CN101279104B - Preparation of collagen protein sponge containing growth factor - Google Patents
Preparation of collagen protein sponge containing growth factor Download PDFInfo
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- CN101279104B CN101279104B CN2007100108444A CN200710010844A CN101279104B CN 101279104 B CN101279104 B CN 101279104B CN 2007100108444 A CN2007100108444 A CN 2007100108444A CN 200710010844 A CN200710010844 A CN 200710010844A CN 101279104 B CN101279104 B CN 101279104B
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Abstract
A preparation method of collagen sponge with growth factor relates to a preparation method of collagen sponge containing epidermic cell growth factor and basic fibroblast growth factor. The invention provides the preparation method of collagen sponge which contains growth factors, has active recovery function and can facilitate the healing of chronic wound. The technical proposal of the inventionis as follows: 1. the skin or the connective tissue of animals is used for producing collagen enzymolysis liquid; 2. the epidermic cell growth factor and the basic fibroblast growth factor are added into the collagen enzymolysis liquid respectively; 3. the collagen enzymolysis liquid containing the epidermic cell growth factor and the basic fibroblast growth factor is frozen in vacuum and dried into collagen sponge; 4. the collagen sponge containing the epidermic cell growth factor and the basic fibroblast growth factor is cut, packaged and disinfected by cobalt-60.
Description
Technical field:
The present invention relates to contain the preparation method of the collagen protein sponge of epithelical cell growth factor and basic fibroblast growth factor.
Background technology:
1, under normal circumstances, healthy people's the wound healing factor (cell growth factor) seems very unimportant.But to suffering from the patient of multiple chronic difficult healing wound surface, the wound healing factor then shows magical effect, so this is only the main application direction of various cell growth factor.
When suffering from some disease, as diabetes, arteriosclerosis, chronic venous expansion and obstruction, decubital ulcer, lupus erythematosus, rheumatoid arthritis, old people etc., may be because of Developmental and Metabolic Disorder, part or nutrition of whole body are not enough and cause The wound refused to heal.Lower limb foot wound as diabetics was several years even disunion decades.The Knighton research group once used the platelet source wound healing factor (the Platelet-derivedwound healing factor of patient from body, PDWHF), be actually the mixture of multiple somatomedin, the wound of the difficult healing of treatment, lower limb foot wound, decubital ulcer, tremulous pulse blood supply insufficiency and the venous return that comprises the diabetes patient be smooth patient's ulcer etc. not.The result obtains unexpected effect, and cure rate reaches 97%, and the mean treatment time was 10.6 weeks.And 14 people recovery from illness is arranged among 19 patients of report with the PDWHF treatment, and still have 9 people not heal after 8 weeks for 12 patients of matched group of placebo.
Some treatment measure also can cause disease more of wound, as accepts the patient of chemotherapy, radiotherapy and steroid therapy, because of some cell in the body is suppressed the effect that loses some somatomedin of secretion, causes patient's The wound refused to heal.As accept the patient of 17-hydroxy-11-dehydrocorticosterone treatment, intravital macrophage is suppressed, and has lost the function of secretion TGF-β, thereby has influenced wound healing, uses the wound healing factor will more apparent necessity in this case.
The damage of some tissues is original just to be difficult to healing than other tissue, and as corneal injury, head injury etc., this also is to use one of first-selected case of the wound healing factor.
2, along with somatomedin in Clinical Application, brought new thinking for treatments of these difficult healing wound surface.Epidermal growth factor (EGF) is the mitogen of epidermis cell, can promote the division and the propagation of epidermis cell.Basic fibroblast growth factor (bFGF) can promote the division and the propagation of fibroblast and vascular endothelial cell effectively.These two kinds of somatomedin play important effect in the wound healing process.The use of uniting of these two kinds of somatomedin has obtained gratifying effect on clinical treatment.People such as Xue Bin have the patient of chronic difficult healing wound surface to use EGF and bFGF to find simultaneously to 58, unite and use these two kinds of somatomedin obviously to promote wound healing.(Xue Bin etc., Chinese clinical rehabilitation the 8th volume the 2nd phase 2004-01-15)
3, the effect of somatomedin such as EGF and bFGF in wound active repair process is familiar with by people more and more widely, but the activity of most of somatomedin can descend rapidly at normal temperatures.Easily run off again in the wound when using as liquid preparation.So, inconvenience is arranged when clinical use, can not give full play to its curative effect.
Summary of the invention:
The present invention is exactly at the problems referred to above, provides a kind of and promotes the chronic wound healing and have the initiatively preparation method of the collagen protein sponge that contains somatomedin of repair function.
The present invention adopts following technical scheme:
1, produces collagen protein enzymolysis liquid with skin or the connective tissue of animal;
2, add respectively in the enzymolysis solution of collagen protein with epithelical cell growth factor and fibroblast growth factor, the concentration of epithelical cell growth factor is 120IU/ml-250IU/ml, and the concentration of fibroblast growth factor is 200AU/ml-350AU/ml;
3, the collagen protein enzymolysis liquid that will contain epithelical cell growth factor, fibroblast growth factor becomes spongy through vacuum lyophilization;
Collagen protein sponge cutting, packing, cobalt 60 radiation sterilizations that 4, will contain epithelical cell growth factor and fibroblast growth factor.
The beneficial effect of the invention:
The present invention provides the new combined material of an epidermal growth factor and fibroblast growth factor collagen protein sponge on the basis of fibroblast growth factor collagen protein sponge, this new material as surgical dressing for wound particularly the healing of chronic wound facilitation is arranged.This biogenic dressing that contains the active repairing activity uses just clinically, and effect is obvious.
Our experiments show that because two kinds of factors using among the present invention complementarity of repair functions initiatively, epithelical cell growth factor and fibroblast growth factor are united to use and made collagen protein sponge better therapeutic is arranged in clinical practice.Especially the reparation to chronic difficult healing wound surface is that other wound dressing institute can not reach.
The specific embodiment:
Embodiment 1:
1, producing of collagen protein enzymolysis liquid:
Fresh Corii Sus domestica that degrease is cleaned out or Corii Bovis seu Bubali or connective tissue are put into stainless steel cask, add soaking in sodium carbonate solution 15-30 minute.After pulling out, wash repeatedly with pure water, the drop branch that anhydrates with animal connective tissue's chopping, is weighed.Putting into meat grinder rubs.Add 75% alcohol solution dipping 10-25 minute.Pulling the back out cleans with pure water.The drop branch that anhydrates is weighed.Ratio in 1kg skin/15-30L water adds pure water, goes in the agitated kettle.Adding appropriate hydrochloric acid solution in homogenate makes solution be acid.Add an amount of pepsin solution, at room temperature, airtight stirring 15-24 hour makes into thickness, the colloid fluid.Add an amount of saturated nacl aqueous solution, stirred 3 hours.7000rpm is centrifugal, collecting precipitation.Be dissolved in the dilute hydrochloric acid solution.Pack in the bag filter, dialysed 48 hours with pure water.Transfer to PH neutrality, obtain collagen protein enzymolysis liquid.
Add somatomedin in enzymolysis solution, wherein the epidermal growth factor final concentration is 200IU/ml, and the fibroblast growth factor final concentration is 250AU/ml; Stir, put into stainless steel disc.
Stainless steel disc is placed vacuum freeze drier, lyophilizing 10-16 hour.Take out the lyophilized protein sponge, cut into the product of different size by clinical needs.The inner packing sealing.The collagen protein sponge that contains somatomedin cobalt 60 radiation sterilizations with package encapsulation.Product after the sterilization uses for clinical experiment.
The somatomedin collagen protein sponge quickens the clinical trial of chronic difficult healing wound healing:
1, materials and methods:
1.1 clinical data: this organizes 28 examples totally 33 wound surface, wherein male 18 examples, women 10 examples; Age 10-90 year; Traumatic ulcer 12 examples, totally 14 wound surface; Pressure ulcer 9 examples, totally 12 wound surface; Diabetic ulcer 4 examples; Radiation ulcer 3 examples; The shortest person of the course of disease 30 days, elder 10 years (table 1).In this research, will cause by a variety of causes, be decided to be chronic difficult healing wound surface through treating the wound surface that do not heal in 4 weeks.33 wound surface of this group all are through healing person not of conventional 4 weeks.
Table 128 example is the related data of totally 33 chronic ulcer in body surfaces
| Group | Wound surface number/routine number | The ulcer size | The course of disease (my god) |
| Traumatic ulcer | 14/12 | 1.0×3.0-5.0×10.0 | 30-90 |
| Pressure ulcer | 12/9 | 2.0×3.0-16.0×18.0 | 38-365 |
| Diabetic ulcer | 4/4 | 1.5×1.5-6.0×8.0 | 45-60 |
| Radiation ulcer | 3/3 | 2.5×3.0-10.0×18.0 | 45-3650 |
1.2 material source:
Somatomedin collagen protein sponge (bioprotein sponge) is prepared by Liaoning Green Valley Medical Apparatus Co., Ltd..
1.3 Therapeutic Method:
All wound surface all carry out debridement before the treatment of bioprotein sponge, routine disinfection needs blood sugar control simultaneously to the diabetic ulcer patient.The bioprotein sponge is with before being stored in 4 ℃ of thermostatic water tanks.Time spent is the complete flap coverage of bioprotein sponge, and slightly exceeds edge of wound 0.5cm.If wound surface oozes out less or inflammatory reaction when heavier, cover again after available injection liquid of gentamicin is moistening.Simultaneously, note the whole body anti-inflammatory treatment.Once a day, it was a course of treatment with 7 days.Simultaneously, visual wound healing situation, proper extension is used the time (2-3 days) instead.
1.4 efficacy determination and statistical analysis:
Because the significant difference that the chronic difficult healing wound surface cause of disease, the course of disease and ulcer distribute, thereby be difficult in the clinical experiment contrast between suitable group.Because this group MethodsThe cases enrolled is for to treat the wound surface that did not heal in 4 weeks through conventional method, still heal fully the required time except that adding up each wound surface under study for action, be curative effect determinate standard also with 4 courses of treatment (28 days).
Table 2 bioprotein sponge is to the curative effect of different wound surface
2, result:
This is organized in 33 wound surface of 28 examples, applying biological protein sponge 20 wound surface of healing person in 2 weeks, and 3 wound surface of healing person in 3 weeks, 8 wound surface of healing person in 4 weeks surpass 2 wound surface of 4 all healing persons.Full group total effective rate total healing rate 93.9% in 100%, 4 week.The curative effect situation of relevant different cause of disease wound surface sees Table 2.
Analyze to adopt the healing ratio of bioprotein sponge to wound surface due to the different causes of disease, as seen 14 traumatic ulcers are at 13 wound surface of treatment early stage (in 2 weeks) healing, account for 92.9%, diabetic ulcer and pressure ulcer then are respectively 75% and 25%, compare significant difference (P＜0.05) with traumatic ulcer.This shows that the bioprotein sponge is different to the chronic healing wound surface curative effect of the different causes of disease.
Claims (2)
1. preparation method that contains the collagen protein sponge of somatomedin, step is:
1), with Corii Sus domestica or Corii Bovis seu Bubali or connective tissue production collagen protein enzymolysis liquid:
2), add respectively in the enzymolysis solution of collagen protein with epithelical cell growth factor and fibroblast growth factor; The concentration of epithelical cell growth factor is 120IU/ml-250IU/ml, and the concentration of fibroblast growth factor is 200AU/ml-350AU/ml;
3), the collagen protein enzymolysis liquid that will contain epithelical cell growth factor, fibroblast growth factor is spongy through vacuum lyophilization;
Collagen protein sponge cutting, packing, cobalt 60 radiation sterilizations that 4), will contain epithelical cell growth factor and fibroblast growth factor;
The process that it is characterized in that enzymolysis is at room temperature carried out, and sealing was stirred 15-24 hour.
2. a kind of preparation method that contains the collagen protein sponge of somatomedin according to claim 1 is characterized in that epithelical cell growth factor concentration is 200IU/ml, and the concentration of fibroblast growth factor is 250AU/ml.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108014370A (en) * | 2017-12-12 | 2018-05-11 | 吕振木 | A kind of promotion, which is press-offed, injures the collagen material that avulsion skin survives |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102416195B (en) * | 2010-09-28 | 2014-02-19 | 北京益而康生物工程开发中心 | Preparation method of collagen sponge |
| CN102357259A (en) | 2011-07-28 | 2012-02-22 | 王珊珊 | Bioprotein sponge and preparation method thereof |
| CN102675473B (en) * | 2012-05-07 | 2013-05-29 | 西安华澳丽康生物工程有限公司 | Gene recombinant human active basic fibroblast growth factor fusion protein, preparation method thereof and application thereof |
| CN104490760B (en) * | 2014-12-12 | 2018-02-16 | 广东医学院 | The preparation method and applications of capsaicine collagen protein sponge |
| CN106139375A (en) * | 2015-04-17 | 2016-11-23 | 天津市赛宁生物工程技术有限公司 | A kind of manufacture method of negative pressure drainage device |
| RU2637634C1 (en) * | 2016-10-21 | 2017-12-05 | Владимир Иосифович Лозинский | Antibacterial protein sponge for chemotherapy of infected wounds and method of its production |
| CN106880871B (en) * | 2017-01-18 | 2019-12-13 | 烟台正海生物科技股份有限公司 | Collagen dermal material for promoting endometrial repair and preparation method thereof |
| CN114632183B (en) * | 2022-03-30 | 2023-02-03 | 温州医科大学 | Wound protein sponge, preparation method thereof and application of wound protein sponge in preparation of medicines for reducing scar formation in skin repair |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5700476A (en) * | 1992-03-25 | 1997-12-23 | Johnson & Johnson Medical, Inc. | Heteromorphic sponges containing active agents |
| CN1228339A (en) * | 1999-02-04 | 1999-09-15 | 李校坤 | Sponge material capable of promoting spinal cord tissue wound healing, its preparing method and use |
| CN1228338A (en) * | 1999-02-04 | 1999-09-15 | 马昭若 | Wound covering material capable of promoting wound healing |
| JP2004035526A (en) * | 2002-07-08 | 2004-02-05 | Minoru Ueda | Wound shrinking agent and method for producing the same |
| CN1511592A (en) * | 2002-12-30 | 2004-07-14 | 中国皮革和制鞋工业研究院 | Construction method for skin tissue engineering rack containing epidermal growth factor |
-
2007
- 2007-04-05 CN CN2007100108444A patent/CN101279104B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5700476A (en) * | 1992-03-25 | 1997-12-23 | Johnson & Johnson Medical, Inc. | Heteromorphic sponges containing active agents |
| CN1228339A (en) * | 1999-02-04 | 1999-09-15 | 李校坤 | Sponge material capable of promoting spinal cord tissue wound healing, its preparing method and use |
| CN1228338A (en) * | 1999-02-04 | 1999-09-15 | 马昭若 | Wound covering material capable of promoting wound healing |
| JP2004035526A (en) * | 2002-07-08 | 2004-02-05 | Minoru Ueda | Wound shrinking agent and method for producing the same |
| CN1511592A (en) * | 2002-12-30 | 2004-07-14 | 中国皮革和制鞋工业研究院 | Construction method for skin tissue engineering rack containing epidermal growth factor |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108014370A (en) * | 2017-12-12 | 2018-05-11 | 吕振木 | A kind of promotion, which is press-offed, injures the collagen material that avulsion skin survives |
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