High-speed vacuum homogenization-based method for preparing high-concentration collagen implant

Abstract

Disclosed in the present invention is a high-speed vacuum homogenization-based method for preparing a high-concentration collagen implant. The method comprises the following steps: S1, preparing a collagen solution: placing collagen and a hydrochloric acid solution in a homogenizer, and uniformly mixing and stirring same under vacuum conditions to obtain a collagen solution; S2, preparing a base solution: uniformly mixing disodium hydrogen phosphate, sodium hydroxide, sodium chloride, glucose, mannitol, and water for injection to obtain a base solution; and S3, mixing: adding the base solution to the collagen solution in batches, then placing same in the homogenizer, uniformly mixing and stirring same under vacuum conditions, and performing filling, to prepare a high-concentration collagen implant. The collagen implant prepared using the method of the present invention has a high concentration, a long duration of action, and uniform fibers; the extrusion of the collagen implant is smooth; the collagen implant can be drawn and stacked; and the collagen implant has good stability and a simple preparation process, and requires a short preparation cycle. Thus, the collagen implant shows broad market prospects.

Description

A method for preparing high-concentration collagen implants based on high-speed vacuum homogenization technology Technical Field

[0001] This invention relates to the field of biomedical materials technology, specifically to a method for preparing high-concentration collagen implants based on high-speed vacuum homogenization technology. Background Technology

[0002] With the rapid development of the medical aesthetics industry, the market size of medical aesthetics has continued to grow. In medical aesthetic settings, injecting collagen products into depressed skin tissue can not only provide support and filling, but also induce the patient's own tissue reconstruction. The gradually generated new tissue works synergistically with the surrounding normal skin to achieve localized correction. At the same time, it can also moisturize, anti-age, whiten, and tighten the skin.

[0003] Currently, collagen products on the market can be divided into two categories: animal-derived collagen and recombinant collagen. Among them, animal-derived collagen, compared with recombinant collagen, has higher biological activity, stronger mechanical properties, and a longer duration of effect due to its triple helix structure and higher molecular weight. It can achieve effects such as whitening, concealing blemishes, nourishing and regenerating.

[0004] Currently, existing collagen-based mesotherapy products generally require a low concentration to meet the requirements for injection and fluidity. However, low-concentration collagen degrades quickly and has limited effects on skin elasticity, radiance restoration, and fine line repair. On the other hand, increasing the concentration of collagen molecules reduces fluidity and causes unevenness, resulting in greater pushing force and making it difficult to meet market application needs.

[0005] Chinese patent application 202210928176.8 discloses a natural, undenatured injectable collagen solution and its preparation method. This collagen solution has a low collagen content, only 2-6 mg / ml, therefore its effects on skin elasticity, radiance restoration, and fine line repair are generally limited. Chinese patent application 202410165029.9 discloses collagen fibers for mesotherapy and their preparation and application. Its preparation process includes steps such as low-concentration collagen pH adjustment, sodium chloride solution treatment, fiber induction treatment for 6 hours, centrifugation concentration, washing, homogenization, central control detection of collagen concentration, dilution to the target concentration, and bottling. This process is complex, time-consuming, and carries a high risk of contamination. Technical issues

[0006] The purpose of this invention is to address the problems existing in the prior art by providing a method for preparing high-concentration collagen implants based on high-speed vacuum homogenization technology. The collagen implants prepared by this method have high concentration, long maintenance time, uniform fibers, stable extrusion, can be drawn and stacked, have good stability, and the preparation process is simple and short, with broad market prospects, thus solving the above problems. Technical solutions

[0007] To achieve the above objectives, the present invention is implemented through the following technical solution:

[0008] This 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. Preparation of collagen solution: Place collagen and hydrochloric acid solution in a homogenizer and mix them evenly under vacuum to obtain collagen solution;

[0010] S2. Preparation of alkaline solution: Mix disodium hydrogen phosphate, sodium hydroxide, sodium chloride, glucose, mannitol and water for injection evenly to obtain an alkaline solution;

[0011] S3. Mixing: The alkaline solution is added to the collagen solution in portions, and then placed in a homogenizer and mixed and stirred evenly under vacuum conditions. After bottling, a high-concentration collagen implant is obtained.

[0012] Furthermore, 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] The homogenizer described in step S1 has a revolution speed of 100-1000 rpm, a rotation speed of 100-1000 rpm, a vacuum degree of -100--80 kPa, and a stirring time of 3-20 minutes.

[0014] Furthermore, a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology: the collagen solution contains 5.0–15.0 wt% collagen, with the remainder being 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 to 12.5.

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

[0017] Furthermore, a method for preparing a high-concentration collagen implant based on high-speed vacuum homogenization technology: the weight ratio of the collagen solution to the alkaline solution in step S3 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 high-concentration collagen implant prepared in step S3 has a pH of 6.5-8.0 and an osmotic pressure of 270-350 mOsmol / kg, and the collagen concentration in the high-concentration collagen implant is 30-120 mg / ml, and the length of the collagen fibers is 10-50 μm.

[0019] Furthermore, a method for preparing high-concentration collagen implants 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. Beneficial effects

[0020] (1) The method for preparing high-concentration collagen implants based on high-speed vacuum homogenization technology provided by the present invention adopts high-speed vacuum homogenization technology to ensure the high swelling rate and uniformity of high-concentration collagen raw materials. After adding alkaline solution, the material is refined by high-speed shearing technology to achieve a highly uniform dispersion state with uniform fiber length and good stability. At the same time, the whole process maintains a certain degree of vacuum, which can efficiently remove a large number of bubbles generated during the mixing process.

[0021] (2) The method for preparing high-concentration collagen implants based on high-speed vacuum homogenization technology provided by the present invention has a high concentration of collagen in the prepared collagen implants, which can reach 30 to 120 mg / ml, and achieve a maintenance time of 2 to 6 months. The fibers are uniform, stable when pushed, can be drawn and stacked, and have strong stability. Moreover, the preparation process is simple, the cycle is short, and the sterility level is high, which has broad market prospects. Attached Figure Description

[0022] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the following description of the embodiments will be briefly introduced. 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 effort.

[0023] Figure 1 shows the extrusion force test results of the collagen implants prepared in Examples 1-3 of the present invention at 0 months;

[0024] Figure 2 is a microscopic image of collagen fibers in the collagen implant prepared in Example 1;

[0025] Figure 3 is a microscopic image of collagen fibers in the collagen implant prepared in Example 2;

[0026] Figure 4 shows a microscopic image of collagen fibers in the collagen implant prepared in Example 3. Embodiments of the present invention

[0027] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the present invention or its application or use. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention. Example

[0028] 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:

[0029] S1. Preparation of collagen solution: Place collagen and hydrochloric acid solution in a homogenizer mixing tank and mix them evenly under vacuum to obtain collagen solution;

[0030] The collagen solution contains 5.0 wt% collagen and 95.0 wt% hydrochloric acid, and the pH of the collagen solution is approximately 3.0; the homogenizer has a revolution speed of 800 rpm, a rotation speed of 500 rpm, a homogenization time of 15 minutes, and a vacuum degree of -95 kPa.

[0031] S2. Preparation of alkaline solution: Mix 6.6 wt% disodium hydrogen phosphate, 0.25 wt% sodium hydroxide, 1.85 wt% sodium chloride, and the balance of water for injection evenly to obtain an alkaline solution with a pH of approximately 11.0.

[0032] S3. Mixing: Add the prepared alkaline solution to the collagen solution in one go, then place it in a homogenizer and mix and stir evenly under vacuum. After bottling, a high-concentration collagen implant is obtained, wherein the collagen concentration is 35mg / ml.

[0033] The weight ratio of the alkaline solution to the collagen solution is 0.32:1; in step S3, the homogenizer has a revolution speed of 600 rpm, a rotation speed of 500 rpm, a stirring time of 15 minutes, and a vacuum degree of -95 kPa. Example

[0034] 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:

[0035] S1. Preparation of collagen solution: Place collagen and hydrochloric acid solution in a homogenizer mixing tank and mix them evenly under vacuum to obtain collagen solution;

[0036] The collagen solution contains 12.0 wt% collagen and 88.0 wt% hydrochloric acid, and the pH of the collagen solution is approximately 3.5. In step S1, the homogenizer has a revolution speed of 350 rpm, a rotation speed of 200 rpm, a homogenization time of 10 minutes, and a vacuum degree of -80 kPa.

[0037] S2. Preparation of alkaline solution: Mix 4.0 wt% disodium hydrogen phosphate, 0.1 wt% sodium hydroxide, 1.3 wt% sodium chloride, 4.0 wt% mannitol, and the balance of water for injection evenly to obtain an alkaline solution.

[0038] S3. Mixing: The alkaline solution prepared above is added to the collagen solution in two portions, and then placed in a homogenizer and mixed and stirred evenly under vacuum. After bottling, a high-concentration collagen implant is obtained, wherein the collagen concentration is 80 mg / ml.

[0039] The weight ratio of the alkaline solution to the collagen solution is 0.45:1; in step S3, the homogenizer has a revolution speed of 500 rpm, a rotation speed of 300 rpm, a stirring time of 10 minutes, and a vacuum degree of -80 kPa. Example

[0040] 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:

[0041] S1. Preparation of collagen solution: Place collagen and hydrochloric acid solution in a homogenizer mixing tank and mix them evenly under vacuum to obtain collagen solution;

[0042] The collagen solution contains 8.0 wt% collagen and 92.0 wt% hydrochloric acid, and the pH of the collagen solution is approximately 2.5. In step S1, the homogenizer has a revolution speed of 900 rpm, a rotation speed of 600 rpm, a homogenization time of 15 minutes, and a vacuum degree of -100 kPa.

[0043] S2. Preparation of alkaline solution: Mix 7.5 wt% disodium hydrogen phosphate, 0.88 wt% sodium hydroxide, 0.5 wt% sodium chloride, 3.0 wt% glucose, 0.5 wt% mannitol, and the balance of water for injection evenly to obtain an alkaline solution.

[0044] S3. Mixing: The alkaline solution prepared above is added to the collagen solution in three portions, and then placed in a homogenizer and mixed and stirred evenly under vacuum. After bottling, a high-concentration collagen implant is obtained, wherein the collagen concentration is 60 mg / ml.

[0045] The weight ratio of the alkaline solution to the collagen solution is 0.29:1; in step S3, the homogenizer has a revolution speed of 800 rpm, a rotation speed of 600 rpm, a stirring time of 7 minutes, and a vacuum degree of -100 kPa.

[0046] test:

[0047] (1) The pH and osmotic pressure of the collagen implants prepared in Examples 1-3 were measured using a pH meter and an osmotic pressure analyzer; at the same time, the extrusion force was tested using a 27G needle at a speed of 30 mm / min. After storing the collagen implants prepared in Examples 1-3 at 2-8℃ for 1 month, 3 months and 6 months, their pH, osmotic pressure and extrusion force were measured again. The results are shown in Table 1 and Figure 1:

[0048] Table 1 shows the pH, osmotic pressure, and extrusion force of the collagen implants obtained in Examples 1-3.

[0049]

[0050] As can be seen from Table 1 and Figure 1, 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 this invention all meet the requirements, their extrusion force is less than 10 N, and they are relatively stable after being stored at 2-8℃ for 6 months, indicating that they have good stability.

[0051] (2) The collagen implants prepared in Examples 1 to 3 were diluted to 0.5 mg / ml and stirred evenly. The collagen fibers were observed using an inverted microscope. The results are shown in Figures 2 to 4. As can be seen from Figures 2 to 4, the collagen fibers in the collagen implants prepared in Examples 1 to 3 are 10 to 50 μm in length and are relatively evenly distributed.

[0052] In summary, the method for preparing high-concentration collagen implants based on high-speed vacuum homogenization technology provided by this invention has a simple preparation process, short cycle, high collagen concentration, uniform fibers, collagen fiber length of 10-50μm, milky white color in pre-filled syringes, stable extrusion, extrusion force of less than 10N, can be drawn into fibers, can be stacked, and has good stability.

[0053] The above-described preferred embodiments of the present invention are for illustrative purposes only and are not intended to limit the scope of the invention. Any obvious variations or modifications derived from the technical solutions 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 a high-speed vacuum homogenization technique, characterized by, The method includes the following steps: S1. Preparation of collagen solution: Place collagen and hydrochloric acid solution in a homogenizer and mix them evenly under vacuum to obtain collagen solution; S2. Preparation of alkaline solution: Mix disodium hydrogen phosphate, sodium hydroxide, sodium chloride, glucose, mannitol and water for injection evenly to obtain an alkaline solution; S3. Mixing: The alkaline solution is added to the collagen solution in portions, and then placed in a homogenizer and mixed and stirred evenly under vacuum conditions. After bottling, a high-concentration collagen implant is obtained.

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

3. The method for preparing a high-concentration collagen implant based on a high-speed vacuum homogenization technique according to claim 1 or 2, characterized in that, The collagen solution contains 5.0–15.0 wt% collagen, with the remainder being hydrochloric acid.

4. The method of claim 1, wherein the high concentration collagen implant is prepared based on a high-speed vacuum homogenization technique. The pH value of the alkaline solution in step S2 is 10.5 to 12.

5.

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

6. The method of claim 1, wherein the high concentration collagen implant is prepared based on a high-speed vacuum homogenization technique. In step S3, the weight ratio of the collagen solution to the alkaline solution is 1:(0.1 to 0.5).

7. The method of claim 1, wherein the high concentration collagen implant is prepared based on a high-speed vacuum homogenization technique. The high-concentration collagen implant prepared in step S3 has a pH of 6.5–8.0 and an osmotic pressure of 270–350 mOsmol / kg. The collagen concentration in the high-concentration collagen implant is 30–120 mg / ml, and the collagen fiber length is 10–50 μm.

8. The method of claim 1, wherein the high concentration collagen implant is prepared based on a high-speed vacuum homogenization technique. The homogenizer described in step S3 has a revolution speed of 500-800 rpm, a rotation speed of 300-600 rpm, a vacuum degree of -100--80 kPa, and a stirring time of 5-15 minutes.

9. The method of claim 1, wherein the high concentration collagen implant is prepared based on a high-speed vacuum homogenization technique. Step S3 involves adding the alkaline solution to the collagen solution in 1 to 3 portions.

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