CN103690995A - Bioabsorbable fiber, as well as preparation method and application thereof - Google Patents
Bioabsorbable fiber, as well as preparation method and application thereof Download PDFInfo
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- CN103690995A CN103690995A CN201310671548.4A CN201310671548A CN103690995A CN 103690995 A CN103690995 A CN 103690995A CN 201310671548 A CN201310671548 A CN 201310671548A CN 103690995 A CN103690995 A CN 103690995A
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Abstract
The invention relates to the technical field of functional materials, and in particular relates to a bioabsorbable fiber. The fiber comprises cellulase and bacterial cellulose, wherein the cellulose is uniformly embedded in the bacterial cellulose material. The invention also provides a preparation method of the bioabsorbable fiber. The bioabsorbable fiber is combined through twice lyophilization by utilizing the enzyme engineering, the cellulase is embedded into the bacterial cellulose, the bacterial cellulose is degraded by utilizing the cellulose, so that in vivo degradation and absorption of cellulose can be realized, the fiber is mildly degraded, the cellulose can be completely degraded, and the product glucose can be completely absorbed; the preparation method of the bioabsorbable fiber is simple, cannot destroy cellulase, does not need chemicals, does not require high-voltage and other conditions, is safe, and cannot pollute the environment.
Description
[technical field]
The present invention relates to technical field of function materials, be specifically related to a kind of can bio-absorbable fiber and its preparation method and application.
[background technology]
At present, macromolecule polyalcohol has become prepares the most important base material of biomaterial, adopts biomaterial prepared by macromolecule polyalcohol to carry out body while being implanted into, and requires it to have higher biocompatibility and biodegradability, absorbability.Macromolecule polyalcohol comprises synthetic polymer and natural polymer according to source.In prior art, the natural macromolecular polymer of cellulose family is widely used in material conventional in biologic medical product, as: Bacterial cellulose is to be synthesized by antibacterial, by β-1, a kind of polysaccharide macromolecule that 4-glycosidic bond couples together glucosyl group, the structure of its structure and plant cellulose is basic identical, but Bacterial cellulose is synthetic more efficient than synthesizing of plant cellulose, and purity is higher, the ultra micro fleece that Bacterial cellulose is comprised of ultra micro cellulose, therefore have the incomparable characteristic of many plant celluloses.Bacterial cellulose all has been widely used at aspects such as food service industry, biomedicine, paper industry and oil exploitations.Bacterial cellulose, as a kind of new bio medical engineering material, has high-purity, high-crystallinity, high polymerization degree, and high-hydroscopicity, high-moisture-retention, to characteristics such as the high permeability of liquids and gases, high wet strengths.But, owing to lacking relevant enzyme in body, in the material bodies of cellulose family, can not degrade, or degradation speed is uncontrollable.
Therefore, prepare the absorbable cellulose of biodegradable in a kind of body and become problem demanding prompt solution.
[summary of the invention]
The object of the invention is to overcome above-mentioned weak point, biodegradable fiber in a kind of body is provided, in solution prior art, bio-medical fiber material can not degradation in vivo or the uncontrollable technical problem of degradation speed.
Another object of the present invention is to provide the preparation method of the fiber that in above-mentioned body, biodegradable absorbs.
The present invention is that to solve the problems of the technologies described above adopted scheme as follows:
Can bio-absorbable in body a fiber, comprising: cellulase and Bacterial cellulose, described cellulase is embedded in bacteria cellulose material equably.
Preferably, the weight ratio of described cellulase and described Bacterial cellulose is 1:8～1:4.
Preferably, described cellulase derives from Trichoderma viride or Trichoderma reesei.
The present invention provides the preparation method of the fiber that in a kind of body, biodegradable absorbs in addition, comprises the steps:
Step 1: prepare Bacterial cellulose thin film;
Step 2: step 1 gained Bacterial cellulose thin film is carried out to lyophilization;
Step 3: preparation cellulase solution, after being mixed with step 2 gained Bacterial cellulose, cellulase solution carries out lyophilization, and the weight ratio of described cellulase and described Bacterial cellulose is 1:8～1:4.
Preferably, described step 1 specifically comprises the steps:
The culture fluid that preparation comprises glucose, peptone and inorganic salt;
By preserving number, be that ATCC700178, preserving number are that ATCC53528, preserving number are that the acetobacter xylinum strain that ATCC53524 or preserving number are ATCC23769 is seeded in described culture fluid, 28～32 ℃ of static culture 5～7 days, in described culture fluid, generate bacterial fibers thin film;
Gained bacterial fibers thin film is processed to 30～60min with 0.1～0.2mol/mL sodium hydroxide solution at 80～100 ℃;
By the bacterial fibers thin film deionized water rinsing after processing, will regulate pH value to neutral.
Preferably, described culture fluid also comprises organic acid, magnesium sulfate, ammonium sulfate and Semen Maydis pulp extracting solution; Described organic acid is acetic acid, lactic acid or citric acid; Described inorganic salt is sodium hydrogen phosphate, Ca
2+salt or Mg
2+salt.
Preferably, in the culture fluid of every 100ml, contain: 10～40g glucose, 3～6g peptone, 1～2g citric acid, 2～3g sodium hydrogen phosphate, 0.5～1.5g magnesium sulfate, 0.5～1.5g ammonium sulfate, 0.5～1.5ml Semen Maydis extracting solution.
Preferably, described cellulase derives from Trichoderma viride or Trichoderma reesei.
Preferably, in step 1, the thickness of gained Bacterial cellulose thin film is 3～6mm.
The present invention also provides the application in preparing wound healing field medicine of fiber that above-mentioned biodegradable absorbs.
Compared with prior art, beneficial effect of the present invention is, biodegradable fiber utilization enzyme engineering of the present invention, by secondary freeze drying, carry out combination, cellulase is embedded in Bacterial cellulose, utilizes cellulose degraded Bacterial cellulose, realize cellulosic vivo degradation; Biodegradable fiber degradation of the present invention is gentle, and cellulose degradation is complete.The preparation method of the biodegradable fiber of the present invention is simple, to cellulase, can not produce destruction; The preparation method of biodegradable fiber of the present invention, does not use chemicals, without conditions such as high voltages, has safety, environment is not produced and polluted, and has higher economic benefit and social benefit during large-scale culture.
[accompanying drawing explanation]
Fig. 1 is the flow chart of the preparation method of biodegradable fiber in body of the present invention;
Fig. 2 is biodegradable fiberoptic fiber element enzymatic activity variation diagram in body of the present invention;
Fig. 3 is biodegradable fiber degradation figure in body of the present invention;
Fig. 4 is biodegradable fiber glucose burst size variation diagram in the experiment of simulation wound body fluid in body of the present invention.
[specific embodiment]
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with the drawings and specific embodiments, the present invention is described in further detail.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
The fiber that in the body that the embodiment of the present invention provides, biodegradable absorbs, comprising: cellulase and Bacterial cellulose, described cellulase is embedded in bacteria cellulose material equably.
Cellulose is as macromolecule polyalcohol, in human body, shortage can make the enzyme of its degraded, the present invention is directly embedded into cellulase in cellulose macromolecule polymer, utilize the enzyme catalysis cellulose degradation of above-mentioned embedding, for by the speed controlling of cellulose degradation in required scope, the weight ratio of above-mentioned biodegradable fibrous material cellulase and Bacterial cellulose is 1:8～1:4; In a preferred version of the present embodiment, cellulase derives from Trichoderma viride or Trichoderma reesei.
The embodiment of the present invention also provides the preparation method of the fiber that in a kind of body, biodegradable absorbs, and as shown in Figure 1, the method comprises:
Step S101: prepare Bacterial cellulose thin film;
Step S102: step S101 gained Bacterial cellulose thin film is carried out to lyophilization;
Step S103: preparation cellulase solution, after being mixed with step S102 gained Bacterial cellulose, cellulase solution carries out lyophilization.
In step S101, first, the culture fluid that preparation comprises glucose, peptone and inorganic salt; Then, by preserving number, be that ATCC700178, preserving number are that ATCC53528, preserving number are that the acetobacter xylinum strain that ATCC53524 or preserving number are ATCC23769 is seeded in described culture fluid, 28～32 ℃ of static culture 5～7 days, in described culture fluid, generate bacterial fibers thin film; Again gained bacterial fibers thin film is processed to 30～60min with 0.1～0.2mol/mL sodium hydroxide solution at 80～100 ℃; Finally, by the bacterial fibers thin film deionized water rinsing after processing, will regulate pH value to neutral.
Wherein, culture fluid is liquid culture medium, meets the required composition of thalli growth and metabolism.In the present embodiment, culture fluid comprises carbon source, nitrogenous source and inorganic salt composition, and carbon source can be glucose, fructose or sucrose; Nitrogenous source can be selected from one or more in peptone, yeast extract, Semen Maydis pulp extracting solution; Culture fluid also comprises organic acid, magnesium sulfate, ammonium sulfate; Wherein, organic acid is acetic acid, lactic acid or citric acid; Inorganic salt is sodium hydrogen phosphate, Ca
2+salt or Mg
2+salt.In a preferred version of the present embodiment, in the culture fluid of every 100ml, contain: 10～40g glucose, 3～6g peptone, 1～2g citric acid, 2～3g sodium hydrogen phosphate, 0.5～1.5g magnesium sulfate, 0.5～1.5g ammonium sulfate, 0.5～1.5ml Semen Maydis extracting solution.According to different strain to nutritional solution without demand, can in culture fluid, add other composition.
Acetobacter xylinum is one of antibacterial that synthetic cellulose is the strongest, and the cellulose of acetobacter xylinum synthesized has higher degree of crystallinity, the degree of polymerization and very consistent molecularly oriented, and exists with single Cellulosed molded article; Purity is high simultaneously, and the cellulose Tensile strength of acquisition is high, has extremely strong retentiveness and permeable, breathability; Also there is biodegradability, belong to environmentally friendly product.In the present embodiment, selecting preserving number is that ATCC700178, preserving number are that ATCC53528, preserving number are the acetobacter xylinum strain that ATCC53524 or preserving number are ATCC23769, as the strain that produces Bacterial cellulose thin film, 28～32 ℃ of static culture 5～7 days, in described culture fluid, generate bacterial fibers thin film, in whole cultivation, process in, keep sterile working, to guarantee to obtain needed antibacterial always.
As a preferred version of the present embodiment, in step S101, the thickness of gained Bacterial cellulose thin film is 3～6mm.
For by the speed controlling of cellulose degradation in required scope, the weight ratio of step S103 cellulase and Bacterial cellulose is 1:8～1:4.Cellulase derives from Trichoderma viride or Trichoderma reesei.
Particularly, the biodegradable fiber utilization enzyme engineering of the embodiment of the present invention, carries out combination by secondary freeze drying, and cellulase is embedded in Bacterial cellulose, utilizes cellulose degraded Bacterial cellulose, realizes cellulosic vivo degradation; The biodegradable fiber degradation of the embodiment of the present invention is gentle, and cellulose degradation is complete.The preparation method of the biodegradable fiber of the embodiment of the present invention is simple, to cellulase, can not produce destruction; The preparation method of biodegradable fiber of the present invention, does not use chemicals, without conditions such as high voltages, has safety, environment is not produced and polluted, and has higher economic benefit and social benefit during large-scale culture.Biodegradable fiber of the present invention can be used for wound suture, in the process of wound healing, the cellulase that Bacterial cellulose is embedded into is degraded gradually, noresidue, and its catabolite glucose, can also be supplied with wound as nutrient substance and recover without any harmfulness cell.
The embodiment of the present invention 1 provides the preparation method of the fiber that in a kind of body, biodegradable absorbs, and comprises the steps:
Step 1: prepare Bacterial cellulose thin film;
Prepare culture fluid, in the culture fluid of every 100ml, contain: 10～40g glucose, 3～6g peptone, 1～2g citric acid, 2～3g sodium hydrogen phosphate, 0.5～1.5g magnesium sulfate, 0.5～1.5g ammonium sulfate, 0.5～1.5ml Semen Maydis extracting solution;
By preserving number, be that ATCC700178, preserving number are that ATCC53528, preserving number are that the acetobacter xylinum strain that ATCC53524 or preserving number are ATCC23769 is seeded in described culture fluid, 28～32 ℃ of static culture 5～7 days, in described culture fluid, generate bacterial fibers thin film, the thickness of Bacterial cellulose thin film is 3～6mm;
Gained bacterial fibers thin film is processed to 30～60min with 0.1～0.2mol/mL sodium hydroxide solution at 80～100 ℃, repeat 3～4 times;
By the bacterial fibers thin film deionized water rinsing after processing, will regulate pH value to neutral.
Step 2: step 1 gained Bacterial cellulose thin film is carried out to lyophilization;
Step 3: the cellulase solution that compound concentration is 6～12mol/mL, after being mixed with step 2 gained Bacterial cellulose 1.44g, cellulase solution 0.3ml carries out lyophilization, and wherein, cellulase is Sigma C0615.
The fiber cellulase that in body prepared by the present embodiment, biodegradable absorbs and the weight ratio of Bacterial cellulose are 1:8～1:4.Cellulase Sigma C0615 derives from Trichoderma viride.
Embodiment 2
The embodiment of the present invention 2 provides the preparation method of the fiber that in a kind of body, biodegradable absorbs, and comprises the steps:
Step 1: prepare Bacterial cellulose thin film;
Prepare culture fluid, in the culture fluid of every 100ml, contain: 10～40g glucose, 3～6g peptone, 1～2g citric acid, 2～3g sodium hydrogen phosphate, 0.5～1.5g magnesium sulfate, 0.5～1.5g ammonium sulfate, 0.5～1.5ml Semen Maydis extracting solution;
By preserving number, be that ATCC700178, preserving number are that ATCC53528, preserving number are that the acetobacter xylinum strain that ATCC53524 or preserving number are ATCC23769 is seeded in described culture fluid, 28～32 ℃ of static culture 5～7 days, in described culture fluid, generate bacterial fibers thin film, the thickness of Bacterial cellulose thin film is 3～6mm;
Gained bacterial fibers thin film is processed to 30～60min with 0.1～0.2mol/mL sodium hydroxide solution at 80～100 ℃, repeat 3～4 times;
By the bacterial fibers thin film deionized water rinsing after processing, will regulate pH value to neutral.
Step 2: step 1 gained Bacterial cellulose thin film is carried out to lyophilization;
Step 3: the cellulase solution that compound concentration is 6～12mol/mL, after being mixed with step 2 gained Bacterial cellulose 1.44g, cellulase solution 0.3ml carries out lyophilization, and wherein, cellulase is Sigma C8546.
Biodegradable fiber cellulase prepared by the present embodiment and the weight ratio of Bacterial cellulose are 1:8～1:4.Cellulase Sigma C8546 derives from Trichoderma reesei.
Embodiment 3
The embodiment of the present invention 3 provides a kind of preparation method of biodegradable fiber, comprises the steps:
Step 1: prepare Bacterial cellulose thin film;
Prepare culture fluid, in the culture fluid of every 100ml, contain: 10～40g glucose, 3～6g peptone, 1～2g citric acid, 2～3g sodium hydrogen phosphate, 0.5～1.5g magnesium sulfate, 0.5～1.5g ammonium sulfate, 0.5～1.5ml Semen Maydis extracting solution;
By preserving number, be that ATCC700178, preserving number are that ATCC53528, preserving number are that the acetobacter xylinum strain that ATCC53524 or preserving number are ATCC23769 is seeded in described culture fluid, 28～32 ℃ of static culture 5～7 days, in described culture fluid, generate bacterial fibers thin film, the thickness of Bacterial cellulose thin film is 3～6mm;
Gained bacterial fibers thin film is processed to 30～60min with 0.1～0.2mol/mL sodium hydroxide solution at 80～100 ℃, repeat 3～4 times;
By the bacterial fibers thin film deionized water rinsing after processing, will regulate pH value to neutral.
Step 2: step 1 gained Bacterial cellulose thin film is carried out to lyophilization;
Step 3: the cellulase solution that compound concentration is 6～12mol/mL, after being mixed with step 2 gained Bacterial cellulose 1.44g, cellulase solution 0.3ml carries out lyophilization, and wherein, cellulase is Sigma C1794.
Biodegradable fiber cellulase prepared by the present embodiment and the weight ratio of Bacterial cellulose are 1:8～1:4.Cellulase Sigma C1794 derives from Trichoderma viride.
the activity experiment of biodegradable fiber cellulase
With normal saline, respectively embodiment 1, embodiment 2, the biodegradable fiber of embodiment 3 gained are soaked, be placed in the environment of pH6.5～7.0;
Detect embodiment 1, embodiment 2, the embodiment biodegradable fiber of the 3 gained cellulase activity of 1～30 day.
Result as shown in Figure 2, the activity of the biodegradable fiber cellulase of embodiment 2 has all kept relatively low and stable activity at 1～30 day, guarantee that biodegradable fiber within a certain period of time can be by slow and lasting degraded and absorbed, be that Sigma C0615, Sigma C8546, Sigma C1794 compare, Sigma C8546 embeds best results in Bacterial cellulose.
biodegradable fiber degradation experiment
With normal saline, respectively embodiment 1, embodiment 2, the biodegradable fiber of embodiment 3 gained are soaked;
As shown in Figure 3, be the degraded situation of the Biodegradable fibers of embodiment 2, under pH7.0, Biodegradable fibers is fast degradation in 7 days.
external buffer simulated body fluid experiment
With normal saline, respectively embodiment 1, embodiment 2, the biodegradable fiber of embodiment 3 gained are soaked;
Biodegradable fiber of the present invention can be used for wound suture, in the process of wound healing, the cellulase that Bacterial cellulose is embedded into is degraded gradually, noresidue, and its catabolite glucose, can also be supplied with wound as nutrient substance and recover without any harmfulness cell.The variation of human body wound pH value is from acidity to alkalescence, finally keep being stabilized near pH7.4, what that is to say human body wound pH value is changed to pH4.5 → pH6.0 → pH7.4, this experiment is simulated wound body fluid in body with external buffer, result as shown in Figure 4, in three kinds of different pH environment, embodiment 1, embodiment 2, in the biodegradable fiber of embodiment 3 gained, enzyme has all kept certain activity, especially in pH4.5, embodiment 1, embodiment 2, cellulase in the biodegradable fiber of embodiment 3 gained has all kept higher activity, along with pH raises, activity decreases.After the variation of this activity and human body are injured, the variation of the pH of wound is consistent.Therefore, can be according to the active variation under condition of different pH of different cellulase Biodegradable fibers be made to certain design according to application choice, as can be seen from Figure 4, glucose burst size in Biodegradable fibers of the present invention is higher, the highlyest can reach more than 90%, the impact of removing the approximately 5% albumen impurity comprising in material, this Biodegradable fibers has reached degradable requirement.
The above the specific embodiment of the present invention, does not form limiting the scope of the present invention.Various other corresponding changes and distortion that any technical conceive according to the present invention is made, all should be included in the protection domain of the claims in the present invention.
Claims (10)
1. can bio-absorbable a fiber, it is characterized in that, comprising: cellulase and Bacterial cellulose, described cellulase is embedded in bacteria cellulose material equably.
According to claim 1 can bio-absorbable fiber, it is characterized in that, the weight ratio of described cellulase and described Bacterial cellulose is 1:8～1:4.
According to claim 1 can bio-absorbable fiber, it is characterized in that, described cellulase derives from Trichoderma viride or Trichoderma reesei.
4. can bio-absorbable the preparation method of fiber, it is characterized in that, comprise the steps:
Step 1: prepare Bacterial cellulose thin film;
Step 2: step 1 gained Bacterial cellulose thin film is carried out to lyophilization;
Step 3: preparation cellulase solution, after being mixed with step 2 gained Bacterial cellulose, cellulase solution carries out lyophilization, and wherein, the weight ratio of described cellulase and described Bacterial cellulose is 1:8～1:4.
5. preparation method according to claim 4, is characterized in that, described step 1 specifically comprises the steps:
The culture fluid that preparation comprises glucose, peptone and inorganic salt;
By preserving number, be that ATCC700178, preserving number are that ATCC53528, preserving number are that the acetobacter xylinum strain that ATCC53524 or preserving number are ATCC23769 is seeded in described culture fluid, 28～32 ℃ of static culture 5～7 days, in described culture fluid, generate bacterial fibers thin film;
Gained bacterial fibers thin film is processed to 30～60min with 0.1～0.2mol/mL sodium hydroxide solution at 80～100 ℃;
By the bacterial fibers thin film deionized water rinsing after processing, will regulate pH value to neutral.
6. preparation method according to claim 5, is characterized in that, described culture fluid also comprises organic acid, magnesium sulfate, ammonium sulfate and Semen Maydis pulp extracting solution; Described organic acid is acetic acid, lactic acid or citric acid; Described inorganic salt is sodium hydrogen phosphate, Ca
2+salt or Mg
2+salt.
7. preparation method according to claim 6, is characterized in that, in the culture fluid of every 100ml, contains: 10～40g glucose, 3～6g peptone, 1～2g citric acid, 2～3g sodium hydrogen phosphate, 0.5～1.5g magnesium sulfate, 0.5～1.5g ammonium sulfate, 0.5～1.5ml Semen Maydis extracting solution.
8. preparation method according to claim 4, is characterized in that, described cellulase derives from Trichoderma viride or Trichoderma reesei.
9. preparation method according to claim 4, is characterized in that, in step 1, the thickness of gained Bacterial cellulose thin film is 3～6mm.
Described in claim 1 or 2 can bio-absorbable the application of fiber in preparing wound healing field medicine.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104587516A (en) * | 2014-12-29 | 2015-05-06 | 深圳先进技术研究院 | Transparent reproductive bacterial cellulose reproductive membrane as well as preparation method and application thereof |
| CN106616323A (en) * | 2016-12-13 | 2017-05-10 | 南京高新工大生物技术研究院有限公司 | Preparation method of high-toughness gelatinization-free noodles |
| CN112138215A (en) * | 2020-09-26 | 2020-12-29 | 江苏大学 | Construction method and application of cell growth factor slow-release anisotropic scaffold based on nano-cellulose |
| CN115382003A (en) * | 2021-05-24 | 2022-11-25 | 海南光宇生物科技有限公司 | Biological cellulose surgical suture without stitches taking off |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104587516A (en) * | 2014-12-29 | 2015-05-06 | 深圳先进技术研究院 | Transparent reproductive bacterial cellulose reproductive membrane as well as preparation method and application thereof |
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| CN112138215A (en) * | 2020-09-26 | 2020-12-29 | 江苏大学 | Construction method and application of cell growth factor slow-release anisotropic scaffold based on nano-cellulose |
| CN115382003A (en) * | 2021-05-24 | 2022-11-25 | 海南光宇生物科技有限公司 | Biological cellulose surgical suture without stitches taking off |
| CN115382003B (en) * | 2021-05-24 | 2024-01-30 | 海南光宇生物科技有限公司 | Biological cellulose operation suture line without disconnecting |
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Application publication date: 20140402 |