CN103316380B - Bone defect repairing material, preparation method and applications thereof - Google Patents
Bone defect repairing material, preparation method and applications thereof Download PDFInfo
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Abstract
The invention relates to a bone defect repairing material. The bone defect repairing material comprises a bacteria cellulose membrane, a biologic ceramic material and cellulase, wherein the biologic ceramic material is deposited on a microfiber surface of the bacteria cellulose membrane, and the cellulase is dispersed in the bacteria cellulose membrane and can degrade the bacteria cellulose membrane. When the bone defect repairing material is implanted into bone defect positions, as osteoblasts are attached on the microfiber surface of the bacteria cellulose membrane for growing and the ceramic material is deposited on the microfiber surface of the bacteria cellulose membrane, the osteoblasts growing on the ceramic material deposited on the microfiber surface are wrapped easily, and an intact bone organization is formed by the osteoblasts combining with collagen generated from the osteoblasts. The cellulose membrane is degraded gradually by the cellulase embedded, leaving no residues. Glucose as a degraded product does no harm to cells, and the glucose can support osteocyte copying and growing as nutrients. Furthermore, the invention also relates to a preparation method for the bone defect repairing material and applications of the bone defect repairing material.
Description
Technical field
The present invention relates to biomedical active Material Field, especially relate to a kind of bone defect repair material and its preparation method and application.
Background technology
Bone defect (bone defects) disease normally due to injured, that the reason of disease as osteoporosis or operation get involved causes part osseous tissue disappearance, thus causes the afunction of deleted areas.In a DAGU, as Thigh bone, if bone rejected region is when 8mm, usual this defect can spontaneous recovery.If but defect is greater than 8mm, or in an ossiculum, as phalanx, defect is even less than 8mm, and this bone defect then can not spontaneous recovery, needs bone material to fill, or utilizes bone material to guide the osseous tissue of fault location and Growth of Cells thus to repair bone defect.Traditional material being used for filling bone defect has from xenotransplantation, homotransplantation, autograft and inertia biomaterial etc. several.First three plant fill method and material owing to easily causing rejection, endotoxin remains and patient caused to the damage at other position of health, therefore utilizes the active biomaterial of Nonimplantation more and more to become to fill bone rejected region the focus that current developing material utilizes.
Because the active biomaterial being used for filling bone defect needs mutually merge with the osseous tissue grown afterwards and cell and wrap up, different with the material be used for fixing after fracture, the not easily separated material merged of second operation, therefore the biological degradability of biomaterial to material being used for treating bone defect requires higher, after material degradation, unharmful substance remains, and preferably finally can enter metabolic cycles, simultaneously, require that material require has the higher osteoblast compatibility, and there is certain bio-mechanical performance.Traditional degradable material is used in the collagen protein of mainly polyesters macromolecule, animality in the treatment of bone defect and inorganic ceramic material.Polyesters macromolecule have excellent can degeneration and controlled degradation time, but due to catabolite be slant acidity, microenvironment osteocyte being copied to neutrality required for growth and meta-alkalescence creates certain negatively influencing.The use of animality collagen protein, owing to producing possible rejection or having endotoxic residual, therefore has very high requirement to the source of material and use.Inorganic ceramic material, as hydroxyapatite, is exactly a part for osseous tissue usually, due to porous and network structure can not reach the same requirement of macromole, is therefore usually combined the bone defect packing material forming compound with other material.Traditional bone defect repair material all has certain limitation, therefore needs to develop new bone defect repair material.
Summary of the invention
Based on this, be necessary to provide a kind of biodegradable and bone defect repair material that catabolite is harmless and its preparation method and application.
A kind of bone defect repair material, comprise bacteria cellulose film, be deposited on the bioceramic material of the microfiber surface of described bacteria cellulose film and be dispersed in the cellulase of bacteria cellulose film described in the degradable in described bacteria cellulose film, described bacteria cellulose film, described bioceramic material and be 10:1:0.1 ~ 0.17 with the mass ratio of described cellulase.
Wherein in an embodiment, described bacteria cellulose film is Acetobacter gluconicum extracellular fiber element film.
Wherein in an embodiment, described bioceramic material is at least one in hydroxyapatite, calcium triphosphate, Calcium monohydrogen phosphate system, dicalcium phosphate dehydrate and calcium phosphate magnesite.
Wherein in an embodiment, described cellulase is come from the cellulase that Trichoderma viride belongs to or the cellulase coming from trichoderma reesei genus.
Above-mentioned bone defect repair material comprises bacteria cellulose film, bioceramic material and cellulase.Bacterial cellulose has good affinity to osteoblast, and Bacterial cellulose portability enough moisture, provide to the growth of osteocyte and grow microenvironment preferably.When by above-mentioned bone defect repair material implantable bone defect, be grown on the microfiber surface of bacteria cellulose film because osteoblast adheres to, and ceramic material deposits is on the microfiber surface of bacteria cellulose film, therefore, the osteoblast be grown on microfiber surface ceramic material is easy to be wrapped and forms complete osseous tissue with the collagen protein that osteoblast produces.Bacteria cellulose film is then decomposed gradually by cellulase, noresidue, and catabolite glucose without any harmfulness to cell can also be supported copying of osteocyte as part nutrient substance and grow.This bone defect repair material can be widely used in preparing bone defect repair drug world, as:
A kind of bone defect repair medicine, comprise above-mentioned bone defect repair material, described medicine is freeze-dried formulation.
A preparation method for bone defect repair material, comprises the steps:
Can the strain inoculation of production of cellulose cultivate until obtaining thickness is disrupt bacteria cell after the bacteria cellulose film of 1 ~ 5mm in culture medium, and remove broken bacterial cell, collect described bacteria cellulose film;
By described bacteria cellulose film after lyophilization process, the calcium chloride solution being placed in 0.1 ~ 0.3mol/L activates 24 ~ 48 hours, rinsing, removes calcium chloride, obtains the bacteria cellulose film activated;
Bioceramic material is joined in the solution of simulated body fluid and prepare the suspension that concentration is the bioceramic material of 1 ~ 5mg/mL, the bacteria cellulose film of described activation is placed in the suspension of described bioceramic material, rotating with the rotating speed of 1 ~ 2 rev/min at 35 ~ 38 DEG C carries out on microfibre that deposition reaction makes described bioceramic material be deposited on described bacteria cellulose film for 24 ~ 48 hours, after deposition reaction terminates, by rinsed with deionized water, lyophilization, obtain Bacterial cellulose ceramic composite, wherein, the mass ratio of described bacteria cellulose film and described bioceramic material is 10:1,
Configuration concentration is the cellulase solution of 0.5 ~ 1mg/mL, the ratio being 0.1 ~ 0.17:10 according to the mass ratio of described cellulase and described bacteria cellulose film is drawn described cellulase solution and is placed in described Bacterial cellulose ceramic composite surface, leaving standstill makes described cellulase be dispersed in described bacteria cellulose film for 15 ~ 20 minutes, obtains described bone defect repair material after lyophilization.
Wherein in an embodiment, described strain is Acetobacter gluconicum; Described culture medium comprises each component of following concentration: the disodium hydrogen phosphate,anhydrous of the glucose of 0.1 ~ 0.4g/mL, the peptone of 0.03 ~ 0.06g/mL, 0.02 ~ 0.03g/mL, the citric acid of 0.01 ~ 0.02g/mL, the magnesium sulfate of 0.005 ~ 0.015g/mL, the ammonium sulfate of 0.005 ~ 0.01g/mL and percentage by volume are the corn syrup extracting solution of 0.005 ~ 0.015%, and the solvent of described culture medium is water.
Wherein in an embodiment, the sodium hydroxide solution of the step of described disrupt bacteria cell to be working concentration be 0.1mol/L vibrates disrupt bacteria cell 30 minutes in 80 DEG C of water-baths, and repeated multiple times until all described bacterial cells are broken.
Wherein in an embodiment, the sodium hydroxide solution disrupt bacteria cell of described disrupt bacteria cell to be working concentration be 0.1mol/L; The described bacterial cell removing fragmentation is the bacterial cell solution after using rinsed with deionized water fragmentation, till the bacterial cell of fragmentation washes out with the sodium hydroxide remained in bacteria cellulose film.
Wherein in an embodiment, described bioceramic material is at least one in hydroxyapatite, calcium triphosphate, Calcium monohydrogen phosphate system, dicalcium phosphate dehydrate and calcium phosphate magnesite.
Wherein in an embodiment, described cellulase is come from the cellulase that Trichoderma viride belongs to or the cellulase coming from trichoderma reesei genus.
The preparation method of above-mentioned bone defect repair material uses bioceramic material to be scattered in the solution of simulated body fluid, the mother solution that the solution of simulated body fluid grows on the microfibre of bacteria cellulose film as bioceramic material, this kind of suspension is compared with single simulated body fluid with single bioceramic material water slurry, and bioceramic material more can be made in the fast deposition of microfiber surface and growth.Simultaneously, adopt Freeze Drying Technique, cellulase is incorporated in Bacterial cellulose ceramic composite, simple, both maintained the integrity of composite, cellulase is made to be dispersed in composite inner smoothly, can keep again the activity of cellulase, when deployed, tissue fluid invades composite, cellulase activity is activated, and starts to exercise degradation function.
Accompanying drawing explanation
Fig. 1 is harvested from bacteria cellulose film in culture fluid in embodiment 1;
Fig. 2 is that Bacterial cellulose (left side) and Bacterial cellulose ceramic composite (right side) compare at optical electron microscope hypograph;
Fig. 3 is that image ratio is comparatively under a scanning electron microscope for Bacterial cellulose (left side) and Bacterial cellulose ceramic composite (right side);
Fig. 4 is the degraded situation schematic diagram in 5 days (Day) that Bacterial cellulose ceramic composite simulates in degradation experiment in vitro;
Fig. 5 is the analysis schematic diagram of glucose content in enzymolysis solution;
Fig. 6 is that after enzymatic degradation, in enzymolysis solution, residue is through centrifugal, rinsed with deionized water, dried infrared spectrogram, and wherein, abscissa is wave number (Wavenumber), unit cm
-1, vertical coordinate is absworption peak (Transmittance);
Fig. 7 be bone defect repair material in vitro cell (human embryo's osteoblast) grow contrast experiment, wherein, left figure is the growing state on human embryo's osteoblast culture bottle surface under an optical microscope, middle figure is the growing state on Bacterial cellulose surface under an optical microscope after the dyeing of human embryo's osteoblast, right figure is under an optical microscope can the growing state on Bacterial cellulose ceramic composite surface of bio-absorbable after the dyeing of human embryo's osteoblast, green represents living cells, and redness represents dead cell.
Detailed description of the invention
Mainly in conjunction with the drawings and the specific embodiments bone defect repair material and its preparation method and application is described in further detail below.
The bone defect repair material of one embodiment, comprises bacteria cellulose film, is deposited on the bioceramic material of the microfiber surface of bacteria cellulose film and is dispersed in the cellulase of the degradable bacteria cellulose film in bacteria cellulose film.
In the present embodiment, bacteria cellulose film is Acetobacter gluconicum (Gluconacetobacter xylinum, also known as acetobacter xylinum) extracellular polysaccharide film.Bioceramic material is at least one in hydroxyapatite (HAP), calcium triphosphate (TCP), Calcium monohydrogen phosphate system (BCP), dicalcium phosphate dehydrate and calcium phosphate magnesite.Cellulase is the cellulase (C0615 or C1794 etc. as Sigma company) in neutral conditions with longer vital stage and more SA Trichoderma viride genus and the one come from the cellulase (C8546 as Sigma company) of trichoderma reesei genus.Bacteria cellulose film, bioceramic material and be 10:1:0.1 ~ 0.17 with the mass ratio of cellulase.
This bone defect repair material comprises bacteria cellulose film, bioceramic material and cellulase.Bacterial cellulose has good affinity to osteoblast, and Bacterial cellulose portability enough moisture, provide to the growth of osteocyte and grow microenvironment preferably.When by above-mentioned bone defect repair material implantable bone defect, be grown on the microfiber surface of bacteria cellulose film because osteoblast adheres to, and ceramic material deposits is on the microfiber surface of bacteria cellulose film, therefore, the osteoblast be grown on microfiber surface ceramic material is easy to be wrapped and forms complete osseous tissue with the collagen protein that osteoblast produces.Bacteria cellulose film is then decomposed gradually by cellulase, noresidue, and catabolite glucose without any harmfulness to cell can also be supported copying of osteocyte as part nutrient substance and grow.This bone defect repair material can be widely used in preparing bone defect repair drug world, as:
Present embodiment additionally provides a kind of bone defect repair medicine, and comprise above-mentioned bone defect repair material, medicine is freeze-dried formulation.
In addition, present embodiment additionally provides a kind of preparation method of bone defect repair material, comprises the steps:
Step one: can the strain inoculation of production of cellulose cultivate until obtaining thickness is disrupt bacteria cell after the bacteria cellulose film of 1 ~ 5mm in culture medium, and remove broken bacterial cell, and collect bacteria cellulose film.
Such as, can choose concentration is 0.80to2.30 × 10
5cFU/mL(CFU:Colony FormingUnits, bacterium colony number) Acetobacter gluconicum strain solution, this strain solution is inoculated in culture medium with the ratio of 1:50 ~ 100, static culture at 28 ~ 32 DEG C.Wherein, the composition of culture medium is as follows: the disodium hydrogen phosphate,anhydrous of the glucose of 0.1 ~ 0.4g/mL, the peptone of 0.03 ~ 0.06g/mL, 0.02 ~ 0.03g/mL, the citric acid of 0.01 ~ 0.02g/mL, the magnesium sulfate of 0.005 ~ 0.015g/mL, the ammonium sulfate of 0.005 ~ 0.01g/mL and percentage by volume are the corn syrup extracting solution of 0.005 ~ 0.015%, and the solvent of culture medium is water.After the bacteria cellulose film of acquisition 1 ~ 5mm thickness, can working concentration be that the sodium hydroxide solution of 0.1mol/L vibrates disrupt bacteria cell in 80 DEG C of water-baths, repeated multiple times, until bacterial cell is completely broken, then broken cell and sodium hydroxide is washed away with deionized water, until the pH on bacteria cellulose film surface is close to neutral.For the bacteria cellulose film collected, after steam sterilization, deionized water can be placed in and save backup in 4 DEG C of environment.
Step 2: by bacteria cellulose film after lyophilization process, the calcium chloride solution being placed in 0.1 ~ 0.3mol/L activates 24 ~ 48 hours, rinsing, removes calcium chloride, obtains the bacteria cellulose film activated.
The object of calcium chloride activation makes a kind of calcium ion layer of bacteria cellulose film Surface Creation, and this calcium ion layer can induce bioceramic material to adsorb thereon, and progressively forms new Ca/P(calcium/phosphorus) layer.
Step 3: bioceramic material is joined in the solution of simulated body fluid (SBF) and prepare the suspension that concentration is the bioceramic material of 1 ~ 5mg/mL, the bacteria cellulose film of activation is placed in the suspension of bioceramic material, with 35 ~ 38 DEG C to rotate with the rotating speed of 1 ~ 2 rev/min and carry out deposition reaction and within 24 ~ 48 hours, make bioceramic material be deposited on the microfibre of bacteria cellulose film, after deposition reaction terminates, by rinsed with deionized water, lyophilization, obtain Bacterial cellulose ceramic composite, wherein, the mass ratio of bacteria cellulose film and bioceramic material is 10:1.
Bioceramic material can be at least one in hydroxyapatite (HAP), calcium triphosphate (TCP), Calcium monohydrogen phosphate system (BCP), dicalcium phosphate dehydrate and calcium phosphate magnesite.
Step 4: configuration concentration is 0.5-1mg/mL cellulase solution, the ratio being 0.1 ~ 0.17:10 according to the mass ratio of cellulase and bacteria cellulose film is drawn cellulase solution and is placed in Bacterial cellulose ceramic composite surface, leaving standstill makes cellulase be dispersed in bacteria cellulose film for 15 ~ 20 minutes, obtains bone defect repair material after lyophilization.
The preparation method of this bone defect repair material uses bioceramic material to be scattered in the solution of simulated body fluid, the mother solution that the solution of simulated body fluid grows on the microfibre of bacteria cellulose film as bioceramic material, this kind of suspension is compared with single simulated body fluid with single bioceramic material water slurry, bioceramic material more can be made in the fast deposition of microfiber surface and growth, such as, when adopting hydroxyapatite, hydroxyapatite is scattered in the solution of simulated body fluid, Ca/P layer very easily deposits at the microfiber surface of bacteria cellulose film and grows and forms layered crystal.Simultaneously, adopt Freeze Drying Technique, cellulase is incorporated in Bacterial cellulose ceramic composite, simple, both maintained the integrity of composite, cellulase is embedded smoothly and is dispersed in composite inner, can keep again the activity of cellulase, when deployed, tissue fluid invades composite, cellulase activity is activated, and starts to exercise degradation function.
Screening to use has longer vital stage and more SA cellulase under near-neutral sulfite deinking, as ... cellulose can be made can to realize in vivo controlledly degrading slowly and absorbing, generally in 3 ~ 7 days, cellulose masterplate in Bacterial cellulose ceramic composite can be degraded completely.In addition, can also by regulating the contamination of cellulase, this bacteria cellulose film can extend in 30 days and degrade.
Be below specific embodiment part:
Embodiment 1
Step one: choosing concentration is 0.80to2.30 × 10
5the Acetobacter gluconicum strain solution of CFU/mL, is inoculated in culture medium by this strain solution with the ratio of 1:100, static culture at 30 DEG C.Wherein, the composition of culture medium is as follows: the disodium hydrogen phosphate,anhydrous of the glucose of 0.1 ~ 0.4g/mL, the peptone of 0.03 ~ 0.06g/mL, 0.02 ~ 0.03g/mL, the citric acid of 0.01 ~ 0.02g/mL, the magnesium sulfate of 0.005 ~ 0.015g/mL, the ammonium sulfate of 0.005 ~ 0.01g/mL and percentage by volume are the corn syrup extracting solution of 0.005 ~ 0.015%, and the solvent of culture medium is water.When the thickness of extracellular fiber element film reaches 1 ~ 5mm, working concentration is that the sodium hydroxide solution of 0.1mol/L vibrates disrupt bacteria cell in 80 DEG C of water-baths, repeated multiple times, until bacterial cell is completely broken, then broken cell and sodium hydroxide is washed away with deionized water, until the pH on bacteria cellulose film surface is close to neutral, collect cellulose membrane, as shown in Figure 1.To the bacteria cellulose film collected, after steam sterilization, deionized water can be placed in and save backup in 4 DEG C of environment.
Step 2: by bacteria cellulose film after lyophilization process, the calcium chloride solution being placed in 0.1mol/L activates 48 hours, rinsing, removes calcium chloride, obtains the bacteria cellulose film activated.
Step 3: hydroxyapatite is joined in the solution of simulated body fluid (SBF) and prepare the suspension that concentration is the hydroxyapatite of 1mg/mL, the bacteria cellulose film of activation is placed in the suspension of hydroxyapatite, with 37 DEG C to rotate with the rotating speed of 2 revs/min and carry out deposition reaction and within 48 hours, make hydroxyapatite deposition on the microfibre of bacteria cellulose film, after deposition reaction terminates, by rinsed with deionized water, after lyophilization, be cut into 2.5cm
2the strip of size, obtains Bacterial cellulose ceramic composite.Wherein, the mass ratio of bacteria cellulose film and hydroxyapatite is 10:1.
Shown in Fig. 2 be Bacterial cellulose (left side) with the image ratio of Bacterial cellulose ceramic composite (right side) under optical electron microscope comparatively, as can be seen from Figure 2, Bacterial cellulose ceramic composite improves a lot compared to single bacteria cellulose material compactness, show that hydroxyapatite has adhered to and grown in the surface of bacteria cellulose film and inside thus, be namely deposited on microfibre.
Shown in Fig. 3 is that image ratio is comparatively under a scanning electron microscope for Bacterial cellulose (left side) and Bacterial cellulose ceramic composite (right side).As can be seen from Figure 3, hydroxyapatite has firmly wrapped up the microfibre of bacteria cellulose film, defines firm crystalline texture around it.
Prove thus, hydroxyapatite, through deposition reaction, is firmly combined with microfibre, and around microfibre, define a porous class ball layer structure.
Step 4: configuration concentration is 0.1% μ g/mL cellulase solution, be the ratio of 0.1 ~ 0.17:10 according to the mass ratio of cellulase and bacteria cellulose film, be placed on small pieces by pipette, extract cellulase solution, leaving standstill makes cellulase be dispersed in bacteria cellulose film for 15 minutes, obtains bone defect repair material after lyophilization.
Show through external degradation experiment, hydroxyapatite layer can't stop the embedding of cellulase.As shown in Figure 4, within 7 day time, Bacterial cellulose successfully, is fully degraded.Liner in Fig. 4 in visible simulated body fluid is consider in experimental design that the environment of material can not be the medium of a whole soln.The environment of bone fault location is a medium be made up of osseous tissue and tissue fluid.Material can not be fully immersed in tissue fluid, but a kind of material encircle by osseous tissue and its surface contact tissue fluid.Therefore, adopt this liner analog bone tissue in experimental design, allow material surface contact to simulated body fluid solution instead of be fully immersed in wherein.
As shown in Figure 5, find the analysis of sugar in degradation solution, in Bacterial cellulose ceramic composite (right side), glucose release does not have the change on statistical significance compared with single bacteria cellulose material (left side).In range of error, the burst size of glucose can up to about 95%.Glucose can not reach 100%, is because the content of Bacterial cellulose in Bacterial cellulose is not 100% yet, and normally 95 ~ 98%, all the other are very small amount of lipid and glycoprotein etc.
As shown in Figure 6, in degradation solution, remaining fine particle shape whiteness is detected as through infrared spectrum the hydroxyapatite be attached to around microfibre.Arrow institute target absworption peak band representative in Fig. 6 is at 800 ~ 1100cm
-1have obviously absworption peak, these absworption peaks represent phosphate anion (PO
4 3-), phosphoric acid hydrogen radical ion (HPO
4 2-) infrared absorption spectroscopy, show in residue containing a large amount of hydroxyapatite.
This experimental result well demonstrates Bacterial cellulose and as a masterplate, can introduce hydroxyapatite, and through enzyme effect, be finally degraded, and hydroxyapatite self any change does not occur and is that deposition gathers together.
As shown in Figure 7, external cytotoxicity experiment, left figure is the growing state on human embryo's osteoblast culture bottle surface under an optical microscope.Middle figure is the growing state on Bacterial cellulose surface under an optical microscope after the dyeing of human embryo's osteoblast.Right figure is the growing state of the bone defect repair material surface of the present embodiment under an optical microscope after the dyeing of human embryo's osteoblast.Green represents living cells, and redness represents dead cell.Result shows, at the bone defect repair material surface of the present embodiment, and the attachment that human embryo's osteoblast equally can be good, copy and grow, thus also demonstrate that Human embryo osteoblast can easier adhere to, copy, be grown in this Bacterial cellulose ceramic composite.
Embodiment 2
Hydroxyapatite is substituted with calcium triphosphate.
Be configured to the calcium triphosphate simulated body fluid suspension of 3mg/mL.
Other steps are with embodiment 1.
Embodiment 3
Hydroxyapatite is replaced with Calcium monohydrogen phosphate system (BCP).
Increase a BCP synthetic reaction.Respectively according to the ratio of mass ratio 3:2, take hydroxyapatite and the calcium triphosphate powder of constant weight, fully after mixing, 800 DEG C of calcinings 5 hours.After cooling, fully grind.The BCP of synthesis is scattered in simulated body fluid with the concentration ratio of 5mg/mL and makes suspension.
Other steps are with embodiment 1.
The above embodiment only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but therefore can not be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (12)
1. a bone defect repair material, it is characterized in that, comprise bacteria cellulose film, be deposited on the bioceramic material of the microfiber surface of described bacteria cellulose film and be dispersed in the cellulase of bacteria cellulose film described in the degradable in described bacteria cellulose film, described bacteria cellulose film, described bioceramic material and be 10:1:0.1 ~ 0.17 with the mass ratio of described cellulase.
2. bone defect repair material as claimed in claim 1, is characterized in that, described bacteria cellulose film is Acetobacter gluconicum extracellular fiber element film.
3. bone defect repair material as claimed in claim 1, it is characterized in that, described bioceramic material is at least one in hydroxyapatite, calcium triphosphate, Calcium monohydrogen phosphate system, dicalcium phosphate dehydrate and calcium phosphate magnesite.
4. bone defect repair material as claimed in claim 1, is characterized in that, described cellulase is come from the cellulase that Trichoderma viride belongs to or the cellulase coming from trichoderma reesei genus.
5. a preparation method for bone defect repair material, is characterized in that, comprises the steps:
Can the strain inoculation of production of cellulose cultivate until obtaining thickness is disrupt bacteria cell after the bacteria cellulose film of 1 ~ 5mm in culture medium, and remove broken bacterial cell, collect described bacteria cellulose film;
By described bacteria cellulose film after lyophilization process, the calcium chloride solution being placed in 0.1 ~ 0.3mol/L activates 24 ~ 48 hours, rinsing, removes calcium chloride, obtains the bacteria cellulose film activated;
Bioceramic material is joined in the solution of simulated body fluid and prepare the suspension that concentration is the bioceramic material of 1 ~ 5mg/mL, the bacteria cellulose film of described activation is placed in the suspension of described bioceramic material, rotating with the rotating speed of 1 ~ 2 rev/min at 35 ~ 38 DEG C carries out on microfibre that deposition reaction makes described bioceramic material be deposited on described bacteria cellulose film for 24 ~ 48 hours, after deposition reaction terminates, by rinsed with deionized water, lyophilization, obtain Bacterial cellulose ceramic composite, wherein, the mass ratio of described bacteria cellulose film and described bioceramic material is 10:1,
Configuration concentration is the cellulase solution of 0.5 ~ 1mg/mL, the ratio being 0.1 ~ 0.17:10 according to the mass ratio of described cellulase and described bacteria cellulose film is drawn described cellulase solution and is placed in described Bacterial cellulose ceramic composite surface, leaving standstill makes described cellulase be dispersed in described bacteria cellulose film for 15 ~ 20 minutes, obtains described bone defect repair material after lyophilization.
6. the preparation method of bone defect repair material as claimed in claim 5, it is characterized in that, described strain is Acetobacter gluconicum; Described culture medium comprises each component of following concentration: the disodium hydrogen phosphate,anhydrous of the glucose of 0.1 ~ 0.4g/mL, the peptone of 0.03 ~ 0.06g/mL, 0.02 ~ 0.03g/mL, the citric acid of 0.01 ~ 0.02g/mL, the magnesium sulfate of 0.005 ~ 0.015g/mL, the ammonium sulfate of 0.005 ~ 0.01g/mL and percentage by volume are the corn syrup extracting solution of 0.005 ~ 0.015%, and the solvent of described culture medium is water.
7. the preparation method of bone defect repair material as claimed in claim 5, it is characterized in that, the sodium hydroxide solution of the step of described disrupt bacteria cell to be working concentration be 0.1 ~ 0.2mol/L vibrates disrupt bacteria cell 30 minutes in 80 DEG C of water-baths, and repeated multiple times until all described bacterial cells are broken.
8. the preparation method of bone defect repair material as claimed in claim 5, is characterized in that, the sodium hydroxide solution disrupt bacteria cell of described disrupt bacteria cell to be working concentration be 0.1mol/L; The described bacterial cell removing fragmentation is the bacterial cell solution after using rinsed with deionized water fragmentation, till the bacterial cell of fragmentation washes out with the sodium hydroxide remained in bacteria cellulose film.
9. the preparation method of bone defect repair material as claimed in claim 5, it is characterized in that, described bioceramic material is at least one in hydroxyapatite, calcium triphosphate, Calcium monohydrogen phosphate system, dicalcium phosphate dehydrate and calcium phosphate magnesite.
10. the preparation method of bone defect repair material as claimed in claim 5, is characterized in that, described cellulase is come from the cellulase that Trichoderma viride belongs to or the cellulase coming from trichoderma reesei genus.
11. bone defect repair materials according to any one of Claims 1 to 4 are in the application preparing bone defect repair drug world.
12. 1 kinds of bone defect repair medicines, comprise the bone defect repair material according to any one of Claims 1 to 4, described medicine is freeze-dried formulation.
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103690995A (en) * | 2013-12-10 | 2014-04-02 | 深圳先进技术研究院 | Bioabsorbable fiber, as well as preparation method and application thereof |
| CN103861153B (en) * | 2014-02-24 | 2015-06-10 | 钟春燕 | Preparation method of carbon nanofiber composite Beta-calcium phosphate with three-dimensional net structure |
| CN104587516B (en) * | 2014-12-29 | 2017-06-06 | 深圳先进技术研究院 | A kind of transparent degradable bacteria cellulose regeneration membrane and its preparation method and application |
| CN105031722B (en) * | 2015-08-18 | 2018-01-30 | 北京在声生物科技有限公司 | Modified bacteria cellulose for bone repairing support and preparation method thereof |
| CN105601318B (en) * | 2015-12-22 | 2018-11-30 | 广东省微生物研究所 | A kind of porous calcium phosphate calcium material and preparation method thereof |
| CN109771693A (en) * | 2018-04-13 | 2019-05-21 | 中国人民解放军第五医院 | A kind of preparation method for the new injectable spontaneous coagulation cmposite artificial bone carrying rhBMP_2 microballoon |
| CN110464876B (en) * | 2019-08-30 | 2022-03-29 | 东营凤起生物科技发展有限公司 | Growth factor-loaded bacterial cellulose/biological ceramic composite membrane |
| CN116370713A (en) * | 2023-01-11 | 2023-07-04 | 华东交通大学 | Ionic crosslinking bacterial cellulose bone tissue engineering scaffold and preparation and application thereof |
-
2013
- 2013-05-23 CN CN201310196824.6A patent/CN103316380B/en active Active
Non-Patent Citations (2)
| Title |
|---|
| In vitro biodegradability and mechanical properties of bioabsorbable bacterial cellulose incorporating cellulases;Yang Hu等;《Acta Biomaterialia》;20110401;第7卷;第2835–2845页 * |
| 羟基磷灰石/纳米纤维素复合材料的制备及表征;翟云敬;《中国优秀硕士学位论文全文数据库医药卫生科技辑》;20120215(第02期);E080-31 * |
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