate buffer by the different proportion mixing, add growth hormone again and lignocaine gets final product.
2,, it is characterized in that pepsic amount is a 0.1-2.9g/100g wet tissue according to the described method for preparing human placental collagen of claim 1.
3, the method for preparing human placental collagen according to claim 1, the concentration that it is characterized in that NaCl is 5-15%.
4, the method for preparing human placental collagen according to claim 1, the concentration that it is characterized in that citrate buffer solution is 0.1-0.4mol/L, PH2.5-4.0.
5, the method for preparing human placental collagen according to claim 1 is characterized in that collagen A and collagen B were in 10: 1,9: 1,8: 1,7: 1,6: 1,5: 1,4: 1,3: 1,2: 1,1: 1 ratio mixing.
6, the method for preparing human placental collagen according to claim 1 is characterized in that the amount of the growth hormone that added is 0.01-0.2 μ/ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN92108780A CN1030528C (en) | 1992-07-29 | 1992-07-29 | Human placental Collagen and the prepn. thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN92108780A CN1030528C (en) | 1992-07-29 | 1992-07-29 | Human placental Collagen and the prepn. thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1081379A true CN1081379A (en) | 1994-02-02 |
| CN1030528C CN1030528C (en) | 1995-12-20 |
Family
ID=4943808
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN92108780A Expired - Fee Related CN1030528C (en) | 1992-07-29 | 1992-07-29 | Human placental Collagen and the prepn. thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1030528C (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1043554C (en) * | 1994-06-04 | 1999-06-09 | 徐念维 | Technology for producing full placenta nutritious agent |
| CN1063327C (en) * | 1995-05-17 | 2001-03-21 | 崔秀云 | Application of RNA enzyme inhibition factor used as tumour inhibitor |
| CN1064970C (en) * | 1996-01-23 | 2001-04-25 | 孙书臣 | Non-densatured extraction of medical collagen protein and denatured application |
| CN1066066C (en) * | 1996-07-09 | 2001-05-23 | 王革生 | Filling injection material for cosmetic face-lifting |
| CN102599401A (en) * | 2011-01-19 | 2012-07-25 | 班云峰 | Fetus cervi collagen beauty food and preparation method thereof |
| CN103525890A (en) * | 2013-09-27 | 2014-01-22 | 四川大学 | Preparation method of human placenta collagen |
| CN104894201A (en) * | 2015-06-26 | 2015-09-09 | 济南唐恩生物技术有限公司 | Method for preparing freeze-dried active placenta powder through water solution enzymolysis |
| CN104195205B (en) * | 2014-04-24 | 2019-08-16 | 吉林金梓源生物科技股份有限公司 | Method and prepared high activity collagen peptide and application thereof from animal placenta preparation high activity collagen peptide |
| CN113402599A (en) * | 2021-05-18 | 2021-09-17 | 铜仁市泛特尔生物技术有限公司 | Optimization method for extracting and purifying type I collagen from human placenta |
-
1992
- 1992-07-29 CN CN92108780A patent/CN1030528C/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1043554C (en) * | 1994-06-04 | 1999-06-09 | 徐念维 | Technology for producing full placenta nutritious agent |
| CN1063327C (en) * | 1995-05-17 | 2001-03-21 | 崔秀云 | Application of RNA enzyme inhibition factor used as tumour inhibitor |
| CN1064970C (en) * | 1996-01-23 | 2001-04-25 | 孙书臣 | Non-densatured extraction of medical collagen protein and denatured application |
| CN1066066C (en) * | 1996-07-09 | 2001-05-23 | 王革生 | Filling injection material for cosmetic face-lifting |
| CN102599401A (en) * | 2011-01-19 | 2012-07-25 | 班云峰 | Fetus cervi collagen beauty food and preparation method thereof |
| CN102599401B (en) * | 2011-01-19 | 2013-06-12 | 班云峰 | Fetus cervi collagen beauty food and preparation method thereof |
| CN103525890A (en) * | 2013-09-27 | 2014-01-22 | 四川大学 | Preparation method of human placenta collagen |
| CN103525890B (en) * | 2013-09-27 | 2016-08-10 | 四川大学 | A kind of preparation method of human placental collagen |
| CN104195205B (en) * | 2014-04-24 | 2019-08-16 | 吉林金梓源生物科技股份有限公司 | Method and prepared high activity collagen peptide and application thereof from animal placenta preparation high activity collagen peptide |
| CN104894201A (en) * | 2015-06-26 | 2015-09-09 | 济南唐恩生物技术有限公司 | Method for preparing freeze-dried active placenta powder through water solution enzymolysis |
| CN113402599A (en) * | 2021-05-18 | 2021-09-17 | 铜仁市泛特尔生物技术有限公司 | Optimization method for extracting and purifying type I collagen from human placenta |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1030528C (en) | 1995-12-20 |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C15 | Extension of patent right duration from 15 to 20 years for appl. with date before 31.12.1992 and still valid on 11.12.2001 (patent law change 1993) | ||
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| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |