CN101513523A - Medicine containing cell polypeptide growth factor and preparation method thereof - Google Patents
Medicine containing cell polypeptide growth factor and preparation method thereof Download PDFInfo
- Publication number
- CN101513523A CN101513523A CNA2009100428776A CN200910042877A CN101513523A CN 101513523 A CN101513523 A CN 101513523A CN A2009100428776 A CNA2009100428776 A CN A2009100428776A CN 200910042877 A CN200910042877 A CN 200910042877A CN 101513523 A CN101513523 A CN 101513523A
- Authority
- CN
- China
- Prior art keywords
- growth factor
- cell polypeptide
- polypeptide growth
- medicine
- titanium dioxide
- 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.)
- Granted
Links
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a medicine containing a polypeptide cell growth factor and a preparation method thereof. The medicine comprises titanium dioxide particles which have the particle diameter of 1-100nm and have an amino (-NH2) group on surfaces, and the surfaces of the titanium dioxide particles are adhered with the polypeptide cell growth factor. The medicine can play photocatalytic sterilization and bacteriostasis action of titanium dioxide while playing sustained-release action on the polypeptide cell growth factor so that the growth factor continuously and stably act on wound/wound surface cells to maintain an effective action concentration in a proper time period; and the nano-titanium dioxide without the growth factor can enhance sterilization and bacteriostasis action and decompose organic substances of the wound/wound surface such as secretion, enzyme, bacterial toxin and the like into carbon dioxide and water so as to facilitate faster wound/wound repair and reduce adverse reaction.
Description
Technical field
The present invention relates to a kind of medicine that contains cell polypeptide growth factor and preparation method thereof.
Background technology
Cell growth factor is to regulate the class protein polypeptides matter of cell proliferation and differentiation, and its molecular weight does not wait to several ten thousand from hundreds of, also often is referred to as cell polypeptide growth factor.Compare with proteohormone with the polypeptide of classics, polypeptide growth factor does not have specific endocrine gland and endocrine cell, but paracrine by some cells and autocrine discharge and be diffused into target cell, thereby coordinate the unification of body self and reaction to external world.Very fast about evaluation, structure and the engineered progress of somatomedin in recent years, part of polypeptide somatomedin class medicine has been widely used in clinical, and obtained clinical efficacy preferably, but this class medicine ubiquity following shortcoming: 1, medicine stability is poor, changeableness inactivation and being eliminated.Infecting on wound/wound surface especially, the secretions of wound/wound surface, enzyme and bacteriotoxin etc. have promoted decomposition, degeneration and the inactivation of somatomedin.This has had a strong impact on contacting of peptide growth factor medicine and wound/wound surface cambium, thereby has had a strong impact on the performance of biological availability.2, directly peptide growth factor coating/spray is shone the continuous action time that reaches enough of when wound/wound surface, being difficult to, because polypeptide at room temperature is very unsettled, all the more so when particularly having water to exist, its half-life was significantly shorter than the synthetic required lag time (being about 8～12 hours) at wound location inducing cell DNA less than 1 hour.3, in the short time that somatomedin is used, somatomedin can only be to the short proliferation function of a part of cell performance.In the middle of outgrowth tissue, be in outgrowth cell and only occupy certain ratio, and be the state of a relative equilibrium, the proliferation function of somatomedin pair cell mainly is to act on to be in the cell proliferation cell of interval, directly peptide growth factor is acted on wound surface and only only acts on intermitotic cell at short notice.4, present peptide growth factor preparation process complexity, price is expensive, and the repeated multiple times administration has caused a large amount of wastes of medicine, has increased the weight of patient's financial burden.And the slow release growth factor preparation can continuous and effective acting on fraction be in intermitotic different cell, thereby changed the poised state of proliferative cell, just allow the cell proportion that is in proliferative phase raise, such result makes hyperplastic tissue grow apace continually and steadily.In sum, the medicine that how will contain somatomedin makes that slow releasing preparation makes it continually and steadily, safety, useful effect is in wound/wound surface, has become research focus that is rich in challenge and has practical value in biomedical engineering and the pharmaceutics field.The nano-particle complex medicine at effective control drug dose, reduce poisonous side effect of medicine, improve that medicine stability and effective rate of utilization, realization drug targeting are carried and the function that alleviates aspects such as patient suffering receives much concern.Generally be free polypeptide growth factor heeling-in to be gone in the materials such as high-molecular gel at present both at home and abroad, make it to be the slow release state, to prolong its action time.Most nano controlled-release medicines are pursued slow release simply, drug effect reaches and does not wait two weeks to half a year, and adopt the intravenously administrable mode more, and topical agent thing slow-release time was suitable with 12～48 hours generally, so the slow release combination drug that present stage has developed can not effectively be applied to wound surface.
Summary of the invention
The purpose of this invention is to provide a kind of medicine that contains cell polypeptide growth factor and preparation method thereof, make this medicine can effectively be applied to wound/wound surface.
In order to achieve the above object, the technical scheme that the present invention contains the medicine of cell polypeptide growth factor is to comprise that the surface has amino (NH
2) group, particle diameter is the nano-titania particle of 1～100nm, in the TiO 2 particles surface attachment cell polypeptide growth factor is arranged.
Described cell polypeptide growth factor is by physical absorption, Electrostatic Absorption and by carboxyl (COOH) and the amino (NH on TiO 2 particles surface attached to the TiO 2 particles surface
2) group generation bonding reaction and combination.
Described cell polypeptide growth factor can be recombinant human epidermal growth factor (rhEGF), nerve growth factor (NGF), basic fibroblast growth factor (bFGF), acid fibroblast growth factor (aFGF), people and M-EGF (hEGF and mEGF), the somatomedin (PDGF) in platelet source, transforming growth factor (TGF), insulin like growth factor (IGFs), insulin like growth factor (IGF), vascular endothelial cell growth factor (VEGF), Connective Tissue Growth Factor (CTGF), keratinocyte growth factor (KGF), multiple effect growth factor (Ptn), among the lymphatic vessel somatomedin (VEGF-C and VEGF-D) etc. one or more.
The technical scheme of the preparation method of the medicine that contains cell polypeptide growth factor of the present invention comprises, comprise that (1) is dispersed in particle diameter in the distilled water at 1～100nm nano titanium oxide, ultimate density 10ppm～10000ppm disperseed 2～12 hours, and regulating pH value is 5.0～8.0; (2) add cell polypeptide growth factor freeze dried powder or cell polypeptide growth factor solution to above-mentioned solution the inside, the ultimate density that makes somatomedin is 0.01 μ g/ml～100 μ g/ml, regulate pH value to 5.0～8.0 once more, the control temperature is carried out compound reaction under 0 ℃～60 ℃, response time is 0.5～24 hour, generates the medicament nano titanium dioxide-cell polypeptide growth factor complex solution that contains cell polypeptide growth factor.
As improvement of the present invention, in described titanium dioxide-cell polypeptide growth factor complex solution, add one or more and mix homogeneously in protective agent, antiseptic, the surfactant.
Described protective agent is human albumin or mannitol, and addition is 0.01%～5% (weight), and described surfactant is Tween 80 or lecithin, and addition is 0.01%～5% (weight).
Described antiseptic is nipalgin fourth vinegar, sodium benzoate, and sorbic acid, nipalgin second vinegar, Metagin vinegar, any one in nipalgin third vinegar, addition is 0.01%～5% (weight).
As another improvement of the present invention, described titanium dioxide-cell polypeptide growth factor complex solution can be drained into lyophilized powder under-100 ℃～60 ℃ and vacuum.
As a further improvement on the present invention, described titanium dioxide-cell polypeptide growth factor complex lyophilized powder is under the condition that has polyethylene of dispersing agent alcohol (PVA), add polyglycolic acid (PGA), polylactic acid (PLA), lactic acid one ethanol copolymer (PLGA), gelatin, chitosan, poly-D-lysine, among the PAH one or more adopt ultrasonic emulsification-solvent evaporation method that complex is sealed again and form the nano controlled-release microsphere.
As a further improvement on the present invention, described titanium dioxide-cell polypeptide growth factor complex lyophilized powder is joined in the medicinal filmogen, make the ultimate density of titanium dioxide-cell polypeptide growth factor complex reach 0.01%～10% (weight), be prepared into the sustained release film formulation of titanium dioxide-cell polypeptide growth factor complex.
Because when the titanium dioxide granule size entered nanometer scale, the small-size effect that the size confinement causes, quantum confined effect and skin effect significantly strengthened, and its sterilizing ability is strengthened greatly, have produced qualitative leap.Only the nano titanium oxide with minute quantity can produce powerful bactericidal action.Nano titanium oxide can effectively be killed multiple pathogen (comprising antibacterial, mycete, infusorian, mycoplasma, chlamydia, virus etc.).The sterilization mechanism of nano titanium oxide mainly is: nano titanium oxide is a kind of photocatalysis antibacterial agent, under illumination condition, produces hydroxyl radical free radical (OH) and elemental oxygen (O).Hydroxyl radical free radical and elemental oxygen have strong oxidizing property, can divide the Organic substance that is deconstructed into pathogen quickly and effectively, and end product is carbon dioxide and water, thereby plays antibacterial action.Simultaneously, the Organic substances such as secretions, enzyme and bacteriotoxin of wound/wound surface also can also be decomposed into carbon dioxide and water, quicken the healing of wound/wound surface.The poisonous complex that effective bacterium for degrading remains discharge, its bactericidal effect is thorough rapidly.And its sterilization characteristics are not limited by the classification of antibacterial, have wide spectrum and press down bactericidal action, rely the antibacterial of medicine to have same purpose to antibiotic yet, do not produce drug dependence, and have advantages such as safe without toxic side effect and relative environmental protection.The medicine of cell polypeptide growth factor that contains of the present invention is in application process, nano titanium oxide is when bacteriostasis is killed in the performance photocatalysis, performance is to the slow releasing function of cell polypeptide growth factor, somatomedin is slowly split away off from titanium dioxide nanoparticle, thereby make somatomedin act on wound/wound surface cell sustainedly and stably, in the appropriate time, keep effective function concentration, simultaneously, the nano titanium oxide of somatomedin of having come off can strengthen it again and kill bacteriostasis, and with the secretions of wound/wound surface, Organic substance such as enzyme and bacteriotoxin is decomposed into carbon dioxide and water, promotes wound/wound surface to repair faster and reduces untoward reaction.In addition, the preparation method of the medicine that contains cell polypeptide growth factor of the present invention is simple, and various parameters are easy to control, and the drug prepared steady quality is convenient to suitability for industrialized production, and production cost is lower.
The specific embodiment
Embodiment 1
1, titanium dioxide-recombinant human epidermal growth factor slow release complex (TiO
2-rhEGF) preparation
According to the following step, can obtain TiO
2-rhEGF complex solution
(1) nanometer titanium dioxide titanium solution preparation: the nano titanium oxide 2mg that particle diameter is modified in the surface amino groups of 100nm is dispersed in the distilled water, and making its ultimate density is 10ppm, ultra-sonic dispersion 10 hours.After phosphate buffer was regulated pH value, making it pH value was 7.0.
(2) add people's epidermal growth factor (rhEGF) of recombinating to above-mentioned nano titanium oxide solution the inside, the ultimate density that makes somatomedin is 10 μ g/ml, regulates pH value to 7.0 once more, and temperature was carried out ultra-sonic dispersion 24 hours at 60 ℃.
(3) at-100 ℃ and drain machine with vacuum and drain into lyophilized powder ,-20 ℃ of preservations.
Can add protective agent, antiseptic, stabilizing agent and saline solution (NaCl) when (4) being configured to solution mixes.
Concrete prescription can be as follows:
TiO
2+ hEGF complex 4mg
Sodium benzoate 15g
Sodium hydrogen phosphate 680.46mg
Mannitol 10g
Tween 80 10g
Glycerol 10g
Distilled water 1L
Embodiment 2
Nano titanium oxide-basic fibroblast growth factor slow release complex (TiO
2-bFGF) preparation
(1) nanometer titanium dioxide titanium solution preparation: the nanometer titanium dioxide titanium valve 2mg that particle diameter is modified in the surface amino groups of 30nm is dispersed in the distilled water, and making its ultimate density is 10000ppm, ultra-sonic dispersion 10 hours.Phosphate buffer makes it pH 5.0 after regulating pH value.
(2) add human fibroblastic growth factor (bFGF) to above-mentioned nano titanium oxide solution the inside, the ultimate density that makes somatomedin is 100 μ g/ml, regulates pH value to 5.0 once more, carries out ultra-sonic dispersion 10 hours under 0 ℃.
(3) at 60 ℃ and drain machine with vacuum and drain into lyophilized powder ,-20 ℃ of preservations.
Can add protective agent, antiseptic, stabilizing agent and saline solution (NaCl) when (4) being configured to solution mixes.
Concrete prescription can be as follows:
TiO
2+ bFGF complex 4mg
Sodium benzoate 15g
Sodium hydrogen phosphate 680.46mg
Mannitol 10g
Tween 80 10g
Glycerol 10g
Distilled water 1L
Embodiment 3
Contain the preparation of the nano titanium oxide-human fibroblastic growth factor slow release complex of Nano silica sol:
(1) nanometer titanium dioxide titanium solution preparation: the nanometer titanium dioxide titanium valve 2mg that particle diameter is modified in the surface amino groups of 1nm is dispersed in the distilled water, and making its ultimate density is 5000ppm, ultra-sonic dispersion 10 hours.Phosphate buffer makes it pH 8.0 after regulating pH value.
(2) add human fibroblastic growth factor (bFGF) to above-mentioned nano titanium oxide solution the inside, regulate pH value to 8.0 once more, under 10 ℃, carried out ultra-sonic dispersion 0.5 hour.
(3) add Nano silica sol in the solution that makes to (2), the ultimate density that makes somatomedin is 0.1 μ g/ml, ultra-sonic dispersion 4 hours.
(4) at 20 ℃ and drain machine with vacuum and drain into lyophilized powder ,-20 ℃ of preservations.
Can add an amount of protective agent, antiseptic, stabilizing agent and saline solution (NaCl) hybrid reaction when (5) being configured to solution.
Concrete prescription can be as follows:
TiO
2+ bFGF complex 4mg
Nano silica sol 10mg
Sodium benzoate 15g
Sodium hydrogen phosphate 680.46mg
Mannitol 10g
Tween 80 10g
Glycerol 10g
Distilled water 1L
Embodiment 4
The preparation of sustained-release micro-spheres colloid solution preparation
(1) nanometer titanium dioxide titanium solution preparation: the nanometer titanium dioxide titanium valve that particle diameter is modified in the 30nm surface amino groups is dispersed in the distilled water, and making its ultimate density is 2000ppm, ultra-sonic dispersion 10 hours.Phosphate buffer makes it pH 6.0 after regulating pH value.
(2) add polypeptide growth factor or polypeptide growth factor solution to above-mentioned solution the inside, make the polypeptide growth factor final concentration, regulate pH value to 6.0 once more, 20 ℃ of following ultra-sonic dispersion 3 hours at 2 μ g/ml.
(3) room temperature gentle agitation or standing and reacting are 8 hours.
(4) at 10 ℃ and drain machine with freezing vacuum and drain into lyophilized powder, take by weighing each component by weight, lyophilized powder is dissolved in the distilled water, make the aqueous phase solution that concentration is 0.1～3% (weight); Polylactic acid is dissolved in dichloromethane: glycerol=3～6: in 1 the mixed solvent, make the oil-phase solution that concentration is 10～60% (weight).
(5) above-mentioned aqueous phase solution, oil-phase solution are mixed, the preparation water-in-oil emulsion, joining then by weight average molecular weight is that 3～250,000 polyvinyl alcohol resin and buffer are made in 10～20% the solution, ultra-sonic dispersion 45 minutes is made microemulsion.
(6) microemulsion that step (5) is obtained stirring at low speed 4 hours at room temperature by volatilization or extracting naturally, is removed organic solvent dichloromethane, promptly gets nano titanium oxide-cell growth factor stable complex Nano microsphere colloid solution.
Embodiment 5
The preparation of sustained release film formulation
A kind of cell growth factor and nano titanium oxide complex membrane, comprise: 1., effective Wound healing and bone regeneration dosage nano titanium oxide-cell growth factor lyophilized powder 0.01%～1% (weight), 2., pharmaceutically useful filmogen 10%～90% (weight), 3., pharmaceutic adjuvant 0%～40% (weight), belong to many skins somatomedin recombinant human epidermal growth factor (rhEGF) of a class together with described hEGF, nerve growth factor (NGF), basic fibroblast growth factor (b FGF), acid fibroblast growth factor (a FGF), M-EGF (mEGF), the somatomedin (PDGF) in platelet source, transforming growth factor (TGF), insulin like growth factor (IGFs), insulin like growth factor (IGF), vascular endothelial cell growth factor (VEGF), Connective Tissue Growth Factor (CTGF), keratinocyte growth factor (KGF), multiple effect growth factor (Ptn), lymphatic vessel somatomedin (VEGF-C and VEGF-D), human growth hormone (GH) all can be used as the alternative bEGF of somatomedin and makes corresponding membrane.
More specifically proportioning is as follows for it
(1) nano titanium oxide that provides by present embodiment three-cell growth factor lyophilized powder 0.01%～1% (weight)
(2) pharmaceutically useful filmogen PVA 10%～90% (weight)
(3) somatomedin protective agent and medicinal filler mannitol 0～10% (weight)
(4) medicinal plasticizer glycerol 0～15% (weight)
(5) pharmaceutical preservative sorbic acid 0.05～0.2% (weight)
(6) medicinal surfactant lecithin 0～2% (weight)
Nano titanium oxide-cell growth factor slow release the complex solution of following three test indications is all for being made by embodiment 1.
Test the experiment of 1 nano titanium oxide-recombinant human epidermal growth factor complex antibacterial effect.
Experiment detects the nanometer titanium dioxide titanium solution that this institute adopts through microbiology, its anti-biocidal property reached more than 99.9% various pathogenic microorganism action effects in 24 hours, the anti-inhibitory effect of the main checking of this experiment nano titanium oxide-recombinant human epidermal growth factor complex, concrete experimental technique is taked to paste aseptic filter paper sheet method on the culture dish surface that is paved with antibacterial, the Quality Control bacterial strain that bacterial strain adopts Central South University's refined three hospital laboratories in Hunan to provide, kind covers Gram-positive, negative bacterium, fungus, anaerobe, coccus, bacillus, mycoplasma, chlamydia etc., experiment divides 5 groups, be respectively nano titanium oxide-epithelical cell growth factor slow release complex group, nano titanium oxide and epithelical cell growth factor use in conjunction group, simple nano titanium oxide group, simple epidermal growth factor group, simple normal saline matched group, drip on filter paper every two hours that respectively to organize two maintenances of corresponding trial target moistening, 37 ℃ of incubators are hatched after 24 hours with big divider and the antibacterial ring size of vernier caliper measurement (mm), keep uitraviolet intensity 100 μ w/cm in the incubator
2Its result for details see attached table 1.
Subordinate list 1 nano titanium oxide-cell growth factor slow release complex resisting pathogenic microbes effect comparison is observed
| The antibacterial name | Somatomedin and nano titanium oxide complex group (bacteriostatic diameter mm) | Somatomedin and nano titanium oxide use in conjunction group (bacteriostatic diameter mm) | Nano titanium oxide group (bacteriostatic diameter mm) | Somatomedin group (bacteriostatic diameter mm) | Normal saline group (bacteriostatic diameter mm) |
| Staphylococcus aureus | 17 | 13 | 12 | 0 | 0 |
| Escherichia coli | 18 | 12 | 14 | 0 | 0 |
| The false single luxuriant bacterium of Aerugo | 14 | 11 | 10 | 0 | 0 |
| Gonorrhea nesa bacterium | 15 | 10 | 11 | 0 | 0 |
| Candida albicans | 21 | 15 | 16 | 0 | 0 |
| Klebsiella Pneumoniae | 18 | 13 | 14 | 0 | 0 |
| The trachoma mycoplasma | 9 | 8 | 8 | 0 | 0 |
| Chlamydia trachomatis | 10 | 8 | 9 | 0 | 0 |
Result of study shows from subordinate list 1, nano titanium oxide-epithelical cell growth factor slow release complex group, nano titanium oxide and epithelical cell growth factor use in conjunction group, simple nano titanium oxide group all has strong inhibitory action to above eight kinds of pathogen, and commute produces the staphylococcus aureus that relies medicine, bacillus pyocyaneus etc. also have good inhibition effect, and simple epidermal growth factor group and simple normal saline matched group are to antibacterial unrestraint effect.Through statistical results show, first three groups obviously is better than two groups (P＜0.01), back on anti-microbial property, nano titanium oxide-epithelical cell growth factor slow release complex group and nano titanium oxide and epithelical cell growth factor use in conjunction group and simple nano titanium oxide group, though do not occur notable difference statistically, on data, can see that still the complex group is better than use in conjunction group and nano titanium oxide group.This shows that the antibiotic property to nano titanium oxide itself behind nano titanium oxide process and the somatomedin compound reaction formation complex is greatly improved, its antibacterial effect may obtain reinforcement.
Test the short cell proliferation experiment of 2 nano titanium oxides-cell growth factor slow release complex
Mtt assay is observed nano titanium oxide-somatomedin complex solution to the effect of human dermis's fibroblast proliferation.Get the Hacat cell by 4 * 10
4/ ml inoculates 96 orifice plates, every hole 100 μ l, establish 9 holes for every group, design 5 groups, be respectively nano titanium oxide-epithelical cell growth factor slow release complex group, nano titanium oxide and epithelical cell growth factor use in conjunction group, simple nano titanium oxide group, simple epidermal growth factor group, simple normal saline matched group, at first with the conventional culture medium DMEM+10%NBS of all cells at 37 ℃ and 5%CO
2Cultivated 6 hours under the condition, go to add 20 μ l respectively after the culture medium and respectively organize corresponding liquid, add 80 μ l again and do not contain blood serum medium DMEM cultivation, respectively take out the cell in three holes during respectively at 12h, 24h and 36h and do the MTT detection, all detect, and sky goes culture medium to add 80 μ l fresh cultures, 20 μ l MTT (concentration 5mg/ml) cultivated 4 hours, go liquid to add DMSO liquid 100 μ l, incubation 5 minutes, vibration makes blue first bank product dissolving, measure the OD value down in microplate reader 570nm wavelength, organize the cell growth with the equal value representation in per 3 holes.The gained data are carried out variance analysis with the SPSS13.0 statistical package and are compared in twos.Somatomedin concentration in its mesocomplex group, use in conjunction group and the epidermal growth factor group is 50 μ g/L.Experimental result is as follows:
Nano titanium oxide-epithelical cell growth factor slow release complex group when (1) complex group and all the other are respectively organized there was no significant difference (P＞0.05) (2) 12h 6h the time, the OD value of nano titanium oxide and epithelical cell growth factor use in conjunction group and simple epidermal growth factor group is higher than simple nano titanium oxide group and simple normal saline matched group (P＜0.05), and nano titanium oxide-epithelical cell growth factor slow release complex group, compare not statistically significant (P＞0.05) between nano titanium oxide and epithelical cell growth factor use in conjunction group and the simple epidermal growth factor group; (3) when 24h and 36h, the OD value of nano titanium oxide-epithelical cell growth factor slow release complex group, nano titanium oxide and epithelical cell growth factor use in conjunction group and simple epidermal growth factor group is apparently higher than simple nano titanium oxide group and simple normal saline matched group (P＜0.01), and nano titanium oxide-epithelical cell growth factor slow release complex group is also apparently higher than nano titanium oxide and epithelical cell growth factor use in conjunction group and simple epidermal growth factor group (P＜0.05).By above experimental result as seen: nano titanium oxide-epithelical cell growth factor slow release complex group can better more effectively promote cell division propagation, in conjunction with slowly separating the developmental biology effect with certain speed from its surface with the somatomedin that is adsorbed on the nano-titania particle surface, play the effect of slow release, thereby strengthened the curative effect of medicine.
Test the promoting healing effect of 3 animal wound healing laboratory observation nano titanium oxide-cell growth factor stable complexes
Get 40 of Wistar rats, be divided into 5 groups at random, be respectively nano titanium oxide-epithelical cell growth factor slow release complex group, nano titanium oxide and epithelical cell growth factor use in conjunction group, simple nano titanium oxide group, simple epidermal growth factor group, simple normal saline matched group, all epidermal growth factor concentration that contain in the group of epidermal growth factor are 2,000IU/ml. 10 every group, coat the sodium sulfide paste earlier, scrape off depilatory gently with Glass rod behind the 5min, the reuse clear water will lose hair or feathers to distinguish to clean to dry and get final product.Intramuscular injection speed is done skin holostrome otch at spinal column lateral symmetry position, back behind the routine disinfection drape after sleeping and newly anaesthetizing, and removes two oval skin chunk and fascias, make minor axis 2cm, the wound surface of major diameter 3cm is used the sterile gauze hemostasis by compression, respectively forms face spray every day and drips corresponding reagent once.Hinder and carried out wound surface measurement and gross examination of skeletal muscle in back 7 days, mainly observe the granulation tissue filling rate; Hinder and carried out wound surface measurement and gross examination of skeletal muscle in back 14 days, mainly observe the residual wound percentage; Note down the complete healing time of wound surface at last.Concrete data see attached list 2.
The short wound healing curative effect comparision of subordinate list 2 nano titanium oxides-cell growth factor slow release complex is observed
| The experiment grouping | Hinder and observed granulation tissue filling rate (%) in back 7 days substantially | Hinder and observed wound healing area (%) in back 14 days substantially | Wound healing time (my god) |
| Nano titanium oxide-cell growth factor complex group | 62.3±4.6 | 82.2±3.5 | 16.4±2.9 |
| Nano titanium oxide and somatomedin use in conjunction group | 60.9±6.5 | 64.7±5.2 | 18.8±2.2 |
| The epidermal growth factor group | 58.5±5.8 | 60.9±2.7 | 18.6±2.4 |
| The nano titanium oxide group | 37.8±6.5 | 52.3±5.4 | 23.1±2.6 |
| The normal saline group | 35.1±7.9 | 50.9±2.5 | 22.2±3.1 |
From subordinate list 2, can draw: show by zoopery, hinder and observed wound surface granulation tissue filling rate in back 7 days substantially, nano titanium oxide-epithelical cell growth factor slow release complex group, nano titanium oxide and epithelical cell growth factor use in conjunction group and simple epidermal growth factor group are better than simple nano titanium oxide group and simple normal saline matched group (P＜0.05), and nano titanium oxide-epithelical cell growth factor slow release complex group, compare not statistically significant (P＞0.05) between nano titanium oxide and epithelical cell growth factor use in conjunction group and the simple epidermal growth factor group; At wound gross examination of skeletal muscle wound healing area in the time of back 14 days, nano titanium oxide-epithelical cell growth factor slow release complex group, nano titanium oxide and epithelical cell growth factor use in conjunction group and simple epidermal growth factor group are better than simple nano titanium oxide group and simple normal saline matched group (P＜0.01), and nano titanium oxide-epithelical cell growth factor slow release complex group also obviously is better than nano titanium oxide and epithelical cell growth factor use in conjunction group and simple epidermal growth factor group (P＜0.05); At final wound healing time relatively, nano titanium oxide-epithelical cell growth factor slow release complex group, nano titanium oxide and epithelical cell growth factor use in conjunction group and simple epidermal growth factor group are better than simple nano titanium oxide group and simple normal saline matched group (P＜0.01), and nano titanium oxide-epithelical cell growth factor slow release complex group also obviously is better than nano titanium oxide and epithelical cell growth factor use in conjunction group and simple epidermal growth factor group (P＜0.05).Zoopery shows that nano titanium oxide-cell growth factor complex can promote the healing of wound surface and be better than single cell growth factor and nano titanium oxide and epithelical cell growth factor use in conjunction used, and shows nano titanium oxide-cell growth factor complex determined curative effect thus.
The present invention can summarize with other concrete forms without prejudice to spirit of the present invention or principal character.Therefore, in any change of implication and the scope suitable, all should think to be included in the scope of claims with claims of the present invention.The medicine that contains cell polypeptide growth factor of the present invention not only can be used for performing the operation and the healing of trauma wound/wound surface is treated, and also can be used for chronic ulcer, corneal wound, oral ulcer, cervical erosion, diabetic foot, the healing treatment of decubital ulcer and various sinus tracts.
Claims (10)
1, a kind of medicine that contains cell polypeptide growth factor is characterized in that, comprises that the surface has amino (NH
2) group, particle diameter is the nano-titania particle of 1～100nm, in the TiO 2 particles surface attachment cell polypeptide growth factor is arranged.
2, according to the medicine that contains cell polypeptide growth factor of claim 1, it is characterized in that described cell polypeptide growth factor is by physical absorption, Electrostatic Absorption and by carboxyl (COOH) and the amino (NH on TiO 2 particles surface attached to the TiO 2 particles surface
2) group generation bonding reaction and combination.
3, the medicine that contains cell polypeptide growth factor according to claim 1, it is characterized in that, described cell polypeptide growth factor can be recombinant human epidermal growth factor (rhEGF), nerve growth factor (NGF), basic fibroblast growth factor (bFGF), acid fibroblast growth factor (aFGF), people and M-EGF (hEGF and mEGF), the somatomedin (PDGF) in platelet source, transforming growth factor (TGF), insulin like growth factor (IGFs), insulin like growth factor (IGF), vascular endothelial cell growth factor (VEGF), Connective Tissue Growth Factor (CTGF), keratinocyte growth factor (KGF), multiple effect growth factor (Ptn), among the lymphatic vessel somatomedin (VEGF-C and VEGF-D) etc. one or more.
4, a kind of preparation method that contains the medicine of cell polypeptide growth factor, it is characterized in that, comprise that (1) is dispersed in the distilled water ultimate density 10ppm～10000ppm with the nano titanium oxide of particle diameter at 1～100nm, disperseed 2～12 hours, regulating pH value is 5.0～8.0; (2) add cell polypeptide growth factor freeze dried powder or cell polypeptide growth factor solution to above-mentioned solution the inside, the ultimate density that makes somatomedin is 0.01 μ g/ml～100 μ g/ml, regulate pH value to 5.0～8.0 once more, the control temperature is carried out compound reaction under 0 ℃～60 ℃, response time is 0.5 hour～24 hours, generates the medicament nano titanium dioxide-cell polypeptide growth factor complex solution that contains cell polypeptide growth factor.
5, according to the preparation method of the medicine that contains cell polypeptide growth factor of claim 4; it is characterized in that, in described titanium dioxide-cell polypeptide growth factor complex solution, add one or more and mix homogeneously in protective agent, antiseptic, the surfactant.
6, according to the preparation method of the medicine that contains cell polypeptide growth factor of claim 5; it is characterized in that; described protective agent; be human albumin or mannitol; addition is 0.01%～5% (weight); described surfactant is Tween 80 or lecithin, and addition is 0.01%～5% (weight).
7, according to the preparation method of the medicine that contains cell polypeptide growth factor of claim 5, it is characterized in that, described antiseptic, be nipalgin fourth vinegar, sodium benzoate, sorbic acid, nipalgin second vinegar, Metagin vinegar, any one in nipalgin third vinegar or multiple, addition is 0.01%～5% (weight).
According to the preparation method of the medicine that contains cell polypeptide growth factor of claim 4, it is characterized in that 8, described titanium dioxide-cell polypeptide growth factor complex solution can be drained into lyophilized powder under one 100 ℃～60 ℃ and vacuum condition.
9, the preparation method of the medicine that contains cell polypeptide growth factor according to Claim 8, it is characterized in that, described titanium dioxide-cell polypeptide growth factor complex lyophilized powder is under the condition that has polyethylene of dispersing agent alcohol (PVA), add polyglycolic acid (PGA), polylactic acid (PLA), lactic acid one ethanol copolymer (PLGA), gelatin, chitosan, poly-D-lysine, among the PAH one or more, make the ultimate density of titanium dioxide-cell polypeptide growth factor complex reach 0.01%～10% (weight), adopt ultrasonic emulsification-solvent evaporation method that complex is sealed again and form the nano controlled-release microsphere.
10, the preparation method of the medicine that contains cell polypeptide growth factor according to Claim 8, it is characterized in that, described nano titanium oxide-cell polypeptide growth factor complex lyophilized powder is joined in the medicinal filmogen, make the ultimate density of titanium dioxide-cell polypeptide growth factor complex reach 0.01%～10%, be prepared into the sustained release film formulation of titanium dioxide-cell polypeptide growth factor complex.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100428776A CN101513523B (en) | 2009-03-18 | 2009-03-18 | Medicine containing cell polypeptide growth factor and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100428776A CN101513523B (en) | 2009-03-18 | 2009-03-18 | Medicine containing cell polypeptide growth factor and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101513523A true CN101513523A (en) | 2009-08-26 |
| CN101513523B CN101513523B (en) | 2011-09-14 |
Family
ID=41038245
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100428776A Expired - Fee Related CN101513523B (en) | 2009-03-18 | 2009-03-18 | Medicine containing cell polypeptide growth factor and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101513523B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104474533A (en) * | 2014-11-24 | 2015-04-01 | 华南理工大学 | Method for preparing compound microspheres for repairing wound |
| CN106798915A (en) * | 2017-03-21 | 2017-06-06 | 郭燕立 | A kind of utilization rhEGF treatment cervical erosion and the gel of damage and preparation method thereof |
| CN107007819A (en) * | 2017-03-30 | 2017-08-04 | 中南大学湘雅三医院 | A kind of spray containing cell polypeptide growth factor |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101371925B (en) * | 2007-12-21 | 2010-12-22 | 周建大 | Nano silver-cell growth factor sustained-release composite body as well as preparation method and use thereof |
-
2009
- 2009-03-18 CN CN2009100428776A patent/CN101513523B/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104474533A (en) * | 2014-11-24 | 2015-04-01 | 华南理工大学 | Method for preparing compound microspheres for repairing wound |
| CN106798915A (en) * | 2017-03-21 | 2017-06-06 | 郭燕立 | A kind of utilization rhEGF treatment cervical erosion and the gel of damage and preparation method thereof |
| CN107007819A (en) * | 2017-03-30 | 2017-08-04 | 中南大学湘雅三医院 | A kind of spray containing cell polypeptide growth factor |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101513523B (en) | 2011-09-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101371925B (en) | Nano silver-cell growth factor sustained-release composite body as well as preparation method and use thereof | |
| Gao et al. | Electrospun nanofibers promote wound healing: Theories, techniques, and perspectives | |
| Oyarzun-Ampuero et al. | Nanoparticles for the treatment of wounds | |
| Smith et al. | Antibiotic delivery strategies to treat skin infections when innate antimicrobial defense fails | |
| Wang et al. | Antibacterial electrospun nanofibrous materials for wound healing | |
| Jiang et al. | Dual delivery of chlorhexidine and platelet-derived growth factor-BB for enhanced wound healing and infection control | |
| Losi et al. | Fibrin-based scaffold incorporating VEGF-and bFGF-loaded nanoparticles stimulates wound healing in diabetic mice | |
| Wang et al. | Antibacterial fluorescent nano-sized lanthanum-doped carbon quantum dot embedded polyvinyl alcohol for accelerated wound healing | |
| Alphonsa et al. | Antimicrobial drugs encapsulated in fibrin nanoparticles for treating microbial infested wounds | |
| Li et al. | Electrospun fibrous membrane containing a cyclodextrin covalent organic framework with antibacterial properties for accelerating wound healing | |
| Nguyen et al. | Enzymatically triggered graphene oxide released from multifunctional carriers boosts anti-pathogenic properties for promising wound-healing applications | |
| CN103330680A (en) | Nano drug transdermal preparation and preparation method thereof | |
| CN101513523B (en) | Medicine containing cell polypeptide growth factor and preparation method thereof | |
| Cheng et al. | Antibacterial and osteoinductive biomacromolecules composite electrospun fiber | |
| Fang et al. | Glucose oxidase loaded thermosensitive hydrogel as an antibacterial wound dressing | |
| Yan et al. | Curcumin-regulated constructing of defective zinc-based polymer-metal-organic framework as long-acting antibacterial platform for efficient wound healing | |
| Mirshekar et al. | Potential antibacterial activity and healing effect of topical administration of bone marrow and adipose mesenchymal stem cells encapsulated in collagen-fibrin hydrogel scaffold on full-thickness burn wound infection caused by Pseudomonas aeruginosa | |
| Yin et al. | Sequential anti-infection and proangiogenesis of DMOG@ ZIF-8/Gelatin-PCL electrospinning dressing for chronic wound healing | |
| Wang et al. | The preparation of lactoferrin/magnesium silicate lithium injectable hydrogel and application in promoting wound healing | |
| Yan et al. | Extracellular vesicle biopotentiated hydrogels for diabetic wound healing: The art of living nanomaterials combined with soft scaffolds | |
| Issaka | State-of-the-art of synthesized exosomes and NPs-based biomimetic nanoparticles for wound rehabilitation: a review | |
| Jamshidifar et al. | Improvement of in vitro osteogenesis and anti-infection properties by GelMA scaffold containing levofloxacin nanoparticles and strontium microspheres for osteomyelitis | |
| Proctor et al. | Furanone loaded aerogels are effective antibiofilm therapeutics in a model of chronic Pseudomonas aeruginosa wound infection | |
| Huang et al. | Sprayable chitosan nanogel with nitric oxide to accelerate diabetic wound healing through bacteria inhibition, biofilm eradication and macrophage polarization | |
| Shen et al. | Multifunctional Injectable Microspheres Containing “Naturally-Derived” Photothermal Transducer for Synergistic Physical and Chemical Treating of Acute Osteomyelitis through Sequential Immunomodulation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C53 | Correction of patent for invention or patent application | ||
| CB03 | Change of inventor or designer information |
Inventor after: Zhou Jianda Inventor after: Xu Dabao Inventor before: Zhou Jianda |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: INVENTOR; FROM: ZHOU JIANDA TO: ZHOU JIANDA XU DABAO |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110914 Termination date: 20170318 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |