Method for preparing high-concentration collagen implant based on high-speed vacuum homogenization technology

Abstract

The invention discloses a method for preparing a high-concentration collagen implant based on a high-speed vacuum homogenization technology, and the method comprises the following steps: S1, preparing a collagen solution: putting collagen and a hydrochloric acid solution into a homogenizer, and uniformly mixing and stirring under a vacuum condition to obtain the collagen solution; s2, preparing an alkali solution: uniformly mixing disodium hydrogen phosphate, sodium hydroxide, sodium chloride, glucose, mannitol and water for injection to obtain the alkali solution; s3, mixing: adding the alkali solution into the collagen solution in batches, then putting the collagen solution into a homogenizer, uniformly mixing and stirring under a vacuum condition, and canning to obtain the high-concentration collagen implant. The collagen implant prepared by the method disclosed by the invention is high in concentration, long in maintenance time, uniform in fiber, stable in pushing, capable of wiredrawing and stacking, good in stability, simple in preparation process, short in period and wide in market prospect.

Description

Technical Field

[0001] The invention relates to the technical field of biomedical materials, and in particular to a method for preparing a high-concentration collagen implant based on a high-speed vacuum homogenization technology. Background Art

[0002] With the rapid development of the current medical beauty industry, the scale of the medical beauty market is also growing continuously. In the medical beauty scene, injecting collagen products into the sunken skin tissue can not only play a supporting and filling role, but also induce the patient's own tissue reconstruction. The gradually generated new tissue and the surrounding normal skin work efficiently together to achieve the effect of local correction; at the same time, it can also play a role in hydrating, anti-aging, whitening, and firming the skin.

[0003] At present, collagen products on the market can be divided into two categories: animal-derived collagen and recombinant collagen. Among them, animal-derived collagen has higher biological activity, stronger mechanical properties, and longer maintenance time than recombinant collagen due to its triple helix structure and high molecular weight, and can achieve the effects of whitening and freckle concealing, nourishing and regenerating.

[0004] At present, the existing collagen hydrating injections generally require a lower concentration to meet the requirements of hydrating injection and fluidity. However, collagen with a lower concentration degrades quickly and has average effects on skin elasticity, radiance restoration and fine line repair. However, after increasing the concentration of collagen molecules, its fluidity will decrease and become uneven, resulting in a large pushing force, which is difficult to meet market application needs.

[0005] Chinese patent application 202210928176.8 discloses a natural undenatured collagen solution for injection and a preparation method thereof. The collagen content in the collagen solution is low, only 2 to 6 mg / ml, so the elasticity and gloss restoration effect on the skin and the repair effect of fine lines are average. Chinese patent application 202410165029.9 discloses collagen fibers for water light needles and their preparation and application. Its preparation process includes low-concentration collagen pH adjustment, sodium chloride solution treatment, fiber induction treatment for 6 hours, centrifugal concentration, washing, homogenization, central control detection of collagen concentration, dilution to target concentration, canning and other steps. It has the problems of complex process, long cycle and high risk of contamination. Summary of the invention

[0006] The purpose of the present invention is to provide a method for preparing a high-concentration collagen implant based on a high-speed vacuum homogenization technology to solve the problems existing in the above-mentioned prior art. The collagen implant prepared by the method has high concentration, long maintenance time, uniform fibers, stable pushing, can be drawn and stacked, has good stability, and has a simple preparation process and a short cycle, and has broad market prospects, thereby solving the above-mentioned problems.

[0007] To achieve the above object, the present invention is implemented by the following technical solutions:

[0008] The present invention provides a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology, the method comprising the following steps:

[0009] S1. preparing a collagen solution: placing collagen and a hydrochloric acid solution in a homogenizer and mixing them evenly under vacuum conditions to obtain a collagen solution;

[0010] S2. preparing an alkaline solution: uniformly mixing disodium hydrogen phosphate, sodium hydroxide, sodium chloride, glucose, mannitol and water for injection to obtain an alkaline solution;

[0011] S3, mixing: adding the alkaline solution to the collagen solution in portions, and then placing the solution in a homogenizer and mixing and stirring the solution evenly under vacuum conditions, and canning the solution to obtain a high-concentration collagen implant.

[0012] Further, a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology: the pH value of the collagen solution in step S1 is 2.0 to 4.0;

[0013] In step S1, the revolution speed of the homogenizer is 100 to 1000 rpm, the rotation speed is 100 to 1000 rpm, the vacuum degree is -100 to -80 kPa, and the stirring time is 3 to 20 minutes.

[0014] Furthermore, a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology: the collagen content in the collagen solution is 5.0-15.0wt%, and the balance is hydrochloric acid.

[0015] Furthermore, a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology: the pH value of the alkaline solution in step S2 is 10.5-12.5.

[0016] Furthermore, a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology is provided: the alkaline solution comprises the following components in mass fractions: 2.0-8.0wt% disodium hydrogen phosphate, 0.05-1.0wt% sodium hydroxide, 0.5-3.0wt% sodium chloride, 0-4.0wt% glucose, 0-4.0wt% mannitol, and the balance water for injection.

[0017] Furthermore, a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology: in step S3, the weight ratio of the collagen solution to the alkaline solution is 1:(0.1-0.5).

[0018] Furthermore, a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology: the pH of the high-concentration collagen implant prepared in step S3 is 6.5-8.0, the osmotic pressure is 270-350mOsmol / kg, the concentration of collagen in the high-concentration collagen implant is 30-120mg / ml, and the length of the collagen fiber is 10-50μm.

[0019] Furthermore, a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology: in step S3, the revolution speed of the homogenizer is 500-800 rpm, the rotation speed is 300-600 rpm, the vacuum degree is -100-80 kPa, and the stirring time is 5-15 minutes.

[0020] Beneficial effects of the present invention:

[0021] (1) The method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology provided by the present invention adopts high-speed vacuum homogenization technology to ensure the fast swelling rate and uniformity of the high-concentration collagen raw material. After adding alkali solution, the material is refined by high-speed shearing technology to achieve a highly uniform dispersion state, uniform fiber length and good stability. At the same time, a certain vacuum degree is maintained throughout the process, which can efficiently remove a large number of bubbles generated during the mixing process.

[0022] (2) The method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology provided by the present invention has a high collagen concentration in the prepared collagen implant, which can reach 30 to 120 mg / ml, and can be maintained for 2 to 6 months. The fibers are uniform, the pushing is stable, the wire drawing and stacking are possible, the stability is strong, and the preparation process is simple, the cycle is short, and the sterility assurance level is high, which has broad market prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative work.

[0024] Figure 1 The pushing force test results of the collagen implants prepared in Examples 1 to 3 of the present invention at 0 month;

[0025] Figure 2 This is a microscopic image of collagen fibers in the collagen implant prepared in Example 1;

[0026] Figure 3 This is a microscopic image of collagen fibers in the collagen implant prepared in Example 2;

[0027] Figure 4 This is a microscopic image of collagen fibers in the collagen implant prepared in Example 3. DETAILED DESCRIPTION

[0028] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. The following description of at least one exemplary embodiment is actually only illustrative and is by no means intended to limit the present invention and its application or use. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] Example 1

[0030] This embodiment 1 provides a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology, the method comprising the following steps:

[0031] S1. preparing a collagen solution: placing collagen and a hydrochloric acid solution in a homogenizer stirring tank and mixing and stirring them evenly under vacuum conditions to obtain a collagen solution;

[0032] The collagen solution contains 5.0wt% collagen and 95.0wt% hydrochloric acid, and the pH of the collagen solution is about 3.0; the revolution speed of the homogenizer is 800rpm, the rotation speed is 500rpm, the homogenization time is 15 minutes, and the vacuum degree is -95kPa;

[0033] S2. preparing an alkaline solution: uniformly mixing 6.6 wt % of disodium hydrogen phosphate, 0.25 wt % of sodium hydroxide, 1.85 wt % of sodium chloride, and the remainder of water for injection, thereby obtaining an alkaline solution having a pH of about 11.0;

[0034] S3, mixing: adding the prepared alkaline solution to the collagen solution in full at one time, and then placing in a homogenizer and mixing and stirring evenly under vacuum conditions, and canning to obtain a high-concentration collagen implant, wherein the concentration of collagen is 35 mg / ml;

[0035] The weight ratio of the alkaline solution to the collagen solution is 0.32:1; in step S3, the revolution speed of the homogenizer is 600 rpm, the rotation speed is 500 rpm, the stirring time is 15 minutes, and the vacuum degree is -95 kPa.

[0036] Example 2

[0037] This embodiment 2 provides a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology, the method comprising the following steps:

[0038] S1. preparing a collagen solution: placing collagen and a hydrochloric acid solution in a homogenizer stirring tank and mixing and stirring them evenly under vacuum conditions to obtain a collagen solution;

[0039] The collagen solution contains 12.0 wt% collagen and 88.0 wt% hydrochloric acid, and the pH of the collagen solution is about 3.5; in step S1, the revolution speed of the homogenizer is 350 rpm, the rotation speed is 200 rpm, the homogenization time is 10 minutes, and the vacuum degree is -80 kPa;

[0040] S2. preparing an alkaline solution: uniformly mixing 4.0 wt % of disodium hydrogen phosphate, 0.1 wt % of sodium hydroxide, 1.3 wt % of sodium chloride, 4.0 wt % of mannitol, and the remainder of water for injection, thereby obtaining an alkaline solution;

[0041] S3, mixing: adding the prepared alkaline solution into the collagen solution twice, and then placing in a homogenizer and mixing and stirring evenly under vacuum conditions, and canning to obtain a high-concentration collagen implant, wherein the concentration of collagen is 80 mg / ml;

[0042] The weight ratio of the alkaline solution to the collagen solution is 0.45:1; in step S3, the revolution speed of the homogenizer is 500 rpm, the rotation speed is 300 rpm, the stirring time is 10 minutes, and the vacuum degree is -80 kPa.

[0043] Example 3

[0044] This embodiment 3 provides a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology, the method comprising the following steps:

[0045] S1. preparing a collagen solution: placing collagen and a hydrochloric acid solution in a homogenizer stirring tank and mixing and stirring them evenly under vacuum conditions to obtain a collagen solution;

[0046] The collagen solution contains 8.0 wt% collagen and 92.0 wt% hydrochloric acid, and the pH of the collagen solution is about 2.5; in step S1, the revolution speed of the homogenizer is 900 rpm, the rotation speed is 600 rpm, the homogenization time is 15 minutes, and the vacuum degree is -100 kPa;

[0047] S2. preparing an alkaline solution: uniformly mixing 7.5 wt % of disodium hydrogen phosphate, 0.88 wt % of sodium hydroxide, 0.5 wt % of sodium chloride, 3.0 wt % of glucose, 0.5 wt % of mannitol, and the remainder of water for injection, thereby obtaining an alkaline solution;

[0048] S3, mixing: adding the prepared alkaline solution into the collagen solution three times, and then placing in a homogenizer and mixing and stirring evenly under vacuum conditions, and canning to obtain a high-concentration collagen implant, wherein the concentration of collagen is 60 mg / ml;

[0049] The weight ratio of the alkaline solution to the collagen solution is 0.29:1; in step S3, the revolution speed of the homogenizer is 800 rpm, the rotation speed is 600 rpm, the stirring time is 7 minutes, and the vacuum degree is -100 kPa.

[0050] test:

[0051] (1) The pH and osmotic pressure of the collagen implants prepared in Examples 1 to 3 were tested using a pH meter and an osmotic pressure detector; a 27G needle was used to test the pushing force at a speed of 30 mm / min. The collagen implants prepared in Examples 1 to 3 were then stored at 2 to 8° C. for 1 month, 3 months, and 6 months, and then the pH, osmotic pressure, and pushing force were tested. The results are shown in Tables 1 and Figure 1 As shown:

[0052] Table 1 shows the pH, osmotic pressure and pushing force of the collagen implants obtained in Examples 1 to 3

[0053]

[0054] From Table 1 and Figure 1 It can be seen that the pH (standard: 6.5-8.0) and osmotic pressure (standard: 270-350 mOsmol / kg) of the collagen implants prepared in Examples 1-3 of the present invention meet the requirements, the pushing force is less than 10 N, and it is relatively stable after being stored at 2-8° C. for 6 months, indicating that it has good stability.

[0055] (2) The collagen implants prepared in Examples 1 to 3 were diluted to 0.5 mg / ml, stirred evenly, and the collagen fibers were observed using an inverted microscope. The results were as follows: Figures 2 to 4 shown by Figures 2 to 4 It can be seen that the length of the collagen fibers in the collagen implants prepared in Examples 1 to 3 is 10 to 50 μm and is evenly distributed.

[0056] In summary, the method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology provided by the present invention has a simple preparation process and a short cycle. The prepared collagen implant has a high collagen concentration and uniform fibers. The collagen fibers are 10 to 50 μm long and are milky white in a canned prefilled syringe. The implant is stable when pushed with a pushing force of less than 10 N. The implant can be drawn and stacked, and has good stability.

[0057] The above are preferred embodiments of the present invention and are only used to explain the present invention, not to limit the present invention. Any obvious changes or modifications derived from the technical solution of the present invention are still within the protection scope of the present invention.

Claims

1. A method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology, characterized in that: The method comprises the following steps: S1. preparing a collagen solution: placing collagen and a hydrochloric acid solution in a homogenizer and mixing them evenly under vacuum conditions to obtain a collagen solution; S2. preparing an alkaline solution: uniformly mixing disodium hydrogen phosphate, sodium hydroxide, sodium chloride, glucose, mannitol and water for injection to obtain an alkaline solution; S3, mixing: adding the alkaline solution to the collagen solution in portions, and then placing the solution in a homogenizer and mixing and stirring the solution evenly under vacuum conditions, and canning the solution to obtain a high-concentration collagen implant.

2. The method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology according to claim 1, characterized in that: The pH value of the collagen solution in step S1 is 2.0-4.0; In step S1, the revolution speed of the homogenizer is 100 to 1000 rpm, the rotation speed is 100 to 1000 rpm, the vacuum degree is -100 to -80 kPa, and the stirring time is 3 to 20 minutes.

3. The method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology according to claim 1 or 2, characterized in that: The content of collagen in the collagen solution is 5.0-15.0 wt %, and the balance is hydrochloric acid.

4. The method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology according to claim 1, characterized in that: The pH value of the alkaline solution in step S2 is 10.5-12.

5.

5. The method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology according to claim 1 or 4, characterized in that: The alkaline solution comprises the following components in mass fractions: 2.0-8.0wt% of disodium hydrogen phosphate, 0.05-1.0wt% of sodium hydroxide, 0.5-3.0wt% of sodium chloride, 0-4.0wt% of glucose, 0-4.0wt% of mannitol, and the balance of water for injection.

6. The method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology according to claim 1, characterized in that: In step S3, the weight ratio of the collagen solution to the alkaline solution is 1:(0.1-0.5).

7. The method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology according to claim 1, characterized in that: The pH of the high-concentration collagen implant prepared in step S3 is 6.5-8.0, and the osmotic pressure is 270-350 mOsmol / kg. The concentration of collagen in the high-concentration collagen implant is 30-120 mg / ml, and the length of the collagen fiber is 10-50 μm.

8. The method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology according to claim 1, characterized in that: In step S3, the revolution speed of the homogenizer is 500 to 800 rpm, the rotation speed is 300 to 600 rpm, the vacuum degree is -100 to -80 kPa, and the stirring time is 5 to 15 minutes.

9. The method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology according to claim 1, characterized in that: Step S3: adding the alkaline solution into the collagen solution in 1 to 3 times.

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