CN108187128B - Absorbable hemostatic bone wax and preparation method thereof - Google Patents
Absorbable hemostatic bone wax and preparation method thereof Download PDFInfo
- Publication number
- CN108187128B CN108187128B CN201810123352.4A CN201810123352A CN108187128B CN 108187128 B CN108187128 B CN 108187128B CN 201810123352 A CN201810123352 A CN 201810123352A CN 108187128 B CN108187128 B CN 108187128B
- Authority
- CN
- China
- Prior art keywords
- gelatin
- bone wax
- poloxamer
- hemostatic bone
- hemostatic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/0047—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
- A61L24/0073—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material with a macromolecular matrix
- A61L24/0094—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material with a macromolecular matrix containing macromolecular fillers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0042—Materials resorbable by the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/04—Materials for stopping bleeding
Abstract
The invention provides absorbable hemostatic bone wax and a preparation method thereof, wherein the hemostatic bone wax is composed of gelatin and poloxamer, and the mass ratio of the gelatin to the poloxamer is 1: 1-9: 1. The hemostatic bone wax provided by the invention has degradability, excellent physical plugging effect and hemostatic performance. In addition, the preparation method of the hemostatic bone wax is that the cross-linked gelatin is fully swelled, then mechanically crushed, freeze-dried and secondarily crushed, and then swelled in poloxamer solution and freeze-dried again to obtain the hemostatic bone wax. The preparation method disclosed by the invention is simple in process, the prepared hemostatic bone wax has a proper volume swelling rate and a proper saturated water absorption rate, the degradation time of the hemostatic bone wax is 8-50 weeks, the bursting pressure is not lower than 60mmHg, the hemostatic bone wax has a good plugging effect, and the hemostatic bone wax can be widely applied to the field of orthopedics hemostasis.
Description
Technical Field
The invention relates to the technical field of hemostatic materials, and particularly relates to absorbable hemostatic bone wax and a preparation method thereof.
Background
The bone wax is a material for blocking capillary blood seepage of a medullary part by a physical method, can effectively block the bleeding of cancellous bone, and can be used for stopping bleeding of various emergency patients during bone blood seepage. The specific mechanism of action of bone wax is: on one hand, the bone wax has very high absorption effect on liquid in the hemostasis process, and promotes the concentration of platelets by rapidly absorbing water in blood on a bleeding wound surface, so that the agglutination speed and the agglutination capacity of the platelets are enhanced; on the other hand, the bone wax forms hydrogel after absorbing water, can be blocked on the surface of the wound, and increases the hemostatic effect. In addition, after the bone wax absorbs water and expands, the bone wax can form a compression effect with a certain pressure on peripheral blood vessels, and further promotes hemostasis.
The bone wax used in clinic is a mixture of 70% of beeswax and 30% of vaseline, has good softening performance, can be shaped after being softened by rubbing with hands, is nontoxic, but cannot be degraded and absorbed by organisms. Since the bone wax material prepared from the mixture of beeswax and vaseline cannot be absorbed by organism, it is left as foreign body in vivo to increase the probability of infection. Meanwhile, because the molecular gap is too small, bone tissues can not pass through bone wax materials, the growth of bones can be hindered, the wound can not be healed after operation, and the operation is required to be debrided if necessary. Thus not only prolonging the treatment time, but also increasing the pain of the patient.
Therefore, the bone wax material which has good hemostatic effect and can be degraded and absorbed is developed, and meanwhile, the bone wax can meet the mechanical performance requirement of clinical use, and has important significance and value.
Disclosure of Invention
The invention aims to provide an absorbable hemostatic bone wax aiming at the defects that bone wax in the prior art cannot be degraded and absorbed and is not beneficial to wound healing and the like in vivo. The bone wax product prepared by utilizing the temperature phase change of poloxamer and the degradability of gelatin not only has excellent physical plugging and filling properties and hemostatic effect, but also can be completely biodegraded and absorbed, and simultaneously has excellent mechanical properties.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
an absorbable hemostatic bone wax is prepared from gelatin and poloxamer, wherein the mass ratio of the gelatin to the poloxamer is 1: 1-9: 1. Preferably, the mass ratio of gelatin to poloxamer is 7: 3.
Preferably, the poloxamer has a hydrophilic-lipophilic balance (HLB) of 20 to 29.
Preferably, the poloxamer is one or more of poloxamer 188, poloxamer 338 and poloxamer 407, and further preferably, the poloxamer is poloxamer 407.
The bone wax is an effective plugging material for cancellous bone bleeding, and the traditional material is a mixture of paraffin and beeswax, so that the bone wax cannot be degraded in vivo, and the growth of a new bone structure can be limited. The hemostatic bone wax is prepared from biodegradable absorbable gelatin and temperature phase-changeable poloxamer, has excellent physical plugging and filling properties and hemostatic effect, can be completely biodegraded and absorbed, does not limit the growth of a new bone structure, and has excellent mechanical properties.
The invention also provides a preparation method of the hemostatic bone wax, which comprises the following steps:
s1, preparing a gelatin water solution;
s2, adding a cross-linking agent into the gelatin aqueous solution for cross-linking treatment;
s3, placing the cross-linked gelatin in water for full swelling;
s4, cutting the swelled gelatin in the step S3 into small blocks, and then crushing the small blocks into micron-sized gelatin particles;
s5, freezing the gelatin particles at low temperature, and freeze-drying after complete freezing to obtain dried gelatin;
s6, carrying out secondary crushing on the frozen and dried gelatin;
s7, placing the gelatin obtained in the step S6 after the secondary crushing into a poloxamer solution for full swelling;
s8, freeze-drying the gelatin swelled in the poloxamer solution in the step S7.
The hemostatic bone wax is prepared by swelling crosslinked gelatin fully with water, then carrying out mechanical crushing, freeze drying and secondary crushing, then placing the crosslinked gelatin in a poloxamer solution for fully swelling, fully fusing poloxamer and gelatin, and finally carrying out freeze drying. The volume swelling behavior of the bone wax depends on the crosslinking degree of the gelatin crosslinking treatment and the swelling degree of the bone wax soaked in water and poloxamer solution, wherein the poloxamer has hydrophilic groups to adjust the swelling rate of the prepared hemostatic bone wax, so that the hemostatic bone wax prepared by the method not only has good water absorption performance and physical plugging and packing performance, but also can perform good plugging and hemostasis on bone hemorrhage.
Preferably, the mass concentration of the gelatin aqueous solution in the step S1 is 5-50%, and the temperature for heating and completely dissolving the gelatin aqueous solution is 40-80 ℃.
Preferably, the amount of the cross-linking agent used in step S2 is 4-20% of the mass of the gelatin. Further preferably, the cross-linking agent is added during stirring of the gelatin solution.
Preferably, the pulverization in step S4 is performed by an aqueous phase material pulverizer.
Preferably, the gelatin is pulverized into particles of 0.1 to 2 μm in step S6.
More preferably, the pulverization speed used in step S6 is 30000r/min, and the pulverization time is 1 min.
Preferably, the mass concentration of the poloxamer solution in the S6 is 10% -50%, and the poloxamer is poloxamer 407.
In order to obtain more uniform and better packing performance of the hemostatic bone wax, the preparation method further comprises the following steps:
s9, crushing the freeze-dried product obtained in the step S8 for the third time. Therefore, the micro particles of the hemostatic bone wax are smaller, the texture is finer, and the plasticity and the plugging and packing performance of the hemostatic bone wax are better.
Preferably, the preparation method further comprises the step of performing irradiation sterilization on the obtained hemostatic bone wax to obtain the hemostatic product capable of being applied to the wound surface.
The bone wax prepared by the preparation method can be directly applied to bleeding wound surfaces for hemostasis, and can also be added with a small amount of water for mixing for use, so that the hemostatic bone wax has good flexibility, can be shaped according to needs, and can achieve better filling and plugging effects.
Preferably, in order to reduce the residual amount of the cross-linking agent in the hemostatic bone wax and improve the safety of the hemostatic bone wax, the invention adopts the cross-linking agent water-soluble carbodiimide (EDC) which does not participate in the reaction, and the cross-linking agent is removed through a specific step in the preparation method, so that the finally obtained hemostatic bone wax product has the residual amount of the cross-linking agent below 100ppm, good safety and no stimulation to tissues.
Specifically, the preparation steps further include the following steps:
(1) in the step S2, the cross-linking agent is water-soluble carbodiimide (EDC), the mass of the EDC is 4-20% of that of gelatin, the EDC is added in the process of stirring the gelatin solution, the stirring speed of the gelatin solution is 100-5000 r/min, preferably 1000r/min, and the adding speed of the EDC is 0.1-10 mL/S, preferably 2 mL/S;
(2) after the step S4, the gelatin is placed in pure water by a centrifugal deposition mode to carry out ultrasonic-centrifugal circulation, preferably, the centrifugal rotating speed is 10000 r/min-20000 r/min, the time of single centrifugation is 5-10 min, more preferably, the centrifugal rotating speed is 15000r/min, the ultrasonic frequency is 25 KHz-130 KHz, and the time of single ultrasonic treatment is 5 min-30 min.
For the low-swelling gelatin material, glutaraldehyde and the like are conventionally selected for crosslinking, but after crosslinking is finished, a crosslinking agent component exists in a final product and is gradually released in the subsequent degradation process of the gelatin material, so that the risk of irritation or potential carcinogenicity is brought. According to the invention, the gelatin material is effectively crosslinked by using the crosslinking agent which does not participate in the reaction, and the crosslinking agent is purified and removed by centrifugal deposition separation, purified water dialysis and other modes after crosslinking, so that the residues and toxicity of irritant substances are effectively avoided.
A hemostatic product comprises the absorbable hemostatic bone wax prepared by the method.
The anti-explosion pressure of the hemostatic bone wax prepared by the invention is not less than 60mmHg, the adjustable range of the volume swelling ratio is 50-500%, the adjustable range of the saturated water absorption ratio is 100-2000%, and the adjustable range of the degradation time is 8-50 weeks, so that the hemostatic product which has a good hemostatic effect, excellent mechanical properties and degradable absorption can be prepared.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides absorbable hemostatic bone wax and a preparation method thereof, and the absorbable hemostatic bone wax consists of gelatin and poloxamer, and has degradability, excellent physical plugging function and hemostatic performance. The preparation method of the hemostatic bone wax provided by the invention comprises the steps of fully swelling crosslinked gelatin, mechanically crushing, freeze-drying, secondarily crushing, then swelling in poloxamer solution, and freeze-drying again to obtain the hemostatic bone wax. The preparation method disclosed by the invention is simple in process, the prepared hemostatic bone wax has a proper volume swelling rate and a proper saturated water absorption rate, the degradation time of the bone wax is 8-50 weeks, the bursting pressure is not lower than 60mmHg, the hemostatic bone wax has a good plugging effect, and the hemostatic bone wax can be widely applied to the field of orthopedics hemostasis.
Drawings
Fig. 1 is a photograph of an absorbable hemostatic bone wax of the present invention.
Fig. 2 is a photograph of the absorbable hemostatic bone wax of the present invention mixed with a small amount of water.
FIG. 3 shows the results of culturing cells in the leaching solution of absorbable hemostatic bone wax of example 1 for 24 hours.
FIG. 4 shows the results of culturing the cells of the leaching solution of absorbable hemostatic bone wax of example 1 for 72 hours.
Detailed Description
The present invention will be further described with reference to specific embodiments, but the present invention is not limited to the examples in any way.
Example 1
An absorbable hemostatic bone wax, wherein the mass ratio of gelatin to poloxamer is 7:3, and the absorbable hemostatic bone wax comprises the following preparation steps:
s1, placing 40g of gelatin in a beaker, adding 60g of water, placing the beaker in a 50 ℃ oven until the gelatin is completely dissolved, and preparing a gelatin water solution with the mass concentration of 40%;
s2, dissolving 1.6g of water-soluble carbodiimide (EDC) in 10mL of purified water (the mass of EDC is 4% of the mass of gelatin), adding a cross-linking agent into the gelatin aqueous solution while stirring, and carrying out cross-linking treatment, wherein the stirring speed is 1000r/min, and the adding speed of the cross-linking agent is 2 mL/S;
s3, placing the cross-linked gelatin in water for full swelling;
s4, cutting the swelled gelatin in the step S3 into small blocks, then crushing the small blocks into micron-sized gelatin particles by using a water-phase material crusher, placing the gelatin in pure water in a centrifugal deposition mode for ultrasonic-centrifugal circulation treatment, wherein the centrifugal rotation speed is 15000r/min, the single centrifugation time is 5min, the ultrasonic frequency is 25KHz, the single ultrasonic treatment time is 5min, and the circulation times are 3 times;
s5, freezing the gelatin particles at low temperature, and freeze-drying after complete freezing to obtain dried gelatin;
s6, carrying out secondary crushing on the frozen and dried gelatin, wherein the crushing speed is 30000r/min, and the crushing time is 1 min;
s7, placing the gelatin obtained in the step S6 after the secondary crushing into a poloxamer 407 solution for full swelling, wherein the mass concentration of the poloxamer 407 solution is 25%;
s8, freeze-drying the gelatin swelled in the step S7, carrying out freeze-drying, then crushing again, filling into an auxiliary device, and irradiating for later use to obtain the absorbable hemostatic bone wax, as shown in figure 1.
The obtained bone wax can be directly applied to the wound surface with hemorrhage for hemostasis, or mixed with small amount of water for use, as shown in FIG. 2. Therefore, the hemostatic bone wax is soft and can be shaped according to the requirement, and better filling and blocking effects can be achieved.
Example 2
An absorbable hemostatic bone wax, wherein the mass ratio of gelatin to poloxamer is 9:1, and the absorbable hemostatic bone wax comprises the following preparation steps:
s1, placing 20g of gelatin in a beaker, adding 80g of water, placing the beaker in an oven at 80 ℃ until the gelatin is completely dissolved, and preparing a gelatin water solution with the mass concentration of 20%;
s2, dissolving 2g of water-soluble carbodiimide (EDC) in 10mL of purified water (the mass of the EDC is 10% of that of the gelatin), adding a cross-linking agent into the gelatin aqueous solution while stirring, and carrying out cross-linking treatment, wherein the stirring speed is 1000r/min, and the adding speed of the cross-linking agent is 2 mL/S;
s3, placing the cross-linked gelatin in water for full swelling;
s4, cutting the swelled gelatin in the step S3 into small blocks, then putting the small blocks into a water phase material crusher to crush the small blocks into micron-sized gelatin particles, putting the gelatin into pure water to perform ultrasonic-centrifugal circulation treatment in a centrifugal deposition mode, wherein the centrifugal rotation speed is 15000r/min, the single centrifugation time is 5min, the ultrasonic frequency is 100KHz, the single ultrasonic treatment time is 10min, and the circulation times are 3 times;
s5, freezing the gelatin particles at low temperature, and freeze-drying after complete freezing to obtain dried gelatin;
s6, carrying out secondary crushing on the frozen and dried gelatin, wherein the crushing speed is 30000r/min, and the crushing time is 1 min;
s7, placing the gelatin obtained in the step S6 after the secondary crushing into a poloxamer 407 solution for full swelling, wherein the mass concentration of the poloxamer 407 solution is 10%;
and S8, freeze-drying the gelatin swelled in the step S7, carrying out freeze-drying, then crushing again, filling into an auxiliary instrument, and carrying out irradiation for later use.
Example 3
An absorbable hemostatic bone wax, wherein the mass ratio of gelatin to poloxamer is 1:1, and the absorbable hemostatic bone wax comprises the following preparation steps:
s1, placing 5g of gelatin in a beaker, adding 95g of water, placing the beaker in a 40 ℃ oven until the gelatin is completely dissolved, and preparing a gelatin water solution with the mass concentration of 5%;
s2, dissolving 0.5g of water-soluble carbodiimide (EDC) in 10mL of purified water (the mass of EDC is 10% of that of gelatin), adding a cross-linking agent into the gelatin aqueous solution while stirring, and carrying out cross-linking treatment, wherein the stirring speed is 1000r/min, and the adding speed of the cross-linking agent is 2 mL/S;
s3, placing the cross-linked gelatin in water for full swelling;
s4, after swelling in the step S3, cutting the gelatin material into small pieces after the gelatin material is completely swelled, placing the small pieces into a water phase material crusher, wherein the crushed gelatin is micron-sized saturated swelling gelatin particles, placing the gelatin into pure water in a centrifugal deposition mode to perform ultrasonic-centrifugal circulation treatment, wherein the centrifugal rotation speed is 15000r/min, the single centrifugation time is 10min, the ultrasonic frequency is 75KHz, the single ultrasonic treatment time is 10min, and the circulation times are 3 times;
s5, freezing the gelatin particles at low temperature, and freeze-drying after complete freezing to obtain dried gelatin;
s6, carrying out secondary crushing on the frozen and dried gelatin, wherein the crushing speed is 30000r/min, and the crushing time is 1 min;
s7, placing the gelatin obtained in the step S6 after the secondary crushing into a poloxamer 407 solution for full swelling, wherein the mass concentration of the poloxamer 407 solution is 50%;
and S8, freeze-drying the gelatin swelled in the step S7, carrying out freeze-drying, then crushing again, filling into an auxiliary instrument, and carrying out irradiation for later use.
Performance detection
(1) Specific surface area measurement
The specific surface area detection method comprises the following steps: a multipoint BET method is adopted, and specifically, a product to be detected is put into a sample tube of an analytical instrument, wherein the analytical instrument is a rapid full-automatic specific surface area and aperture analyzer with the model of American Congta NOVA 4200 e. Under the condition of low temperature (liquid nitrogen bath), a certain amount of adsorbate gas (N) is introduced into the sample tube2) Determining adsorbate molecules (N) of the sample to be detected according to the change of gas volume before and after adsorption2) The adsorption amount of (c); the specific surface area of the solid substance is determined by referring to the national standard GB/T243358-2009-gas adsorption BET principle.
The specific surface area was calculated as: in a sample placed in a gaseous environment, the surface of its material (surface area of the external and internal voids of the particles) will be physisorbed at low temperatures. When the adsorption reaches the equilibrium, measuring the amount of adsorbed gas of the equilibrium adsorption pressure, and calculating the adsorption amount of the monomolecular layer of the sample according to the BET equation so as to calculate the specific surface area of the sample. Wherein the BET equation is:
in the formula:
p-adsorbate partial pressure in Pa;
P0-the adsorbent saturation vapour pressure in Pa;
v-actual adsorption amount of sampleIn units of cm3；
VmThe single layer saturated adsorption capacity in cm3；
C-constant related to the adsorption capacity of the sample.
The gelatin granules obtained after the second pulverization in examples 1 to 3 were subjected to the test, and the test results are shown in Table 1.
(2) Measurement of volume swelling ratio
The volume test method adopts a liquid discharge method, the hemostatic bone wax material is soaked in the measuring cylinder, and then the liquid level rising value is read. The volume swelling ratio calculation method comprises the following steps: volume V after saturation swelling1With the initial volume V0The difference of (A) accounts for the initial volume V0Percentage of (c). The results are shown in Table 1.
(3) Measurement of Mass swelling Rate
The method for testing the mass swelling rate adopts a weighing method, the hemostatic bone wax material is completely soaked, removed, and weighed after removing the excess water which is not absorbed. The mass swelling ratio calculation method comprises the following steps: mass M after saturation swelling1With an initial mass M0The difference of (D) accounts for the initial mass M0Percentage of (c). The results are shown in Table 1.
(4) Resistance to compression bursting test
The in vitro compression resistant blasting test experimental method comprises the following steps: a pressure testing method is adopted, specifically, an acrylic circular plate with a porous structure and a thickness of 1cm is prepared, the circular plate can be hermetically fixed at a port of a pressure tester, the pressure tester is an acrylic box with a circular plate interface, a pressure detection device and a pressure applying device are connected to the opposite side of the circular plate interface, after the sealed acrylic box is connected with the circular plate, the hole position of the circular plate is coated with the hemostatic bone wax material, the whole hole is filled with the material, the direction of the circular plate is downward, the box is filled with water, and pressure is applied and a leakage value is detected.
The hemostatic bone wax obtained in examples 1 to 3 was subjected to in vitro compression burst test, and the test results are shown in table 1.
Table 1 hemostatic bone wax performance parameter test results
| Examples | BET surface area of gelatin particles | Volume swell ratio of bone wax | Mass swelling ratio of bone wax | Bone wax in vitro compression resistance blasting test value |
| 1 | 30 m2/g | 296% | 530% | 60.23±11.74 mmHg |
| 2 | 40 m2/g | 396% | 1260% | 80.35±20.61 mmHg |
| 3 | 70 m2/g | 512% | 1800% | 85.95±15.23 mmHg |
(5) Cytotoxicity assay
Cells were cultured by contact of the extract, and the toxic effect of the bone wax prepared in example 1 on in vitro cells was evaluated by observing cell proliferation using L929 as an experimental cell.
The specific procedure of the experiment is as follows: the hemostatic bone wax material was extracted at a ratio of 0.1g/mL using complete medium (DMEM medium +10% Fetal Bovine Serum (FBS) +1% diabody (mixed solution of penicillin and streptomycin)) to obtain 100% extract, and diluted with 100% extract to obtain 75% extract and 25% extract.
Experimental groups: complete medium (DMEM medium +10% Fetal Bovine Serum (FBS) +1% diabody (mixed solution of penicillin and streptomycin)) was used, and the leaching solution was added to the cells, followed by cell culture.
Control group: the cell suspension was added with complete medium (DMEM medium +10% Fetal Bovine Serum (FBS) +1% diabody (mixture of penicillin and streptomycin)) to perform cell culture. The control group was not added with the leaching solution, and the rest of the cell culture conditions were the same as those of the experimental group.
Blank group: complete medium (DMEM medium +10% Fetal Bovine Serum (FBS) +1% diabody (mixed solution of penicillin and streptomycin)) is adopted, and the blank group is not added with leaching liquor and cell suspension and is placed in the same environment as the experimental group and the control group at the same time to be used as a reference when the experimental group and the control group measure the absorbance values.
The hemostatic bone wax of example 1 is adopted to prepare leaching liquor, the leaching temperature is (37 +/-1) DEG C, the leaching time is (24 +/-2) h, and 100%, 75% and 25% leaching liquor with concentration gradient is obtained. Then, the L929 cells were resuspended using a complete medium to a concentration of 4X 104Individual cells/mL of cell suspension. Taking a 96-well plate as a culture plate, wherein the experimental group and the control group are obtained by adding the cell suspension into the culture plate, and adding 100 mu L of the cell suspension into each well; blank set is plate with 100. mu.L of complete medium per well and no cell suspension added.
The plates of the experimental, control and blank groups were pre-incubated in an incubator for 24 hours (at 37 ℃ C., 5% CO)2Under conditions). Adding the leaching solution with the above concentration gradient into experimental group, adding 100 μ L per well, adding no leaching solution into control group and blank group, and culturingThe plates were incubated in an incubator for 24 hours, 72 hours (5% CO at 37 ℃)2Under conditions). The culture medium was then carefully aspirated from the wells, 100. mu.L of CCK-8 mixture (prepared by mixing 90. mu.L of complete medium + 10. mu.L of CCK-8) was added to each well, and the plates were incubated in an incubator for 2 hours. The absorbance at 450nm was then measured with a microplate reader.
FIGS. 3 and 4 show absorbance values (OD values) of the control and experimental groups, respectively, minus the blank group in the cytotoxicity test. As can be seen from the figure, the cell proliferation rate after the culture by the leaching liquor is close to that of the control group, so the hemostatic bone wax has good safety and no toxic or side effect on the cell growth.
(6) Plugging effectiveness test
The plugging effectiveness is proved by an in-vitro compression-resistant blasting test experiment, and the specific experimental method comprises the following steps: preparing an acrylic circular plate with a porous structure and a thickness of 1cm, wherein the circular plate can be hermetically fixed at a port of a pressure tester, the pressure tester is an acrylic box with a circular plate interface, the opposite side of the circular plate interface is connected with a pressure detection device and a pressure application device, after the sealed acrylic box is connected with the circular plate, the material is coated at the position of a circular plate hole, the whole hole is filled with the material, the circular plate is downward, the box is filled with water, and the pressure application device applies pressure and detects a leakage value.
(7) Hemostasis effectiveness test
After the experimental dog is anesthetized, the skin of the hind limb of the dog is incised by adopting the conventional aseptic operation to make bone defect bleeding. The experimental group was treated with the hemostatic bone wax of example 1, and the control group was not treated with hemostasis. The result experiment group can effectively stop bleeding within 20 seconds, has no bleeding phenomenon and has obvious hemostatic effect. Bleeding continued from the bleeding sites in the control group.
Claims (9)
1. An absorbable hemostatic bone wax is characterized in that the hemostatic bone wax is prepared from gelatin and poloxamer, wherein the mass ratio of the gelatin to the poloxamer is 1: 1-9: 1; the poloxamer is one or more of poloxamer 188, poloxamer 338 and poloxamer 407.
2. A method of preparing the resorbable hemostatic bone wax of claim 1, comprising the steps of:
s1, preparing a gelatin water solution;
s2, adding a cross-linking agent into the gelatin aqueous solution for cross-linking treatment;
s3, placing the crosslinked gelatin in water for full swelling;
s4, cutting the swelled gelatin in the step S3 into small blocks, and then crushing the small blocks into micron-sized gelatin particles;
s5, freezing the gelatin particles at a low temperature, and freeze-drying after complete freezing to obtain dried gelatin;
s6, carrying out secondary crushing on the frozen and dried gelatin;
s7, placing the gelatin obtained in the step S6 after the secondary crushing into a poloxamer solution for full swelling;
and S8, freeze-drying the gelatin swelled in the poloxamer solution in the step S7.
3. The method for preparing absorbable hemostatic bone wax of claim 2, wherein the amount of the cross-linking agent used in step S2 is 4-20% by mass of gelatin.
4. The method for preparing absorbable hemostatic bone wax of claim 2, wherein the gelatin is pulverized into particles of 0.1 μm to 2 μm in step S6.
5. The method for preparing the absorbable hemostatic bone wax of claim 2, wherein the mass concentration of the poloxamer solution in the S7 is 10% -50%, and the poloxamer is poloxamer 407.
6. The method of preparing an absorbable hemostatic bone wax of claim 2, further comprising the steps of: and S9, crushing the freeze-dried product obtained in the step S8 for the third time.
7. The method of preparing an absorbable hemostatic bone wax of claim 2, further comprising radiation sterilizing the resulting hemostatic bone wax.
8. The method for preparing an absorbable hemostatic bone wax of any one of claims 2-7, further comprising:
in the step S2, the cross-linking agent is water-soluble carbodiimide, the mass of the water-soluble carbodiimide is 4-20% of that of gelatin, the water-soluble carbodiimide is added in the process of stirring the gelatin solution, the stirring speed of the gelatin solution is 100-5000 r/min, and the adding speed of the water-soluble carbodiimide is 0.1-10 mL/S; between step S4 and step S5, the resulting gelatin particles are subjected to an ultrasonic-centrifugal circulation treatment in pure water to remove the crosslinking agent.
9. A hemostatic article comprising the absorbable hemostatic bone wax of claim 1, or the absorbable hemostatic bone wax obtained by the method of any one of claims 2 to 8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810123352.4A CN108187128B (en) | 2018-02-07 | 2018-02-07 | Absorbable hemostatic bone wax and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810123352.4A CN108187128B (en) | 2018-02-07 | 2018-02-07 | Absorbable hemostatic bone wax and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108187128A CN108187128A (en) | 2018-06-22 |
| CN108187128B true CN108187128B (en) | 2021-08-31 |
Family
ID=62593265
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810123352.4A Active CN108187128B (en) | 2018-02-07 | 2018-02-07 | Absorbable hemostatic bone wax and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108187128B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109172858A (en) * | 2018-11-23 | 2019-01-11 | 广州新诚生物科技有限公司 | Absorbable hemostasis bone wax of one kind and preparation method thereof |
| CN109453419A (en) * | 2018-11-26 | 2019-03-12 | 广州新诚生物科技有限公司 | Absorbable hemostasis bone wax of one kind and preparation method thereof |
| CN109200332A (en) * | 2018-11-26 | 2019-01-15 | 广州新诚生物科技有限公司 | A kind of biodegradable hemostasis bone wax and preparation method thereof |
| CN109908397A (en) * | 2019-03-01 | 2019-06-21 | 广州锐澄医疗技术有限公司 | A kind of absorbable hemostatic bone wax and preparation method thereof |
| CN109893677A (en) * | 2019-03-01 | 2019-06-18 | 广州锐澄医疗技术有限公司 | A kind of absorbable bone wax and preparation method thereof |
| CN115887741B (en) * | 2022-11-10 | 2024-03-19 | 东莞博捷生物科技有限公司 | Absorbable bone wax and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101161298A (en) * | 2007-11-20 | 2008-04-16 | 郑隆泗 | A technique for preparing absorbable gelatin sponge particle suppository with glutin as raw material |
| WO2009123903A1 (en) * | 2008-04-03 | 2009-10-08 | Zymogenetics, Inc. | Hemostatic microspheres |
| CN104353106A (en) * | 2014-11-05 | 2015-02-18 | 张自强 | Composition of fast hemostatic sponge gel and preparation method of composition |
| CN105688269A (en) * | 2016-02-03 | 2016-06-22 | 广州迈普再生医学科技有限公司 | Modified gelatin hemostatic material and preparation method thereof |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4439420A (en) * | 1982-11-16 | 1984-03-27 | Ethicon, Inc. | Absorbable hemostatic composition |
| KR101100045B1 (en) * | 2003-09-23 | 2011-12-29 | 오소콘 인코퍼레이티드 | Absorbable implants and methods for their use in hemostasis and in the treatment of osseous defects |
| RU2013156437A (en) * | 2011-06-07 | 2015-07-20 | СПАЙ Груп Лтд. | COMPOSITION AND METHODS FOR IMPROVING STABILITY AND EXTENDING THE PERIOD OF STORAGE OF SENSITIVE FOOD ADDITIVES AND FOOD PRODUCTS FROM THEM |
| MX346958B (en) * | 2011-10-11 | 2017-04-06 | Baxter Int | Hemostatic compositions. |
| CN106310348B (en) * | 2015-07-10 | 2020-09-01 | 山东美泰医药有限公司 | Starch/poloxamer composite styptic powder and preparation method thereof |
| CN106581737B (en) * | 2017-01-06 | 2019-09-06 | 哈尔滨鼓润生物技术有限公司 | A kind of hemostatic composition and preparation method thereof |
-
2018
- 2018-02-07 CN CN201810123352.4A patent/CN108187128B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101161298A (en) * | 2007-11-20 | 2008-04-16 | 郑隆泗 | A technique for preparing absorbable gelatin sponge particle suppository with glutin as raw material |
| WO2009123903A1 (en) * | 2008-04-03 | 2009-10-08 | Zymogenetics, Inc. | Hemostatic microspheres |
| CN104353106A (en) * | 2014-11-05 | 2015-02-18 | 张自强 | Composition of fast hemostatic sponge gel and preparation method of composition |
| CN105688269A (en) * | 2016-02-03 | 2016-06-22 | 广州迈普再生医学科技有限公司 | Modified gelatin hemostatic material and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| Gelatine Based ‘Bone Wax’;Katrin Van Loock等;《NEUROSURGERY》;20100801;第67卷(第2期);第E521页左栏第2-3段 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108187128A (en) | 2018-06-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108187128B (en) | Absorbable hemostatic bone wax and preparation method thereof | |
| Huang et al. | Using absorbable chitosan hemostatic sponges as a promising surgical dressing | |
| Maver et al. | Functional wound dressing materials with highly tunable drug release properties | |
| CA1146469A (en) | Lyophilized hydrocolloid foam | |
| Quan et al. | Black hemostatic sponge based on facile prepared cross-linked graphene | |
| KR20190004735A (en) | MEDICAL HYDROGEN GEL COMPOSITION, MEDICAL HYDRAULIC GEL, METHOD FOR PRODUCING THE SAME, AND USES | |
| CN108310448B (en) | Preparation method of fluid gelatin hemostatic material | |
| CN108187129B (en) | Absorbable gelatin styptic powder and preparation method thereof | |
| Kazimierczak et al. | Novel synthesis method combining a foaming agent with freeze-drying to obtain hybrid highly macroporous bone scaffolds | |
| JPS6134829B2 (en) | ||
| WO2006012218A1 (en) | Aluminophosphate-based materials for the treatment of wounds | |
| CN108530671B (en) | Porous gelatin sponge and preparation method thereof | |
| WO2016176186A1 (en) | Hemostatic composition and device | |
| WO2011106646A1 (en) | Recycling of superabsorbent polymer particles | |
| CN113633817A (en) | In-situ polymerization strongly-adhered antibacterial hemostatic hydrogel and preparation method and application thereof | |
| Echizenya et al. | Mineralization and biodegradation of CSF shunting systems | |
| Jungprasertchai et al. | Freeze-dried carboxymethyl chitosan/starch foam for use as a haemostatic wound dressing | |
| US8766032B2 (en) | Recycled superabsorbent polymer particles | |
| Liu et al. | Studies of silk fibroin/poly (lactic-co-glycolic acid) scaffold, prepared by thermally induced phase separation, as a possible wound dressing | |
| Wang et al. | A contact-polymerizable hemostatic powder for rapid hemostasis | |
| Linder et al. | Acute Local Tissue Effects of Polymerizing Acrylic Bone Cement: An Intravital Microscopic Study in the Hamster's Cheek Pouch on the Chemically Induced Microvascular Changes. | |
| Nguyen et al. | Novel gelatin–graphene oxide crosslinking induced by nonthermal atmospheric pressure plasma for alendronate delivery system | |
| Singh et al. | Radiation formation of functionalized polysaccharide-protein based skin mimicking semi-inter penetrating network for biomedical application | |
| Caili et al. | Antibacterial microspheres with a bionic red-blood-cell like hollow structure and superior swelling recovery capacity for efficient traumatic hemostasis | |
| CN107261199A (en) | A kind of preparation method of the gel dressing containing Zeolite hemostatic |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: 510663 Guangdong Guangzhou high tech Industrial Development Zone, Science City, 80 E third district. Applicant after: Guangzhou Maple regenerative medicine Polytron Technologies Inc Address before: 510663 Guangzhou District, Guangdong, new high tech Industrial Development Zone, Science City, 80 E third district. Applicant before: Medprin Regenerative Medical Technologies Co., Ltd. |
|
| CB02 | Change of applicant information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |