阅读说明：本技术 一种长效抗菌促愈合角蛋白敷料的制备方法 (A kind of preparation method of long acting antibiotic promoting healing keratin dressing ) 是由 汤佳鹏 葛彦 刘希文 朱俐 于 2019-09-20 设计创作，主要内容包括：本发明公开了一种长效抗菌促愈合角蛋白敷料的制备方法,包括(1)将一定量再生人发角蛋白完全溶解于98v/v％甲酸溶液,搅拌均匀,角蛋白溶液；(2)采用冷冻干燥,得到角蛋白冻干海绵；(3)将角蛋白冻干海绵经等离子处理器处理活化；(4)将活化处理的角蛋白冻干海绵浸泡在重组人表皮生长因子hEGF、二硫苏糖醇和N,N-二(癸基丙胺基)-巯基甘氨酸的溶液中进行负压闪爆,之后进行接枝反应；(5)将完成接枝反应的角蛋白海绵离心冻干。本发明制备的角蛋白海绵是一种能够诱导皮肤组织损伤部的再生的医疗用材料,能够抑制伤口微生物的生长,刺激皮肤成纤维细胞的增殖,促进伤口愈合。(The invention discloses a kind of preparation methods of long acting antibiotic promoting healing keratin dressing, including (1), and a certain amount of regeneration human hair keratin is dissolved completely in 98v/v% formic acid solution, stirs evenly, keratin solution；(2) using freeze-drying, keratin freeze-drying sponge is obtained；(3) by the plasma treated device processing activation of keratin freeze-drying sponge；(4) the keratin freeze-drying sponge of activation processing is immersed in recombinant human epidermal growth factor hEGF, dithiothreitol (DTT) and N, progress negative pressure is dodged quick-fried in N- bis- (decyl Propylamino)-sulfydryl glycine solution, carries out graft reaction later；(5) the keratin sponge centrifugation freeze-drying of graft reaction will be completed.Keratin sponge prepared by the present invention is a kind of regenerated medical material for capableing of induced skin tissue damage portion, is able to suppress the growth of wound microbes, stimulates the proliferation of skin fibroblasts, promotes wound healing.)

1. a kind of preparation method of antibacterial heal-promoting close angle albumen dressing, which comprises the steps of:

S1: regeneration human hair keratin is dissolved completely in 98v/v% formic acid solution, stirs evenly, obtains keratin solution；

S2: the keratin solution is freeze-dried, and obtains keratin freeze-drying sponge；

S3: by the plasma treated device processing activation of keratin freeze-drying sponge, keratin freeze-drying sponge must be activated；

S4: activation keratin freeze-drying sponge is immersed in containing recombinant human epidermal growth factor, dithiothreitol (DTT) and N, N- It is quick-fried that negative pressure sudden strain of a muscle is carried out in the mixed solution of two (decyl Propylamino)-sulfydryl glycine, carries out graft reaction later, and centrifugation is lyophilized Antibacterial heal-promoting close angle albumen dressing.

2. a kind of preparation method of antibacterial heal-promoting close angle albumen dressing according to claim 1, which is characterized in that step S1 In, the concentration that human hair keratin is regenerated in the keratin solution is 10~30g/L.

3. a kind of preparation method of antibacterial heal-promoting close angle albumen dressing according to claim 1, which is characterized in that step S2 In, the temperature of the freeze-drying is -30~-20 DEG C, and vacuum degree is 0.100~0.024mBar, and freeze-drying time is 3~5d.

4. a kind of preparation method of antibacterial heal-promoting close angle albumen dressing according to claim 1, which is characterized in that step S3 In, the condition of the plasma processor processing are as follows: gas uses nitrogen or oxygen, and processing power is 250~300W, pressure 50 ~60Pa, processing time are 10~15min.

5. a kind of preparation method of antibacterial heal-promoting close angle albumen dressing according to claim 1, which is characterized in that step S4 In, the concentration of recombinant human epidermal growth factor is 40~80mg/L in the mixed solution, the concentration of dithiothreitol (DTT) is 4~ 8g/L, N, N- bis- (decyl Propylamino)-sulfydryl glycine concentration are 5~10g/L.

6. a kind of preparation method of antibacterial heal-promoting close angle albumen dressing according to claim 1, which is characterized in that step S4 In, it is 0.100~0.024mBar that the negative pressure, which dodges quick-fried vacuum degree,.

7. a kind of preparation method of antibacterial heal-promoting close angle albumen dressing according to claim 1, which is characterized in that step S4 In, the soaking bath ratio of the graft reaction is 1:100~300, and reaction temperature is 0~4 DEG C, the reaction time is 12~for 24 hours.

8. a kind of preparation method of antibacterial heal-promoting close angle albumen dressing according to claim 1, which is characterized in that step S5 In, the acceleration of gravity of the centrifugation is 10000g, and time 10min, the temperature of the freeze-drying is -30~-20 DEG C, vacuum degree For 0.100~0.024mBar, freeze-drying time is 3~5d.

Technical field

The present invention relates to biomedical engineering fields, and in particular to a kind of preparation of long acting antibiotic promoting healing keratin dressing Method.

Background technique

Clinically, due to the environment that wound surface is a warm and damp, staphylococcus aureus, Escherichia coli and The pathogen such as Candida albicans are quickly bred on wound, are caused wound easy infection and are not easy to heal.In order to contain that microorganism is numerous Grow, control wound infection, a large amount of medical dressing be widely used in wound covering protection, and medical dressing using while examine Consider and antibiotic or other antibacterial agents are added, to assist control to stop blooding, protection wound, reduces infection, absorbs secretion, holding body temperature With promotion wound healing.

In antibacterial agent, Ag-carried antibacterial dressing is because of its excellent antibiotic property and has no drug resistance, in medical dressing Using being paid attention to extensively, as being applied to skin ulcer caused by burn and scald surgery, traumatology department, a variety of diseases, especially use In cureless bedsore or other chronic infection surface of a wound.But nano silver is to human body and environment that there are certain risk.Have Research report nano silver has larger impact to animals' reproduction system.Therefore, it is necessary to a kind of novel medical dressing, can persistently resist Bacterium, and there is biocompatibility and eco-compatibility.

Keratin has the advantage biodegradable, bioactivity is high and compatibility is good, the keratin extracted from human hair Also there is low immune rejection.Relative to other bulk materials, keratin source is almost unrestricted, thus in cosmetic field Research using more and more being paid attention to.In recent years, human hair keratin is with gel, sponge bracket, coating, film and compound The form of material is applied to neural material, dressing for skin, cell culture, pharmaceutical carrier and soft tissue repair etc., display Its development potentiality in regenerative medicine field out.Keratin is the major structural protein of various types epithelial cell, is cured to wound Conjunction plays an important role.Therefore, the matrix that regeneration human hair keratin is used for facial mask can be weakened into its sensitization, bio-compatible significantly Property is more preferably.But due to hydrophobic, acidproof, the alkaline-resisting characteristic of keratin, chemical process is carried out to it and is not easy, therefore intermolecular interaction Power is weaker, and loosely, mechanical strength is high, poor flexibility for keratin material.

Summary of the invention

In view of this, the purpose of the present invention is to provide a kind of preparation method of long acting antibiotic promoting healing keratin dressing, The keratin dressing that method provided by the invention is prepared is able to suppress microorganism growth, repairs Skin Cell.

In order to solve the above technical problems, the technical scheme adopted by the invention is that:

A kind of preparation method of long acting antibiotic promoting healing keratin dressing, includes the following steps:

S1: regeneration human hair keratin is dissolved completely in 98v/v% formic acid solution, stirs evenly, obtains keratin solution；

S2: the keratin solution is freeze-dried, and obtains keratin freeze-drying sponge；

S3: by the plasma treated device processing activation of keratin freeze-drying sponge, keratin freeze-drying sponge must be activated；

S4: by the activation keratin freeze-drying sponge be immersed in containing recombinant human epidermal growth factor, dithiothreitol (DTT) and It is quick-fried that negative pressure sudden strain of a muscle is carried out in N, N- bis- (decyl Propylamino)-sulfydryl glycine mixed solution, carries out graft reaction later, and centrifugation is frozen Do to obtain long acting antibiotic promoting healing keratin dressing.

Preferably, in step S1, the concentration that human hair keratin is regenerated in the keratin solution is 10~30g/L.

Preferably, in step S2, the temperature of freeze-drying is -30~-20 DEG C, and vacuum degree is 0.100~0.024mBar, Freeze-drying time is 3~5d.

Preferably, in step S3, the condition of the plasma processor processing are as follows: gas uses nitrogen or oxygen, processing Power is 250~300W, and 50~60Pa of pressure, the processing time is 10~15min.

Preferably, in step S4, in the mixed solution concentration of recombinant human epidermal growth factor be 40~80mg/L, two The concentration of sulphur threitol is 4~8g/L, and N, N- bis- (decyl Propylamino)-sulfydryl glycine concentration are 5~10g/L.

Preferably, in step S4, it is 0.100~0.024mBar that the negative pressure, which dodges quick-fried vacuum degree,.

Preferably, in step S4, the soaking bath ratio of the graft reaction is 1:100~300, and reaction temperature is 0~4 DEG C, Reaction time be 12~for 24 hours.

Preferably, in step S5, the acceleration of gravity of the centrifugation is 10000g, time 10min, the temperature of the freeze-drying Degree is -30~-20 DEG C, and vacuum degree is 0.100~0.024mBar, and freeze-drying time is 3~5d.

Compared with prior art, the present invention having the following advantages that and effect:

1) present invention is added dithiothreitol (DTT) and carries out reformation company to its disulfide bond using a large amount of cystine in keratin It connects, greatly optimizes its mechanical property, increase keratin dressing tensile strength and elongation at break.

2) simultaneously, antibacterial agent N, N- bis- (decyl Propylamino)-sulfydryl glycine also have sulfydryl can be by plasma at Reason is grafted on keratin and recombinant human epidermal growth factor, and recombinant human epidermal growth factor is enabled to have certain antibacterial Activity.Dressing use has stretching and deformation in skin surface, skin.So dressing will have certain intensity and flexibility.In addition, Antibacterial agent, which is grafted on keratin, can make the long-term antibacterial of dressing, and it is rotten that antibacterial moleucles are not susceptible to degradation；Antibacterial agent grafting Tissue exudates there are many capable of infiltrating into antibacterial action on to recombinant human epidermal growth factor inside wound internal wounds, hold Bacterium is easily bred, but antibacterial agent discharges meeting stimulating wound healing too much, antibacterial agent has very little is not achieved antibacterial effect, passes through weight Group hEGF, which carries antibacterial agent, can effectively control antibacterial agent burst size.

Detailed description of the invention

L929 mouse fibroblast cells proliferation rate under Fig. 1 embodiment of the present invention and comparative example processing

Specific embodiment

For a further understanding of the present invention, the preferred embodiment of the invention is described below with reference to embodiment, still It should be appreciated that these descriptions are intended merely to further illustrate the features and advantages of the present invention, rather than to the claims in the present invention Limitation.

All raw materials of the present invention, are not particularly limited its source, buying on the market or according to those skilled in the art The preparation of conventional method known to member.

The present invention provides a kind of preparation methods of long acting antibiotic promoting healing keratin dressing, include the following steps:

S1: regeneration human hair keratin is dissolved completely in 98v/v% formic acid solution, stirs evenly, obtains keratin solution；

S2: the keratin solution is freeze-dried, and obtains keratin freeze-drying sponge；

S3: by the plasma treated device processing activation of keratin freeze-drying sponge, keratin freeze-drying sponge must be activated；

S4: by the activation keratin freeze-drying sponge be immersed in containing recombinant human epidermal growth factor, dithiothreitol (DTT) and It is quick-fried that negative pressure sudden strain of a muscle is carried out in N, N- bis- (decyl Propylamino)-sulfydryl glycine mixed solution, carries out graft reaction later, and centrifugation is frozen Do to obtain long acting antibiotic promoting healing keratin dressing.

Specifically, regeneration human hair keratin is dissolved completely in 98v/v% formic acid solution first, stirs evenly, obtain angle Protein solution.It is preferably 10~30g/L that the concentration of human hair keratin is regenerated in keratin solution, more preferably 20g/L.

After obtaining keratin solution, keratin solution is freeze-dried, obtains keratin freeze-drying sponge.It is freezed in the present invention Dry temperature is preferably -30~-20 DEG C, and more preferably -30 DEG C, vacuum degree is preferably 0.100~0.024mBar, more preferably For 0.024mBar, freeze-drying time is preferably 3~5d, more preferably 4d.

It after obtaining keratin freeze-drying sponge, handles and activates through plasma device, obtain activation keratin freeze-drying sponge.The present invention The condition of middle plasma device processing are as follows: gas preferably uses nitrogen or oxygen, more preferably oxygen, and processing power is preferably 250 ~300W, more preferably 280W, pressure are preferably 50~60Pa, more preferably 55Pa, and the processing time is preferably 10~15min, More preferably 15min.

After obtaining activation keratin freeze-drying sponge, activation keratin freeze-drying sponge is immersed in and is grown containing recombinant human epidermal The factor, dithiothreitol (DTT) and N, carry out in N- bis- (decyl Propylamino)-sulfydryl glycine mixed solution negative pressure dodge it is quick-fried, it is laggard Long acting antibiotic promoting healing keratin dressing is lyophilized to obtain in row graft reaction, centrifugation.Recombinant human epidermal is grown in mixed solution of the present invention The concentration of the factor is preferably 40~80mg/L, more preferably 60mg/L, and the concentration of dithiothreitol (DTT) is preferably 4~8g/L, more excellent It is selected as 6g/L, N, N- bis- (decyl Propylamino)-sulfydryl glycine dense preferably 5~10g/L, more preferably 6g/L.

For a further understanding of the present invention, below with reference to embodiment to a kind of long acting antibiotic promoting healing angle provided by the invention The preparation method of albumen dressing is described in detail, and protection scope of the present invention is not limited by the following examples.