CN107137784A - Deep-sea fish collagen peptide hydrogel - Google Patents

Abstract

The invention provides a deep sea fish collagen peptide hydrogel, the preparation method of which is: (1), dissolving 3-5 grams of collagen peptide powder in 100 milliliters of PBS buffer; (2), adjusting the pH value to 6 ‑7.4; (3), add 30ml of nitrogen nitrogen dimethyl metamidamide to 100ml of collagen peptide solution, fully stir to dissolve; (4), then add 4‑5ml of triethylamine and stir until fully dissolved; (5), Then add 8-10 milliliters of methacrylic anhydride, stir overnight until fully dissolved; (6), stir intermittently, react for 3 days; (7), dialyze with a 1000 Dalton molecular weight dialysis bag for three days, and change the water intermittently; (8), pre-freeze the dialyzed peptide solution in a dish; (9), freeze-dry it in a lyophilizer; (10), finally collect the powder for preservation; the invention is mainly based on fish extraction, through acid-base, Collagen peptides whose molecular weight is controlled within 6000 Daltons by enzymatic digestion and other technologies have good solubility and water retention capacity, as well as low immunogenicity and biodegradability.

Description

Deep sea fish collagen peptide hydrogel

technical field

The invention relates to the field of hydrogel technology, in particular to a deep-sea fish collagen white peptide hydrogel.

Background technique

Skin is a natural barrier and bridge between the human body and the environment. Skin tissue exposed to the external environment in large areas may be damaged or lose function due to burns, mechanical trauma, and chronic diseases (such as diabetes). There are tens of millions of patients with various skin defects every year in our country. The key to the treatment of skin defects is to seal the wound as soon as possible and reduce the complications caused by infection. So far, autologous skin grafting remains the most effective method for treating full-thickness skin defects. For patients with large skin defects, there are problems such as insufficient skin donor sites in autologous skin transplantation, and multiple transplants are required to finally heal. The high mortality rate of patients with large full-thickness skin defects is still one of the clinical problems to be solved urgently. The development of tissue engineering and regenerative medicine has made it possible to completely regenerate and repair skin defects. The research content of skin tissue engineering mainly includes three aspects, namely skin seed cell culture, dermal scaffold material and in vitro construction of active composite skin.

One of the key steps in tissue engineering skin is to construct the scaffold structure of missing skin. The extracellular matrix macromolecules derived from animal tissue can provide the spatial structure for cell survival, have ideal histocompatibility and have the characteristics of cell attachment points. , is currently the main material used in tissue engineering skin scaffolds (Clark et al., 2007). Due to the limited absorption rate of collagen extracted from animals, and the fear of highly pathogenic infectious diseases such as mad cow disease, foot-and-mouth disease and bird flu, this more traditional method is on the verge of becoming obsolete. The US Food and Drug Administration (FDA) has restricted and gradually banned its clinical use. And collagen can be subdivided into: macromolecular collagen and small molecular collagen peptides. The collagen in the trotters that we usually eat is a macromolecular protein with a molecular weight of more than 300,000 Daltons, which cannot be directly absorbed by the human body, and the absorption rate is very low.

Contents of the invention

The purpose of the present invention is to address the defects and deficiencies of the prior art, and provide a deep-sea fish collagen white peptide hydrogel, which is mainly extracted from fish, and the molecular weight is controlled at 6000 Daltons by acid-base, enzyme cutting and other technologies Collagen peptides within have good solubility and water retention capacity, as well as low immunogenicity and biodegradability.

In order to achieve the above object, the technical scheme adopted in the present invention is:

A deep-sea fish collagen white peptide hydrogel, the preparation method of which is:

1. Dissolve 3-5 grams of collagen peptide powder in 100 ml of PBS buffer;

2. Adjust the pH value to 6-7.4;

3. Add 30ml of nitrogen-nitrogen-dimethylmethane to 100ml of collagen peptide solution, stir well to dissolve;

4. Then add 4-5 ml of triethylamine and stir until fully dissolved;

5. Add 8-10 ml of methacrylic anhydride and stir overnight until fully dissolved;

6. Stir intermittently and react for 3 days;

7. Dialyze with a 1000 Dalton molecular weight dialysis bag for three days, changing the water intermittently;

8. Pre-freeze the dialyzed peptide solution in a dish;

9. Freeze-dry in a freeze dryer;

10. Finally, collect the powder and save it.

The beneficial effects of the present invention are as follows: the present invention is mainly based on fish extraction, and the collagen peptide whose molecular weight is controlled within 6000 Daltons through acid-base, enzyme digestion and other technologies has good solubility and water retention capacity, and low immunity Originality and biodegradability.

detailed description

In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, and are not intended to limit the present invention.

Example:

A deep-sea fish collagen white peptide hydrogel, the preparation method of which is:

1. Dissolve 5 grams of collagen peptide powder in 100 ml of PBS buffer;

2. Adjust the pH value to 7.4;

3. Add 30ml of nitrogen-nitrogen-dimethylmethane to 100ml of collagen peptide solution, stir well to dissolve;

4. Then add 5 ml of triethylamine and stir until fully dissolved;

5. Add 10 ml of methacrylic anhydride and stir overnight until fully dissolved;

6. Stir intermittently and react for 3 days;

7. Dialyze with a 1000 Dalton molecular weight dialysis bag for three days, changing the water intermittently;

8. Pre-freeze the dialyzed peptide solution in a dish;

9. Freeze-dry in a freeze dryer;

10. Finally, collect the powder and save it.

The usage method of this embodiment:

1. Prepare fifteen percent collagen solution with culture medium;

2. Adjust the pH value to 7.4;

3. Sterilize by filtration;

4. Digest cells, add medium to stop digestion, and collect by centrifugation;

5. Resuspend the cells with a small amount of medium;

6. Add a small amount of density cell suspension to the peptide solution;

7. Add 10 microliters of 10% I2959 solution per milliliter;

8. Quickly inoculate the well plate and irradiate with 365 nm wavelength ultraviolet light for 10 to 20 minutes.

The collagen used in this specific embodiment is mainly extracted from fish, and the collagen whose molecular weight is controlled within 6000 Daltons by techniques such as acid-base and enzymatic digestion is called collagen peptide; the larger molecule of collagen peptide has Certain solubility and water retention capacity, as well as low immunogenicity and biodegradability, will be scaffold materials with superior performance.

It will be apparent to those skilled in the art that the invention is not limited to the details of the above-described exemplary embodiments, but that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Accordingly, the embodiments should be regarded in all points of view as exemplary and not restrictive, the scope of the invention being defined by the appended claims rather than the foregoing description, and it is therefore intended that the scope of the invention be defined by the appended claims rather than by the foregoing description. All changes within the meaning and range of equivalents of the elements are embraced in the present invention.

In addition, it should be understood that although this specification is described according to implementation modes, not each implementation mode only includes an independent technical solution, and this description in the specification is only for clarity, and those skilled in the art should take the specification as a whole , the technical solutions in the various embodiments can also be properly combined to form other implementations that can be understood by those skilled in the art.

Claims (1)

1. A deep-sea fish collagen white peptide hydrogel, characterized in that: the preparation method is: (1), dissolve 3-5 grams of collagen peptide powder in 100 milliliters of PBS buffer; (2), adjust the pH value to 6-7.4; (3), in 100ml collagen peptide solution, add 30ml nitrogen nitrogen dimethyl metamidamide, fully stir to dissolve; (4), then add 4-5 milliliters of triethylamine and stir until fully dissolved; (5), add 8-10 milliliters of methacrylic anhydride again, stir overnight, until fully dissolving; (6), intermittent stirring, reacted for 3 days; (7), dialysis with 1000 Dalton molecular weight size dialysis bag for three days, change water intermittently; (8), pre-freezing the dialyzed peptide solution in a dish; (9), put into lyophilizer freeze-drying; (10), finally collect the powder for preservation.