CN108543115B - Osteoinductive collagen-based composite hydrogel and preparation method thereof - Google Patents
Osteoinductive collagen-based composite hydrogel and preparation method thereof Download PDFInfo
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Abstract
The invention discloses an osteoinductive collagen-based composite hydrogel and a preparation method thereof, wherein an acidic buffer solution is adopted to prepare a collagen solution, the solution is fully stirred to assist dissolution and dispersion, then an alkali liquor is adopted to slowly adjust the pH value of the solution to 6.0-8.0, and then the solution is placed into a constant-temperature incubator to be incubated to obtain a collagen hydrogel substrate; immersing the skin collagen hydrogel substrate into a chemical modifier solution, adding dopamine, and adding an oxidant to fully react to obtain a dopamine surface-modified skin collagen hydrogel substrate; adding the nano fishbone, adjusting the temperature and the pH value, fully washing by adopting ultrapure water after reaction, and then performing irradiation sterilization to obtain the osteoinductive collagen-based composite hydrogel. The invention has the advantages that: the interface compatibility between the granular bone and the organic phase hydrogel in the prepared hydrogel is greatly improved; so that the prepared hydrogel has excellent osteoinductivity; opens up a new field for the high-value transformation of the skin collagen and the fishbone.
Description
Technical Field
The invention relates to the technical field of biological materials, in particular to an osteoinductive collagen-based composite hydrogel and a preparation method thereof.
Background
The repair of bone defects caused by trauma, tumor, inflammation, surgical operation and the like is a great problem for orthopedists and materials scientists. In clinical treatment nowadays, the material exchange and signal conduction between materials and cells in bone repair are delayed due to the structure of large bone limited in transplanted bone, the design and development of granular bone repair materials are produced as soon as possible, the main components of natural bone are collagen fiber and hydroxyapatite nanocrystal, and the granular bone mainly comes from animal bone tissue (bovine bone, porcine bone, coral bone, fish bone, etc.) or hydroxyapatite, calcium silicate, calcium phosphate, calcium carbonate, etc. with similar components due to limited availability of autologous bone tissue. However, the particulate bone has inherent drawbacks of insufficient dimensional stability (loose structure, readily flushable by blood or washing fluids) during its application (Kim J.A., Yun H.S., Choi Y.A., Kim J.E., Choi S.Y., Kwon T.G., Kim Y.K., Kwon T.Y., Bae M.A., Kim N.J., Bae Y.C., Shin H.I., Park E.K.Magnesium phosphate ceramics incorporation a novel inorganic synthesis and modification of bone cement and bone regeneration in vivo [ J.biomaterials, 2018,157: 51-61; Sunzhen, Xiaoyu, Hongsong, Zhang.
Thus, the research focus has been on the study of osteoinductive hydrogels, which are expected to impart better steric stability to the particulate bone by means of hydrogel loading (Chang C.W., Wu Y.R., Chang K.C., Ko C.L., Lin D.J., Chen W.C. in vitro characteristics of bone in the cavity calcium phosphate coated with crystalline hydrogel in association with inorganic organic inorganic organic hybrid [ J ]. ceramic International,2018,44(2): 1575-1582; Hosselndehydrah H., Ramin S.F. structural starch-poly (acrylic amide)/graphene oxide/hydrogel inorganic organic hydrogel composition [ 2018,106 ] Biological inorganic complex [ 31J ]. 25). For example, the chitosan is modified by carboxymethyl and then compounded with tricalcium silicate, so that the space stability of the granular bone material can be improved to a certain degree; or preparing the hyaluronic acid with different molecular weights into hydrogel for loading calcium phosphate or calcium carbonate and the like; for another example, a bone repair hydrogel based on a synthetic polymer is prepared by changing the chemical structures of the main chain and side groups of the polylactic acid-glycolic acid copolymer and the molecular weight of the polymer for calcium carbonate material loading (zhao ningbo. hyaluronic acid composite bone substitute material promotes osteogenesis effect and mechanism research [ D ].2016, fourth university of military medical science, doctor's paper). Whereas natural Bone itself is a highly complex composed of inorganic and organic substances, the basic constituent unit of which is hydroxyapatite nanocrystals containing collagen fibers, from the viewpoint of compositional biomimetic, composite hydrogels based on collagen and hydroxyapatite have received much attention, including organically combining collagen hydrogels with hydroxyapatite by means of biomimetic mineralization, or directly loading nano-or micro-hydroxyapatite into collagen hydrogels, for preparing osteoinductive hydrogels (go t, Xie J, Liao J, Zhang t, Lin s, Lin y. nanomaterials and Bone regeneration [ J ] Bone Research,2015,3: 1-7; skinnfam m., Bertoldi s, albanene n, Cerri a, Tanzi m.c., immuno r, baheir n, fak m, surface s.n., polyurethane/porous material J.), 2018,82:130-140). In general, hydrogels are used as excipients for organic compounding with particulate bone material (on a micron or nanometer scale), or are expected to address the inherent lack of spatial stability during particulate bone repair. However, the poor compatibility between the organic phase (polymer hydrogel) and the particle bone interface makes the synergistic effect between the two undesirable. At present, a high-molecular bionic method is used for organically combining mussel adhesive protein-imitated functional elements (3, 4-dihydroxyamphetamine, dopamine) with a high-molecular material, so that the excellent interface adhesive property of mussel adhesive protein is reproduced, and great attention is paid to people due to the strong structure designability and the simplicity and convenience of operation (Li Hui Hua. polylactic acid modified based on polydopamine adhesive layer biology and cell compatibility research thereof [ D ].2016, university in river and south, university of Master university, Bie). However, the current research is mostly limited to the interaction between synthetic macromolecules and dopamine, the biological induction effect of the final product is limited, and the research is still insufficient in the aspect of carrying out mussel bionic design on osteoinductive hydrogel based on bioactive macromolecules.
In summary, the prior art is mostly limited to simple loading of hydrogel and particulate bone, and poor interfacial compatibility between an organic phase and the particulate bone, and although there are reports related to the use of dopamine for interfacial adhesion, there are many limitations to interaction between synthetic macromolecules and dopamine, and osteoinductivity is significantly insufficient, and especially there are no reports on biomimetic construction of osteoinductive hydrogel loaded with nano fishbone based on "collagen chemical modification" and "dopamine self-assembly" and a preparation method thereof.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a nano fishbone-loaded osteoinductive hydrogel biomimetically constructed based on 'collagen chemical modification' and 'dopamine self-assembly' and a preparation method thereof. The method not only greatly improves the interface compatibility between the granular bone and the organic phase hydrogel in the prepared hydrogel, but also ensures that the prepared hydrogel has excellent osteoinductivity. This not only develops a new bone repair hydrogel material, but also opens up a new field for high value transformation of collagen and fishbone.
In order to realize the purpose, the technical scheme adopted by the invention is as follows:
an osteoinductive collagen-based composite hydrogel and a preparation method thereof, the preparation steps are as follows:
(1) preparing a collagen solution by adopting an acidic buffer solution at the dissolving temperature of 0-10 ℃, wherein the concentration range of the buffer solution is 0.1-0.5 mol/L, and the pH value is 2.0-5.0; the concentration of the collagen solution is 1-15 mg/mL; the skin collagen is one or more of pig skin, cow skin, sheep skin, donkey skin and fish skin type I collagen;
fully stirring to assist dissolution and dispersion, slowly adjusting the pH value of the collagen solution to 6.0-8.0 by adopting alkali liquor with the concentration of 0.1mol/L, and then putting the collagen solution into a constant-temperature incubator at the temperature of 30-40 ℃ for incubation for 4-12 hours to obtain a collagen hydrogel substrate;
(2) immersing the skin collagen hydrogel base material into a chemical modifier solution for 4-24 hours at the temperature of 20-37 ℃, wherein the chemical modifier is one of oxidized hyaluronic acid and condensed tannin, and the using amount of the chemical modifier is 1-8 wt% of the dry weight of the skin collagen hydrogel base material;
then adding dopamine, wherein the dosage of the dopamine is 5-10 wt% of the dry weight of the collagen hydrogel substrate, adding an oxidant accounting for 0.1-3 wt% of the weight of the dopamine, and fully reacting for 1-4 hours to allow the dopamine to perform molecular self-polymerization and assembly on the surface of the collagen hydrogel substrate, so as to obtain the dopamine surface-modified collagen hydrogel substrate;
(3) adding pre-prepared nano fishbone after dopamine reaction, wherein the using amount of the nano fishbone is 1-10 wt% of the dry weight of the collagen hydrogel substrate, adjusting the loading temperature to be 20-37 ℃, the pH value to be 5.0-8.0, fully washing by using ultrapure water after reacting for 1-12 h, then bagging and sterilizing by adopting cobalt-60 ray irradiation to obtain the osteoinductive collagen-based composite hydrogel.
Preferably, the collagen of step 1 is bovine hide type I collagen having an intact triple helix structure, other than gelatin or collagen small peptides which are degradation products thereof; the acidic buffer solution is acetic acid-sodium acetate acidic buffer solution, the concentration of the acidic buffer solution is 0.25mol/L, the pH value is 2.5, the dissolving temperature of the collagen solution prepared by the acidic buffer solution is 4 ℃, and the concentration of the collagen solution is 10 mg/mL.
Preferably, the alkali liquor in the step 1 is NaOH solution with the concentration of 0.1mol/L, the temperature of the constant-temperature incubator is preferably 37 ℃, and the incubation time is 8 h.
Preferably, the chemical modifier in step 2 is oxidized hyaluronic acid, and the amount of the chemical modifier is 5 wt% of the dry weight of the collagen hydrogel substrate.
Preferably, when the collagen hydrogel substrate is immersed in the chemical modifier solution in the step 2, the reaction temperature is 30 ℃ and the reaction time is 8 hours.
Preferably, the dopamine is used in an amount of 8 wt% of the dry weight of the collagen hydrogel substrate,
preferably, the oxidant in step 2 is H2O2The dosage is 0.5 wt% of the weight of dopamine, and the reaction time is 3h after the oxidant is added.
Preferably, the nano fishbone in the step 3 is one or a mixture of tilapia fishbone, salmon fishbone, grass carp fishbone, silver carp fishbone, green fish fishbone and the like prepared by high-energy wet ball milling, and the particle size of the nano fishbone is 20-100 nm; the dosage of the nano fishbone is 5 wt% of the dry weight of the collagen hydrogel substrate, the loading temperature is adjusted to be 30 ℃, the pH value is 7.0, and the fishbone is fully washed for 3 times by adopting ultrapure water after 4 hours of reaction.
Preferably, the adding time of the nano fishbone in the step 3 is replaced by synchronizing with the dopamine adding.
Preferably, the nano fishbone and the collagen hydrogel are loaded and fixed by dopamine, so that the phase separation of an organic phase and an inorganic phase is avoided, the effective loading rate of the nano fishbone is 80-95% (based on the weight of the added nano fishbone), the nano fishbone is a light gray or black gel in appearance, no impurities are visible to naked eyes, the hydrogel has the average porosity of 50-90%, the pH value of 7.0-7.5, the cytotoxicity is not more than 0 grade, the water retention rate is 80-95%, the swelling rate is 50-300%, and the elastic modulus is 100 Pa-3000 Pa.
Compared with the prior art, the invention has the advantages that:
1. compared with the pure granular bone material, the osteoinductive collagen-based composite hydrogel prepared by the invention has good space stability and is not easy to be dispersed by blood or washing fluid.
2. The addition of the cross-linking agent improves the mechanical property of the hydrogel, and simultaneously, compared with a glutaraldehyde cross-linking agent adopted by the traditional collagen hydrogel, the cross-linking agent used by the invention belongs to a biological cross-linking agent, so that the potential biological toxicity introduced from the source by the traditional chemical cross-linking agent (glutaraldehyde) is avoided.
3. Compared with the method for directly extracting the mussel adhesive protein from the natural mussels, the method has the advantages that the sources of the raw materials used in the method are wide, and the cost of the obtained product is relatively lower.
4. Compared with the traditional synthetic polymer hydrogel loaded granular bone, the osteoinductive collagen-based composite hydrogel prepared by the invention has excellent tissue induction effect and better interface compatibility between an organic phase and an inorganic phase.
Drawings
FIG. 1 is a line graph showing a set of elastic modulus measurements of an osteoinductive collagen-based composite hydrogel according to an embodiment of the present invention;
FIG. 2 is a line graph showing a set of viscous modulus measurements of an osteoinductive collagen-based composite hydrogel according to an embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and examples.
Example 1
Dissolving pigskin collagen by adopting 0.1mol/L acetic acid-sodium acetate solution with pH of 2.0, fully stirring at 4 ℃ to assist dissolution and dispersion to obtain collagen solution with concentration of 15mg/mL, then adjusting the pH value of the solution to 7.0 by adopting 0.1mol/L NaOH solution, and then incubating at constant temperature of 30 ℃ for 4 hours to obtain a skin collagen hydrogel substrate; the prepared collagen hydrogel substrate is immersed into a chemical modifier containing oxidized hyaluronic acid (the dosage is 5 wt% of the dry weight of the collagen hydrogel substrate) at the temperature of 20 ℃ for reaction for 8 hours. Followed by dopamine and H addition2O2The dosage of dopamine is 5 wt% of the dry weight of the collagen hydrogel substrate, H2O2The dosage is 0.5 wt% of the weight of dopamine, and the reaction lasts for 3 hours. Then adding nano tilapia mossambica bone (the dosage is 5 wt% of dry weight of the collagen hydrogel substrate), and adjusting the loading temperature to 30%Reacting at the pH of 7.0 for 4h, fully washing with ultrapure water for 3 times, bagging, and sterilizing by cobalt-60 ray irradiation to obtain the osteoinductive collagen-based composite hydrogel; a group of performance characterization results of the osteoinductive collagen-based composite hydrogel prepared by the invention are shown in Table 1.
TABLE 1
Example 2
Dissolving sheepskin collagen by adopting 0.1mol/L acetic acid-sodium acetate solution with pH of 2.0, fully stirring at 10 ℃ to assist dissolution and dispersion to obtain collagen solution with concentration of 10mg/mL, then adjusting the pH value of the solution to 7.5 by adopting 0.1mol/L NaOH solution, and then incubating at constant temperature of 37 ℃ for 8 hours to obtain a skin collagen hydrogel substrate; the prepared collagen hydrogel substrate is immersed into a chemical modifier containing oxidized hyaluronic acid (the dosage is 6 wt% of the dry weight of the collagen hydrogel substrate) at the temperature of 30 ℃ for reaction for 8 hours. Followed by dopamine and H addition2O2The dosage of dopamine is 10 wt% of the dry weight of the collagen hydrogel substrate, H2O2The amount of the solution is 1.0 wt% of the weight of dopamine, and the reaction is carried out for 3 hours. Adding nano salmon fish bones (the dosage is 10 wt% of the dry weight of the collagen hydrogel substrate), adjusting the loading temperature to 30 ℃, adjusting the pH to 7.5, reacting for 8 hours, fully washing for 3 times by using ultrapure water, bagging, and sterilizing by using cobalt-60 ray irradiation to obtain the osteoinductive collagen-based composite hydrogel, wherein a linear graph of a group of elastic modulus measurement results of the osteoinductive collagen-based composite hydrogel in the embodiment of the invention is shown in fig. 1.
Example 3
Dissolving bovine collagen by adopting 0.15mol/L acetic acid-sodium acetate solution with pH of 2.5, fully stirring at 4 ℃ to assist dissolution and dispersion to obtain collagen solution with the concentration of 5mg/mL, then adjusting the pH value of the solution to 8.0 by adopting 0.1mol/L NaOH solution, and then incubating at the constant temperature of 37 ℃ for 6 hours to obtain a collagen hydrogel substrate; immersing the prepared collagen hydrogel substrate at 30 deg.C in a solution containing condensed tannin in an amount of 5 wt% based on the dry weight of the collagen hydrogel substrate%) for 8 h. Followed by dopamine and H addition2O2The dosage of dopamine is 6 wt% of the dry weight of the collagen hydrogel substrate, H2O2The amount of the solution is 2.0 wt% of the weight of dopamine, and the reaction time is 2 hours. Then adding nano grass carp fishbone (the dosage is 8 wt% of the dry weight of the skin collagen hydrogel base material), adjusting the loading temperature to 30 ℃, adjusting the pH to 6.5, reacting for 12 hours, fully washing for 3 times by using ultrapure water, bagging, and sterilizing by using cobalt-60 ray irradiation to obtain the osteoinductive collagen-based composite hydrogel, wherein a linear graph of a group of viscous modulus measurement results of the osteoinductive collagen-based composite hydrogel in the embodiment of the invention is shown in fig. 2.
Example 4
Dissolving donkey skin collagen by using 0.2mol/L acetic acid-sodium acetate solution with pH of 3.0, fully stirring at 4 ℃ to assist dissolution and dispersion to obtain collagen solution with the concentration of 12mg/mL, then adjusting the pH value of the solution to 7.0 by using 0.1mol/L NaOH solution, and then incubating at the constant temperature of 37 ℃ for 8 hours to obtain a collagen hydrogel substrate; the prepared collagen hydrogel substrate is immersed into a chemical modifier containing condensed tannin (the dosage is 4 wt% of the dry weight of the collagen hydrogel substrate) at the temperature of 30 ℃ for reaction for 8 hours. Followed by dopamine and H addition2O2The dosage of dopamine is 5 wt% of the dry weight of the collagen hydrogel substrate, H2O2The amount of the solution is 3.0 wt% of the weight of dopamine, and the reaction is carried out for 3 hours. Then adding nano silver carp bones (the dosage is 5 wt% of the dry weight of the collagen hydrogel substrate), adjusting the loading temperature to 30 ℃, adjusting the pH to 7.0, fully washing for 3 times by adopting ultrapure water after reacting for 4 hours, then bagging and sterilizing by adopting cobalt-60 ray irradiation to obtain the osteoinductive collagen-based composite hydrogel.
Example 5
Dissolving fish skin collagen by adopting 0.1mol/L acetic acid-sodium acetate solution with pH of 2.0, fully stirring at 10 ℃ to assist dissolution and dispersion to obtain collagen solution with the concentration of 10mg/mL, then adjusting the pH value of the solution to 7.0 by adopting 0.1mol/L NaOH solution, and then incubating at the constant temperature of 37 ℃ for 8 hours to obtain a skin collagen hydrogel substrate; soaking the prepared collagen hydrogel substrate at 30 deg.C in a solution containing condensed tannin (the amount is dry weight of the collagen hydrogel substrate)5 wt%) of a chemical modifier for 8 h. Then simultaneously adding dopamine, nano black carp bone and H2O2The dosage of dopamine is 5 wt% of the dry weight of the collagen hydrogel substrate, H2O2The dosage of the nano black carp fishbone is 3.0 wt% of dopamine, the dosage of the nano black carp fishbone is 5 wt% of the dry weight of the collagen hydrogel substrate, the loading temperature is adjusted to be 30 ℃, the pH value is 7.0, after reaction for 4 hours, the nano black carp fishbone is fully washed for 3 times by adopting ultrapure water, and then the nano black carp fishbone is bagged and sterilized by adopting cobalt-60 ray irradiation, so that the osteoinductive collagen-based composite hydrogel is obtained.
Example 6
Dissolving fish skin collagen by adopting 0.1mol/L acetic acid-sodium acetate solution with pH of 2.0, fully stirring at 4 ℃ to assist dissolution and dispersion to obtain collagen solution with the concentration of 8mg/mL, then adjusting the pH value of the solution to 7.5 by adopting 0.1mol/L NaOH solution, and then incubating at constant temperature of 30 ℃ for 8 hours to obtain a skin collagen hydrogel substrate; the prepared collagen hydrogel substrate is immersed into a chemical modifier containing oxidized hyaluronic acid (the dosage is 5 wt% of the dry weight of the collagen hydrogel substrate) at the temperature of 30 ℃ for reaction for 8 hours. Then simultaneously adding dopamine, nano grass carp fishbone and H2O2The dosage of dopamine is 6 wt% of the dry weight of the collagen hydrogel substrate, H2O2The dosage of the nano grass carp fishbone is 1.0 wt% of the weight of dopamine, the dosage of the nano grass carp fishbone is 8 wt% of the dry weight of the collagen hydrogel substrate, the loading temperature is adjusted to 37 ℃, the pH value is 6.5, after reaction for 3 hours, the nano grass carp fishbone is fully washed for 3 times by using ultrapure water, and then the nano grass carp fishbone is bagged and sterilized by using cobalt-60 ray irradiation, so that the osteoinductive collagen-based composite hydrogel is obtained.
Those skilled in the art will appreciate that, in the embodiments of the methods of the present invention, the sequence numbers of the steps are not used to limit the sequence of the steps, and it is within the scope of the present invention for those skilled in the art to change the sequence of the steps without inventive work. The examples described herein are intended to aid the reader in understanding the practice of the invention and it is to be understood that the scope of the invention is not limited to such specific statements and examples. Those skilled in the art can make various other specific changes and combinations based on the teachings of the present invention without departing from the spirit of the invention, and these changes and combinations are within the scope of the invention.
Claims (10)
1. A preparation method of the osteoinductive collagen-based composite hydrogel is characterized by comprising the following steps: the preparation steps are as follows:
(1) preparing a collagen solution by adopting an acidic buffer solution at the dissolving temperature of 0-10 ℃, wherein the concentration range of the buffer solution is 0.1-0.5 mol/L, and the pH value is 2.0-5.0; the concentration of the collagen solution is 1-15 mg/mL; the skin collagen is one or more of pig skin, cow skin, sheep skin, donkey skin and fish skin type I collagen;
fully stirring to assist dissolution and dispersion, slowly adjusting the pH value of the collagen solution to 6.0-8.0 by adopting alkali liquor with the concentration of 0.1mol/L, and then putting the collagen solution into a constant-temperature incubator at the temperature of 30-40 ℃ for incubation for 4-12 hours to obtain a collagen hydrogel substrate;
(2) immersing the skin collagen hydrogel base material into a chemical modifier solution for 4-24 hours at the temperature of 20-37 ℃, wherein the chemical modifier is one of oxidized hyaluronic acid and condensed tannin, and the using amount of the chemical modifier is 1-8 wt% of the dry weight of the skin collagen hydrogel base material;
then adding dopamine, wherein the dosage of the dopamine is 5-10 wt% of the dry weight of the collagen hydrogel substrate, adding an oxidant accounting for 0.1-3 wt% of the weight of the dopamine, and fully reacting for 1-4 hours to allow the dopamine to perform molecular self-polymerization and assembly on the surface of the collagen hydrogel substrate, so as to obtain the dopamine surface-modified collagen hydrogel substrate;
(3) adding pre-prepared nano fishbone after dopamine reaction, wherein the using amount of the nano fishbone is 1-10 wt% of the dry weight of the collagen hydrogel substrate, adjusting the loading temperature to be 20-37 ℃, the pH value to be 5.0-8.0, fully washing by using ultrapure water after reacting for 1-12 h, then bagging and sterilizing by adopting cobalt-60 ray irradiation to obtain the osteoinductive collagen-based composite hydrogel.
2. The method of claim 1, wherein: the hide collagen in the step 1 is cow leather type I collagen; the acidic buffer solution is acetic acid-sodium acetate acidic buffer solution, the concentration of the acidic buffer solution is 0.25mol/L, the pH value is 2.5, the dissolving temperature of the collagen solution prepared by the acidic buffer solution is 4 ℃, and the concentration of the collagen solution is 10 mg/mL.
3. The method of claim 1, wherein: the alkali liquor in the step 1 is NaOH solution with the concentration of 0.1mol/L, the temperature of the constant-temperature incubator is 37 ℃, and the incubation time is 8 hours.
4. The method of claim 1, wherein: the chemical modifier in the step 2 is oxidized hyaluronic acid, and the dosage of the chemical modifier is 5 wt% of the dry weight of the collagen hydrogel substrate.
5. The method of claim 1, wherein: and (2) when the collagen hydrogel substrate is immersed in the chemical modifier solution in the step 2, the reaction temperature is 30 ℃, and the reaction time is 8 hours.
6. The method of claim 1, wherein: the dopamine in the step 2 accounts for 8 wt% of the dry weight of the collagen hydrogel substrate.
7. The method of claim 1, wherein: the oxidant in the step 2 is H2O2The dosage is 0.5 wt% of the weight of dopamine, and the reaction time is 3h after the oxidant is added.
8. The method of claim 1, wherein: the nano fishbone in the step 3 is one or a mixture of tilapia fishbone, salmon fishbone, grass carp fishbone, silver carp fishbone and green fishbone prepared by high-energy wet ball milling, and the particle size of the nano fishbone is 20-100 nm; the dosage of the nano fishbone is 5 wt% of the dry weight of the collagen hydrogel substrate, the loading temperature is adjusted to be 30 ℃, the pH value is 7.0, and the fishbone is fully washed for 3 times by adopting ultrapure water after 4 hours of reaction.
9. The method of claim 1, wherein: and 4, replacing the adding time of the nano fishbone in the step 3 with the adding time of the dopamine.
10. The method of claim 1, wherein: according to the osteoinductive collagen-based composite hydrogel, the nano fishbone and the collagen hydrogel are loaded and fixed through dopamine, so that the phase separation of an organic phase and an inorganic phase is avoided, the effective loading rate of the nano fishbone is 80-95%, the nano fishbone is a light gray or black gel in appearance, no visible impurities exist, the average porosity of the hydrogel is 50-90%, the pH value is 7.0-7.5, the cytotoxicity is not more than 0 grade, the water retention rate is 80-95%, the swelling rate is 50-300%, and the elastic modulus is 100-3000 Pa.
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| CN113336972B (en) * | 2021-06-21 | 2022-08-30 | 陕西科技大学 | Multifunctional collagen-based nano composite hydrogel and preparation method thereof |
| CN113403846A (en) * | 2021-07-30 | 2021-09-17 | 南昌大学附属口腔医院(江西省口腔医院) | Polylactic acid nanofiber composite membrane with biomimetic mineralization and antibacterial functions and preparation method and application thereof |
| CN115304775A (en) * | 2022-06-27 | 2022-11-08 | 天津大学 | Preparation method of collagen matrix modified by modified hyaluronic acid |
| CN115501122B (en) * | 2022-09-21 | 2023-09-22 | 四川大学 | Antibacterial mineralized hydrogel and precursor material, and preparation and application methods thereof |
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