CN114874468A - Hyaluronic acid composite gel for filling lacrimal canals and preparation method thereof - Google Patents
Hyaluronic acid composite gel for filling lacrimal canals and preparation method thereof Download PDFInfo
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Abstract
The invention provides a hyaluronic acid composite gel for filling lacrimal canals and a preparation process thereof, wherein the composite gel is prepared by crosslinking medium-molecular-weight sodium hyaluronate, low-molecular-weight sodium hyaluronate and recombinant collagen, the mass percentages of the medium-molecular-weight sodium hyaluronate and the low-molecular-weight sodium hyaluronate in the composite gel are respectively 1.2-2.0%, the mass percentage of the recombinant collagen in the composite gel is 0.4-0.7%, and the balance of the composite gel is phosphate buffer solution. The preparation method comprises the steps of crosslinking through a physical means of rapid heating, heat preservation and rapid cooling. The composite gel provided by the invention has the advantages that through the matching of the sodium hyaluronate with medium molecular weight, the sodium hyaluronate with low molecular weight and the recombinant collagen, the filling effect is ensured, meanwhile, the duration of the filling effect can be prolonged, the tyndall effect is effectively avoided, the safety is high, and the skin nutrition can be repaired and supplemented; the physical crosslinking method of rapid heating-heat preservation-rapid cooling does not need a crosslinking agent and has no residual risk.
Description
Technical Field
The invention relates to the technical field of high polymer materials, in particular to a preparation method of hyaluronic acid composite gel and the hyaluronic acid composite gel prepared by the same.
Background
With the improvement of living standard, the pursuit of beauty is gradually improved, and filling products aiming at wrinkles on the face and the neck become pets of the market. The products for filling and repairing lacrimal canals on the market at present mainly comprise cross-linked sodium hyaluronate gel and sodium hyaluronate composite solution containing amino acid and other nutrient substances. The cross-linked sodium hyaluronate gel has a certain risk in lacrimal duct filling, and has very high requirements on injection manipulation of doctors due to the fact that the skin of an injection part is thin, the injection doctors cannot control the injection depth, angle, force, dosage and injection method accurately, or the anatomical structure of lacrimal ducts is unclear, so that poor injection effect is caused, the lacrimal duct part is bluish, a raised tyndall effect is generated at the lacrimal duct part when the injection is too shallow, and the lacrimal duct cannot be supported when the injection is too deep, so that the effect is not good. The sodium hyaluronate compound solution has less risk, but the selected sodium hyaluronate has low molecular weight, low sodium hyaluronate content in the solution and the sodium hyaluronate exists in a free form, so that the sodium hyaluronate compound solution has good instant effect and nutrition supplement effect during injection, has poor continuous effect and is degraded within a few days. In addition, a chemical crosslinking method is often adopted in the preparation process of the gel, and the safety of the gel is affected by the residue of the chemical crosslinking agent.
Chinese patent CN110016152A provides a method for preparing sodium hyaluronate gel for cross-linking filling, which comprises dissolving sodium hyaluronate gel in sodium hydroxide solution, adding cross-linking agent 1, 4-butanediol diglycidyl ether for cross-linking reaction, swelling to obtain sodium hyaluronate gel, dehydrating in organic solvent, and repeating for several times to remove cross-linking agent. The preparation process of the patent is complex, an additional step is needed to remove the cross-linking agent, the molecular weight of the hyaluronic acid is not described, and the problems of difficult injection, excessive metabolism and the like are inevitable.
Therefore, the development of a lacrimal passage filling gel which is convenient for injection, long in maintenance time and high in safety is an urgent technical problem to be solved in the field.
Disclosure of Invention
In view of this, the present invention provides a hyaluronic acid complex gel and a preparation method thereof.
The technical scheme of the invention is as follows:
a hyaluronic acid composite gel for filling lacrimal canals is prepared by cross-linking sodium hyaluronate with medium molecular weight, sodium hyaluronate with low molecular weight and recombinant collagen;
wherein the mass percentages of the medium molecular weight sodium hyaluronate and the low molecular weight sodium hyaluronate in the composite gel are both 1.2-2.0%; the mass percentage of the recombinant collagen in the composite gel is 0.4-0.7%; the balance of the composite gel is phosphate buffer solution.
Further, the mass ratio of the medium molecular weight sodium hyaluronate, the low molecular weight sodium hyaluronate and the recombinant collagen in the hyaluronic acid composite gel is 3:3: 1.
Further, in the hyaluronic acid composite gel, the mass percentages of the medium-molecular-weight sodium hyaluronate and the low-molecular-weight sodium hyaluronate in the composite gel are both 1.6%; the mass percentage of the recombinant collagen in the composite gel is 0.5 percent; the balance of the composite gel is phosphate buffer solution.
Furthermore, the molecular weight of the medium molecular weight sodium hyaluronate is 1300kDa of 1000-.
The invention also provides a preparation method of the hyaluronic acid composite gel, which comprises the following steps:
s1, mixing and dispersing medium-molecular-weight sodium hyaluronate, low-molecular-weight sodium hyaluronate and recombinant collagen into a phosphate buffer solution, stirring the mixed solution, and simultaneously adjusting the pH of the mixed solution until a homogeneous viscous solution is formed;
s2, stirring and degassing the viscous solution obtained in the step S1 in vacuum until no obvious bubbles exist in the viscous solution, and filling the viscous solution into a pre-filled syringe;
and S3, carrying out rapid heating-heat preservation-rapid cooling operation on the filled pre-filled syringe obtained in the step S2 to crosslink the medium molecular weight sodium hyaluronate, the low molecular weight sodium hyaluronate and the recombinant collagen.
Further, the above preparation method, S1, is carried out at 20-35 deg.C, and the pH of the viscous solution obtained after pH adjustment is 6-8.
Further, in the above preparation method, the step of adjusting pH in S1 is as follows:
measuring the pH value of the mixed solution;
if the pH value is less than 6, adding 0.1mol/L sodium hydroxide solution into the mixed solution until the pH value of the mixed solution is more than or equal to 6;
if the pH value is more than 8, adding 0.1mol/L hydrochloric acid solution into the mixed solution until the pH value of the mixed solution is less than or equal to 8.
Further, in the preparation method, when the vacuum stirring and degassing of S2 are carried out, the vacuum degree of a vacuum stirrer is less than or equal to-900 mbar, and the stirring and degassing are carried out for 30-50 minutes at the rotating speed of 20 rpm.
Further, in the preparation method, the steps of S3 rapid heating-heat preservation-rapid cooling are as follows:
heating the steam-air mixed gas medium, pressurizing and heating to ensure that the filled pre-encapsulation injector obtained in the step S3 is heated to 110-130 ℃ within 10 minutes;
preserving the temperature of the prefilled syringe in the gas medium for 5-10 minutes;
after the heat preservation is finished, spraying chilled water to the pre-filled injector to ensure that the temperature is reduced to 20-35 ℃ within 10 minutes.
The invention has the following beneficial effects: the hyaluronic acid composite gel provided by the invention has the advantages that through the matching of the sodium hyaluronate with medium molecular weight, the sodium hyaluronate with low molecular weight and the recombinant collagen, the filling effect is ensured, the duration of the filling effect can be prolonged, the tyndall effect is effectively avoided, the safety is high, and the skin nutrition can be repaired and supplemented. According to the preparation method of the hyaluronic acid composite gel, the components are crosslinked by physical means of rapid heating, heat preservation and rapid cooling, a crosslinking agent is not needed, and no residual risk exists.
Drawings
The invention is further described with the aid of the accompanying drawings, in which the embodiments do not constitute any limitation of the invention, and in which further drawings may be derived by a person skilled in the art, without inventive effort, from the following figures.
FIG. 1 is a diagram showing a state in which hyaluronic acid complex gels prepared in examples 1 to 3 of the present invention and comparative examples 1 to 4 are left standing on a plastic plate;
FIG. 2 is a view showing the state of hyaluronic acid complex gels prepared in examples 1-3 and comparative examples 1-4 of the present invention on a plastic plate inclined at 30 °.
Detailed Description
The experimental methods used in the following examples are all conventional methods unless otherwise specified; the reagents, materials and the like used in the following examples are all commercially available products or can be prepared by known methods unless otherwise specified. The medium and low molecular weight sodium hyaluronate used in the examples below were purchased from Huaxi Biotech Inc.; the recombinant collagen was purchased from Jiangsu Jiangshan Convergence Biotech, Inc. Experimental procedures without specific conditions noted in the following examples, molecular cloning is generally performed according to conventional conditions such as Sambrook et al: the conditions described in the Laboratory Manual (New York: Cold spring harbor Laboratory Press,1989), or according to the manufacturer's recommendations.
Example 1
Mixing and dispersing 0.16g of sodium hyaluronate with the molecular weight of 1200kDa, 0.16g of sodium hyaluronate with the molecular weight of 250kDa and 0.05g of recombinant collagen with the molecular weight of 50kDa into Phosphate Buffer Solution (PBS) until the total weight is 10g, stirring and dissolving, meanwhile, measuring the pH value of the mixed solution to be 5.5, adding 0.1mol/L sodium hydroxide solution to adjust the pH value of the mixed solution to be 7, fully and uniformly dissolving the mixed solution until a clear homogeneous viscous solution is obtained, stirring and degassing in a vacuum stirrer at vacuum degree of-1000 mbar and rotation speed of 20rpm for 40 min, degassing to obtain bubble-free viscous solution, filling the degassed viscous solution into a pre-filled syringe under the conditions of negative pressure and sterility, heating a steam-air mixed medium under 110kPa to enable the filled pre-filled syringe to reach 125 ℃ within 8 minutes, preserving the temperature for 8min, and then spraying chilled water on the pre-filled syringe to enable the temperature to be reduced to 25 ℃ within 10 minutes.
Example 2
Mixing and dispersing 0.2g of sodium hyaluronate with molecular weight of 1300kDa, 0.2g of sodium hyaluronate with molecular weight of 500kDa and 0.07g of recombinant collagen with molecular weight of 60kDa into Phosphate Buffer Solution (PBS) until the total weight is 10g, stirring and dissolving, simultaneously measuring the pH value of the mixed solution to be 8.2, adding 0.1mol/L hydrochloric acid solution to adjust the pH value of the mixed solution to be 8, fully and uniformly dissolving the mixed solution until clear homogeneous viscous solution is obtained, stirring and degassing in a vacuum stirrer at vacuum degree of-1200 mbar and rotation speed of 20rpm for 50 min, degassing to obtain bubble-free viscous solution, filling the degassed viscous solution into a pre-encapsulation injector under the conditions of negative pressure and sterility, heating a steam-air mixed medium under 110kPa to enable the filled pre-encapsulation injector to reach 130 ℃ within 10 minutes, preserving the temperature for 10min, and then spraying chilled water to the pre-encapsulation injector to enable the pre-encapsulation injector to reduce the temperature to 25 ℃ within 10 minutes.
Example 3
Mixing 0.12g of sodium hyaluronate with molecular weight of 1000kDa, 0.12g of sodium hyaluronate with molecular weight of 200kDa and 0.04g of recombinant collagen with molecular weight of 40kDa, dispersing into Phosphate Buffer Solution (PBS) until the total weight is 10g, stirring for dissolving, meanwhile, measuring the pH value of the mixed solution to be 5.2, adding 0.1mol/L sodium hydroxide solution to adjust the pH value of the mixed solution to be 6, fully and uniformly dissolving the mixed solution until clear homogeneous viscous solution is obtained, stirring and degassing in a vacuum stirrer at vacuum degree of-900 mbar and rotation speed of 20rpm for 30 min, degassing to obtain bubble-free viscous solution, filling the degassed viscous solution into a pre-filled syringe under the conditions of negative pressure and sterility, heating a steam-air mixed medium under 110kPa to ensure that the filled pre-filled syringe reaches 110 ℃ within 5 minutes, preserving the temperature for 5 minutes, and then spraying chilled water on the pre-filled syringe to ensure that the temperature is reduced to 25 ℃ within 10 minutes.
Comparative example 1
Mixing and dispersing 0.32g of sodium hyaluronate with the molecular weight of 1200kDa and 0.06g of recombinant collagen with the molecular weight of 50kDa into Phosphate Buffer Solution (PBS) until the total weight is 10g, stirring and dissolving, measuring the pH value of the mixed solution to be 5.7, adding 0.1mol/L of sodium hydroxide solution to adjust the pH value of the mixed solution to be 7, fully and uniformly dissolving until clear homogeneous viscous solution is obtained, stirring and degassing in a vacuum stirrer at the vacuum degree of-1000 mbar and the rotating speed of 20rpm for 40 minutes, degassing until bubble-free viscous solution is obtained, filling the degassed viscous solution into a pre-filling syringe under the conditions of negative pressure and sterility, heating steam and air mixed medium under 110kPa to enable the filled pre-filling syringe to reach 125 ℃ within 8 minutes, preserving the temperature for 8 minutes, and spraying chilled water on the pre-filling syringe to enable the pre-filling syringe to be reduced to 25 ℃ within 10 minutes.
Comparative example 2
Mixing and dispersing 0.5g of sodium hyaluronate with molecular weight of 1200kDa, 0.5g of sodium hyaluronate with molecular weight of 250kDa and 0.15g of recombinant collagen with molecular weight of 50kDa into Phosphate Buffer Solution (PBS) until the total weight is 10g, stirring and dissolving, simultaneously measuring the pH value of the mixed solution to be 8.2, adding 0.1mol/L hydrochloric acid to adjust the pH value of the mixed solution to be 7, fully and uniformly dissolving the mixed solution until clear homogeneous viscous solution is obtained, stirring and degassing in a vacuum stirrer at vacuum degree of-1000 mbar and rotation speed of 20rpm for 40 min, degassing to obtain bubble-free viscous solution, filling the degassed viscous solution into a pre-filled syringe under the conditions of negative pressure and sterility, heating a steam-air mixed medium under 110kPa to enable the filled pre-filled syringe to reach 125 ℃ within 8 minutes, preserving the temperature for 8min, and then spraying chilled water on the pre-filled syringe to enable the temperature to be reduced to 25 ℃ within 10 minutes.
Comparative example 3
Mixing 0.32g of sodium hyaluronate with the molecular weight of 250kDa and 0.06g of recombinant collagen with the molecular weight of 50kDa, dispersing the mixture into Phosphate Buffer Solution (PBS) until the total weight is 10g, stirring and dissolving, measuring the pH value of the mixed solution to be 5.6, adding 0.1mol/L sodium hydroxide solution to adjust the pH value of the mixed solution to be 7, fully and uniformly dissolving until clear homogeneous viscous solution is obtained, stirring and degassing in a vacuum stirrer at the vacuum degree of-1000 mbar and the rotating speed of 20rpm for 40 minutes, degassing until bubble-free viscous solution is obtained, filling the degassed viscous solution into a pre-filling syringe under the conditions of negative pressure and sterility, heating steam and air mixed medium under 110kPa to enable the filled pre-filling syringe to reach 125 ℃ within 8 minutes, preserving the temperature for 8 minutes, and spraying chilled water on the pre-filling syringe to enable the pre-filling syringe to be reduced to 25 ℃ within 10 minutes.
Comparative example 4
Mixing and dispersing 0.1g of sodium hyaluronate with molecular weight of 1200kDa, 0.1g of sodium hyaluronate with molecular weight of 250kDa and 0.03g of recombinant collagen with molecular weight of 50kDa into Phosphate Buffer Solution (PBS) until the total weight is 10g, stirring and dissolving, simultaneously measuring the pH value of the mixed solution to be 8.4, adding 0.1mol/L hydrochloric acid to adjust the pH value of the mixed solution to be 7, fully and uniformly dissolving the mixed solution until clear homogeneous viscous solution is obtained, stirring and degassing in a vacuum stirrer at vacuum degree of-1000 mbar and rotation speed of 20rpm for 40 min, degassing to obtain bubble-free viscous solution, filling the degassed viscous solution into a pre-filled syringe under the conditions of negative pressure and sterility, heating a steam-air mixed medium under 110kPa to enable the filled pre-filled syringe to reach 125 ℃ within 8 minutes, preserving the temperature for 8min, and then spraying chilled water on the pre-filled syringe to enable the temperature to be reduced to 25 ℃ within 10 minutes.
Comparative example 5
Mixing and dispersing 0.16g of sodium hyaluronate with the molecular weight of 1200kDa, 0.16g of sodium hyaluronate with the molecular weight of 250kDa and 0.05g of recombinant collagen with the molecular weight of 50kDa into Phosphate Buffer Solution (PBS) until the total weight is 10g, stirring and dissolving, meanwhile, measuring the pH value of the mixed solution to be 5.7, adding 0.1mol/L sodium hydroxide solution to adjust the pH value of the mixed solution to be 7, fully and uniformly dissolving the mixed solution until a clear homogeneous viscous solution is obtained, stirring and degassing in a vacuum stirrer at vacuum degree of-1000 mbar and rotation speed of 20rpm for 40 min, degassing to obtain bubble-free viscous solution, filling the degassed viscous solution into a pre-filled syringe under the conditions of negative pressure and sterility, heating a steam-air mixed medium under 110kPa to ensure that the filled pre-filled syringe reaches 100 ℃ within 8 minutes, preserving the temperature for 8min, and then spraying chilled water on the pre-filled syringe to ensure that the temperature is reduced to 25 ℃ within 10 minutes.
Comparative example 6
Mixing and dispersing 0.16g of sodium hyaluronate with the molecular weight of 1200kDa, 0.16g of sodium hyaluronate with the molecular weight of 250kDa and 0.05g of recombinant collagen with the molecular weight of 50kDa into Phosphate Buffer Solution (PBS) until the total weight is 10g, stirring and dissolving, meanwhile, measuring the pH value of the mixed solution to be 5.7, adding 0.1mol/L sodium hydroxide solution to adjust the pH value of the mixed solution to be 7, fully and uniformly dissolving the mixed solution until a clear homogeneous viscous solution is obtained, stirring and degassing in a vacuum stirrer at vacuum degree of-1000 mbar and rotation speed of 20rpm for 40 min, degassing to obtain bubble-free viscous solution, filling the degassed viscous solution into a pre-filled syringe under the conditions of negative pressure and sterility, heating a steam-air mixed medium under 110kPa to enable the filled pre-filled syringe to reach 125 ℃ within 8 minutes, preserving the temperature for 3min, and then spraying chilled water on the pre-filled syringe to enable the temperature to be reduced to 25 ℃ within 10 minutes.
Performance test
(1) Appearance of the product
The hyaluronic acid complex gels prepared in examples 1 to 3 and comparative examples 1 to 4 were left standing on a plastic plate at room temperature, and the state of the complex gel when laid flat was as shown in FIG. 1; the state after tilting the plastic plate by 30 ° is shown in fig. 2.
TABLE 1 preparation of the samples
As shown in fig. 1 and 2, when the hyaluronic acid composite gels prepared in the examples and the comparative examples are statically placed on a plastic plate, the appearance characters are similar and are transparent liquid drops; after the plastic plate is inclined by 30 degrees, the hyaluronic acid composite gels prepared in the comparative examples 3 and 4 flow downwards, and the hyaluronic acid composite gel prepared in the comparative example 4 has higher fluidity than that of the comparative example 3; the hyaluronic acid composite gel prepared in example 3 also has a certain fluidity, but the fluidity is less than that of comparative examples 3 and 4; the hyaluronic acid complex gel obtained by other preparation methods does not flow. Comparative examples 5, 6 (not shown in the figure) also flow on plastic plates inclined at 30 °, to a similar extent as comparative example 4.
(2) Shear viscosity
At 25 ℃ using a rotational rheometer at shear rates from 0.001s -1 ~1000s -1 Flow scanning was performed down and 2.5s was taken -1 The shear viscosity versus shear rate is compared.
TABLE 2 shear viscosity of gels obtained by different preparation methods
The shear viscosity reflects the viscosity of the fluid. Examples 1-3 and comparative examples 1-2 had shear viscosities greater than 30 and comparative examples 3-6 had shear viscosities less than 20. The above appearance and shear viscosity test results show that, compared to examples 1 to 3 and comparative examples 1 and 2, comparative examples 3 to 6 are more likely to be displaced and absorbed after injection, and do not satisfy the requirement of maintaining a long-term filling effect.
(3) Modulus of elasticity
At 25 ℃, an oscillation test was performed at 5% strain using a rotational rheometer, and the elastic modulus values corresponding to a frequency of 1Hz were compared.
TABLE 3 elastic modulus of gels obtained by different preparation methods
The elastic modulus is an index for measuring the difficulty of the material in generating elastic deformation, and the larger the value is, the larger the rigidity of the material is; for gels, the greater the stiffness, the more resistant to degradation. Examples 1-3 and comparative examples 3-6 have an elastic modulus of less than 140Pa, whereas comparative examples 1, 2 have an elastic modulus exceeding 250 Pa. The test results show that compared with other preparation processes, the gels prepared in the comparative examples 1 and 2 are easy to have adverse reactions such as hard touch, Tyndall effect, difficult degradation and the like in use.
(4) Pushing force
Taking the prepared hyaluronic acid composite gel, pushing a syringe push rod at a constant speed, wherein the needle head of the syringe is 27G, the pushing speed is 30mm/min, and detecting the pushing force.
TABLE 4 extrusion force of gels obtained by different preparation processes
Preparation process | Pushing force (N) |
Comparative example 1 | 18.41 |
Comparative example 2 | 17.23 |
Example 1 | 10.09 |
Example 2 | 12.87 |
Example 3 | 8.21 |
Comparative example 3 | 6.65 |
Comparative example 4 | 2.68 |
Comparative example 5 | 4.56 |
Comparative example 6 | 3.47 |
The requirement of the surgical operation aptitude on the pushing force is less than 15N, the pushing force of the examples 1-3 and the comparative examples 3-6 is less than 15N, and the pushing force of the comparative examples 1 and 2 is more than 16N. The above results show that the gel obtained in comparative examples 1 and 2 had too high a pushing force to satisfy the requirement of the surgical operation.
(5) In vivo experiments
The hyaluronic acid complex gel prepared by the preparation methods of examples and comparative examples was injected into a seven-week-old male nude mouse, the appearance of the injection site was observed, the injection site was sacrificed 60 days later, the skin of the injection site was incised, the remaining gel was extracted, and the percentage of the remaining gel was measured by weighing.
TABLE 5 in vivo gel test results obtained by different preparation methods
On the day of injection, the appearance of the injection site of each group of mice is similar, and the mice all have bulges with the size of soybean grains; within one week after injection, subcutaneous projections disappeared from the mice of the comparative examples 3-6, which were smooth to the touch and free of a sense of projection. The subcutaneous bulges of the mice of the groups of comparative examples 1-2 and the mice of the groups of examples 1-3 are reduced to different degrees, the touch bulges, the texture of the bulges of the mice of the groups of comparative examples 1-2 is hard, and the texture of the bulges of the mice of the groups of examples 1-3 is soft and natural. The above phenomena show that the gel obtained in examples 1 to 3 has a better filling effect than the comparative example.
Although embodiments of the present application have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present application, and that changes, modifications, substitutions and alterations in the above embodiments may be made by those of ordinary skill in the art within the scope of the present application.
Claims (9)
1. A hyaluronic acid composite gel for lacrimal duct filling, which is prepared by cross-linking medium molecular weight sodium hyaluronate, low molecular weight sodium hyaluronate and recombinant collagen, and is characterized in that:
the mass percentages of the medium molecular weight sodium hyaluronate and the low molecular weight sodium hyaluronate in the composite gel are both 1.2-2.0%;
the mass percentage of the recombinant collagen in the composite gel is 0.4-0.7%;
the balance of the composite gel is phosphate buffer solution.
2. The hyaluronic acid composite gel of claim 1, wherein the mass ratio of the medium-molecular-weight sodium hyaluronate, the low-molecular-weight sodium hyaluronate and the recombinant collagen in the composite gel is 3:3: 1.
3. The hyaluronic acid complex gel of claim 1, wherein the mass percentages of the medium-molecular-weight sodium hyaluronate and the low-molecular-weight sodium hyaluronate in the complex gel are both 1.6%;
the mass percentage of the recombinant collagen in the composite gel is 0.5%;
the balance of the composite gel is phosphate buffer solution.
4. The hyaluronic acid composite gel of any of claims 1-3, wherein the molecular weight of the medium-molecular-weight sodium hyaluronate is 1000-1300kDa, the molecular weight of the low-molecular-weight sodium hyaluronate is 200-500kDa, and the molecular weight of the recombinant collagen is 40-60 kDa.
5. A method for preparing the hyaluronic acid complex gel of claim 1, comprising the steps of:
s1, mixing and dispersing medium-molecular-weight sodium hyaluronate, low-molecular-weight sodium hyaluronate and recombinant collagen into a phosphate buffer solution, stirring the mixed solution, and simultaneously adjusting the pH of the mixed solution until a homogeneous viscous solution is formed;
s2, stirring and degassing the viscous solution obtained in the step S1 in vacuum until no obvious bubbles exist in the viscous solution, and filling the viscous solution into a pre-filled syringe;
and S3, carrying out rapid heating-heat preservation-rapid cooling operation on the filled pre-filled syringe obtained in the step S2 to crosslink the medium molecular weight sodium hyaluronate, the low molecular weight sodium hyaluronate and the recombinant collagen.
6. The method according to claim 5, wherein S1 is carried out at 20-35 ℃, and the pH of the viscous solution obtained after the pH adjustment is 6-8.
7. The method according to claim 5 or 6, wherein the step of adjusting the pH in S1 is as follows: measuring the pH value of the mixed solution;
if the pH value is less than 6, adding 0.1mol/L sodium hydroxide solution into the mixed solution until the pH value of the mixed solution is more than or equal to 6;
if the pH value is more than 8, adding 0.1mol/L hydrochloric acid solution into the mixed solution until the pH value of the mixed solution is less than or equal to 8.
8. The preparation method of claim 5, wherein the vacuum stirring and degassing of S2 is performed at a vacuum degree of less than or equal to-900 mbar and a rotation speed of 20rpm for 30-50 min.
9. The preparation method according to claim 5, wherein the steps of S3 rapid heating-holding-rapid cooling are as follows:
pressurizing and heating the steam-air mixed gas medium to enable the filled pre-filling and sealing injector obtained in the step S3 to be heated to 110-130 ℃ within 10 minutes;
the prefilled syringe is kept warm in the gas medium for 5 to 10 minutes;
after the heat preservation is finished, spraying chilled water to the pre-filled injector to ensure that the pre-filled injector is cooled to 20-35 ℃ within 10 minutes.
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CN115721778A (en) * | 2022-12-15 | 2023-03-03 | 西安德诺海思医疗科技有限公司 | Collagen/hyaluronic acid composite gel for skin injection and preparation method thereof |
CN116019978A (en) * | 2023-02-09 | 2023-04-28 | 云南贝泰妮生物科技集团股份有限公司 | Micro-crosslinked sodium hyaluronate-recombinant collagen composite gel and preparation method and application thereof |
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Cited By (3)
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CN115721778A (en) * | 2022-12-15 | 2023-03-03 | 西安德诺海思医疗科技有限公司 | Collagen/hyaluronic acid composite gel for skin injection and preparation method thereof |
CN116019978A (en) * | 2023-02-09 | 2023-04-28 | 云南贝泰妮生物科技集团股份有限公司 | Micro-crosslinked sodium hyaluronate-recombinant collagen composite gel and preparation method and application thereof |
CN116019978B (en) * | 2023-02-09 | 2024-01-23 | 云南贝泰妮生物科技集团股份有限公司 | Micro-crosslinked sodium hyaluronate-recombinant collagen composite gel and preparation method and application thereof |
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