CN114569789A - Preparation method and application of BPN-DFO gel scaffold - Google Patents
Preparation method and application of BPN-DFO gel scaffold Download PDFInfo
- Publication number
- CN114569789A CN114569789A CN202210287535.6A CN202210287535A CN114569789A CN 114569789 A CN114569789 A CN 114569789A CN 202210287535 A CN202210287535 A CN 202210287535A CN 114569789 A CN114569789 A CN 114569789A
- Authority
- CN
- China
- Prior art keywords
- bpn
- dfo
- black phosphorus
- chitosan
- precursor solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/222—Gelatin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/02—Inorganic materials
- A61L27/12—Phosphorus-containing materials, e.g. apatite
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/52—Hydrogels or hydrocolloids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/10—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing inorganic materials
- A61L2300/112—Phosphorus-containing compounds, e.g. phosphates, phosphonates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/216—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials with other specific functional groups, e.g. aldehydes, ketones, phenols, quaternary phosphonium groups
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
- A61L2300/602—Type of release, e.g. controlled, sustained, slow
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Dermatology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Inorganic Chemistry (AREA)
- Dispersion Chemistry (AREA)
- Medicinal Preparation (AREA)
- Materials For Medical Uses (AREA)
Abstract
The invention relates to the technical field of biomedical materials, in particular to a preparation method and application of a BPN-DFO gel scaffold; the invention takes the black phosphorus nano-sheet (BPN) and the Deferoxamine (DFO) as the bioactive components for promoting the regeneration of blood vessels and bones and is used for repairing ischemic bone defects, wherein the BPN can release controllable phosphorus, and the DFO can be used as an iron chelating agent and can promote the angiogenesis by activating key angiogenesis promoting genes such as Vascular Endothelial Growth Factor (VEGF) and the like. BPN-DFO (black phosphorus nanoplate and deferoxamine) gel scaffolds showed significant upregulation of mRNA expression associated with bone regeneration and cell proliferation. In vivo, BPN-DFO gel scaffold can significantly improve osteogenesis and neovascularization at ischemic bone defect site.
Description
Technical Field
The invention relates to the technical field of biomedical materials, in particular to a preparation method and application of a BPN-DFO gel scaffold.
Background
It is generally accepted that delayed healing or nonunion in the bone healing process is due to insufficient blood supply. Although bone tissue has good regenerative ability, most of ischemic bone defects are not completely restored due to insufficient blood supply, which is one of the main causes of bone nonunion. When bone nonunion exists, the patient may suffer from bone loss, bone deformity, and persistent infection. To date, the extensive clinical strategies for treating nonunion include small defect autologous bone grafting, bone transport, and induced membrane techniques. The above-described regimen is prone to a series of complications that are difficult to avoid.
Recently, hydrogel scaffolds have been considered as an effective material with a variety of medical values, including simple cell or tissue scaffolds, tissue engineering platforms, advanced drug and/or growth factor carriers. Many researchers report that hydrogel scaffolds achieve clinically desirable regenerative effects in peripheral nerves, spinal cord, wounds, bone and cartilage due to encapsulation of cells, growth factors and/or bioactive molecules that can be controllably released to improve differentiation and regeneration. The hydrogel has good biocompatibility, drug loading property and osteoconductivity, so that the hydrogel is considered to be the best scaffold material for bone defect repair.
Notably, in most studies to date, osteogenesis in normal bone defects is more important than osteogenesis in ischemic bone defects. In ischemic conditions, these scaffolds may induce fibrous tissue formation in the bone defect, failing to treat the bone defect. In the process of bone defect healing, due to the lack of bone blood vessels, fibroplasia, delayed bone healing, nonunion and necrosis occur. Therefore, in order to promote bone regeneration at an ischemic site, the gel scaffold must have various functions including promotion of osteogenesis and angiogenesis.
Disclosure of Invention
The present invention is directed, at least in part, to overcoming the above-mentioned and/or other potential problems in the art: provides a preparation method and application of BPN-DFO gel scaffold for promoting osteogenesis and angiogenesis.
The technical solution of the invention is as follows: a preparation method of a BPN-DFO gel scaffold comprises the following steps:
1) preparing black phosphorus nanosheet-chitosan: dispersing the black phosphorus nanosheet and chitosan in water, stirring and uniformly mixing in a dark place to obtain a suspension, centrifuging the suspension to obtain a precipitate, and drying the precipitate in vacuum to obtain a product, namely the black phosphorus nanosheet-chitosan; wherein the content of the black phosphorus nanosheet in the suspension is 0.5-1mg/mL, and the content of the chitosan is 5-10 mg/mL;
2) dissolving a photoinitiator (LAP) and methacrylic acid acylated gelatin (GelMA) into PBS buffer solution, and heating at the water bath temperature of 30-50 ℃ for 10-20min to obtain hydrogel precursor solution, wherein the concentration of the photoinitiator (LAP) in the hydrogel precursor solution is 1.25-2.5mg/mL, and the concentration of the methacrylic acid acylated gelatin (GelMA) is 40-60 mg/mL;
3) adding black phosphorus nanosheet-chitosan (BPN-QCS) and Desferrioxamine (DFO) into the hydrogel precursor solution, and heating at a water bath temperature of 50-70 ℃ for 25-40 min; the concentration of the black phosphorus nanosheet-chitosan in the hydrogel precursor solution is 75-125 mug/mL, and the concentration of the deferoxamine is 0.75-1.25 mg/mL;
4) Irradiating the prepared hydrogel precursor solution prepared in the step 3) by using a long-wave ultraviolet lamp to prepare the BPN-DFO gel scaffold.
As an optimization, in step 1), the black phosphorus nanosheet is prepared by the following steps: grinding the Black Phosphorus (BP) crystal powder, transferring the powder to a centrifuge tube, fixing the volume with water until the concentration of the Black Phosphorus (BP) is 1mg/mL, stirring and uniformly mixing the powder to form a suspension, centrifuging the suspension, removing the bottom precipitate, taking the supernatant, and drying the supernatant in vacuum to obtain the Black Phosphorus Nanosheets (BPNs).
In the step 4), the wavelength of the long-wave ultraviolet lamp is 65-365nm, the power is 300-400W, and the irradiation time is 5-10 s.
Preferably, in the step 1), the chitosan is quaternized chitosan.
The invention provides application of a BPN-DFO gel scaffold, and particularly relates to application of the BPN-DFO gel scaffold in repairing ischemic bone defects.
The beneficial effects of the invention are: the invention takes the black phosphorus nano-sheet (BPN) and the Desferrioxamine (DFO) as the bioactive components for promoting the regeneration of blood vessels and bones, and is used for repairing ischemic bone defects, wherein the BPN can release controllable phosphorus, and the DFO is taken as an iron chelator, and can promote the angiogenesis by activating key angiogenesis promoting genes such as Vascular Endothelial Growth Factor (VEGF) and the like. The gel scaffold prepared by the invention has good swelling, degrading and releasing rates and good biocompatibility. BPN-DFO (black phosphorus nanoplate and deferoxamine) gel scaffolds showed significant upregulation of mRNA expression associated with bone regeneration and cell proliferation. In vivo, BPN-DFO gel scaffold can significantly improve osteogenesis and neovascularization at ischemic bone defect site.
Drawings
FIG. 1 is a graph showing the comparison of the swelling rate and degradation rate of BPN-DFO gel scaffolds prepared in example 1 and a hydrogel in ultrapure water at room temperature, FIG. 1a is a graph showing the comparison of the swelling rate, and FIG. 1b is a graph showing the comparison of the degradation rate.
FIG. 2 is a graph showing the cumulative percentage of phosphorus and iron ion released in vitro from BPN-DFO gel scaffolds prepared in example 1. Fig. 2a is a graph of the cumulative release percentage of phosphorus ions, and fig. 2b is a graph of the cumulative release percentage of iron ions.
FIG. 3 shows live/dead staining fluorescence images of two separate hBMSCs incubations. Panel a, b live/dead staining fluorescence image of BPN-DFO group. Panels c, d live/dead stain fluorescence images of the hydrogel set.
Detailed Description
The present invention will be described in further detail with reference to the following examples, but the present invention is not limited to the following examples.
Example 1
BPN-DFO gel scaffolds were prepared according to the following procedure:
1) preparing black phosphorus nanosheet-chitosan: dispersing the black phosphorus nanosheet and chitosan in water, stirring and uniformly mixing in a dark place to obtain a suspension, then centrifuging the suspension to obtain a precipitate, and drying the precipitate in vacuum to obtain a product, namely the black phosphorus nanosheet-chitosan; wherein the content of the black phosphorus nanosheet in the suspension is 0.7mg/mL, and the content of the chitosan is 6 mg/mL;
2) Dissolving a photoinitiator (LAP) and methacrylic acid acylated gelatin (GelMA) into PBS buffer solution, and heating at the water bath temperature of 40 ℃ for 15min to obtain hydrogel precursor solution, wherein the concentration of the photoinitiator (LAP) in the hydrogel precursor solution is 2.5mg/mL, and the concentration of the methacrylic acid acylated gelatin (GelMA) in the hydrogel precursor solution is 50 mg/mL;
3) adding black phosphorus nanosheet-chitosan (BPN-QCS) and Desferrioxamine (DFO) into the hydrogel precursor solution, and heating at a water bath temperature of 60 ℃ for 30 min; the concentration of the black phosphorus nanosheet-chitosan in the hydrogel precursor solution is 100 mug/mL, and the concentration of the deferoxamine is 1 mg/mL;
4) irradiating the prepared hydrogel precursor solution prepared in the step 3) for 10s by using a long-wave ultraviolet lamp to prepare the BPN-DFO gel scaffold. The wavelength of the long-wave ultraviolet lamp is 365nm, and the power is 400W.
Example 2
BPN-DFO gel scaffolds were prepared according to the following procedure:
1) preparing black phosphorus nanosheet-chitosan: dispersing the black phosphorus nanosheet and the quaternized chitosan in water, stirring and uniformly mixing in a dark place to obtain a suspension, centrifuging the suspension to obtain a precipitate, and drying the precipitate in vacuum to obtain a product, namely the black phosphorus nanosheet-chitosan; wherein the content of the black phosphorus nanosheet in the suspension is 0.5mg/mL, and the content of the chitosan is 5 mg/mL;
2) Dissolving a photoinitiator (LAP) and methacrylic acid acylated gelatin (GelMA) into PBS buffer solution, and heating at the water bath temperature of 40 ℃ for 15min to obtain hydrogel precursor solution, wherein the concentration of the photoinitiator (LAP) in the hydrogel precursor solution is 2.5mg/mL, and the concentration of the methacrylic acid acylated gelatin (GelMA) in the hydrogel precursor solution is 50 mg/mL;
3) adding black phosphorus nanosheet-chitosan (BPN-QCS) and Desferrioxamine (DFO) into the hydrogel precursor solution, and heating at a water bath temperature of 60 ℃ for 30 min; the concentration of the black phosphorus nanosheet-chitosan in the hydrogel precursor solution is 100 mug/mL, and the concentration of the deferoxamine is 1 mg/mL;
4) irradiating the prepared hydrogel precursor solution prepared in the step 3) for 10s by using a long-wave ultraviolet lamp to prepare the BPN-DFO gel scaffold. The wavelength of the long-wave ultraviolet lamp is 365nm, and the power is 400W.
Example 3
BPN-DFO gel scaffolds were prepared according to the following procedure:
1) preparing black phosphorus nanosheet-chitosan: dispersing the black phosphorus nanosheet and the quaternized chitosan in water, stirring and uniformly mixing in a dark place to obtain a suspension, centrifuging the suspension to obtain a precipitate, and drying the precipitate in vacuum to obtain a product, namely the black phosphorus nanosheet-chitosan; wherein the content of the black phosphorus nanosheet in the suspension is 0.5mg/mL, and the content of the chitosan is 5 mg/mL;
2) Dissolving a photoinitiator (LAP) and methacrylic acid acylated gelatin (GelMA) into a PBS buffer solution, and heating at the water bath temperature of 50 ℃ for 10min to obtain a hydrogel precursor solution, wherein the concentration of the photoinitiator (LAP) in the hydrogel precursor solution is 1.25mg/mL, and the concentration of the methacrylic acid acylated gelatin (GelMA) in the hydrogel precursor solution is 40 mg/mL;
3) adding black phosphorus nanosheet-chitosan (BPN-QCS) and Desferrioxamine (DFO) into the hydrogel precursor solution, and heating at a water bath temperature of 60 ℃ for 30 min; the concentration of the black phosphorus nanosheet-chitosan in the hydrogel precursor solution is 100 mug/mL, and the concentration of the deferoxamine is 1 mg/mL;
4) irradiating the prepared hydrogel precursor solution prepared in the step 3) for 10s by using a long-wave ultraviolet lamp to prepare the BPN-DFO gel scaffold. The wavelength of the long-wave ultraviolet lamp is 365nm, and the power is 400W.
Example 4
BPN-DFO gel scaffolds were prepared according to the following procedure:
1) preparing black phosphorus nanosheet-chitosan: dispersing the black phosphorus nanosheet and the quaternized chitosan in water, stirring and uniformly mixing in a dark place to obtain a suspension, centrifuging the suspension to obtain a precipitate, and drying the precipitate in vacuum to obtain a product, namely the black phosphorus nanosheet-chitosan; wherein the content of the black phosphorus nanosheet in the suspension is 1mg/mL, and the content of the chitosan is 10 mg/mL;
2) Dissolving a photoinitiator (LAP) and methacrylic acid acylated gelatin (GelMA) into a PBS buffer solution, and heating at the water bath temperature of 50 ℃ for 10min to obtain a hydrogel precursor solution, wherein the concentration of the photoinitiator (LAP) in the hydrogel precursor solution is 1.25mg/mL, and the concentration of the methacrylic acid acylated gelatin (GelMA) in the hydrogel precursor solution is 40 mg/mL;
3) adding black phosphorus nanosheet-chitosan (BPN-QCS) and Desferrioxamine (DFO) into the hydrogel precursor solution, and heating at a water bath temperature of 60 ℃ for 30 min; the concentration of the black phosphorus nanosheet-chitosan in the hydrogel precursor solution is 100 mug/mL, and the concentration of the deferoxamine is 1 mg/mL;
4) irradiating the prepared hydrogel precursor solution prepared in the step 3) for 10s by using a long-wave ultraviolet lamp to prepare the BPN-DFO gel scaffold. The wavelength of the long-wave ultraviolet lamp is 365nm, and the power is 400W.
Example 5
BPN-DFO gel scaffolds were prepared according to the following procedure:
1) preparing black phosphorus nanosheet-chitosan: dispersing the black phosphorus nanosheet and the quaternized chitosan in water, stirring and uniformly mixing in a dark place to obtain a suspension, centrifuging the suspension to obtain a precipitate, and drying the precipitate in vacuum to obtain a product, namely the black phosphorus nanosheet-chitosan; wherein the content of the black phosphorus nanosheet in the suspension is 1mg/mL, and the content of the chitosan is 10 mg/mL;
2) Dissolving a photoinitiator (LAP) and methacrylic acid acylated gelatin (GelMA) into PBS buffer solution, and heating at the water bath temperature of 50 ℃ for 10min to obtain hydrogel precursor solution, wherein the concentration of the photoinitiator (LAP) in the hydrogel precursor solution is 2.5mg/mL, and the concentration of the methacrylic acid acylated gelatin (GelMA) in the hydrogel precursor solution is 60 mg/mL;
3) adding black phosphorus nanosheet-chitosan (BPN-QCS) and Desferrioxamine (DFO) into the hydrogel precursor solution, and heating at a water bath temperature of 60 ℃ for 30 min; the concentration of the black phosphorus nanosheet-chitosan in the hydrogel precursor solution is 75 mug/mL, and the concentration of the deferoxamine is 0.75 mg/mL;
4) irradiating the prepared hydrogel precursor solution prepared in the step 3) for 10s by using a long-wave ultraviolet lamp to prepare the BPN-DFO gel scaffold. The wavelength of the long-wave ultraviolet lamp is 365nm, and the power is 400W.
Example 6
BPN-DFO gel scaffolds were prepared according to the following procedure:
1) preparing black phosphorus nanosheets: grinding the Black Phosphorus (BP) crystal powder, transferring the powder to a centrifuge tube, fixing the volume with water until the concentration of the Black Phosphorus (BP) is 1mg/mL, stirring and uniformly mixing the powder to form a suspension, centrifuging the suspension, removing the bottom precipitate, taking the supernatant, and drying the supernatant in vacuum to obtain the Black Phosphorus Nanosheets (BPNs).
2) Preparing black phosphorus nanosheet-chitosan: dispersing the black phosphorus nanosheet and the quaternized chitosan in water, stirring and uniformly mixing in a dark place to obtain a suspension, centrifuging the suspension to obtain a precipitate, and drying the precipitate in vacuum to obtain a product, namely the black phosphorus nanosheet-chitosan; wherein the content of the black phosphorus nanosheet in the suspension is 1mg/mL, and the content of the chitosan is 10 mg/mL;
3) Dissolving a photoinitiator (LAP) and methacrylic acid acylated gelatin (GelMA) into PBS buffer solution, and heating at the water bath temperature of 50 ℃ for 10min to obtain hydrogel precursor solution, wherein the concentration of the photoinitiator (LAP) in the hydrogel precursor solution is 2.5mg/mL, and the concentration of the methacrylic acid acylated gelatin (GelMA) in the hydrogel precursor solution is 60 mg/mL;
4) adding black phosphorus nanosheet-chitosan (BPN-QCS) and Desferrioxamine (DFO) into the hydrogel precursor solution, and heating at a water bath temperature of 60 ℃ for 30 min; the concentration of the black phosphorus nanosheet-chitosan in the hydrogel precursor solution is 125 mug/mL, and the concentration of the deferoxamine is 1.25 mg/mL;
5) irradiating the prepared hydrogel precursor solution prepared in the step 4) for 10s by using a long-wave ultraviolet lamp to prepare the BPN-DFO gel scaffold. The wavelength of the long-wave ultraviolet lamp is 365nm, and the power is 400W.
The following experiment was carried out using the BPN-DFO gel prepared in example 1:
swelling: the dried BPN-DFO hydrogel was weighed (W0) and then soaked in 10ml of ultrapure water.
The water on the surface of the sample was removed before weighing at each measurement time (Wt).
The swelling ratio was calculated by the formula SR = (Wt-W0)/W0X 100%.
In vitro degradation: the dried BPN-DFO hydrogel was weighed (W0), and each sample was washed with ultrapure water at each measurement time, dried for 24 hours, incubated in collagenase IIPBS solution at 37 ℃ and then weighed at the following time nodes (1, 2, 3, 5, 7, 9, 11, 14 days). After drying the samples at 40 ℃, they were washed with PBS and weighed (Wt). The degradation rate of BPN-DFO hydrogel was calculated by the formula DR = (W0-Wt)/W0X 100%. The swelling rates of BPN-DFO gels and hydrogels in ultrapure water at room temperature are shown in FIG. 1 a. The degradation rates of BPN-DFO gels and hydrogels are shown in FIG. 1 b.
Releasing the medicine: in order to study the release rate of BPN-DFO gel to phosphorus and iron ions, the in vitro release test method is adopted to study the release rate of BPN-DFO gel to phosphorus ions (P)5+) And placing the hydrogel sample in 1ml of phosphate-free PBS buffer solution for incubation at 37 ℃, collecting 100 mu L of supernatant on days 1, 2, 3, 4, 5, 7, 14 and 21 respectively, and then adding 100 mu L of fresh buffer solution. All samples were 3 replicates (n = 3). The supernatant samples were analyzed for phosphorus ion concentration using a phosphorus colorimetric kit. Iron ion (Fe) under the same conditions2+) The release rates of (c) were analyzed using an iron colorimetric assay kit at (1, 3, 5, 10, 24, 48, 72, 96 h). FIG. 2a shows the cumulative percentage of phosphate ion release in vitro from BPN-DFO gels. The cumulative release percentage of phosphorus ions is sustained within 21 d. Figure 2b is the cumulative percentage release of iron ions, which is explosive during the first 24 h and then slowly released after 48 h.
As shown in FIG. 3, the live/dead staining fluorescence images of the incubation of hBMSCs on the BPN-DFO gel and the hydrogel, respectively, were obtained, and no dead cells were observed in both groups. Panel a, b live/dead staining fluorescence image of BPN-DFO group. Panels c, d live/dead stain fluorescence images of the hydrogel set.
It can be seen that the BPN-DFO gel scaffold prepared in example 1 of the present invention has good swelling, degradation and release rates, and good biocompatibility. The BPN-DFO gel scaffold can obviously improve osteogenesis and neovascularization of ischemic bone defect parts.
The above are merely exemplary embodiments of the features of the present invention, and do not limit the scope of the present invention in any way. All technical solutions formed by adopting the equivalent exchange or the equivalent substitution fall within the protection scope of the present invention.
Claims (6)
1. A preparation method of a BPN-DFO gel scaffold is characterized by comprising the following steps:
1) preparing black phosphorus nanosheet-chitosan: dispersing the black phosphorus nanosheet and chitosan in water, stirring and uniformly mixing in a dark place to obtain a suspension, then centrifuging the suspension to obtain a precipitate, and drying the precipitate in vacuum to obtain a product, namely the black phosphorus nanosheet-chitosan; wherein the content of the black phosphorus nanosheets in the suspension is 0.5-1mg/mL, and the content of the chitosan is 5-10 mg/mL;
2) dissolving a photoinitiator and methacrylic acid acylated gelatin into PBS buffer solution, and heating at the water bath temperature of 30-50 ℃ for 10-20min to obtain hydrogel precursor solution, wherein the concentration of the photoinitiator in the hydrogel precursor solution is 1.25-2.5mg/mL, and the concentration of the methacrylic acid acylated gelatin is 40-60 mg/mL;
3) Adding the black phosphorus nanosheet-chitosan and the deferoxamine into the hydrogel precursor solution, and heating at the water bath temperature of 50-70 ℃ for 25-40 min; the concentration of the black phosphorus nanosheet-chitosan in the hydrogel precursor solution is 75-125 mug/mL, and the concentration of the deferoxamine is 0.75-1.25 mg/mL;
4) irradiating the prepared hydrogel precursor solution prepared in the step 3) by using a long-wave ultraviolet lamp to prepare the BPN-DFO gel scaffold.
2. The method for preparing the BPN-DFO gel scaffold according to claim 1, wherein in the step 1), the black phosphorus nanosheet is prepared by: grinding the black phosphorus crystal powder, transferring the ground black phosphorus crystal powder into a centrifuge tube, fixing the volume with water until the black phosphorus concentration is 1mg/mL, stirring and uniformly mixing to obtain a suspension, centrifuging, removing bottom sediment, taking the supernatant, and drying in vacuum to obtain the black phosphorus nanosheet.
3. The method for preparing the BPN-DFO gel scaffold according to claim 1, wherein, in step 4), the wavelength of the long-wave UV lamp is 65-365nm, the power is 300-400W, and the irradiation time is 5-10 s.
4. The method for preparing a BPN-DFO gel scaffold according to claim 1, wherein in step 1), the chitosan is quaternized chitosan.
5. The method for preparing the BPN-DFO gel scaffold according to claim 1, wherein in step 1), the photoinitiator is a photoinitiator LAP.
6. Use of a BPN-DFO gel scaffold prepared according to any of claims 1-5, in particular for the repair of ischemic bone defects.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210287535.6A CN114569789A (en) | 2022-03-23 | 2022-03-23 | Preparation method and application of BPN-DFO gel scaffold |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210287535.6A CN114569789A (en) | 2022-03-23 | 2022-03-23 | Preparation method and application of BPN-DFO gel scaffold |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114569789A true CN114569789A (en) | 2022-06-03 |
Family
ID=81776060
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210287535.6A Withdrawn CN114569789A (en) | 2022-03-23 | 2022-03-23 | Preparation method and application of BPN-DFO gel scaffold |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114569789A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115607683A (en) * | 2022-10-24 | 2023-01-17 | 浙江大学 | Chitosan-deferoxamine composite nano suspension and preparation method and application thereof |
| CN116059400A (en) * | 2023-01-11 | 2023-05-05 | 苏州大学附属第一医院 | Preparation method and application of hydrogel microsphere for regulating nucleus pulposus oxygen metabolism balance |
| CN117065087A (en) * | 2023-10-16 | 2023-11-17 | 中国人民解放军总医院第四医学中心 | Preparation method and application of black phosphorus chitosan DFO temperature-sensitive hydrogel |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20180133368A1 (en) * | 2016-11-15 | 2018-05-17 | The Board Of Regents Of The University Of Texas System | 3D Printed Ti-6Al-4V Scaffolds with Hydrogel Matrix |
| CN108653809A (en) * | 2018-05-23 | 2018-10-16 | 中山大学 | A kind of composite hydrogel based on black phosphorus and gelatin and its application in terms of bone tissue engineer |
| CN110101903A (en) * | 2019-04-09 | 2019-08-09 | 温州医科大学 | A kind of BG compound rest of inducible hypoxemia and its application |
| CN112980009A (en) * | 2021-03-16 | 2021-06-18 | 华南理工大学 | Nano composite porous gel scaffold and construction method and application thereof |
| CN113274550A (en) * | 2021-05-31 | 2021-08-20 | 福州大学 | Vascularized bone bionic multifunctional tissue engineering scaffold with anti-inflammatory effect and preparation method thereof |
| CN113349988A (en) * | 2021-05-31 | 2021-09-07 | 浙江大学 | Tissue engineering bone for repairing jaw cleft palate defect and preparation method thereof |
-
2022
- 2022-03-23 CN CN202210287535.6A patent/CN114569789A/en not_active Withdrawn
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20180133368A1 (en) * | 2016-11-15 | 2018-05-17 | The Board Of Regents Of The University Of Texas System | 3D Printed Ti-6Al-4V Scaffolds with Hydrogel Matrix |
| CN108653809A (en) * | 2018-05-23 | 2018-10-16 | 中山大学 | A kind of composite hydrogel based on black phosphorus and gelatin and its application in terms of bone tissue engineer |
| CN110101903A (en) * | 2019-04-09 | 2019-08-09 | 温州医科大学 | A kind of BG compound rest of inducible hypoxemia and its application |
| CN112980009A (en) * | 2021-03-16 | 2021-06-18 | 华南理工大学 | Nano composite porous gel scaffold and construction method and application thereof |
| CN113274550A (en) * | 2021-05-31 | 2021-08-20 | 福州大学 | Vascularized bone bionic multifunctional tissue engineering scaffold with anti-inflammatory effect and preparation method thereof |
| CN113349988A (en) * | 2021-05-31 | 2021-09-07 | 浙江大学 | Tissue engineering bone for repairing jaw cleft palate defect and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| DINGLI XU等: "A Composite Deferoxamine/Black Phosphorus Nanosheet/Gelatin Hydrogel Scaffold for Ischemic Tibial Bone Repair", 《INTERNATIONAL JOURNAL OF NANOMEDICINE》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115607683A (en) * | 2022-10-24 | 2023-01-17 | 浙江大学 | Chitosan-deferoxamine composite nano suspension and preparation method and application thereof |
| CN115607683B (en) * | 2022-10-24 | 2023-10-27 | 浙江大学 | Chitosan-deferoxamine composite nano suspension, and preparation method and application thereof |
| CN116059400A (en) * | 2023-01-11 | 2023-05-05 | 苏州大学附属第一医院 | Preparation method and application of hydrogel microsphere for regulating nucleus pulposus oxygen metabolism balance |
| CN116059400B (en) * | 2023-01-11 | 2024-04-16 | 苏州大学附属第一医院 | Preparation method and application of hydrogel microsphere for regulating nucleus pulposus oxygen metabolism balance |
| CN117065087A (en) * | 2023-10-16 | 2023-11-17 | 中国人民解放军总医院第四医学中心 | Preparation method and application of black phosphorus chitosan DFO temperature-sensitive hydrogel |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114569789A (en) | Preparation method and application of BPN-DFO gel scaffold | |
| Li et al. | Alkaline phosphatase enzyme-induced biomineralization of chitosan scaffolds with enhanced osteogenesis for bone tissue engineering | |
| Yang et al. | Degradable photothermal bioactive glass composite hydrogel for the sequential treatment of tumor-related bone defects: From anti-tumor to repairing bone defects | |
| Huang et al. | Biofabrication of natural Au/bacterial cellulose hydrogel for bone tissue regeneration via in-situ fermentation | |
| CN111518755A (en) | Bionic periosteum, periosteum-bone substitute and preparation method | |
| Xiao et al. | Synthesis and characterization of cell-laden double-network hydrogels based on silk fibroin and methacrylated hyaluronic acid | |
| Bushkalova et al. | Alginate-chitosan PEC scaffolds: A useful tool for soft tissues cell therapy | |
| CN113924122A (en) | Tunable degradation in hydrogel microparticles | |
| Li et al. | Fabrication and evaluation of bone morphogenetic protein-2 microspheres coated black phosphorus nanosheets@ polylactic-glycolic acid copolymers scaffold: A multifunctional antibacterial photothermal scaffold for bone regeneration | |
| CN115536866A (en) | Injectable hydrogel for cartilage defect repair and preparation method thereof | |
| Ullah et al. | An osteogenic, antibacterial, and anti-inflammatory nanocomposite hydrogel platform to accelerate bone reconstruction | |
| Huang et al. | Biomimetic hydrogels loaded with nanofibers mediate sustained release of pDNA and promote in situ bone regeneration | |
| Shi et al. | A graphene oxide-loaded processed pyritum composite hydrogel for accelerated bone regeneration via mediation of M2 macrophage polarization | |
| Chen et al. | Biocompatible octacalcium phosphate/sodium alginate/silk fibroin composite scaffolds for bone regeneration | |
| Amirazad et al. | A dual synergistic effect of titanium and curcumin co-embedded on extracellular matrix hydrogels of decellularized bone: Potential application in osteoblastic differentiation of adipose-derived mesenchymal stem cells | |
| CN114713150A (en) | Novel crosslinking method and application of sodium alginate hydrogel containing graphene oxide | |
| CN108003360B (en) | Preparation method of II type collagen hydrogel for inducing stem cell chondrogenic differentiation | |
| Zhou et al. | A Self‐Adaptive Biomimetic Periosteum Employing Nitric Oxide Release for Augmenting Angiogenesis in Bone Defect Regeneration | |
| Huang et al. | Harnessing immunomodulation for efficient bone Regeneration: Bioengineered black phosphorus-incorporated Self-Healing hydrogel | |
| CN115475283A (en) | Tissue engineering bone constructed based on hydrogel material and preparation method and application thereof | |
| Wang et al. | Effect of covalent-binding modes of osteogenic-related peptides with artificial carriers on their biological activities in vivo | |
| CN115887772A (en) | Gelatin/sodium alginate hydrogel-based 3D printing biological ink and application thereof | |
| Tang et al. | A biomimetic platelet-rich plasma-based interpenetrating network printable hydrogel for bone regeneration | |
| CN112156230B (en) | Photo-thermal bioactive hydrogel and preparation method and application thereof | |
| CN107412860A (en) | A kind of preparation method based on hydroxyapatite compound support frame material |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20220603 |
|
| WW01 | Invention patent application withdrawn after publication |