CN105169491B - A kind of method for preparing fungi highly -branched polysaccharide xanthan gum hydrogel scaffold - Google Patents
A kind of method for preparing fungi highly -branched polysaccharide xanthan gum hydrogel scaffold Download PDFInfo
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Abstract
The invention discloses a kind of method for preparing fungi highly -branched polysaccharide xanthan gum hydrogel scaffold, belong to natural macromolecular material technical field.Preparation method use by different quality than Pleurotus tuber-regium highly -branched polysaccharide and xanthans be dissolved in the NaOH aqueous solution, gained Pleurotus tuber-regium highly -branched polysaccharide and xanthan gum solution and the sodium trimetaphosphate aqueous solution after the h of the min of cross-linking reaction 10~48, obtain fungi highly -branched polysaccharide xanthan gum hydrogel scaffold at a temperature of 37 DEG C.The inventive method is easy to operate, raw materials abundance, used crosslinking agent has water-soluble and nontoxic, and obtained timbering material has medicine controlled release, good mechanical properties and have biocompatibility concurrently, the highly -branched polysaccharide xanthan gum hydrogel scaffold can be used for preparing artificial organ support, food or nutriment and drug controlled release carrier.
Description
Technical field
The present invention relates to a kind of method for preparing fungi highly -branched polysaccharide-xanthans hydrogel scaffold.Belong to natural polymer
Sub- field of material technology, this fungi highly -branched polysaccharide-xanthans hydrogel three-dimensional insertion porous support can be widely applied to group
Knit the industry such as engineering material and bio-medical material and medicine controlled release.
Background technology
Macromolecule hydrogel has necessarily elasticity, three-dimensional insertion loose structure, is adapted to analog cell epimatrix, provides suitable
The biophysics signal between three-dimensional microenvironment and cell and extracellular matrix needed for cell growth, maintains cell normal phenotype
With physiological function.Polysaccharide is the large biological molecule being widely present in nature, and it participates in the various vital movements of cell and produced
Various biological function, it has close ties with a variety of physiological functions sustained life.In recent years, fungi polysaccharide is as there is life
One important sources of the polysaccharide of thing activity, have caused increasing concern.Numerous studies show that fungi polysaccharide has
Anticancer, liver protection, immune, anticoagulation, hypoglycemic, antiviral and anti-oxidant etc. abundant bioactivity, in health food and medicine
New resources field is widely applied, thus as a unusual active research field.
Fungi polysaccharide abundance, needs further to strengthen the depth and range of its research and development.Pleurotus tuber-regium is growth
A kind of edible mushroom in subtropical and tropical zones.It has the taste and abundant nutritive value of deliciousness；To some diseases, such as roar
Breathe heavily, smallpox and hypertension etc. have certain drug effect；Development of fetus can be promoted, survival rate is improved.In recent years find to carry from Pleurotus tuber-regium
The many carbohydrates and their derivatives taken have antitumor activity, and immunoregulation effect is played to body.The polysaccharide extracted from Pleurotus tuber-regium has
Highly -branched structure, the highly -branched polysaccharide chain conformation spherical in shape, its outer is with can largely carry out the hydroxyl of functionalization, and they can occur
Reaction kinetic interacts with its ambient substance.From the point of view of material, these are spherical, residual with a large amount of sugar
The polysaccharide molecule of base is conducive to occurring many bonding actions and group Shu Xiaoying with other polysaccharide or protein.However, Pleurotus tuber-regium high branch
Change polysaccharide solution viscosity relatively low, be not easy to form hydrogel.Polysaccharide can pass through with other polysaccharide or other biological macromolecular
Hydrogen bond or interionic electrostatic attraction are physical crosslinking, or by being directly chemically crosslinked, and chemical modification post-crosslinking formation water
Gel.
Xanthans is " the pentasaccharides repetition being made up of D- glucans, D-MANNOSE, D-Glucose aldehydic acid, acetic acid and pyruvic acid
The linear water-soluble natural polysaccharide that unit " is formed by connecting.Using xanthans as the hydrogel hydrophily prepared by raw material it is strong, nontoxic,
Degradable, good biocompatibility, is commonly used for super absorbent resin, pharmaceutical carrier and microcapsules etc., has in bio-medical field
It is widely applied prospect.Xanthans large biological molecule is easy to by hydrogen bond formation double-spiral structure, and these double-spiral structures pass through
Intermolecular force, such as electrostatic force, hydrogen bond and inter-chain entanglement further form network-like physical cross-linking hydrogel.However,
The problem of xanthans physical hydrogel is present be：Soluble in water, water resistance and poor mechanical property are easily damaged.Xanthans side chain
On glucuronic acid and pyruvic acid group and whole molecular backbone structure in great amount of hydroxy group, be conducive to the chemistry of xanthans to repair
Adorn and modified.Sodium trimetaphosphate is a kind of food additives, with water-soluble and nontoxic, in physiological temp (37 DEG C) and alkalescent
Under the conditions of, it can be used as esterifying reagent and form water insoluble and mechanical property significantly with the generation chemical crosslink reaction of the hydroxyl on polysaccharide
The hydrogel of raising.Xanthan hydrogel rigidity matter is crisp, frangible, can be by introducing another polymerization in xanthan molecules network
Thing formation interpenetrating networks, dissolve each other and cooperative effect by forcing, so as to further improve the mechanical property of xanthan hydrogel.
(the Carbohydrate Polymers, 2010,79 such as Alireza Shalviri:898-907) using sodium trimetaphosphate as crosslinking
Agent, prepares starch/xanthans intercrossed network type hydrogel, its good film-forming property and gel mould can be entered according to medicine institute is electrically charged
Row selectively penetrating, can be used as multi-medicament carrier.Analyzed based on more than, with reference to the excellent of Pleurotus tuber-regium highly -branched polysaccharide and xanthans
Point, the preparation for the complex polysaccharide bio-medical hydrogel scaffold material being crosslinked through sodium trimetaphosphate has great advantage.Should
Hydrogel can not only provide the microenvironment for maintaining cell growth and have specific bioactivity, so as to be expected to be applied to biological doctor
Learn and field of tissue engineering technology.Fungi highly -branched polysaccharide-xanthan hydrogel not only depends on as a kind of good timbering material
In its biocompatibility and biodegradable, it is often more important that the characteristic such as its unique chemical moieties and bioactivity, because
This is better than synthesis macromolecule hydrogel or other polysaccharide hydrogels.Fungi highly -branched polysaccharide-xanthan hydrogel organizational project,
The fields such as drug controlled release, food and nutrient carriers have broad application prospects.
Just because of natural polysaccharide hydrogel has great application value, therefore it is prepared and application and development turns into current state
One of inside and outside study hotspot.Preferable man-made support material is developed instead of organ transfer operation repair tissue defect or lesion, is
One of biomaterial science and the important topic of medical domain.Current polysaccharide hydrogel prepares main poly- using sodium alginate, shell
Sugar etc. makees raw material.For example：China Patent Publication No. is CN103087334A, and publication date is on May 8th, 2013, entitled
The application case of " preparation method of composite hydrogel of sodium alginate and artemisia desertorum seed gum ".This application case is disclosed by molten in sodium alginate
Artemisia Glue is introduced in liquid system, Ca is slowly released in glucolactone solution using micropore calcium carbonate2+It is formed in situ sea
Alginic acid-Artemisia Glue composite aquogel so that sodium alginate and the performance complement of Artemisia Glue, so as to strengthen sodium alginate based aquagel
Water absorbing properties and mechanical performance.The shortcoming of this method is：The crosslinking agent used is Ca2+, dependence is electrostatic force
Cross-linked network is formed, what is obtained is physical hydrogel.Therefore during drug controlled release, the crosslinking agent Ca in hydrogel2+
It is easy to be replaced by other ions in diffusion system so that the problems such as hydrogel is dissolved in water.
The content of the invention
The deficiency existed for above-mentioned technology, it is easy it is an object of the invention to provide a kind of technique, pollute small, products obtained therefrom
Water-setting with good mechanical property, the release of medicine controllability, good biocompatibility, biological degradability and bioactivity
Glue support preparation method.
To achieve the above object, the technical scheme that provides of the present invention is：
A kind of method for preparing fungi highly -branched polysaccharide-xanthans hydrogel scaffold, described preparation method includes following
Step：
A crushes dry Sclerotium of Pleurotus tuber regium, carries out surname extraction with ethyl acetate, acetone successively and removes fat, then
The Sclerotium of Pleurotus tuber regium after fat will be gone to be immersed in physiological saline, wherein soaked per 100g Sclerotium of Pleurotus tuber regium with 1L physiological saline,
Extracted at a temperature of 120 DEG C of high pressure, centrifuge to obtain and centrifuged again after extract solution, cooling extract solution and collect residue；Residue deionization
Water eccentric cleaning and freeze-drying obtain Pleurotus tuber-regium highly -branched polysaccharide.
Pleurotus tuber-regium highly -branched polysaccharide is dissolved in the NaOH aqueous solution that pH value is 12~14 by b, is prepared into after stirring 2h dense
Spend the Pleurotus tuber-regium highly -branched polysaccharide solution for 2%w/v.
Xanthans is added in the Pleurotus tuber-regium highly -branched polysaccharide solution obtained through b step by c, is stirred 12h, is obtained well mixed
Pleurotus tuber-regium highly -branched polysaccharide-xanthan gum solution, the concentration of xanthans is 0.5%~5%w/v, Pleurotus tuber-regium highly -branched polysaccharide with
The mass ratio of xanthans is 1:0.25~1:2.5.
D dissolves sodium trimetaphosphate in deionized water, stirs to being completely dissolved, and it is 75~262.5mg/mL's to obtain concentration
The sodium trimetaphosphate aqueous solution.
The concentration obtained through Step d is added to through step c and obtained by e for the 75~262.5mg/mL sodium trimetaphosphate aqueous solution
To Pleurotus tuber-regium highly -branched polysaccharide-xanthan gum solution in, wherein Pleurotus tuber-regium highly -branched polysaccharide-xanthan gum solution and sodium trimetaphosphate
The volume ratio of the aqueous solution is 25:8,5min is quickly stirred, cross-linking reaction 10min~48h, obtains fungi highly -branched at a temperature of 37 DEG C
Polysaccharide-xanthans hydrogel scaffold.
As a result of above technical scheme, technical scheme is for Pleurotus tuber-regium highly -branched polysaccharide and xanthans
Design feature, medicine controlled release and good mechanical properties are prepared using water-soluble, nontoxic sodium trimetaphosphate esterification and crosslinking
Highly -branched polysaccharide-xanthans hydrogel scaffold, medicine or Porcine HGF are embedded in the hydrogel scaffold, utilize high branch
Change the content ratio of polysaccharide and xanthans to adjust the mechanical property of hydrogel and the pore size of loose structure.Fungi highly -branched
The bioactivity of polysaccharide is conducive to the adhesion and growth of cell.Hydroxyl, the xanthan glycocoll ring of highly -branched polysaccharide periphery in hydrogel
On hydroxyl and carboxyl, and the sodium metaphosphate group that introduces of crosslinking agent have to medicine or Porcine HGF it is certain mutual
Effect, is discharged with reference to the aperture of hydrogel and the electric charge controllable medicine or Porcine HGF of institute's band with different speed, from
And simulate control release behavior and controllable induced cell proliferation differentiation of the growth factor in extracellular matrix.In addition, xanthans
Exist with the rigid aggregation conformation of double helix chain formation, highly -branched polysaccharide is evenly distributed on these double helix chain ordered arrangements
At the hole of aggregation, esterification and crosslinking formation highly -branched polysaccharide and xanthans water-setting occur along around coiled strand for sodium trimetaphosphate
Glue support, the presence of highly -branched polysaccharide and cross-linking reaction do not make significant difference to the double-spiral structure of xanthans, are crosslinked to hydrogel
Intensity have good enhancing effect.
The present invention prepares the method for highly -branched polysaccharide-xanthans hydrogel scaffold compared with the prior art with following excellent
Point：
Preparation method of the present invention have it is simple to operate, it is with low cost, raw material sources enrich, one of used raw material tiger milk
Mushroom highly -branched polysaccharide has bioactivity, can prepare hydrogel in original position in the case where being adapted to the physiological condition of cell growth, thus prepare
Highly -branched polysaccharide-xanthans hydrogel scaffold that method is obtained, can control medicine and slowly discharges, improve medicine during as pharmaceutical carrier
Effect；As can be loaded during tissue engineering bracket material Porcine HGF and analog cell epimatrix control Porcine HGF delay
The function of On The Drug Release, so that induced cell proliferation is divided into regenerating tissues, and the presence of fungi highly -branched polysaccharide, can assign water
The bioactivity of gel stent.Experiment shows that highly -branched polysaccharide-xanthans hydrogel scaffold obtained by this method is passed through with three-dimensional
Logical loose structure, Model Molecule bovine serum albumin embedding amount in the hydrogel is big, and with good controlled release behavior.
Therefore, this method can be widely applied to prepare artificial organ timbering material, and in the field such as drug controlled release and food
Have broad application prospects.It is worth noting that：Pleurotus tuber-regium highly -branched polysaccharide-xanthan hydrogel is not only in structure and thin
Extracellular matrix is similar, it is often more important that, it wherein contains fungi highly -branched polysaccharide, with specific bioactivity, and conduct
A kind of highly -branched polysaccharide, its structure is easy to flexible modulation physical property or carries various chemical signal molecules, so as to optimize cell
Survive and the specific differentiation behavior of inducing cell.Therefore, Porcine HGF is embedded into Pleurotus tuber-regium highly -branched polysaccharide-xanthans
Regulating and controlling its release behavior in hydrogel and by highly -branched polysaccharide unique chemical moieties can more preferable its releasing in vivo of simulation
Put and induce being formed uniformly for cambium.In addition, in hydrogel scaffold, xanthans maintains rigid double helix aggregation
The Ca added in carboxyl energy inducing cell nutrient solution in structure, and sugared ring2+And PO4 3-Absorption deposition so that support ore deposit
Change, be conducive to adhesion and propagation of osteocyte etc..
Brief description of the drawings
Fig. 1 is the scanning electron microscopic picture of highly -branched polysaccharide-xanthans hydrogel scaffold of embodiment 6.
Fig. 2 is bent to bovine serum albumin control release for highly -branched polysaccharide-xanthans hydrogel scaffold of embodiment 3,4,5,6
Line.
Embodiment
Technical scheme is described further below in conjunction with specific embodiment.
A kind of method for preparing fungi highly -branched polysaccharide-xanthans hydrogel scaffold, described preparation method includes following
Step：
A crushes dry Sclerotium of Pleurotus tuber regium, carries out surname extraction 6h with ethyl acetate, acetone successively and removes fat, institute
It is chemically pure reagent with ethyl acetate and acetone.Then the life that the Sclerotium of Pleurotus tuber regium after fat will be gone to be immersed at a temperature of 80 DEG C
Manage 2h in salt solution, centrifugation；Residue soaks 30min at a temperature of 120 DEG C of high pressure, wherein per 100g Sclerotium of Pleurotus tuber regium 1L physiology salts
Water is soaked, and 20min is centrifuged under 8000 turns of rotating speed and obtains extract solution, is cooled down and is centrifuged again after extract solution and collect residue；Residue is used
Deionized water eccentric cleaning and freeze-drying obtain Pleurotus tuber-regium highly -branched polysaccharide, also can be dried to obtain Pleurotus tuber-regium using other methods
Highly -branched polysaccharide.
B at room temperature, Pleurotus tuber-regium highly -branched polysaccharide is dissolved in the NaOH aqueous solution that pH value is 12~14, magnetic agitation 2h
The Pleurotus tuber-regium highly -branched polysaccharide solution that concentration is 2%w/v is prepared into afterwards, and NaOH concentration is 0.01~1mol/L, 2%w/ herein
V representation quality volumetric concentrations, represent to dissolve 2 grams of solutes in 100 grams of solvents.
At room temperature, xanthans is added in the Pleurotus tuber-regium highly -branched polysaccharide solution obtained through b step by c, is continued magnetic force and is stirred
12h is mixed, well mixed Pleurotus tuber-regium highly -branched polysaccharide-xanthan gum solution is obtained, the concentration of xanthans is 0.5%~5%w/v, tiger
The mass ratio of milk mushroom highly -branched polysaccharide and xanthans is 1:0.25~1:2.5.
D at room temperature, by sodium trimetaphosphate dissolving in deionized water, is stirred to being completely dissolved, can also shaken up, obtaining concentration is
75~262.5mg/mL sodium trimetaphosphate the aqueous solution.
At room temperature, the concentration obtained through Step d is added to warp for the 75~262.5mg/mL sodium trimetaphosphate aqueous solution by e
In Pleurotus tuber-regium highly -branched polysaccharide-xanthan gum solution that step c is obtained, wherein Pleurotus tuber-regium highly -branched polysaccharide-xanthan gum solution and three
The volume ratio of the sodium metaphosphate aqueous solution is 25:8, the mol ratio of the repetition sugar unit of sodium trimetaphosphate and xanthans for 1.46~
14.6,5min is quickly stirred, magnetic agitation can be used herein, mechanical agitation, the cross-linking reaction at a temperature of 37 DEG C can be also used
10min~48h, obtains fungi highly -branched polysaccharide-xanthans hydrogel scaffold, and the rheological behaviour of hydrogel is studied using rheometer,
The three-dimensional insertion porous support of highly -branched polysaccharide-xanthans is cleaned and be freeze-dried and to obtain with deionized water, utilizes scanning electron microscopic observation
The pattern of after-poppet is freeze-dried, is tested in phosphate buffered saline solution and dries the swelling ratio of after-poppet material and to cow's serum egg
White release behavior.
Embodiment 1
The Sclerotium of Pleurotus tuber regium that 500g is dried is crushed, and is carried out surname extraction 6h with ethyl acetate, acetone successively and is removed fat,
Then by go the Sclerotium of Pleurotus tuber regium after fat be immersed in 80 DEG C of 5L at a temperature of physiological saline in 2h, centrifugation, residue is in high pressure
30min is soaked in 5L physiological saline at a temperature of 120 DEG C, is centrifuged to obtain and is centrifuged again after extract solution, cooling extract solution and collect residue, it is residual
Slag deionized water eccentric cleaning and freeze-drying obtain Pleurotus tuber-regium highly -branched polysaccharide.By the dissolving of 0.5g Pleurotus tuber-regium highly -brancheds polysaccharide
In the 25mL pH=12 NaOH aqueous solution, the Pleurotus tuber-regium highly -branched polysaccharide that concentration is 2%w/v is prepared into after magnetic agitation 2h
Solution, 0.125g xanthans is added in the Pleurotus tuber-regium highly -branched polysaccharide solution that above-mentioned concentration is 2%w/v, continues magnetic force
12h is stirred, well mixed Pleurotus tuber-regium highly -branched polysaccharide-xanthan gum solution is obtained, 8mL 75mg/mL sodium trimetaphosphate is added
In above-mentioned highly -branched polysaccharide-xanthan gum solution, quick stirring 5min obtains pre-gel solution, by pre-gel solution at a temperature of 37 DEG C
After cross-linking reaction 10min, fungi highly -branched polysaccharide-xanthans hydrogel scaffold is obtained, the stream of hydrogel scaffold is studied with rheometer
Change behavior, cleans and is freeze-dried with deionized water and to obtain the three-dimensional insertion porous support of highly -branched polysaccharide-xanthans, use ESEM
The pattern of observation freeze-drying after-poppet, tests in phosphate buffered saline solution and dries the swelling ratio of after-poppet material and to ox blood
Albuminised release behavior.
Embodiment 2
The Sclerotium of Pleurotus tuber regium that 500g is dried is crushed, and is carried out surname extraction 6h with ethyl acetate, acetone successively and is removed fat,
Then by go the Sclerotium of Pleurotus tuber regium after fat be immersed in 80 DEG C of 5L at a temperature of physiological saline in 2h, centrifugation, residue is in high pressure
30min is soaked in 5L physiological saline at a temperature of 120 DEG C, is centrifuged to obtain and is centrifuged again after extract solution, cooling extract solution and collect residue, it is residual
Slag deionized water eccentric cleaning and freeze-drying obtain highly -branched Pleurotus tuber-regium polysaccharide.By the dissolving of 0.5g Pleurotus tuber-regium highly -brancheds polysaccharide
In the 25mL pH=13 NaOH aqueous solution, the Pleurotus tuber-regium highly -branched polysaccharide that concentration is 2%w/v is prepared into after magnetic agitation 2h
Solution, 0.25g xanthans is added in the Pleurotus tuber-regium highly -branched polysaccharide solution that above-mentioned concentration is 2%w/v, is continued magnetic force and is stirred
12h is mixed, well mixed highly -branched polysaccharide-xanthan gum solution is obtained, 8mL 112.5mg/mL sodium trimetaphosphate is added above-mentioned
In highly -branched polysaccharide-xanthan gum solution, quick stirring 5min obtains pre-gel solution, and pre-gel solution is crosslinked at a temperature of 37 DEG C
React after 1h, obtain fungi highly -branched polysaccharide-xanthans hydrogel scaffold, the rheological behaviour of hydrogel scaffold is studied with rheometer,
The three-dimensional insertion porous support of highly -branched polysaccharide-xanthans is cleaned and is freeze-dried and to obtain with deionized water, it is cold with scanning electron microscopic observation
The pattern of dry after-poppet is freezed, is tested in phosphate buffered saline solution and dries the swelling ratio of after-poppet material and to bovine serum albumin
Release behavior.
Embodiment 3
The Sclerotium of Pleurotus tuber regium that 500g is dried is crushed, and is carried out surname extraction 6h with ethyl acetate, acetone successively and is removed fat,
Then by go the Sclerotium of Pleurotus tuber regium after fat be immersed in 80 DEG C of 5L at a temperature of physiological saline in 2h, centrifugation, residue is in high pressure
30min is soaked in 5L physiological saline at a temperature of 120 DEG C, is centrifuged to obtain and is centrifuged again after extract solution, cooling extract solution and collect residue, it is residual
Slag deionized water eccentric cleaning and freeze-drying obtain highly -branched Pleurotus tuber-regium polysaccharide.By the dissolving of 0.5g Pleurotus tuber-regium highly -brancheds polysaccharide
In the 25mL pH=14 NaOH aqueous solution, the Pleurotus tuber-regium highly -branched polysaccharide that concentration is 2%w/v is prepared into after magnetic agitation 2h
Solution, 0.5g xanthans is added in the Pleurotus tuber-regium highly -branched polysaccharide solution that above-mentioned concentration is 2%w/v, is continued magnetic force and is stirred
12h is mixed, well mixed highly -branched polysaccharide-xanthan gum solution is obtained, 8mL 150mg/mL sodium trimetaphosphate is added into above-mentioned height
In branched polysaccharide-xanthan gum solution, quick stirring 5min obtains pre-gel solution, and pre-gel solution is crosslinked instead at a temperature of 37 DEG C
Answer after 10min, obtain fungi highly -branched polysaccharide-xanthans hydrogel scaffold, the rheological behaviour of hydrogel scaffold is studied with rheometer,
The three-dimensional insertion porous support of highly -branched polysaccharide-xanthans is cleaned and is freeze-dried and to obtain with deionized water, it is cold with scanning electron microscopic observation
The pattern of dry after-poppet is freezed, is tested in phosphate buffered saline solution and dries the swelling ratio of after-poppet material and to bovine serum albumin
Release behavior.
Embodiment 4
The Sclerotium of Pleurotus tuber regium that 500g is dried is crushed, and is carried out surname extraction 6h with ethyl acetate, acetone successively and is removed fat,
Then by go the Sclerotium of Pleurotus tuber regium after fat be immersed in 80 DEG C of 5L at a temperature of physiological saline in 2h, centrifugation, residue is in high pressure
30min is soaked in 5L physiological saline at a temperature of 120 DEG C, is centrifuged to obtain and is centrifuged again after extract solution, cooling extract solution and collect residue, it is residual
Slag deionized water eccentric cleaning and freeze-drying obtain highly -branched Pleurotus tuber-regium polysaccharide.By the dissolving of 0.5g Pleurotus tuber-regium highly -brancheds polysaccharide
In the 25mL pH=13 NaOH aqueous solution, the Pleurotus tuber-regium highly -branched polysaccharide that concentration is 2%w/v is prepared into after magnetic agitation 2h
Solution, 0.75g xanthans is added in the Pleurotus tuber-regium highly -branched polysaccharide solution that above-mentioned concentration is 2%w/v, is continued magnetic force and is stirred
12h is mixed, well mixed highly -branched polysaccharide-xanthan gum solution is obtained, 8mL 187.5mg/mL sodium trimetaphosphate is added above-mentioned
In highly -branched polysaccharide-xanthan gum solution, quick stirring 5min obtains pre-gel solution, and pre-gel solution is crosslinked at a temperature of 37 DEG C
React after 24h, obtain fungi highly -branched polysaccharide-xanthans hydrogel scaffold, the rheological behaviour of hydrogel scaffold is studied with rheometer,
The three-dimensional insertion porous support of highly -branched polysaccharide-xanthans is cleaned and is freeze-dried and to obtain with deionized water, it is cold with scanning electron microscopic observation
The pattern of dry after-poppet is freezed, is tested in phosphate buffered saline solution and dries the swelling ratio of after-poppet material and to bovine serum albumin
Release behavior.
Embodiment 5
The Sclerotium of Pleurotus tuber regium that 500g is dried is crushed, and is carried out surname extraction 6h with ethyl acetate, acetone successively and is removed fat,
Then by go the Sclerotium of Pleurotus tuber regium after fat be immersed in 80 DEG C of 5L at a temperature of physiological saline in 2h, centrifugation, residue is in high pressure
30min is soaked in 5L physiological saline at a temperature of 120 DEG C, is centrifuged to obtain and is centrifuged again after extract solution, cooling extract solution and collect residue, it is residual
Slag deionized water eccentric cleaning and freeze-drying obtain highly -branched Pleurotus tuber-regium polysaccharide.By the dissolving of 0.5g Pleurotus tuber-regium highly -brancheds polysaccharide
In the 25mL pH=13 NaOH aqueous solution, the Pleurotus tuber-regium highly -branched polysaccharide that concentration is 2%w/v is prepared into after magnetic agitation 2h
Solution, 1.0g xanthans is added in the Pleurotus tuber-regium highly -branched polysaccharide solution that above-mentioned concentration is 2%w/v, is continued magnetic force and is stirred
12h is mixed, well mixed highly -branched polysaccharide-xanthan gum solution is obtained, 8mL 225mg/mL sodium trimetaphosphate is added into above-mentioned height
In branched polysaccharide-xanthan gum solution, quick stirring 5min obtains pre-gel solution, and pre-gel solution is crosslinked instead at a temperature of 37 DEG C
Answer after 48h, obtain fungi highly -branched polysaccharide-xanthans hydrogel scaffold, the rheological behaviour of hydrogel scaffold is studied with rheometer, use
The three-dimensional insertion porous support of highly -branched polysaccharide-xanthans is cleaned and be freeze-dried and to be obtained to deionized water, is freezed with scanning electron microscopic observation
The pattern of after-poppet is dried, is tested in phosphate buffered saline solution and dries the swelling ratio of after-poppet material and to bovine serum albumin
Release behavior.
Embodiment 6
The Sclerotium of Pleurotus tuber regium that 500g is dried is crushed, and is carried out surname extraction 6h with ethyl acetate, acetone successively and is removed fat,
Then by go the Sclerotium of Pleurotus tuber regium after fat be immersed in 80 DEG C of 5L at a temperature of physiological saline in 2h, centrifugation, residue is in high pressure
30min is soaked in 5L physiological saline at a temperature of 120 DEG C, is centrifuged to obtain and is centrifuged again after extract solution, cooling extract solution and collect residue, it is residual
Slag deionized water eccentric cleaning and freeze-drying obtain highly -branched Pleurotus tuber-regium polysaccharide.By the dissolving of 0.5g Pleurotus tuber-regium highly -brancheds polysaccharide
In the 25mL pH=13 NaOH aqueous solution, the Pleurotus tuber-regium highly -branched polysaccharide that concentration is 2%w/v is prepared into after magnetic agitation 2h
Solution, 1.25g xanthans is added in the Pleurotus tuber-regium highly -branched polysaccharide solution that above-mentioned concentration is 2%w/v, is continued magnetic force and is stirred
12h is mixed, well mixed highly -branched polysaccharide-xanthan gum solution is obtained, 8mL 262.5mg/mL sodium trimetaphosphate is added above-mentioned
In highly -branched polysaccharide-xanthan gum solution, quick stirring 5min obtains pre-gel solution, and pre-gel solution is crosslinked at a temperature of 37 DEG C
React after 3h, obtain fungi highly -branched polysaccharide-xanthans hydrogel scaffold, the rheological behaviour of hydrogel scaffold is studied with rheometer,
The three-dimensional insertion porous support of highly -branched polysaccharide-xanthans is cleaned and is freeze-dried and to obtain with deionized water, it is cold with scanning electron microscopic observation
The pattern of dry after-poppet is freezed, is tested in phosphate buffered saline solution and dries the swelling ratio of after-poppet material and to bovine serum albumin
Release behavior.(this embodiment is most preferred embodiment)
Embodiment 7
The Sclerotium of Pleurotus tuber regium that 500g is dried is crushed, and is carried out surname extraction 6h with ethyl acetate, acetone successively and is removed fat,
Then by go the Sclerotium of Pleurotus tuber regium after fat be immersed in 80 DEG C of 5L at a temperature of physiological saline in 2h, centrifugation, residue is in high pressure
30min is soaked in 5L physiological saline at a temperature of 120 DEG C, is centrifuged to obtain and is centrifuged again after extract solution, cooling extract solution and collect residue, it is residual
Slag deionized water eccentric cleaning and freeze-drying obtain highly -branched Pleurotus tuber-regium polysaccharide.By the dissolving of 0.5g Pleurotus tuber-regium highly -brancheds polysaccharide
In the 25mL pH=13 NaOH aqueous solution, the Pleurotus tuber-regium highly -branched polysaccharide that concentration is 2%w/v is prepared into after magnetic agitation 2h
Solution, 1.25g xanthans is added in the Pleurotus tuber-regium highly -branched polysaccharide solution that above-mentioned concentration is 2%w/v, is continued magnetic force and is stirred
12h is mixed, well mixed highly -branched polysaccharide-xanthan gum solution is obtained, 8mL 75mg/mL sodium trimetaphosphate is added into above-mentioned high branch
Change in polysaccharide-xanthan gum solution, quick stirring 5min obtains pre-gel solution, by pre-gel solution at a temperature of 37 DEG C cross-linking reaction
After 3h, fungi highly -branched polysaccharide-xanthans hydrogel scaffold is obtained, the rheological behaviour of hydrogel scaffold is studied with rheometer, is spent
Ionized water cleans and is freeze-dried and to obtain the three-dimensional insertion porous support of highly -branched polysaccharide-xanthans, is freezed with scanning electron microscopic observation dry
The pattern of dry after-poppet, tests in phosphate buffered saline solution and dries the swelling ratio of after-poppet material and bovine serum albumin is released
Let pass and be.
The performance of fungi highly -branched polysaccharide-xanthans hydrogel scaffold of embodiment 3~7 is shown in Table one
Table one
Claims (1)
1. a kind of method for preparing fungi highly -branched polysaccharide-xanthans hydrogel scaffold, it is characterised in that：Described preparation method
Comprise the following steps：
A crushes dry Sclerotium of Pleurotus tuber regium, carries out surname extraction with ethyl acetate, acetone successively and removes fat, then will go
Sclerotium of Pleurotus tuber regium after fat is immersed in physiological saline, wherein being soaked per 100g Sclerotium of Pleurotus tuber regium with 1L physiological saline, in height
Extracted at a temperature of 120 DEG C of pressure, centrifuge to obtain extract solution, centrifuged again after cooling extract solution and collect residue, residue deionized water from
The heart is cleaned and freeze-drying obtains Pleurotus tuber-regium highly -branched polysaccharide；
Pleurotus tuber-regium highly -branched polysaccharide is dissolved in the NaOH aqueous solution that pH value is 12~14 by b, and being prepared into concentration after stirring 2h is
2%w/v Pleurotus tuber-regium highly -branched polysaccharide solution；
Xanthans is added in the Pleurotus tuber-regium highly -branched polysaccharide solution obtained through b step by c, is stirred 12h, is obtained well mixed tiger
Milk mushroom highly -branched polysaccharide-xanthan gum solution, the concentration of xanthans is 0.5%~5%w/v, Pleurotus tuber-regium highly -branched polysaccharide and xanthan
The mass ratio of glue is 1:0.25~1:2.5；
D dissolves sodium trimetaphosphate in deionized water, stirs to being completely dissolved, and it is the three inclined of 75~262.5mg/mL to obtain concentration
Sodium phosphate aqueous solution；
E adds the concentration obtained through Step d to being obtained through step c for the 75~262.5mg/mL sodium trimetaphosphate aqueous solution
In Pleurotus tuber-regium highly -branched polysaccharide-xanthan gum solution, wherein Pleurotus tuber-regium highly -branched polysaccharide-xanthan gum solution and sodium trimetaphosphate are water-soluble
The volume ratio of liquid is 25:8,5min is quickly stirred, cross-linking reaction 10min~48h, obtains fungi highly -branched many at a temperature of 37 DEG C
Sugar-xanthans hydrogel scaffold.
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| CN102895258A (en) * | 2011-07-25 | 2013-01-30 | 香港理工大学 | Pleurotus tuber-regium polysaccharide functionalized nanometer selenium hydrosol having anti-tumor activity and preparation method thereof |
| CN104548200A (en) * | 2015-02-06 | 2015-04-29 | 武汉纺织大学 | Method of preparing highly branched polysaccharide-fibroin hydrogel bracket |
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| CN102895258A (en) * | 2011-07-25 | 2013-01-30 | 香港理工大学 | Pleurotus tuber-regium polysaccharide functionalized nanometer selenium hydrosol having anti-tumor activity and preparation method thereof |
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