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Preparation method and application of DN hydrogel (double network hydrogel) containing squid II gelatin

A hydrogel and double-network technology, applied in prostheses, tissue regeneration, medical science, etc., can solve problems such as the compression strength not meeting the requirements for costal cartilage repair, poor mechanical properties of hydrogel, discomfort for costal cartilage defects, etc. Achieve the effects of preventing collapse, good biocompatibility, and repairing costal cartilage defects

Active Publication Date: 2020-06-26
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the mechanical properties of this material and the induction of cartilage stem / precursor cells derived from costal perichondrium, especially whether it can exert a chondrogenic induction effect under complex factors such as surgical trauma and implantation of exogenous materials are still unclear. don't know
[0006] In summary, the DN hydrogel reported in the current research still has the following deficiencies and is not suitable for the repair of costal cartilage defects: ①The raw materials for preparation are mostly chemically synthesized polymers, which have risks such as slow degradation and degradation products causing inflammation in the body; See DN hydrogel prepared entirely based on natural biomaterials, while the existing reported hydrogels prepared from natural biomaterials have relatively poor mechanical properties, especially the compressive strength cannot meet the requirements for repairing costal cartilage; ③These hydrogels themselves There is still a lack of research on whether there is an effect on cartilage repair, and this effect is precisely the aspect that should be focused on in the repair of costal cartilage defects in the donor site

Method used

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  • Preparation method and application of DN hydrogel (double network hydrogel) containing squid II gelatin
  • Preparation method and application of DN hydrogel (double network hydrogel) containing squid II gelatin
  • Preparation method and application of DN hydrogel (double network hydrogel) containing squid II gelatin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1. Preparation of squid type II gelatin-[4-Arm PEG-SS]-HAMA double network hydrogel.

[0040] After removing the attached muscles of squid cartilage, it is vacuum freeze-dried and crushed into cartilage powder. Add EDTA solution at room temperature, stir slowly for 24 hours, centrifuge and wash the precipitate with deionized water, then add NaOH solution and stir slowly for 24 hours. Centrifuge and completely wash the precipitate with deionized water, add 0.5M acetic acid solution and pepsin, slowly stir and hydrolyze at 37°C, then heat to 50°C for about half an hour, then filter, and put it into a dialysis bag with a molecular weight cutoff of 10kDa to pure Water was used as the external fluid for dialysis and dialyzed for 3 days at room temperature. The dialyzed glue was vacuum freeze-dried to obtain squid type II gelatin.

[0041] Dissolve 1g hyaluronic acid (48kDa) in 100mL deionized water, cool to 0~4℃, add 4mL methacrylic anhydride, and then slowly add 2mL 5M N...

Embodiment 2

[0048] Example 2. Test of mechanical strength of squid type II gelatin-[4-Arm PEG-SS]-HAMA double network hydrogel

[0049] The mechanical performance analysis adopts GT-TCS-2000 polymer tensile machine, and the prepared hydrogel sample is placed on the test platform for testing. This example studied the influence of the concentration of squid type II gelatin and PEG on the mechanical strength of the hydrogel. figure 2 A is the compressive strength of the hydrogel prepared in each ratio in Table 1. It can be seen that when the final concentration of gelatin is 20% or 30%, and the final concentration of PEG is 5%, the prepared squid type II gelatin-[4-Arm PEG-SS]-HAMA double network hydrogel The strength is the best, and when the final concentration of gelatin is 15%, even if the final concentration of PEG reaches 5%, the mechanical strength of the resulting hydrogel is still poor.

Embodiment 3

[0050] Example 3. Degradability detection of squid type II gelatin-[4-Arm PEG-SS]-HAMA double network hydrogel

[0051] Prepare 20% gelatin-5% PEG hydrogel and 30% gelatin-5% PEG hydrogel as described in Example 1. Before preparing the hydrogel, use irradiation to sterilize squid type II gelatin. Aseptic operation was maintained during the hydrogel process, and the HAMA / 12959 mixture and PEG solution were subjected to suction filtration and sterilization with a 0.22 μm filter, and finally a sterile hydrogel was obtained. The hydrogel sample was weighed and immersed in sterile PBS, sealed and placed in a 37°C environment, and the residual rate of the hydrogel was tested after 2, 4, and 8 weeks. From figure 2 It can be seen from B that the degradation rate of hydrogels containing 30% gelatin is higher than that of hydrogels containing 20% ​​gelatin (** means p<0.01), which may be because 20% gelatin-5%PEG-1%HAMA- The ratio of amino and carboxyl groups in the 0.1% I2959 hydrogel m...

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Abstract

The invention provides a preparation method and an application of DN hydrogel (double network hydrogel) containing squid II gelatin. The preparation method comprises steps as follows: (1) HAMA is dissolved in a biocompatible medium, a photosensitizer I2959 and the squid II gelatin are sequentially added, and a mixed solution A is obtained; (2) 4-Arm PEG-SS is dissolved in a biocompatible medium, and a PEG solution is obtained; (3) the PEG solution and the mixed solution A are mixed, and a crosslinking reaction of a first network in the DN hydrogel is completed at room temperature; and (4) crosslinking of a second network of the hydrogel is completed through illumination, and the DN hydrogel containing the squid II gelatin is obtained. The prepared DN hydrogel has high compression strengthclose to rib cartilage of a human body, and collapse of a thoracic cage is avoided; and the hydrogel has degradability and can induce differentiation of cartilage stem / precursor cells derived from ribperichondria into cartilage cells, the expression level of a cartilage matrix is improved, regeneration and repair of defective rib cartilage are promoted, and repair of rib cartilage defects causedby various causes such as wound or plastic surgery is realized.

Description

Technical field [0001] The invention belongs to the field of biological materials, and more specifically, relates to a squid type II gelatin-[4-Arm PEG-SS]-HAMA double network hydrogel material that can be used to repair costal cartilage defects and a preparation method thereof. Background technique [0002] Autogenous costal cartilage is often used clinically to repair deformed ears, nose plastic surgery, and temporomandibular joint reconstruction. Usually, after cost cartilage transplantation is performed clinically, there is no corresponding repair treatment for the defect of cost cartilage at the donor site. Cartilage regeneration ability is poor, cost cartilage defect is difficult to repair on its own, it will cause chest collapse, deformity and other complications, which not only affects the appearance of the body, but also makes the rib cage lose the ability to protect internal organs to a certain extent. It can also cause discomfort to the patient. Therefore, the repair...

Claims

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Application Information

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IPC IPC(8): A61L27/22A61L27/20A61L27/18A61L27/52A61L27/50C08J3/075C08L89/00C08L5/08C08L71/02
CPCA61L27/222A61L27/20A61L27/18A61L27/52A61L27/50C08J3/075C08J2389/00C08J2405/08C08J2471/02A61L2430/06C08L5/08C08L71/02
Inventor 孙皎戴美璐朱麟勇华宇杰
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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