CN111686298A - Tissue engineering gel scaffold for promoting bone healing and preparation method thereof - Google Patents
Tissue engineering gel scaffold for promoting bone healing and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a preparation method of a tissue engineering gel scaffold for promoting bone healing, which comprises the following steps: step 1, preparing a PVA-GMA/g-C3N4 precursor; step 2, preparing an ICA solution; and 3, mixing the PVA-GMA/g-C3N4 precursor with the ICA solution according to the volume ratio of (4-6) to 1, uniformly stirring, injecting the mixture into a bone injury part, and carrying out photo-crosslinking curing to obtain the tissue engineering gel scaffold carrying the drug. The PVA-GMA/g-C3N4 precursor is selected as the material of the tissue engineering epidermis, the mechanical strength is strong after the solidification, a good healing environment can be provided for the bone injury part in the early stage, and the gel scaffold can be slowly degraded along with the bone healing process. The icariin is carried, so that the proliferation and osteogenic differentiation of the mesenchymal stem cells are promoted. The gel scaffold is formed by a photo-crosslinking method and can be cured into any shape to realize close fit with the defective tissue.
Description
Technical Field
The invention belongs to the technical field of tissue engineering bone regeneration, relates to a drug-loaded bone scaffold, and particularly relates to a tissue engineering gel scaffold for promoting bone healing and a preparation method thereof.
Background
As the most rigid organs in the vertebrate body, the skeleton performs a series of important functions during daily life activities, such as protecting the internal organs, supporting movement, etc. In clinic, bone defects are one of the difficult problems in orthopedic treatment, and the traditional methods for treating bone defects include bone grafting, the Ilizanow technology and the like, but the treatment period of the methods is long, the cost is high, and the methods are easy to be accompanied with various complications such as organism immune response, bone hyperplasia, secondary fracture and the like. Therefore, it is very important and urgent to solve the bone healing problem, and finding a new method for effectively promoting bone regeneration is of great significance to the development of modern medicine.
With the rise and the vigorous development of tissue engineering, the treatment of bone defects has a new direction. The bone tissue engineering aims to provide a good cell scaffold for cell growth in a healing process, the biomaterial scaffold can provide a living three-dimensional space for cells, and the differentiation of the cells can be promoted through pharmaceutical intervention.
Icariin (ICA) is an extract of a traditional Chinese medicine epimedium commonly used in China, and at present, a plurality of studies prove that the ICA can promote the proliferation and osteogenic differentiation of mesenchymal stem cells and has wide application prospect in the clinical treatment of bone defects. But the limited administration mode leads to low utilization rate of the drug, which also limits the further application of the drug in the bone repair field.
Disclosure of Invention
The invention aims to overcome the defects in the prior art, and constructs a gel scaffold capable of promoting bone healing by using related principles of tissue engineering, bionics and the like.
In order to achieve the above objects, the present invention provides a method for preparing a tissue engineering gel scaffold for promoting bone healing, the method comprising:
step 1, preparing a PVA-GMA/g-C3N4 precursor;
step 2, preparing an ICA solution;
and 3, mixing the PVA-GMA/g-C3N4 precursor with the ICA solution according to the volume ratio of (4-6) to 1, uniformly stirring, injecting the mixture into a bone injury part, and carrying out photo-crosslinking curing to obtain the tissue engineering gel scaffold carrying the drug.
Preferably, step 1 comprises:
step 1.1, dissolving PVA in DMSO, adding GMA and DMA, and uniformly stirring to obtain PVA-GMA;
and step 1.2, dissolving the PVA-GMA, g-C3N4 powder and a photoinitiator D-2959 in PBS to obtain the PVA-GMA/g-C3N4 precursor.
Preferably, in step 1.1, the PVA dissolved in 100ml of DMSO has a mass of 5.0g to 8.0g, GMA is added in an amount of 0.32ml to 0.79ml, DMA is added in an amount of 6.5ml to 8.1ml, and the resulting solution is stirred at 60 ℃ to 70 ℃ for 12 hours to 24 hours, then the mixture is precipitated in acetone and dried under vacuum for 7 days or more.
Preferably, in step 1.2, PVA-GMA with the mass fraction of 25-35% and g-C3N4 with the mass fraction of 6.25-7.25% are dissolved in PBS, then stirred for 30min-2h at the speed of 50r/min-150r/min, and then D-2959 with the concentration of 0.3-0.6 wt% is added into the mixed solution.
Preferably, in the step 2, the solvent of the ICA solution is 10% to 20% of ethanol water solution, and the mass fraction of the prepared ICA solution is 10% to 15%.
Preferably, the photo-crosslinking curing method in step 3 is curing and molding for 8s-10s under blue light 400-450 nm.
The invention also provides a tissue engineering gel scaffold for promoting bone healing, which is prepared by the preparation method.
Compared with the prior art, the invention has the following beneficial effects:
(1) the PVA-GMA/g-C3N4 precursor is selected as the material of the tissue engineering epidermis, the mechanical strength is strong after the solidification, a good healing environment can be provided for the bone injury part in the early stage, and the gel scaffold can be slowly degraded along with the bone healing process.
(2) The invention is beneficial to promoting the proliferation and osteogenic differentiation of mesenchymal stem cells by carrying the icariin.
(3) The gel scaffold is formed by a photo-crosslinking method and can be cured into any shape to realize close fit with the defective tissue.
Detailed Description
The technical solutions of the present invention are described clearly and completely below, and it is obvious that the described embodiments are some, not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention discloses a preparation method of a tissue engineering gel scaffold for promoting bone healing, which comprises the following steps: a photo-crosslinking scaffold for simulating bone tissues and a medicament for inducing osteogenic differentiation of mesenchymal stem cells; the main component of the photocrosslinking bracket for simulating bone tissue is PVA-GMA/g-C3N4 gel compounded by polyvinyl alcohol (PVA-GMA) modified by Glycidyl Methacrylate (GMA) and hydroxylated carbon nitride (g-C3N4) nano particles; icariin (icarin, ICA) is selected as a drug for inducing osteogenic differentiation of the mesenchymal stem cells.
PVA-GMA/g-C3N4 precursor for constructing tissue engineering epidermis and photoinitiator Darocur 2959(D2959) rapidly break double bonds on PVA-GMA after absorbing photon energy and rapidly chemically bond with hydroxyl on the surface of g-C3N4 and broken double bonds on other nearby PVA-GMAs to form a nano composite structure, so that the mechanical strength of the gel is greatly enhanced.
ICA is an extract of epimedium herb which is a commonly used Chinese medicine in China, and can promote the proliferation of mesenchymal stem cells, and the mesenchymal stem cells can be differentiated into osteoblasts under specific conditions, so that the differentiation and maturation of the osteoblasts can be promoted by the intervention of ICA, and the regeneration of bones is induced.
Examples
Step 1: preparation of the precursor
Step 1.1: 8.0g of polyvinyl alcohol (PVA) was dissolved in 100ml of dimethyl sulfoxide (DMSO). Adding 0.32ml GMA and 6.5ml DMA, stirring at 60 deg.C for 12 hr, precipitating the mixture in acetone, filtering, and vacuum drying for more than 7 days;
step 1.2: taking Phosphate Buffer Solution (PBS) as a solvent, sequentially adding 25% by mass of PVA-GMA and 6.25% by mass of g-C3N4, stirring at a low speed of 50r/min-150r/min for 30min, adding 0.3 wt% by weight of D-2959 into the mixed solution, and storing in a dark place at 4 ℃ in an aseptic environment;
the PVA-GMA/g-C3N4 precursor prepared in the step 1 is used for constructing tissue engineering epidermis, has strong mechanical strength, can provide a good healing environment for a bone injury part in the early stage, and can realize slow degradation along with the process of bone healing.
Step 2: preparation of ICA solution
Weighing 1.504mg ICA, dissolving in 10mL 10% ethanol water solution to obtain icariin solution with mass fraction of 15%, packaging, and storing at 4 deg.C under sterile condition.
And step 3: photo-crosslinking and curing
Uniformly stirring the PVA-GMA/g-C3N4 precursor obtained in the step 1 and the ICA solution obtained in the step 2 according to the volume ratio of 4:1, injecting the mixture into the defect part of the medial condyle of the femur of the rabbit, and curing and molding the mixture for 8s under the blue light of 405 nm.
In the above examples, the gel formed at the bone defect had good mechanical strength, and no adverse reaction was observed after 30 days; after 90 days, the growth state of the neogenetic callus tissue was confirmed to be good by direct observation through the second operation.
In conclusion, the tissue engineering gel scaffold for promoting bone healing and the preparation method thereof provided by the invention have the advantages that the PVA-GMA/g-C3N4 precursor is used for constructing the tissue engineering epidermis, the icariin is used as a medicine for inducing osteogenic differentiation of the mesenchymal stem cells, the gel scaffold is fixedly molded by adopting the photocrosslinking method, the proliferation and osteogenic differentiation of the mesenchymal stem cells are promoted, a good healing environment can be provided for a bone injury part in the early stage, and the gel scaffold is tightly attached to a defective tissue. And the gel scaffold can be slowly degraded along with the process of bone healing.
While the present invention has been described in detail with reference to the preferred embodiments, it should be understood that the above description should not be taken as limiting the invention. Various modifications and alterations to this invention will become apparent to those skilled in the art upon reading the foregoing description. Accordingly, the scope of the invention should be determined from the following claims.
Claims (7)
1. A method for preparing a tissue engineering gel scaffold for promoting bone healing, the method comprising:
step 1, preparing a PVA-GMA/g-C3N4 precursor;
step 2, preparing an ICA solution;
and 3, mixing the PVA-GMA/g-C3N4 precursor with the ICA solution according to the volume ratio of (4-6) to 1, uniformly stirring, injecting the mixture into a bone injury part, and carrying out photo-crosslinking curing to obtain the tissue engineering gel scaffold carrying the drug.
2. The method for preparing a tissue engineering gel scaffold for promoting bone healing as claimed in claim 1, wherein the step 1 comprises:
step 1.1, dissolving PVA in DMSO, adding GMA and DMA, and uniformly stirring to obtain PVA-GMA;
and step 1.2, dissolving the PVA-GMA, g-C3N4 powder and a photoinitiator D-2959 in PBS to obtain the PVA-GMA/g-C3N4 precursor.
3. The method of preparing a tissue engineering gel scaffold for promoting bone healing according to claim 2, wherein in step 1.1, the mass of PVA dissolved in 100ml of DMSO is 5.0g to 8.0g, the amount of GMA added is 0.32ml to 0.79ml, the amount of DMA added is 6.5ml to 8.1ml, the resulting solution is stirred at 60 ℃ to 70 ℃ for 12h to 24h, the mixture is precipitated in acetone, and vacuum-dried for 7 days or more.
4. The method for preparing a tissue engineering gel scaffold for promoting bone healing as claimed in claim 2, wherein in step 1.2, after 25% -35% by mass of PVA-GMA and 6.25% -7.25% by mass of g-C3N4 are dissolved in PBS, stirring is carried out at a speed of 50r/min-150r/min for 30min-2h, and then D-2959 with a concentration of 0.3% -0.6% by weight is added to the mixed solution.
5. The method for preparing a tissue engineering gel scaffold for promoting bone healing as claimed in claim 1, wherein the solvent of the ICA solution is 10 to 20% aqueous ethanol solution to prepare the ICA solution with a mass fraction of 10 to 15% in step 2.
6. The method for preparing a tissue engineering gel scaffold for promoting bone healing as claimed in claim 1, wherein the photo cross-linking curing method in step 3 is curing under blue light of 400nm-450nm for 8s-10 s.
7. A tissue engineering gel scaffold for promoting bone healing, which is prepared by the preparation method of any one of claims 1 to 6.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115054729A (en) * | 2022-07-07 | 2022-09-16 | 广州贝奥吉因生物科技股份有限公司 | Double-network hydrogel and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115054729A (en) * | 2022-07-07 | 2022-09-16 | 广州贝奥吉因生物科技股份有限公司 | Double-network hydrogel and preparation method and application thereof |
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