CN102397586A - Preparation method of bone repair hydrogel and fiber pipe composite material loading growth factors - Google Patents
Preparation method of bone repair hydrogel and fiber pipe composite material loading growth factors Download PDFInfo
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- CN102397586A CN102397586A CN2011103448938A CN201110344893A CN102397586A CN 102397586 A CN102397586 A CN 102397586A CN 2011103448938 A CN2011103448938 A CN 2011103448938A CN 201110344893 A CN201110344893 A CN 201110344893A CN 102397586 A CN102397586 A CN 102397586A
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Abstract
The invention discloses a preparation method of a bone repair hydrogel and fiber pipe composite material loading growth factors. Firstly, a spinning solution is prepared, the spinning solution is arranged in a syringe, a nanometer fiber reticular layer is prepared by high-voltage electrospinning, sodium alginate modified by RGD (Arg-Gly-Asp) is irradiated with gamma rays, and after irradiation, the RGD-sodium alginate is filtered to be degermed and is stored at the room temperature of -20 DEG C so as to prepare hydrogel. According to the method disclosed by the invention, the sodium alginate degraded by gamma ray irradiation can be combined with adhesive peptide in covalence to promote cell attachment; a nanometer fiber pipe is prepared through electrostatic spinning, the fiber has a diameter being the same as that of an extracellular matrix (ECM) and a large specific surface area, can improve cell attachment and migration, and is suitable for osteoblast differentiation and stem cell in vitro regeneration; and the composite material consisting of the bone repair hydrogel and the fiber pipe can guide bone regeneration by hydrogel injected with the sodium alginate and the protein electrostatic spinning nanometer fiber reticular film, promotes the regeneration and repair functions of vessels, can be used as a bone repair transmission system, has a simple production process and is easy to operate.
Description
Technical field:
The present invention relates to a kind of Biocomposite material, relate in particular to a kind of growth factor-loaded bone repair hydrogel and the method for preparing of fiber pipe composite.
Background technology:
From body and allogenic bone transplantation is the treatment fracture and the usual manner of bone defective on a large scale, yet this technology has many drawbacks, limited comprising available bone grafting material, to transplant fault rate high or the like.Through organizational project/regenerative medicine (TE/RM); Researched and developed some potential bone transplantation substitute article,, used three-dimensional rack to load in the support in conjunction with biological biochemistry and physics scaffolding structure; Yet; These scaffolding structures are difficult to design and make the high porosity product, and they can't provide the environment an of the best for damaged cell usually, thereby have hindered the recovery of damaged tissues function.
The electrostatic spinning nano fibrous membrane has multinomial function and is fit to tissue regeneration, and their fibre diameter is big or small identical with extracellular matrix (ECM); Scale composition (fibre diameter is from the nanometer to the submicron) and high surface area; Can improve cell attachment, migration and functionalization, nanofiber grid; Be proved to be and can have supported osteoblast differentiation and stem cell, and tested in the defect of skull model in vivo at external regeneration.
Though frame provides the template of a guiding osteanagenesis, in order to promote the skeleton quickly-healing, also somatomedin need be provided usually; Picture rhBMP-2 can successful induced osteogenesis by clinical proof, promotes knitting, but needs heavy dose; So need to improve delivery system, slow release is provided, lower dosage; Thereby farthest reduce complication, reduce the treatment cost; The sodium alginate hydrogel; The fucoidin derivant; Be a kind of protein that is widely used as frame material and vehicle; But mammalian cell lacks the receptor of sodium alginate polymer, promotes the material of cell and tissue regeneration so we are badly in need of preparing a kind of suitable TE/RM, thereby develops and produce the growth factor-loaded transmission system that bone is repaired that is used for.
Summary of the invention:
The present invention is directed to the deficiency of prior art, a kind of growth factor-loaded bone repair hydrogel and the method for preparing of fiber pipe composite are provided.
Growth factor-loaded bone repair hydrogel and fiber pipe composite material and preparation method thereof, its step is following:
(1) preparation spinning solution; Water-soluble or the organic solvent with macromolecular material is mixed with the macromolecular solution that mass percentage concentration is 10-30%.
(2) the netted pipe of preparation nanofiber; Spinning solution is installed in the 1-5mL syringe, and connects the rustless steel syringe needle at tail end, and syringe links to each other with syringe pump; Its flow velocity be the 0.5-1 milliliter/hour, the fiber that obtains is collected with a plane copper foil plate apart from needle point 20-23cm, spins 3-6h with the high pressure of 13-20kv incoming call; With the netted layer of the thicker nanofiber that obtains, the web-like layer that obtains evaporates in drying baker and spends the night to remove residual solvent, and the nanometer fiber net that makes above using is made implantable tubular material; Cut out the rectangle sample of a 13-19mm from network, in some samples, use diameter to beat the hole of some spacings as 1.5-2.5mm as the pin of 0.5-1mm; It is the pipe of a diameter 5-15 millimeter and length 10-20 millimeter that the rectangular mesh sample uses the steel core axle to twine; Eclipsed part is fixed together with UV glue, and the ethanol flushing twice of the effective 50-80% of nanometer fiber net is invaded bubble then in ethanol; Band can guarantee after the solvent evaporation aseptic, preserves up to implanting.
(3) contain the preparation of the sodium alginate hydrogel of somatomedin; Sodium alginate with radiation gamma R6D modification; Make polymer molecular weight reduce to be suitable for carrying out material in the body, postradiation sodium alginate uses the covalently bound G4RGDASSP peptide sequence of having gone up of carbodiimide chemistry of standard, and its density is that every polymer chain has 1-3 sequence; The RGD-sodium alginate filtration sterilization that obtains; Lyophilizing and be-20 ℃ of storages in room temperature is dissolved in the solution that MEM obtains 1-3%W/V with the RGD-alginate, with the preparation hydrogel; Freeze dried rhBMP-2 is dissolved in and contains in the albuminous 4mM hydrochloric acid of 0.1% rat blood serum; Its final concentration is 100-400mg/mL, and sodium alginate soln and rhBMP-2 solution are with 5: 1-8: 1 mixed, and contain the calcium sulfate slurry 15 of sodium alginate soln and the 0.2-0.4% of rhBMP-2: 1-25: 1 ratio is carried out crosslinked; Sodium alginate soln room temperature in syringe keeps 30 minutes to become gel, transfers to 4 ℃ of preservations then.
The pairing solvent of said macromolecular material is respectively: polycaprolactone (PCL) is dissolved in the mixed liquor of acetone or hexafluoroisopropanol (HFP) and dimethyl formamide (DMF); Polyvinyl acetate (PVAc) is dissolved in dimethyl formamide (DMF); PEO is water-soluble; The mixed liquor of PVA water-soluble or acetone or water and acetone; Polylactic acid (PLA) is dissolved in the mixed liquor of dimethyl formamide (DMF) or dimethyl formamide (DMF) and acetone; Chloroform that polyurethane (PU) is dissolved in and alcoholic acid mixed liquor.
The present invention adopts the sodium alginate of radiation gamma degraded, can promote cell attachment with the adhesin polypeptide covalent bond; Prepare the nanofiber pipe through electrostatic spinning, fibre diameter is big or small identical with extracellular matrix (ECM), and specific surface area is big, can improve cell attachment and migration, is fit to osteoblast differentiation and stem cell at external regeneration; Both form composite for this; Can guide osteanagenesis with injection sodium alginate hydrogel and protein electrostatic spinning nano web-like film, promote the regeneration and the repair function of blood vessel, can do the transmission system that bone is repaired; Production technology is simple, easy operating, can realize extensive industrialization.
The specific embodiment:
In order to deepen that understanding of the present invention is made further detailed description below in conjunction with embodiment to the present invention.
Growth factor-loaded bone repair hydrogel and fiber pipe composite material and preparation method thereof, its step is following:
(1) preparation spinning solution; Polycaprolactone (PCL) is dissolved in the mixed liquor (volume ratio 90: 10) of hexafluoroisopropanol (HFP) and dimethyl formamide (DMF), is mixed with mass percentage concentration and is 12% macromolecular solution.
(2) the netted pipe of preparation nanofiber; Spinning solution is installed in the 3mL syringe, and connects the rustless steel syringe needle at tail end, and syringe links to each other with syringe pump, and its flow velocity is 0.7 milliliter/hour.The fiber that obtains is collected with a plane copper foil plate apart from needle point 22cm, sends a telegram here with the high pressure of 15kv and spins 5h, with the netted layer of the thicker nanofiber that obtains; The web-like layer that obtains evaporates in drying baker and spends the night to remove residual solvent, and the nanometer fiber net that makes above using is made implantable tubular material, cuts out the rectangle sample of a 15mm from network; In some samples; Use diameter to beat the hole of some spacings as 2.5mm as the pin of 1mm, the rectangular mesh sample uses the steel core axle to twine to be the pipe of 10 millimeters of diameters and 15 millimeters of length, and eclipsed part is fixed together with UV glue; Twice of the ethanol flushing of nanometer fiber net effective 70%; Invade bubble then in ethanol, band can guarantee after the solvent evaporation aseptic, preserves up to implanting.
(3) contain the preparation of the sodium alginate hydrogel of somatomedin; Sodium alginate with radiation gamma RGD modification; Make polymer molecular weight reduce to be suitable for carrying out material in the body, postradiation sodium alginate uses the covalently bound G4RGDASSP peptide sequence of having gone up of carbodiimide chemistry of standard, and its density is that every polymer chain has 2 sequences; The RGD-sodium alginate filtration sterilization that obtains; Lyophilizing and be-20 ℃ of storages in room temperature is dissolved in the solution that MEM obtains 2%W/V with the RGD-alginate, with the preparation hydrogel.Freeze dried rhBMP-2 is dissolved in and contains in the albuminous 4mM hydrochloric acid of 0.1% rat blood serum; Its final concentration is 200mg/mL; Sodium alginate soln and rhBMP-2 solution were with 5: 1 mixed; Sodium alginate soln and 0.3% 25: 1 the ratio of calcium sulfate slurry that contains rhBMP-2 carried out crosslinked, and sodium alginate soln room temperature in syringe keeps 30 minutes to become gel, transfers to 4 ℃ of preservations then.
Claims (2)
1. growth factor-loaded bone repair hydrogel and fiber pipe composite material and preparation method thereof is characterized in that: its step is following:
(1) preparation spinning solution; Water-soluble or the organic solvent with macromolecular material is mixed with the macromolecular solution that mass percentage concentration is 10-30%.
(2) the netted pipe of preparation nanofiber; Spinning solution is installed in the 1-5mL syringe, and connects the rustless steel syringe needle at tail end, and syringe links to each other with syringe pump; Its flow velocity be the 0.5-1 milliliter/hour, the fiber that obtains is collected with a plane copper foil plate apart from needle point 20-23cm, spins 3-6h with the high pressure of 13-20kv incoming call; With the netted layer of the thicker nanofiber that obtains, the web-like layer that obtains evaporates in drying baker and spends the night to remove residual solvent, and the nanometer fiber net that makes above using is made implantable tubular material; Cut out the rectangle sample of a 13-19mm from network, in some samples, use diameter to beat the hole of some spacings as 1.5-2.5mm as the pin of 0.5-1mm; It is the pipe of a diameter 5-15 millimeter and length 10-20 millimeter that the rectangular mesh sample uses the steel core axle to twine; Eclipsed part is fixed together with UV glue, and the ethanol flushing twice of the effective 50-80% of nanometer fiber net is invaded bubble then in ethanol; Band can guarantee after the solvent evaporation aseptic, preserves up to implanting.
(3) contain the preparation of the sodium alginate hydrogel of somatomedin; Sodium alginate with radiation gamma RGD modification; Make polymer molecular weight reduce to be suitable for carrying out material in the body, postradiation sodium alginate uses the covalently bound G4RGDASSP peptide sequence of having gone up of carbodiimide chemistry of standard, and its density is that every polymer chain has 1-3 sequence; The RGD-sodium alginate filtration sterilization that obtains; Lyophilizing and be-20 ℃ of storages in room temperature is dissolved in the solution that MEM obtains 1-3%W/V with the RGD-alginate, with the preparation hydrogel; Freeze dried rhBMP-2 is dissolved in and contains in the albuminous 4mM hydrochloric acid of 0.1% rat blood serum; Its final concentration is 100-400mg/mL, and sodium alginate soln and rhBMP-2 solution are with 5: 1-8: 1 mixed, and contain the calcium sulfate slurry 15 of sodium alginate soln and the 0.2-0.4% of rhBMP-2: 1-25: 1 ratio is carried out crosslinked; Sodium alginate soln room temperature in syringe keeps 30 minutes to become gel, transfers to 4 ℃ of preservations then.
2. growth factor-loaded bone repair hydrogel according to claim 1 and fiber pipe composite material and preparation method thereof is characterized in that: the pairing solvent of said macromolecular material is respectively: polycaprolactone is dissolved in the mixed liquor of acetone or hexafluoroisopropanol and dimethyl formamide; Polyvinyl acetate is dissolved in dimethyl formamide; PEO is water-soluble; The mixed liquor of PVA water-soluble or acetone or water and acetone; Polylactic acid is dissolved in the mixed liquor of dimethyl formamide or dimethyl formamide and acetone; Chloroform that polyurethane is dissolved in and alcoholic acid mixed liquor.
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Cited By (6)
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| CN104162186A (en) * | 2014-07-18 | 2014-11-26 | 中国人民解放军***南京总医院 | Biomimetic peritoneum for abdominal cavity open treatment and repairing, and preparation method thereof |
| CN104162186B (en) * | 2014-07-18 | 2017-01-04 | 中国人民解放军***南京总医院 | Bionical peritoneum for abdominal cavity opening treatment and reparation and preparation method thereof |
| CN107236134A (en) * | 2017-04-10 | 2017-10-10 | 魏泓 | Giant salamander secretion hydrogel and its preparation method and application |
| CN109091452A (en) * | 2018-09-17 | 2018-12-28 | 郑州大学 | A kind of partial size based on sodium alginate can be changed the preparation and application of door-control type anti-tumor drug delivery system |
| CN109700576A (en) * | 2019-01-18 | 2019-05-03 | 孟兵 | A technique for skull repairing is carried out using stem cell, natural fiber |
| CN113091776A (en) * | 2021-03-30 | 2021-07-09 | 华中科技大学 | Piezoelectric sensor, preparation method and recycling and degrading method thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104162186A (en) * | 2014-07-18 | 2014-11-26 | 中国人民解放军***南京总医院 | Biomimetic peritoneum for abdominal cavity open treatment and repairing, and preparation method thereof |
| CN104162186B (en) * | 2014-07-18 | 2017-01-04 | 中国人民解放军***南京总医院 | Bionical peritoneum for abdominal cavity opening treatment and reparation and preparation method thereof |
| CN107236134A (en) * | 2017-04-10 | 2017-10-10 | 魏泓 | Giant salamander secretion hydrogel and its preparation method and application |
| CN107236134B (en) * | 2017-04-10 | 2020-01-17 | 魏泓 | Giant salamander secretion hydrogel and preparation method and application thereof |
| CN109091452A (en) * | 2018-09-17 | 2018-12-28 | 郑州大学 | A kind of partial size based on sodium alginate can be changed the preparation and application of door-control type anti-tumor drug delivery system |
| CN109091452B (en) * | 2018-09-17 | 2021-04-30 | 郑州大学 | Preparation and application of particle size-variable gating type antitumor drug delivery system based on sodium alginate |
| CN109700576A (en) * | 2019-01-18 | 2019-05-03 | 孟兵 | A technique for skull repairing is carried out using stem cell, natural fiber |
| CN113091776A (en) * | 2021-03-30 | 2021-07-09 | 华中科技大学 | Piezoelectric sensor, preparation method and recycling and degrading method thereof |
| CN113091776B (en) * | 2021-03-30 | 2022-09-27 | 华中科技大学 | Piezoelectric sensor and preparation method and recycling and degrading method thereof |
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Application publication date: 20120404 |