CN105169491A - Method for preparing fungus hyperbranched polysaccharide-xanthan gum hydrogel bracket - Google Patents
Method for preparing fungus hyperbranched polysaccharide-xanthan gum hydrogel bracket Download PDFInfo
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Abstract
The invention discloses a method for preparing a fungus hyperbranched polysaccharide-xanthan gum hydrogel bracket, and belongs to the technical field of natural macromolecular materials. The preparation method comprises the following steps: dissolving pleurotus tuber-regium hyperbranched polysaccharide and xanthan gum into a NaOH aqueous solution according to different mass ratios, generating crosslinking reaction by an obtained pleurotus tuber-regium hyperbranched polysaccharide and xanthan gum solution and a sodium trimetaphosphate aqueous solution under the temperature of 37 DEG C for 10 minutes to 48 hours, and obtaining the fungus hyperbranched polysaccharide-xanthan gum hydrogel bracket. The method disclosed by the invention is easy and convenient to operate; the adopted raw materials are abundant in source; an adopted crosslinking agent is water-soluble and nontoxic; furthermore, the prepared bracket material has medicament controlled release performance, and is high in mechanical property and biocompatibility. The fungus hyperbranched polysaccharide-xanthan gum hydrogel bracket can be used for preparing an artificial tissue bracket and a food or nutritional substance and medicament controlled release carrier.
Description
Technical field
The present invention relates to a kind of method preparing fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold.Belong to natural macromolecular material technical field, this fungus highly-branched polysaccharide-through porous support of xanthan gum hydrogel three-dimensional can be widely used in tissue engineering material and the industry such as bio-medical material and drug controllable release.
Background technology
Macromolecule hydrogel has certain elasticity, three-dimensional through loose structure, be applicable to analog cell epimatrix, the three-dimensional microenvironment needed for applicable Growth of Cells and the biophysics signal between cell and extracellular matrix are provided, maintain cell normal phenotype and physiological function.Polysaccharide is the biomacromolecule that occurring in nature extensively exists, and it participates in the various vital movement of cell and produces various biological function, and it and all physiological functions sustained life have close ties.In recent years, fungus polysaccharide, as the important sources having bioactive polysaccharide, has caused increasing concern.Large quantity research show fungus polysaccharide have anticancer, protect the liver, biological activity that immunity, anticoagulation, blood sugar lowering, antiviral and antioxidation etc. are abundant; be widely applied at health food and medicine new resources field, thus become a research field enlivened very much.
Fungus polysaccharide abundance, needs to strengthen its depth & wideth of researching and developing further.Pleurotus tuber-regium is a kind of edible fungi of growth in subtropical and tropical zones.It has delicious taste and abundant nutritive value; To some diseases, as asthma, variola and hypertension etc. have certain drug effect; Can fetal development be promoted, improve survival rate.In recent years find that the many carbohydrates and their derivative extracted from Pleurotus tuber-regium have anti-tumor activity, immunoregulation effect is risen to body.The polysaccharide extracted from Pleurotus tuber-regium has highly-branched structure, this highly-branched polysaccharide chain conformation spherical in shape, is with the hydroxyl that can carry out functionalization in a large number outside it, and they generating functionization can react or interact with its ambient substance.From the angle of material, these are spherical, be conducive to many bonding actions occurring with other polysaccharide or protein and rolling into a ball Shu Xiaoying with the polysaccharide molecule of a large amount of saccharide residue.But Pleurotus tuber-regium highly-branched polysaccharide solution viscosity is lower, be not easy to form hydrogel.Polysaccharide can carry out physical crosslinking with other polysaccharide or other biological macromole by hydrogen bond or interionic electrostatic attraction, or by direct chemical crosslinking, and be cross-linked to form hydrogel after chemical modification.
Xanthan gum forms by D-glucosan, D-MANNOSE, D-Glucose aldehydic acid, acetic acid and acetone acid the linear water-soluble natural polysaccharide that " pentasaccharides repetitive " be formed by connecting.Hydrogel hydrophilic with xanthan gum prepared by raw material is strong, nontoxic, degradable, good biocompatibility, is commonly used for super absorbent resin, pharmaceutical carrier and microcapsule etc., is with a wide range of applications in bio-medical field.Xanthan gum biomacromolecule forms double-spiral structure easily through hydrogen bond, and these double-spiral structures pass through intermolecular force, as electrostatic force, hydrogen bond and inter-chain entanglement form network-like physical cross-linking hydrogel further.But xanthan gum physical hydrogel Problems existing is: soluble in water, and resistance to water and poor mechanical property, easily damage.Great amount of hydroxy group in glucuronic acid on xanthan gum side chain and pyruvic acid group and whole molecular backbone structure, is conducive to chemical modification and the modification of xanthan gum.Sodium trimetaphosphate is a kind of food additive, there is water solublity and nontoxic, under physiological temp (37 DEG C) and weak basic condition, the hydroxyl generation chemical crosslink reaction that can be used as on esterifying reagent and polysaccharide forms the hydrogel that water insoluble and mechanical property improves greatly.Xanthan gum hydrogel rigidity matter is crisp, frangible, forming interpenetrating networks, dissolving each other and cooperative effect, thus improve the mechanical property of xanthan gum hydrogel further by forcing by introducing another kind of polymer in xanthan molecules network.(the CarbohydratePolymers such as AlirezaShalviri, 2010,79:898-907) take sodium trimetaphosphate as cross-linking agent, prepare starch/xanthan gum intercrossed network type hydrogel, its good film-forming property and electrically chargedly can carry out selectively penetrating to gel film according to medicine institute, can be used as multi-medicament carrier.Based on above analysis, in conjunction with the advantage of Pleurotus tuber-regium highly-branched polysaccharide and xanthan gum, the preparation of the complex polysaccharide bio-medical hydrogel scaffold material be cross-linked through sodium trimetaphosphate also exists great advantage.This hydrogel not only can provide the microenvironment of maintenance Growth of Cells and have specific biological activity, thus is expected to be applied to biomedicine and field of tissue engineering technology.Fungus highly-branched polysaccharide-xanthan gum hydrogel is as a kind of good timbering material, not only depend on its biocompatibility and biodegradable, the more important thing is the characteristics such as its unique chemical moieties and biological activity, be therefore better than synthesis macromolecule hydrogel or other polysaccharide hydrogel.Fungus highly-branched polysaccharide-xanthan gum hydrogel has broad application prospects in fields such as organizational project, drug controlled release, food and nutrient carriers.
Just because of natural polysaccharide hydrogel exists great using value, therefore the exploitation of its preparations and applicatio becomes one of current home and abroad study hotspot.Developing desirable man-made support material and replace organ transfer operation repair tissue defect or pathological changes, is one of important topic of biomaterial science and medical domain.Current polysaccharide hydrogel preparation mainly adopts sodium alginate, chitosan etc. to make raw material.Such as: China Patent Publication No. is CN103087334A, publication date is on May 8th, 2013, and denomination of invention is the application case of " preparation method of sodium alginate-artemisia glue composite aquogel ".This application case discloses by introducing artemisia glue in sodium alginate soln system, utilizes micropore calcium carbonate to slowly release Ca in gluconic acid lactone solution
2+original position forms alginic acid-artemisia glue composite aquogel, makes the performance complement of sodium alginate and artemisia glue, thus strengthens water absorbing properties and the mechanical performance of sodium alginate based aquagel.The shortcoming of the method is: the cross-linking agent adopted is Ca
2+, dependence be electrostatic force formed cross-linked network, what obtain is physical hydrogel.Therefore in drug controlled release process, the cross-linking agent Ca in hydrogel
2+other ion being easy to be diffused in system is replaced, the problem such as make hydrogel water-soluble.
Summary of the invention
For the deficiency that above-mentioned technology exists, the object of this invention is to provide a kind of simple process, pollute little, products obtained therefrom has good mechanical property, the release of medicine controllability, good biocompatibility, biological degradability and bioactive hydrogel scaffold preparation method.
For achieving the above object, technical scheme provided by the invention is:
Prepare a method for fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, described preparation method comprises the following steps:
The Sclerotium of Pleurotus tuber regium of drying is pulverized by a, carry out surname extraction with ethyl acetate, acetone successively and remove fat, then the Sclerotium of Pleurotus tuber regium after degrease is immersed in normal saline, wherein every 100g Sclerotium of Pleurotus tuber regium 1L normal saline soaks, extract at high pressure 120 DEG C of temperature, centrifugal extracting solution, centrifugal again and collect residue after cooling extracting solution; Residue deionized water eccentric cleaning and lyophilization obtains Pleurotus tuber-regium highly-branched polysaccharide.
It is in the NaOH aqueous solution of 12 ~ 14 that Pleurotus tuber-regium highly-branched polysaccharide is dissolved in pH value by b, is prepared into the Pleurotus tuber-regium highly-branched polysaccharide solution that concentration is 2%w/v after stirring 2h.
Xanthan gum joins in the Pleurotus tuber-regium highly-branched polysaccharide solution obtained through b step by c, stir 12h, obtain the highly-branched polysaccharide-xanthan gum solution of mix homogeneously, the concentration of xanthan gum is 0.5% ~ 5%w/v, and the mass ratio of highly-branched polysaccharide and xanthan gum is 1:0.25 ~ 1:2.5.
Sodium trimetaphosphate dissolves in deionized water by d, is stirred to and dissolves completely, obtain the sodium trimetaphosphate aqueous solution that concentration is 75 ~ 262.5mg/mL.
The concentration obtained through Step d is that the sodium trimetaphosphate aqueous solution of 75 ~ 262.5mg/mL is added in the Pleurotus tuber-regium highly-branched polysaccharide-xanthan gum solution obtained through step c by e, wherein the volume ratio of Pleurotus tuber-regium highly-branched polysaccharide-xanthan gum solution and sodium trimetaphosphate aqueous solution is 25:8, rapid stirring 5min, cross-linking reaction 10min ~ 48h at 37 DEG C of temperature, obtains fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold.
Owing to have employed above technical scheme, technical scheme of the present invention is for the construction features of Pleurotus tuber-regium highly-branched polysaccharide and xanthan gum, employing water solublity, nontoxic sodium trimetaphosphate esterification and crosslinking prepare drug controllable release and the highly-branched of good mechanical properties polysaccharide-xanthan gum hydrogel scaffold, medicine or cell growth factor are embedded in this hydrogel scaffold, utilize the content ratio of highly-branched polysaccharide and xanthan gum to the pore size of the mechanical property and loose structure that regulate hydrogel.The biological activity of fungus highly-branched polysaccharide is conducive to adhesion and the growth of cell.Hydroxyl in hydrogel on the hydroxyl of highly-branched polysaccharide periphery, xanthan gum sugar ring and carboxyl, and the Polymeric sodium metaphosphate. group that cross-linking agent is introduced has certain interaction to medicine or cell growth factor, the aperture of Bound moisture gel and with electric charge controllable medicine or cell growth factor with the release of different speed, thus the Co ntrolled release behavior of the simulate growth factor in extracellular matrix and the differentiation of controlled induced cell proliferation.In addition, xanthan gum exists with the rigidity aggregation conformation of Double helix chain formation, highly-branched polysaccharide is evenly distributed on the hole place of the aggregation of these Double helix chain ordered arrangement, esterification and crosslinking is there is and forms highly-branched polysaccharide and xanthan gum hydrogel scaffold in sodium trimetaphosphate around coiled strand, the existence of highly-branched polysaccharide and the double-spiral structure of cross-linking reaction to xanthan gum do not make significant difference, and crosslinked have good reinforced effects to the intensity of hydrogel.
Compared with the prior art the method that the present invention prepares highly-branched polysaccharide-fibroin hydrogel scaffold has the following advantages:
Preparation method of the present invention has simple to operate, with low cost, abundant raw material source, one of raw material used Pleurotus tuber-regium highly-branched polysaccharide has biological activity, original position hydrogel can be prepared under the physiological condition of applicable Growth of Cells, highly-branched polysaccharide-xanthan gum hydrogel scaffold of obtaining of preparation method thus, as pharmacy slow releasing controlled during pharmaceutical carrier, improves drug effect; As loading cell growth factor during tissue engineering bracket material and the function of analog cell epimatrix control cell growth factor slow releasing, thus induced cell proliferation is divided into regenerating tissues, and the existence of fungus highly-branched polysaccharide, can give the biological activity of hydrogel scaffold.Experiment shows, the highly-branched polysaccharide-xanthan gum hydrogel scaffold of this method gained has three-dimensional through loose structure, and Model Molecule bovine serum albumin embedding amount in this hydrogel is large, and has good controllable release behavior.Therefore, the method can be widely used in preparing artificial organ timbering material, and also has broad application prospects in the field such as drug controlled release and food.It should be noted that: Pleurotus tuber-regium highly-branched polysaccharide-xanthan gum hydrogel is not only structurally similar to extracellular matrix, the more important thing is, it is wherein containing fungus highly-branched polysaccharide, there is specific biological activity, and as a kind of highly-branched polysaccharide, its structure is easy to flexible modulation physical property or carries various chemical signal molecule, thus optimizes cell survival and the specific differentiation behavior of inducing cell.Therefore, cell growth factor to be embedded in Pleurotus tuber-regium highly-branched polysaccharide-xanthan gum hydrogel and to regulate and control its release behavior by highly-branched polysaccharide unique chemical moieties and can simulate its release in vivo and the even formation of induction cambium better.In addition, in hydrogel scaffold, xanthan gum maintains the Double helix aggregate structure of rigidity, and the Ca added in carboxyl energy inducing cell culture fluid on sugared ring
2+and PO
4 3-absorption deposition, make support mineralising, be conducive to the adhesion of osteocyte and propagation etc.
Accompanying drawing explanation
Fig. 1 is the scanning electron microscopic picture of the highly-branched polysaccharide-xanthan gum hydrogel scaffold of embodiment 6.
Fig. 2 is embodiment 3, and the highly-branched polysaccharide-xanthan gum hydrogel scaffold of 4,5,6 is to bovine serum albumin Co ntrolled release curve.
Detailed description of the invention
Below in conjunction with specific embodiment, technical scheme of the present invention is described further.
Prepare a method for fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, described preparation method comprises the following steps:
The Sclerotium of Pleurotus tuber regium of drying is pulverized by a, and carry out surname extraction 6h with ethyl acetate, acetone successively and remove fat, ethyl acetate used and acetone are chemically pure reagent.Then the Sclerotium of Pleurotus tuber regium after degrease is immersed in 2h in the normal saline at 80 DEG C of temperature, centrifugal; Residue soaks 30min at high pressure 120 DEG C of temperature, and wherein every 100g Sclerotium of Pleurotus tuber regium 1L normal saline soaks, and under the rotating speed of 8000 turns, centrifugal 20min obtains extracting solution, centrifugal again and collect residue after cooling extracting solution; Residue deionized water eccentric cleaning and lyophilization obtains Pleurotus tuber-regium highly-branched polysaccharide, also can adopt other method drying to obtain Pleurotus tuber-regium highly-branched polysaccharide.
Under b room temperature, Pleurotus tuber-regium highly-branched polysaccharide being dissolved in pH value is in the NaOH aqueous solution of 12 ~ 14, the Pleurotus tuber-regium highly-branched polysaccharide solution that concentration is 2%w/v is prepared into after magnetic agitation 2h, the concentration of NaOH is 0.01 ~ 1mol/L herein, 2%w/v representation quality volumetric concentration, represents in 100 grams of solvents and dissolves 2 grams of solutes.
Under c room temperature, xanthan gum is joined in the Pleurotus tuber-regium highly-branched polysaccharide solution obtained through b step, continue magnetic agitation 12h, obtain the Pleurotus tuber-regium highly-branched polysaccharide-xanthan gum solution of mix homogeneously, the concentration of xanthan gum is 0.5% ~ 5%w/v, and the mass ratio of Pleurotus tuber-regium highly-branched polysaccharide and xanthan gum is 1:0.25 ~ 1:2.5.
Under d room temperature, sodium trimetaphosphate is dissolved in deionized water, is stirred to and dissolves completely, also can shake up, obtain the sodium trimetaphosphate aqueous solution that concentration is 75 ~ 262.5mg/mL.
Under e room temperature, be that the sodium trimetaphosphate aqueous solution of 75 ~ 262.5mg/mL is added in the Pleurotus tuber-regium highly-branched polysaccharide-xanthan gum solution obtained through step c by the concentration obtained through Step d, wherein the volume ratio of Pleurotus tuber-regium highly-branched polysaccharide-xanthan gum solution and sodium trimetaphosphate aqueous solution is 25:8, the mol ratio of the repetition sugar unit of sodium trimetaphosphate and xanthan gum is 1.46 ~ 14.6, rapid stirring 5min, magnetic agitation can be adopted herein, also mechanical agitation can be adopted, cross-linking reaction 10min ~ 48h at 37 DEG C of temperature, obtain fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, adopt the rheological behaviour of flow graph research hydrogel, by washed with de-ionized water and lyophilization obtains the three-dimensional through porous support of highly-branched polysaccharide-xanthan gum, utilize the pattern of scanning electron microscopic observation lyophilization after-poppet, the swelling ratio of dry after-poppet material and the release behavior to bovine serum albumin is tested in phosphate buffered saline(PBS).
embodiment 1
The Sclerotium of Pleurotus tuber regium of 500g drying is pulverized, carry out surname extraction 6h with ethyl acetate, acetone successively and remove fat, then the Sclerotium of Pleurotus tuber regium after degrease is immersed in 2h in the normal saline at 5L80 DEG C of temperature, centrifugal, residue soaks 30min in 5L normal saline at high pressure 120 DEG C of temperature, centrifugal extracting solution, centrifugal again and collect residue after cooling extracting solution, residue deionized water eccentric cleaning and lyophilization obtains Pleurotus tuber-regium highly-branched polysaccharide.0.5g Pleurotus tuber-regium highly-branched polysaccharide is dissolved in the NaOH aqueous solution of 25mLpH=12, the Pleurotus tuber-regium highly-branched polysaccharide solution that concentration is 2%w/v is prepared into after magnetic agitation 2h, 0.125g xanthan gum being joined above-mentioned concentration is in the Pleurotus tuber-regium highly-branched polysaccharide solution of 2%w/v, continue magnetic agitation 12h, obtain the Pleurotus tuber-regium highly-branched polysaccharide-xanthan gum solution of mix homogeneously, the sodium trimetaphosphate of 8mL75mg/mL is added in above-mentioned highly-branched polysaccharide-xanthan gum solution, rapid stirring 5min obtains pre-gel solution, by pre-gel solution at 37 DEG C of temperature after cross-linking reaction 10min, obtain fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, by the rheological behaviour of flow graph research hydrogel scaffold, by washed with de-ionized water and lyophilization obtains the three-dimensional through porous support of highly-branched polysaccharide-xanthan gum, with the pattern of scanning electron microscopic observation lyophilization after-poppet, the swelling ratio of dry after-poppet material and the release behavior to bovine serum albumin is tested in phosphate buffered saline(PBS).
embodiment 2
The Sclerotium of Pleurotus tuber regium of 500g drying is pulverized, carry out surname extraction 6h with ethyl acetate, acetone successively and remove fat, then the Sclerotium of Pleurotus tuber regium after degrease is immersed in 2h in the normal saline at 5L80 DEG C of temperature, centrifugal, residue soaks 30min in 5L normal saline at high pressure 120 DEG C of temperature, centrifugal extracting solution, centrifugal again and collect residue after cooling extracting solution, residue deionized water eccentric cleaning and lyophilization obtains highly-branched Pleurotus tuber-regium polysaccharide.0.5g Pleurotus tuber-regium highly-branched polysaccharide is dissolved in the NaOH aqueous solution of 25mLpH=13, the Pleurotus tuber-regium highly-branched polysaccharide solution that concentration is 2%w/v is prepared into after magnetic agitation 2h, 0.25g xanthan gum being joined above-mentioned concentration is in the Pleurotus tuber-regium highly-branched polysaccharide solution of 2%w/v, continue magnetic agitation 12h, obtain the highly-branched polysaccharide-xanthan gum solution of mix homogeneously, the sodium trimetaphosphate of 8mL112.5mg/mL is added in above-mentioned highly-branched polysaccharide-xanthan gum solution, rapid stirring 5min obtains pre-gel solution, by pre-gel solution at 37 DEG C of temperature after cross-linking reaction 1h, obtain fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, by the rheological behaviour of flow graph research hydrogel scaffold, by washed with de-ionized water and lyophilization obtains the three-dimensional through porous support of highly-branched polysaccharide-xanthan gum, with the pattern of scanning electron microscopic observation lyophilization after-poppet, the swelling ratio of dry after-poppet material and the release behavior to bovine serum albumin is tested in phosphate buffered saline(PBS).
embodiment 3
The Sclerotium of Pleurotus tuber regium of 500g drying is pulverized, carry out surname extraction 6h with ethyl acetate, acetone successively and remove fat, then the Sclerotium of Pleurotus tuber regium after degrease is immersed in 2h in the normal saline at 5L80 DEG C of temperature, centrifugal, residue soaks 30min in 5L normal saline at high pressure 120 DEG C of temperature, centrifugal extracting solution, centrifugal again and collect residue after cooling extracting solution, residue deionized water eccentric cleaning and lyophilization obtains highly-branched Pleurotus tuber-regium polysaccharide.0.5g Pleurotus tuber-regium highly-branched polysaccharide is dissolved in the NaOH aqueous solution of 25mLpH=14, the Pleurotus tuber-regium highly-branched polysaccharide solution that concentration is 2%w/v is prepared into after magnetic agitation 2h, 0.5g xanthan gum being joined above-mentioned concentration is in the Pleurotus tuber-regium highly-branched polysaccharide solution of 2%w/v, continue magnetic agitation 12h, obtain the highly-branched polysaccharide-xanthan gum solution of mix homogeneously, the sodium trimetaphosphate of 8mL150mg/mL is added in above-mentioned highly-branched polysaccharide-xanthan gum solution, rapid stirring 5min obtains pre-gel solution, by pre-gel solution at 37 DEG C of temperature after cross-linking reaction 10min, obtain fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, by the rheological behaviour of flow graph research hydrogel scaffold, by washed with de-ionized water and lyophilization obtains the three-dimensional through porous support of highly-branched polysaccharide-xanthan gum, with the pattern of scanning electron microscopic observation lyophilization after-poppet, the swelling ratio of dry after-poppet material and the release behavior to bovine serum albumin is tested in phosphate buffered saline(PBS).
embodiment 4
The Sclerotium of Pleurotus tuber regium of 500g drying is pulverized, carry out surname extraction 6h with ethyl acetate, acetone successively and remove fat, then the Sclerotium of Pleurotus tuber regium after degrease is immersed in 2h in the normal saline at 5L80 DEG C of temperature, centrifugal, residue soaks 30min in 5L normal saline at high pressure 120 DEG C of temperature, centrifugal extracting solution, centrifugal again and collect residue after cooling extracting solution, residue deionized water eccentric cleaning and lyophilization obtains highly-branched Pleurotus tuber-regium polysaccharide.0.5g Pleurotus tuber-regium highly-branched polysaccharide is dissolved in the NaOH aqueous solution of 25mLpH=13, the Pleurotus tuber-regium highly-branched polysaccharide solution that concentration is 2%w/v is prepared into after magnetic agitation 2h, 0.75g xanthan gum being joined above-mentioned concentration is in the Pleurotus tuber-regium highly-branched polysaccharide solution of 2%w/v, continue magnetic agitation 12h, obtain the highly-branched polysaccharide-xanthan gum solution of mix homogeneously, the sodium trimetaphosphate of 8mL187.5mg/mL is added in above-mentioned highly-branched polysaccharide-xanthan gum solution, rapid stirring 5min obtains pre-gel solution, by pre-gel solution at 37 DEG C of temperature after cross-linking reaction 24h, obtain fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, by the rheological behaviour of flow graph research hydrogel scaffold, by washed with de-ionized water and lyophilization obtains the three-dimensional through porous support of highly-branched polysaccharide-xanthan gum, with the pattern of scanning electron microscopic observation lyophilization after-poppet, the swelling ratio of dry after-poppet material and the release behavior to bovine serum albumin is tested in phosphate buffered saline(PBS).
embodiment 5
The Sclerotium of Pleurotus tuber regium of 500g drying is pulverized, carry out surname extraction 6h with ethyl acetate, acetone successively and remove fat, then the Sclerotium of Pleurotus tuber regium after degrease is immersed in 2h in the normal saline at 5L80 DEG C of temperature, centrifugal, residue soaks 30min in 5L normal saline at high pressure 120 DEG C of temperature, centrifugal extracting solution, centrifugal again and collect residue after cooling extracting solution, residue deionized water eccentric cleaning and lyophilization obtains highly-branched Pleurotus tuber-regium polysaccharide.0.5g Pleurotus tuber-regium highly-branched polysaccharide is dissolved in the NaOH aqueous solution of 25mLpH=13, the Pleurotus tuber-regium highly-branched polysaccharide solution that concentration is 2%w/v is prepared into after magnetic agitation 2h, 1.0g xanthan gum being joined above-mentioned concentration is in the Pleurotus tuber-regium highly-branched polysaccharide solution of 2%w/v, continue magnetic agitation 12h, obtain the highly-branched polysaccharide-xanthan gum solution of mix homogeneously, the sodium trimetaphosphate of 8mL225mg/mL is added in above-mentioned highly-branched polysaccharide-xanthan gum solution, rapid stirring 5min obtains pre-gel solution, by pre-gel solution at 37 DEG C of temperature after cross-linking reaction 48h, obtain fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, by the rheological behaviour of flow graph research hydrogel scaffold, by washed with de-ionized water and lyophilization obtains the three-dimensional through porous support of highly-branched polysaccharide-xanthan gum, with the pattern of scanning electron microscopic observation lyophilization after-poppet, the swelling ratio of dry after-poppet material and the release behavior to bovine serum albumin is tested in phosphate buffered saline(PBS).
embodiment 6
The Sclerotium of Pleurotus tuber regium of 500g drying is pulverized, carry out surname extraction 6h with ethyl acetate, acetone successively and remove fat, then the Sclerotium of Pleurotus tuber regium after degrease is immersed in 2h in the normal saline at 5L80 DEG C of temperature, centrifugal, residue soaks 30min in 5L normal saline at high pressure 120 DEG C of temperature, centrifugal extracting solution, centrifugal again and collect residue after cooling extracting solution, residue deionized water eccentric cleaning and lyophilization obtains highly-branched Pleurotus tuber-regium polysaccharide.0.5g Pleurotus tuber-regium highly-branched polysaccharide is dissolved in the NaOH aqueous solution of 25mLpH=13, the Pleurotus tuber-regium highly-branched polysaccharide solution that concentration is 2%w/v is prepared into after magnetic agitation 2h, 1.25g xanthan gum being joined above-mentioned concentration is in the Pleurotus tuber-regium highly-branched polysaccharide solution of 2%w/v, continue magnetic agitation 12h, obtain the highly-branched polysaccharide-xanthan gum solution of mix homogeneously, the sodium trimetaphosphate of 8mL262.5mg/mL is added in above-mentioned highly-branched polysaccharide-xanthan gum solution, rapid stirring 5min obtains pre-gel solution, by pre-gel solution at 37 DEG C of temperature after cross-linking reaction 3h, obtain fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, by the rheological behaviour of flow graph research hydrogel scaffold, by washed with de-ionized water and lyophilization obtains the three-dimensional through porous support of highly-branched polysaccharide-xanthan gum, with the pattern of scanning electron microscopic observation lyophilization after-poppet, the swelling ratio of dry after-poppet material and the release behavior to bovine serum albumin is tested in phosphate buffered saline(PBS).(this embodiment is most preferred embodiment)
embodiment 7
The Sclerotium of Pleurotus tuber regium of 500g drying is pulverized, carry out surname extraction 6h with ethyl acetate, acetone successively and remove fat, then the Sclerotium of Pleurotus tuber regium after degrease is immersed in 2h in the normal saline at 5L80 DEG C of temperature, centrifugal, residue soaks 30min in 5L normal saline at high pressure 120 DEG C of temperature, centrifugal extracting solution, centrifugal again and collect residue after cooling extracting solution, residue deionized water eccentric cleaning and lyophilization obtains highly-branched Pleurotus tuber-regium polysaccharide.0.5g Pleurotus tuber-regium highly-branched polysaccharide is dissolved in the NaOH aqueous solution of 25mLpH=13, the Pleurotus tuber-regium highly-branched polysaccharide solution that concentration is 2%w/v is prepared into after magnetic agitation 2h, 1.25g xanthan gum being joined above-mentioned concentration is in the Pleurotus tuber-regium highly-branched polysaccharide solution of 2%w/v, continue magnetic agitation 12h, obtain the highly-branched polysaccharide-xanthan gum solution of mix homogeneously, the sodium trimetaphosphate of 8mL75mg/mL is added in above-mentioned highly-branched polysaccharide-xanthan gum solution, rapid stirring 5min obtains pre-gel solution, by pre-gel solution at 37 DEG C of temperature after cross-linking reaction 3h, obtain fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, by the rheological behaviour of flow graph research hydrogel scaffold, by washed with de-ionized water and lyophilization obtains the three-dimensional through porous support of highly-branched polysaccharide-xanthan gum, with the pattern of scanning electron microscopic observation lyophilization after-poppet, the swelling ratio of dry after-poppet material and the release behavior to bovine serum albumin is tested in phosphate buffered saline(PBS).
The performance of the fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold of embodiment 3 ~ 7 is in table one
Table one
Claims (1)
1. prepare a method for fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold, it is characterized in that: described preparation method comprises the following steps:
The Sclerotium of Pleurotus tuber regium of drying is pulverized by a, carry out surname extraction with ethyl acetate, acetone successively and remove fat, then the Sclerotium of Pleurotus tuber regium after degrease is immersed in normal saline, wherein every 100g Sclerotium of Pleurotus tuber regium 1L normal saline soaks, extract at high pressure 120 DEG C of temperature, centrifugal extracting solution, centrifugal again and collect residue after cooling extracting solution, residue deionized water eccentric cleaning and lyophilization obtains Pleurotus tuber-regium highly-branched polysaccharide;
It is in the NaOH aqueous solution of 12 ~ 14 that Pleurotus tuber-regium highly-branched polysaccharide is dissolved in pH value by b, is prepared into the Pleurotus tuber-regium highly-branched polysaccharide solution that concentration is 2%w/v after stirring 2h;
Xanthan gum joins in the Pleurotus tuber-regium highly-branched polysaccharide solution obtained through b step by c, stir 12h, obtain the highly-branched polysaccharide-xanthan gum solution of mix homogeneously, the concentration of xanthan gum is 0.5% ~ 5%w/v, and the mass ratio of highly-branched polysaccharide and xanthan gum is 1:0.25 ~ 1:2.5;
Sodium trimetaphosphate dissolves in deionized water by d, is stirred to and dissolves completely, obtain the sodium trimetaphosphate aqueous solution that concentration is 75 ~ 262.5mg/mL;
The concentration obtained through Step d is that the sodium trimetaphosphate aqueous solution of 75 ~ 262.5mg/mL is added in the Pleurotus tuber-regium highly-branched polysaccharide-xanthan gum solution obtained through step c by e, wherein the volume ratio of Pleurotus tuber-regium highly-branched polysaccharide-xanthan gum solution and sodium trimetaphosphate aqueous solution is 25:8, rapid stirring 5min, cross-linking reaction 10min ~ 48h at 37 DEG C of temperature, obtains fungus highly-branched polysaccharide-xanthan gum hydrogel scaffold.
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| CN106362208A (en) * | 2016-09-23 | 2017-02-01 | 武汉纺织大学 | Method for preparing fibroin-xanthan gum aquogel bracket |
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| CN114796603A (en) * | 2022-05-30 | 2022-07-29 | 大连理工大学 | Conductive hydrogel based on chitosan/xanthan gum interpenetrating network and preparation method thereof |
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| CN106362208A (en) * | 2016-09-23 | 2017-02-01 | 武汉纺织大学 | Method for preparing fibroin-xanthan gum aquogel bracket |
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| CN117530929A (en) * | 2024-01-10 | 2024-02-09 | 东华大学 | Weight-losing capsule |
| CN117551302A (en) * | 2024-01-10 | 2024-02-13 | 东华大学 | Preparation method of high-hydroscopicity hydrolysis-resistant polysaccharide-based porous material |
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| CN117551302B (en) * | 2024-01-10 | 2024-03-29 | 东华大学 | Preparation method of high-hydroscopicity hydrolysis-resistant polysaccharide-based porous material |
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