A kind of injectable biological glue with high-strength adhesion property and preparation method thereof with
Using
Technical field
The present invention relates to a kind of biological glues, and in particular to a kind of injectable biological glue with high-strength adhesion property.
Background technology
The wound suture problem of surgical operation has had been to be concerned by more and more people.Traditional mode generally uses suture
Suture.But suture suture can introduce new suture wound, can cause secondary damage during taking out stitches to wound, and suture holds
Easily cause infection.
So being developed biological glue at present to replace the suture in operation.Biological glue is generally divided into two major classes:
Natural class and synthesis class biological glue.For wherein natural class biological glue using Fibrin Glue as representative, usual intensity is weaker, and
Easy to produce the cross-infection problem of the hematologic diseases such as AIDS.Synthetic glue is with the derivative of cyanoacrylate
Main, intensity is higher, but biocompatibility is bad, and catabolite is toxic.
People's also some new biological glues in Persisting exploitation, such as the bi-component biological glue based on PEG at present
(Coseal and Duraseal), the biological glue based on PEG and chitosan.These biological glues have stronger adhesion property, and
And biocompatibility is good, but these biological glues are usually all bi-component biological glue, when use, must be noted using duplex
Emitter, equipment is bigger, and operation is complex, and the tissue adhesion operation especially for certain profound levels is more difficult.Simultaneously
Most of these glue are mutually crosslinked using chemical action, in use in case of cracking, lead to the irreversible breaking of chemical bond,
These glue can lose its effect, will usually be disposed of and coat new biological glue, be greatly improved the operating time
And cost.
Invention content
The object of the present invention is to provide a kind of injectable biological glue with high-strength adhesion property and preparation method thereof with
Using.
Injectable biological glue provided by the present invention is by polyethyleneglycol derivative and tannic acid through interaction of hydrogen bond shape
At;
The polyethyleneglycol derivative is polyethylene glycol succinimide ester.
The polyethyleneglycol derivative can be any in Formulas I, Formula II, formula III and formula IV:
In various, n is the natural number between 2~10000, can be 14~56,14,28 or 56;
Group R is formula a or formula b, in formula a and formula b, M indicate nitrogen-atoms, carbon atom, oxygen atom, sulphur atom, phosphorus atoms or
Phenyl ring, preferably oxygen atom, nitrogen-atoms or sulphur atom are connected to polyethylene glycol end, and Z is the integer between 0~5, and such as 0,2 or 3;
Group R1、R2、R3、R4、R5、R6、R7、R8、R9、R10、R11、R12、R13、R14、R15And R16Independently be hydrogen atom,
Hydroxyl, alkyl (preferably methyl), aryl or the sulfonic group that carbon atom number is 1~10.
Polyethyleneglycol derivative of the present invention concretely following 1) -8) in it is any:
1) shown in formula I, wherein R is formula a, and in formula a, Z 2, M are oxygen atom, n 56, R1、R2、R3And R4It is hydrogen
Atom;
2) as shown in Formula II, wherein R is formula b, and in formula b, Z is that 0~1, M is oxygen atom, n 56, R1、R2、R3And R4
For hydrogen atom, R5、R6、R7And R8It is hydroxyl;
3) as shown in Formula II, wherein R is formula b, and in formula b, Z 0, M are oxygen atom, n 56, R1、R2、R3And R4It is hydrogen
Atom, R5、R6、R7And R8It is hydroxyl;
4) as shown in Formula II, wherein R is formula b, and in formula b, Z 1, M are oxygen atom, n 56, R1、R2、R3And R4It is hydrogen
Atom, R5、R6、R7And R8It is hydroxyl;
5) as shown in formula III, wherein R is formula a, and in formula a, Z 3, M are nitrogen-atoms, n 28, R1、R2、R3And R4It is
Hydrogen atom, R5、R6、R7And R8It is methyl, R9、R10、R11And R12It is phenyl ring;
6) as shown in formula IV, wherein R is formula b, and in formula b, Z is that 2~3, M is sulphur atom, and n is 14~16, R1、R3、R7With
R10It is sulfonic group, R2、R4、R5、R6、R8、R9、R11、R12、R13、R14、R15And R16It is hydrogen atom;
7) as shown in formula IV, wherein R is formula b, and in formula b, Z 2, M are sulphur atom, n 14, R1、R3、R7And R10It is sulphur
Acidic group, R2、R4、R5、R6、R8、R9、R11、R12、R13、R14、R15And R16It is hydrogen atom;
8) as shown in formula IV, wherein R is formula b, and in formula b, Z 3, M are sulphur atom, n 16, R1、R3、R7And R10It is sulphur
Acidic group, R2、R4、R5、R6、R8、R9、R11、R12、R13、R14、R15And R16It is hydrogen atom.
In the injectable biological glue, the mass ratio of the polyethyleneglycol derivative and the tannic acid can be 1:
0.001~1000, concretely 1:0.4~4,1:0.4~1,1:0.4~0.5,1:0.4、1:0.5、1:1 or 1:4.
Invention further provides the preparation methods of the injectable biological glue, include the following steps:
(1) solution 1 of the polyethyleneglycol derivative is prepared;
(2) solution 2 of the tannic acid is prepared;
(3) solution 1 and the solution 2 are mixed, is the biological glue through centrifuged pellet.
In above-mentioned preparation method, in step (1), quality-volume of polyethyleneglycol derivative described in the solution 1 is dense
Degree can be 0.01~10000mg/ml, concretely 400~600mg/ml, 400mg/ml, 500mg/ml or 600mg/ml;
The solvent of the solution 1 can be the phosphate buffer solution that secondary water, ultra-pure water, physiological saline or pH are 7.4.
In above-mentioned preparation method, in step (2), quality-volumetric concentration of tannic acid described in the solution 2 can be
0.01~10000mg/ml, concretely 400~600mg/ml, 400mg/ml, 500mg/ml or 600mg/ml;
The solvent of the solution 1 is the phosphate buffer solution that secondary water, ultra-pure water, physiological saline or pH are 7.4.
In above-mentioned preparation method, in step (3), the volume ratio of the solution 1 and the solution 2 can be 1:0.001~
1000, concretely 1:0.4~4,1:0.4~1,1:0.4~0.5,1:0.4、1:0.5、1:1 or 1:4;
The rate of the centrifugation can be 100~20000r/min, and the time can be 1~60min, such as in the item of 3000r/min
5min is centrifuged under part.
Injectable biological glue of the present invention with high-strength adhesion property is in following field with potential application:
(1) wound bonds;
(2) drug carrier delivery systme;
(3) medical inner matter;
(4) face coat of medical inner matter.
Compared with prior art, the present invention has the advantages that:
(1) present invention has good biocompatibility using the substance of PEG derivatives and tannic acid two kinds of FDA approvals.
(2) two groups of substances of PEG derivatives and tannic acid interact by physical action (i.e. hydrogen bond), form injectable
Biological glue.
(3) biological glue of the present invention has higher adhesion ability.
(4) biological glue of the present invention can be used by single syringes, more convenient to use.
Description of the drawings
Fig. 1 is the contrast schematic diagram of the adhesion intensity and Fibrin Glue of biological glue prepared by the embodiment of the present invention 1.
Specific implementation mode
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Adhesion property measures by the following method in following embodiments:
The method stretched using pigskin bonding:Directly biological glue is coated on two panels pigskin using syringe, coating
Area is 1cm × 1cm, and then coating position coincides together, after the power pressing 30s of 30 newton, using tensilon
Measure shear strength when two pieces of pigskins separate, as adhesion strength.
Embodiment 1,
Weighing two-arm polyethylene glycol succinimide 5000mg, (shown in formula I, R is formula a, Z 2, and M is oxygen atom, n values
It is 56, R1、R2、R3And R4All it is hydrogen atom) it is dissolved in the PBS buffer solutions that 10mL pH are 7.4 that (quality-volumetric concentration is
500mg/ml), it weighs tannic acid 5000mg and is dissolved in the PBS solution that 10ml pH are 7.4 that (quality-volumetric concentration is 500mg/
Ml), it is 1 according to volume ratio by above two solution:1 (mass ratio of two-arm polyethylene glycol succinimide and tannic acid be 1:
1) after mixing, after the speed centrifugation 5min of 3000r/min, supernatant is outwelled, gained precipitation is biological glue of the present invention.
Biological glue is put into syringe, remains to use in next step.
Through measuring, the adhesion property of biological glue manufactured in the present embodiment is 249kPa, has higher adhesion ability, much
Better than commercially available Fibrin Glue (15.2kPa), as shown in Figure 1.
After two pieces of pigskins are cemented using the present embodiment biological glue, artificially partitioned two pieces of pigskins, then again by two pieces of pigs
Skin merges, and after 2min, remeasures adhesion strength, adhesion strength can be restored to 135kPa, still better than commercially available
Fibrin Glue, show biological glue of the present invention have be repeated as many times adhesion ability, even during use wound due to
External force cracks again, can also be directly by wound pairing, and biological glue will again cement wound after a period of time.
Embodiment 2,
Weighing four arm polyethylene glycol succinimide 4000mg, (as shown in Formula II, R is formula b, Z 0, and M is oxygen atom, n values
It is 56, R1、R2、R3And R4For hydrogen atom, R5、R6、R7And R8All it is hydroxyl) it is dissolved in the PBS buffer solutions that 10mL pH are 7.4
(quality-volumetric concentration is 400mg/ml) weighs tannic acid 4000mg and is dissolved in (quality-body in the PBS solution that 10ml pH are 7.4
A concentration of 400mg/ml of product), according to volume ratio it is 2 by above two solution:1 (four arm polyethylene glycol succinimides and tannin
The mass ratio of acid is 2:1) after mixing, after the speed centrifugation 5min of 3000r/min, supernatant is outwelled, gained precipitation is
Biological glue of the present invention.Biological glue is put into syringe, remains to use in next step.
Through measuring, the adhesion property of biological glue manufactured in the present embodiment is 265kPa, has higher adhesion ability, much
Better than commercially available Fibrin Glue (15.2kPa).
After two pieces of pigskins are cemented using the present embodiment biological glue, artificially partitioned two pieces of pigskins, then again by two pieces of pigs
Skin merges, and after 2min, remeasures adhesion strength, adhesion strength can be restored to 150Pa, still better than commercially available fibre
Fibrillarin glue shows that biological glue of the present invention has the ability for being repeated as many times adhesion, and wound is due to outer even during use
Power cracks again, can also be directly by wound pairing, and biological glue will again cement wound after a period of time.
Embodiment 3,
Weighing six arm polyethylene glycol succinimide 4000mg, (as shown in formula III, R is formula a, Z 3, and M is nitrogen-atoms, n
Value is 28, R1、R2、R3And R4It is hydrogen atom, R5、R6、R7And R8It is methyl, R9、R10、R11And R12It is phenyl ring) it is dissolved in
In the PBS buffer solutions that 10mL pH are 7.4 (quality-volumetric concentration is 400mg/ml), weighs tannic acid 4000mg and be dissolved in 10ml
In the PBS solution that pH is 7.4 (quality-volumetric concentration is 400mg/ml), by above two solution according to volume ratio be 1:4 (six
The mass ratio of arm polyethylene glycol succinimide and tannic acid is 1:4) after mixing, 5min is centrifuged using the speed of 3000r/min
Afterwards, supernatant is outwelled, gained precipitation is biological glue of the present invention.Biological glue is put into syringe, remains to make in next step
With.
Through measuring, the adhesion property of biological glue manufactured in the present embodiment is 299kPa, has higher adhesion ability, much
Better than commercially available Fibrin Glue (15.2kPa).
After two pieces of pigskins are cemented using the present embodiment biological glue, artificially partitioned two pieces of pigskins, then again by two pieces of pigs
Skin merges, and after 2min, remeasures adhesion strength, adhesion strength can be restored to 170Pa, still better than commercially available fibre
Fibrillarin glue shows that biological glue of the present invention has the ability for being repeated as many times adhesion, and wound is due to outer even during use
Power cracks again, can also be directly by wound pairing, and biological glue will again cement wound after a period of time.
Embodiment 4,
Weighing eight arm polyethylene glycol succinimide 6000mg, (as shown in formula IV, it be 2, M is sulphur atom, n that R, which is formula b, Z value,
Value is 14, R1、R3、R7And R10It is sulfonic group, R2、R4、R5、R6、R8、R9、R11、R12、R13、R14、R15And R16It is hydrogen atom)
It is dissolved in the PBS buffer solutions that 10mL pH are 7.4 (quality-volumetric concentration is 600mg/ml), tannic acid 6000mg is weighed and is dissolved in
In the PBS solution that 10ml pH are 7.4 (quality-volumetric concentration is 600mg/ml), by above two solution according to volume ratio be 7:
3 (mass ratio of eight arm polyethylene glycol succinimides and tannic acid be 7:3) it after mixing, is centrifuged using the speed of 3000r/min
After 5min, supernatant is outwelled, gained precipitation is biological glue of the present invention.Biological glue is put into syringe, is remained next
Step uses.
Through measuring, the adhesion property of biological glue manufactured in the present embodiment is 432kPa, has higher adhesion ability, much
Better than commercially available Fibrin Glue (15.2kPa).
After two pieces of pigskins are cemented using the present embodiment biological glue, artificially partitioned two pieces of pigskins, then again by two pieces of pigs
Skin merges, and after 2min, remeasures adhesion strength, adhesion strength can be restored to 245Pa, still better than commercially available fibre
Fibrillarin glue shows that biological glue of the present invention has the ability for being repeated as many times adhesion, and wound is due to outer even during use
Power cracks again, can also be directly by wound pairing, and biological glue will again cement wound after a period of time.
Embodiment 5,
Weighing eight arm polyethylene glycol succinimide 5000mg, (as shown in formula IV, it be 3, M is sulphur atom, n that R, which is formula b, Z value,
Value is 16, R1、R3、R7And R10It is sulfonic group, R2、R4、R5、R6、R8、R9、R11、R12、R13、R14、R15And R16It is hydrogen atom)
It is dissolved in the PBS buffer solutions that 10mL pH are 7.4 (quality-volumetric concentration is 500mg/ml), tannic acid 6000mg is weighed and is dissolved in
In the PBS solution that 10ml pH are 7.4 (quality-volumetric concentration is 600mg/ml), by above two solution according to volume ratio be 7:
3 (mass ratio of eight arm polyethylene glycol succinimides and tannic acid be 7:3) after mixing, taxol drug is added thereto
200mg outwells supernatant after the speed centrifugation 5min of 3000r/min, and gained precipitation is that the present invention is paclitaxel loaded
Biological glue.Paclitaxel loaded biological glue is put into syringe, can be injected into human body, as drug release body
System.
Embodiment 6,
Weighing four arm polyethylene glycol succinimide 4000mg, (as shown in Formula II, R is formula b, Z 1, and M is oxygen atom, n values
It is 56, R1、R2、R3And R4For hydrogen atom, R5、R6、R7And R8All it is hydroxyl) it is dissolved in the PBS buffer solutions that 10mL pH are 7.4
(quality-volumetric concentration is 400mg/ml) weighs tannic acid 4000mg and is dissolved in (quality-body in the PBS solution that 10ml pH are 7.4
A concentration of 400mg/ml of product), according to volume ratio it is 2 by above two solution:1 (four arm polyethylene glycol succinimides and tannin
The mass ratio of acid is 2:1) after mixing, after the speed centrifugation 5min of 3000r/min, supernatant is outwelled, gained precipitation is
Biological glue of the present invention.Biological glue is put into syringe, remains to use in next step.
Biological glue is coated in the PLGA rack surfaces for Bone Defect Repari, enhance PLGA holders and is put into around position
Adhesive attraction, it was demonstrated that biological glue of the present invention can be used as the face coat of medical inner matter.