CN1557478A - Skin, connective tissue and bone growth promoting composition - Google Patents
Skin, connective tissue and bone growth promoting composition Download PDFInfo
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- CN1557478A CN1557478A CNA2004100158539A CN200410015853A CN1557478A CN 1557478 A CN1557478 A CN 1557478A CN A2004100158539 A CNA2004100158539 A CN A2004100158539A CN 200410015853 A CN200410015853 A CN 200410015853A CN 1557478 A CN1557478 A CN 1557478A
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Abstract
The present invention relates to composition capable of promoting the growth of skin, connective tissue and bone and its preparation, and is especially one kind of growth factor-polypeptide composition capable of promoting the growth of skin, connective tissue and bone and its application. The present invention features that growth factor colony is extracted from cow colostrums via ultrafiltering, concentrated and freeze dried to obtain powder as the growth factor preparation. The growth factor preparation may be applied alone or through combination with gene recombinant growth factor to promote the growth of skin, connective tissue and bone and the healing of wound. The specific scheme is designed for applying the composition in treating wound and promoting growth of bone and teeth and in cosmetics.
Description
Affiliated technical field
The present invention relates to a kind of composition and method of making the same that promotes skin, connective tissue, osteogenesis, particularly a kind of preparation method and application that promotes the somatomedin class peptide composition of skin, connective tissue, osteogenesis.
The invention technical background
Somatomedin is that a class is the compositions of ultimate unit with the peptide, as the term suggests it can promote the growth of various cell masses in the body, segmentation is got up, and in fact its function is diversified, and its composition and effect are similar, often complement each other.
Platelet derived growth factor (PDGF) is a kind of cation of finding in the circulation granule of platelet, thermally-stabilised material, having stimulates the dawn white matter synthetic and stimulate the white effect that generates of collagen dawn by fibroblast, and it also rises fibroblast, smooth muscle cell and neurogliocyte and promotes cell division and chemotaxis.Quasi-insulin growthing factor I and II (GIF-I, II) on cell proliferation, differentiation have facilitation, and wound healing is had obvious benefit.Epidermal growth factor (EGF) can promote epidermal growth, and undermined skin repair is had facilitation.Transforming growth factor and β (TGF α, β) are significant to mucosa growth and tissue injury repair process.Fibroblast growth factor (FGF) can promote to organize normal growth, regeneration, accelerates old and feeble injured muscle, skin collagen matter, cartilage and nervous tissue and repairs.Nerve growth factor (NGF) can stimulate the sympathetic ganglion growth.
There has been researcher to propose to utilize platelet derived growth factor (PDGF) to promote wound healing, GrotendorstG. (J.Trauma 24, No.9,549-552 (1984)) having introduced PDGF improves new white synthetic quantity of collagen dawn, but proposes method and the specified operational procedure used.In addition, also there are a plurality of researcheres to report quasi-insulin growthing factor I and can obviously promote wound healing, but also do not propose concrete approach and the method for using.Known studies show that, other multiple somatomedin all have the effect that promotes tissue growth.
Multiple somatomedin comprises that PDGF and GIF all can be by the mode artificial preparation of gene recombinaton, but price is very expensive, preservation, stores also difficulty.
Contain multiple somatomedin in the cattle colostrums, comprise the somatomedin group that insulin like growth factor, platelet derived growth factor, epidermal growth factor, nerve growth factor, SGF, fibroblast growth factor, transforming growth factor etc. are abundant.
The disclosed technology of Chinese invention patent (application number 96116428), introduced from cattle colostrums the method for extracting the insulin like growth factor that combines in vain in conjunction with the dawn with insulin like growth factor, but this method can not be extracted highly purified insulin like growth factor.
Technical problem to be solved by this invention is, preparation has the somatomedin group preparation of facilitation to skin, connective tissue, osteogenesis from cattle colostrums, reduce the preparation cost of growth factor preparation, by above-mentioned preparation of independent application or use in conjunction gene recombinaton somatomedin, design effectively promotes skin, connective tissue, osteogenesis, promote the compositions of wound healing, and be applied to clinical.
Summary of the invention
The technical solution adopted for the present invention to solve the technical problems is that the application hyperfiltration technique extracts the somatomedin group from cattle colostrums, prepare powder through the ultrafiltration and concentration postlyophilization.Use the growth factor preparation for preparing in the cattle colostrums, or use in conjunction gene recombinaton somatomedin, promote skin, connective tissue, osteogenesis, promote wound healing, and design concrete application scheme these compositionss are applied to treat wound, promote osteogenesis or dental growth, and be applied in the cosmetics.
Technical characterictic of the present invention is, the method of extracting the somatomedin group from cattle colostrums is, collect the cattle colostrums of fresh or freezing preservation, adopt centrifugation to remove fat, after the degrease cattle colostrums dilutes with aseptic distilled water, with low-molecular-weight supermicro filtration membrane separating and filtering, concentrate moisture content with ultrafilter membrane after collecting supernatant, concentrate lyophilization powder process, said preparation includes abundant somatomedin group, in order further to increase yield rate, with the residue of ultra-filtration and separation, transfer PH to highly acid with acid supplement, promote somatomedin and in conjunction with the dawn connection between white interrupt, make somatomedin dissociate out, and then with low-molecular-weight ultrafilter membrane separating and filtering, regulate PH to faintly acid or neutrality with alkali liquor after collecting supernatant, concentrate the moisture content postlyophilization and prepare dry powder.
Technical characterictic of the present invention is, from cattle colostrums, extract the somatomedin group and also can adopt another kind of method, the defat cattle colostrums is directly regulated PH to highly acid with acid solution, make somatomedin from white, dissociate out,, collect supernatant with the direct separating and filtering of low-molecular-weight supermicro filtration membrane in conjunction with the dawn, transfer PH to faintly acid or neutrality with alkaline agent, prepare dry powder through concentrating removal moisture content postlyophilization, this preparation includes the higher somatomedin group of purity, especially IGF content height.
Technical characterictic of the present invention is, adopts pH value that acid supplement adjusts skimmed colostrum to highly acid, and the acid supplement of employing can be HCL or other acid supplements, adjust pH value to 2.2～3.5, preferred 2.8～2.9, adjust pH value to the interval of regulation, be 8 with molecular weight, 000～20,000 ultrafilter membrane adopts conventional ultrafiltration equipment, it would be desirable to adopt the staggered fluid ultrafiltration apparatus of tangent plane to separate, collect filtrate, adjust pH value to 5.5～7.5 of filtrate with basic formulations, preferred 5.8～6.2, adjusting filtrate behind the pH value, to adopt molecular weight be 3,000～6,000 ultrafilter membrane is preferably molecular weight 4,000～5,000 ultrafilter membrane, adopt conventional ultrafiltration equipment to be concentrated into initial capacity 10%～60%, after preferred 20%～30%, deliver to freeze drying equipment, the preparation lyophilized powder includes abundant insulin like growth factor.After adjusting pH value, also can in filtrate, add and be equivalent to 1 times～5 times of filtrate, concentrate behind preferred 2 times～3 times aseptic distilled water dilute filtration liquid, more should adopt back one step when especially adopting tangent plane to interlock the fluid ultrafiltration apparatus.
Technical characterictic of the present invention is, also can adopt another approaches and methods to prepare somatomedin group in the cattle colostrums, its method is a defat cattle colostrums molecular weight 8,000～15,000 supermicro filtration membrane separating and filtering, collect supernatant, with molecular weight is that 4000～6000 supermicro filtration membrane concentrates moisture content to original volume 10%～20%, send into freeze drying plant and prepare dry powder, the residue of ultra-filtration and separation is regulated pH value to 2.7～2.9 with 2N HCL, impels free with the somatomedin that combines white bonding state of dawn, with molecular weight is 10,000～20,000 supermicro filtration membrane separating and filtering is collected supernatant, behind 2N NaOH adjusting pH value to 5.8～7.0, with molecular weight is that 4000～6000 supermicro filtration membrane concentrates moisture content to original volume 20%～30%, and lyophilization prepares dry powder, is rich in multiple somatomedin in the preparation.
Technical characterictic of the present invention is, the preparation for preparing by the inventive method with effective dose puts on the wound, cure patient's external wounds, these external wounds include but not limited to decubital ulcer, laceration, burn, scalpel edge, skin ulcer, the cureless wound of diabetes patient etc., above-mentioned preparation is by the growth of promotion epithelium and connective tissue and synthesizing of promotion dawn white matter and collagen, help healing or healing of wound at least partly, the preparation for treating wound of the inventive method preparation can reach better effect than not handling (promptly not applying the external source agent).
Technical characterictic of the present invention is, by patient being applied the preparation of the present invention's preparation, preferably apply the above-mentioned preparation of effective dose to the zone of injured zone or scarce bone, promote the regeneration of patient's sclerotin, above-mentioned preparation helps inostosis by promoting connective tissue, sclerotin and cemental growth and stimulating the synthetic of dawn white matter and collagen, or at least partly regeneration, adopt preparation of the present invention to promote inostosis, reach better effect than not handling (promptly handling) without the external source agent.
Technical characterictic of the present invention is, the somatomedin that in the above-mentioned preparation of the present invention, adds some gene recombinaton, include but not limited to platelet derived growth factor (PDGG) and insulin like growth factor (GIF), can strengthen the effect that preparation of the present invention promotes skin, connective tissue, osteogenesis, be used for the treatment of patient's wound and can obtain better therapeutic.Can in preparation of the present invention, add platelet derived growth factor (PDGF) separately, also can in preparation of the present invention, add insulin like growth factor (GIF) separately, the independent adding of above-mentioned component can improve the therapeutic effect of preparation of the present invention, preparation of the present invention is internal energy to reach better therapeutic effect but PDGF and GIF add simultaneously, the ratio that PDGF and GIF add is that optimum is that ratio between two is between 1: 1～2: 1 in 1: 5 to 10: 1 the scope.
Technical characterictic of the present invention is, during in the independent application of the above-mentioned preparation of the present invention or with other growth factor preparation use in conjunction, compositions is by acceptable carrier material on a kind of materia medica, for example inert gel that can buy at pharmaceuticals or pharmacy or liquid are (for example, be added with the saline of white or methylcellulose of dawn clearly), other growth factor preparations (for example gene recombinaton growth factor preparation) of preparation of the present invention and/or combination are combined produce.
Above-mentioned inert gel or liquid, and the somatomedin of gene recombinaton can buy commercial prod in biotech firm, pharmaceuticals, pharmaceutical factory.
Technical characterictic of the present invention is, above-mentioned preparation of the present invention can be used separately, also can add other forms of growth factor preparation (for example gene recombinaton growth factor preparation) use in conjunction, these somatomedin include but not limited to insulin like growth factor, epidermal growth factor, platelet derived growth factor etc.Above-mentioned composition can add in the cosmetics, can stimulate the growth of normal skin and the reparation of cell, and the protection and the maintenance of skin had very important function.Above-mentioned composition has obvious effects to the protection and the maintenance of skin than the cosmetics that do not add above-mentioned composition.
The invention has the beneficial effects as follows, the above-mentioned composition of above-mentioned preparation of the present invention and/or use in conjunction provides a safety, method fast and effectively for treatment patient's external wounds and bone tissue growth are not good, this compositions with treat (promptly not adding the external source agent) naturally and compare the formation that has strengthened skin, connective tissue, sclerotin.Promote new connective tissue to form, promote the epithelial tissue growth fraction not add external source agent treatment and increase by 80% that the epithelial tissue layer that obtains is thicker, and comprises the epithelium projection that more and new connective tissue is connected than not adding the about increase by 20% of external source agent treatment.Therefore, it is more firm in conjunction with getting, and protectiveness is more arranged, and in addition, cicatrization is reduced to minimum.Above-mentioned composition also provides a kind of method fast and effectively for patient's that the periodontal history is arranged connective tissue and inostosis, compares with treatment naturally, and compositions has strengthened the formation of connective tissue and sclerotin.In addition, above-mentioned composition can obviously improve the nutrition of skin for the protection and the maintenance of skin provide an effective method, promotes the epiderm skin cellular metabolism, strengthens the subcutaneous connective tissue hypertrophy, increases skin elasticity, reduces wrinkle and produces.
The specific embodiment
The invention will be further described below by embodiment
Embodiment 1
Preparation somatomedin group from cattle colostrums
Collect cattle colostrums, with 3000 rev/mins of speed centrifugal degreasings, with 2 times of amount distilled water dilution defat cattle colostrumss, the employing molecular weight is 12000 supermicro filtration membrane plate and frame ultrafilter isolated by filtration, collect supernatant, adopting molecular weight is that 5000 supermicro filtration membrane is concentrated into original volume 20%, and concentrate is sent into freeze drying plant, lyophilization is prepared into white powder, and preparation contains multiple somatomedin group.
Embodiment 2
Preparation somatomedin group from cattle colostrums
The defat cattle colostrums is transferred PH to 2.8 with 2N HCL, it with molecular weight 15000 supermicro filtration membrane, staggered tangent plane fluid ultrafiltration apparatus (TFF) separating and filtering that adopts U.S. Pall company to produce, collect supernatant, transfer PH to 6.2 with 2N NaOH, use same sampling device, but adopting molecular weight is that 4000 supermicro filtration membrane is concentrated into original volume 30%, concentrate is sent into freezer dryer and is prepared dry powder, be rich in various somatomedin in the preparation, especially insulin like growth factor (GIF) content is the highest, measures through insulin like growth factor radioimmunity medicine box, contains 0.5 microgram insulin like growth factor in every gram dry powder.
Embodiment 3
Preparation promotes the gel of wound healing
Dry powder 3 grams of getting embodiment 1 preparation add in the methyl cellulose gel, use the magnetic stirrer mixing.
Embodiment 4
Preparation promotes the growth factor combination of wound healing
Get dry powder 2 grams of embodiment 2 preparations, get recombination platelet derived growth factor purification preparation (PDGF) 10 micrograms, (from Amgen company's T housand oaks, CA, USA buys), get recombination insulin like growth factor purification preparation 10 micrograms (buying), combinations thereof is suspended in the biocompatible gel (contains white saline of dawn clearly) from above-mentioned same company, standby with even 10 minutes of magnetic stirrer.
Embodiment 5
Yorkshire Xiao Bai pig anesthesia with 10～15 kilograms, chest and back is shaved hair, local 70% alcohol disinfecting of using, leaving 1.5cm * 2.5cm degree of depth with electric knife is the 0.5mm otch, and make epithelium and be embedded in following corium dawn white matter removal, same a kerf is done at the 4cm place being separated by, organize in contrast, the preparation of getting embodiment 4 preparations is applied to wound, gathered biopsy samples in the 3rd day～the tenth day, observe at microscopically, find that the wound use preparation group of the present invention has thicker connective tissue and epithelial layer, and have widely that the epithelium projection links to each other with epithelial layer.
Embodiment 6
Select number to have only the beagle of spontaneous generation periodontal disease, detect the damaged tooth of 50%-70% bone with ultrasonic instrument, carry out the root planification, the preparation of making of embodiment 4 discharges on cacodontia, demonstration is observed by periodontal tissue and bone, root surface exists new osteoplastic obvious zone, and in the root surface with new bone adjacency the similar cemental deposit of one deck is arranged.
Claims (10)
1. the present invention relates to a kind of composition and method of making the same that promotes skin, connective tissue, osteogenesis, be particularly related to a kind of preparation method and application that promotes the somatomedin class peptide composition of skin, connective tissue, osteogenesis, it is characterized in that, use hyperfiltration technique and from cattle colostrums, extract the somatomedin group, prepare powder through the ultrafiltration and concentration postlyophilization.Use the growth factor preparation for preparing in the cattle colostrums, or use in conjunction gene recombinaton somatomedin, promote skin, connective tissue, osteogenesis, promote wound healing, and design concrete application scheme these compositionss are applied to treat wound, promote osteogenesis or dental growth, and be applied in the cosmetics.
2. in accordance with the method for claim 1, technical characterictic of the present invention is, the method of extracting the somatomedin group from cattle colostrums is, collect the cattle colostrums of fresh or freezing preservation, adopt centrifugation to remove fat, after the degrease cattle colostrums dilutes with aseptic distilled water, with low-molecular-weight supermicro filtration membrane separating and filtering, concentrate moisture content with ultrafilter membrane after collecting supernatant, concentrate lyophilization powder process, said preparation includes abundant somatomedin group, in order further to increase yield rate, with the residue of ultra-filtration and separation, transfer PH to highly acid with acid supplement, promote somatomedin and in conjunction with the dawn connection between white interrupt, make somatomedin dissociate out, and then, regulate PH to faintly acid or neutrality with alkali liquor behind the collection supernatant with low-molecular-weight ultrafilter membrane separating and filtering, concentrate the moisture content postlyophilization and prepare dry powder.
In accordance with the method for claim 1 in, technical characterictic of the present invention is, from cattle colostrums, extract the somatomedin group and also can adopt another kind of method, the defat cattle colostrums is directly regulated PH to highly acid with acid solution, make somatomedin from white, dissociate out in conjunction with the dawn, with the direct separating and filtering of low-molecular-weight supermicro filtration membrane, collect supernatant, transfer PH to faintly acid or neutrality with alkaline agent, prepare dry powder through concentrating removal moisture content postlyophilization, this preparation includes the higher somatomedin group of purity, especially IGF content height.
In accordance with the method for claim 1 in, technical characterictic of the present invention is, adopt acid supplement to adjust the pH value of skimmed colostrum to highly acid, the acid supplement that adopts can be HCL or other acid supplements, adjusts pH value to 2.2～3.5, preferred 2.8～2.9, adjust pH value to the interval of regulation, it with molecular weight 8,000～20,000 ultrafilter membrane, adopt conventional ultrafiltration equipment, it would be desirable and adopt the staggered fluid ultrafiltration apparatus of tangent plane to separate, collect filtrate, adjust pH value to 5.5～7.5 of filtrate with basic formulations, preferred 5.8～6.2, filtrate employing molecular weight behind the adjustment pH value is 3,000～6,000 ultrafilter membrane, be preferably molecular weight 4,000～5,000 ultrafilter membrane adopts conventional ultrafiltration equipment to be concentrated into initial capacity 10%～60%, after preferred 20%～30%, deliver to freeze drying equipment, the preparation lyophilized powder includes abundant insulin like growth factor.After adjusting pH value, also can in filtrate, add and be equivalent to 1 times～5 times of filtrate, concentrate behind preferred 2 times～3 times aseptic distilled water dilute filtration liquid, more should adopt back one step when especially adopting tangent plane to interlock the fluid ultrafiltration apparatus.
In accordance with the method for claim 1 in, technical characterictic of the present invention is, also can adopt another approaches and methods to prepare somatomedin group in the cattle colostrums, its method is a defat cattle colostrums molecular weight 8,000～15,000 supermicro filtration membrane separating and filtering, collect supernatant, with molecular weight is that 4000～6000 supermicro filtration membrane concentrates moisture content to original volume 10%～20%, send into freeze drying plant and prepare dry powder, the residue of ultra-filtration and separation is regulated pH value to 2.7～2.9 with 2N HCL, impelling with the somatomedin that combines white bonding state of dawn and dissociate, is 10,000～20 with molecular weight, 000 supermicro filtration membrane separating and filtering, collect supernatant, regulate pH value to 5.8～7.0 with 2N NaOH after, be that 4000～6000 the concentrated moisture content of supermicro filtration membrane is to original volume 20%～30% with molecular weight, lyophilization prepares dry powder, is rich in multiple somatomedin in the preparation.
In accordance with the method for claim 1 in, technical characterictic of the present invention is, the preparation for preparing by the inventive method with effective dose puts on the wound, cure patient's external wounds, these external wounds include but not limited to decubital ulcer, laceration, burn, scalpel edge, skin ulcer, the cureless wound of diabetes patient etc., above-mentioned preparation is by the growth of promotion epithelium and connective tissue and synthesizing of promotion dawn white matter and collagen, help healing or healing of wound at least partly, the preparation for treating wound of the inventive method preparation can reach better effect than not handling (promptly not applying the external source agent).
In accordance with the method for claim 1 in, technical characterictic of the present invention is, by patient being applied the preparation of the present invention's preparation, preferably apply the above-mentioned preparation of effective dose to the zone of injured zone or scarce bone, promote the regeneration of patient's sclerotin, above-mentioned preparation is by promoting connective tissue, sclerotin and cemental growth and stimulating the synthetic of dawn white matter and collagen, help inostosis, or at least partly regeneration, adopt preparation of the present invention to promote inostosis, reach better effect than not handling (promptly handling) without the external source agent.
In accordance with the method for claim 1 in, technical characterictic of the present invention is, the somatomedin that in the above-mentioned preparation of the present invention, adds some gene recombinaton, include but not limited to platelet derived growth factor (PDGG) and insulin like growth factor (GIF), can strengthen the effect that preparation of the present invention promotes skin, connective tissue, osteogenesis, be used for the treatment of patient's wound and can obtain better therapeutic.Can in preparation of the present invention, add platelet derived growth factor (PDGF) separately, also can in preparation of the present invention, add insulin like growth factor (GIF) separately, the independent adding of above-mentioned component can improve the therapeutic effect of preparation of the present invention, preparation of the present invention is internal energy to reach better therapeutic effect but PDGF and GIF add simultaneously, the ratio that PDGF and GIF add is that optimum is that ratio between two is between 1: 1～2: 1 in 1: 5 to 10: 1 the scope.
9. technical characterictic of the present invention is, during in the independent application of the above-mentioned preparation of the present invention or with other growth factor preparation use in conjunction, compositions is by acceptable carrier material on a kind of materia medica, for example inert gel that can buy at pharmaceuticals or pharmacy or liquid are (for example, be added with the saline of white or methylcellulose of dawn clearly), other growth factor preparations (for example gene recombinaton growth factor preparation) of preparation of the present invention and/or combination are combined produce.
10. technical characterictic of the present invention is, above-mentioned preparation of the present invention can be used separately, also can add other forms of growth factor preparation (for example gene recombinaton growth factor preparation) use in conjunction, these somatomedin include but not limited to insulin like growth factor, epidermal growth factor, platelet derived growth factor etc.Above-mentioned composition can add in the cosmetics, can stimulate the growth of normal skin and the reparation of cell, and the protection and the maintenance of skin had very important function.Above-mentioned composition has obvious effects to the protection and the maintenance of skin than the cosmetics that do not add above-mentioned composition.
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| CNA2004100158539A CN1557478A (en) | 2004-01-15 | 2004-01-15 | Skin, connective tissue and bone growth promoting composition |
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| CNA2004100158539A CN1557478A (en) | 2004-01-15 | 2004-01-15 | Skin, connective tissue and bone growth promoting composition |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104434974A (en) * | 2014-12-09 | 2015-03-25 | 东北农业大学 | Bovine coloctrum growth factor crude extract as well as preparation method and application thereof |
| CN105457102A (en) * | 2014-09-05 | 2016-04-06 | 陕西艾美雅生物科技有限公司 | Purifying and concentrating method for composite bioactive factors and concentrated solution |
-
2004
- 2004-01-15 CN CNA2004100158539A patent/CN1557478A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105457102A (en) * | 2014-09-05 | 2016-04-06 | 陕西艾美雅生物科技有限公司 | Purifying and concentrating method for composite bioactive factors and concentrated solution |
| CN105457102B (en) * | 2014-09-05 | 2019-02-26 | 陕西艾美雅生物科技有限公司 | The purification method for concentration and concentrate of the composite bio-active factor |
| CN104434974A (en) * | 2014-12-09 | 2015-03-25 | 东北农业大学 | Bovine coloctrum growth factor crude extract as well as preparation method and application thereof |
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