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CN-122005932-A - GelMA-HA-CGF composite hydrogel and preparation method and application thereof

CN122005932ACN 122005932 ACN122005932 ACN 122005932ACN-122005932-A

Abstract

The invention belongs to the technical field of biological materials, relates to injectable hydrogel for tissue engineering and regenerative medicine, and in particular relates to GelMA-HA-CGF composite hydrogel for the fields of cell culture, tissue repair, regeneration and the like, and a preparation method and application thereof. Compared with GelMA-HA hydrogel, the GelMA-HA-CGF composite hydrogel HAs a reduced pore diameter, and the density of matrix among pores is more uniform, so that a three-dimensional network structure with higher stability is finally formed. Observations under a high-power microscope show that the freeze-dried CGF is uniformly dispersed in the network structure of the hydrogel, and the distribution characteristic is beneficial to slow release of the growth factors in subsequent applications. The composite hydrogel prepared by the invention has proper rheological property, swelling and degradation property, mechanical property, CGF growth factor slow release property and PH responsiveness (acid degradation), and the acid condition can accelerate the degradation rate of the composite hydrogel and the release of the growth factor, thus having acid degradation characteristic.

Inventors

  • DUAN XUEJING
  • FU XIAO
  • LI HUI

Assignees

  • 山东第一医科大学附属省立医院(山东省立医院)

Dates

Publication Date
20260512
Application Date
20260130

Claims (10)

  1. 1. The preparation method of the GelMA-HA-CGF composite hydrogel is characterized by comprising the following steps of: (1) Dissolving oxidized hyaluronic acid (CHO-HA) and methacryloylated gelatin (GelMA) in PBS solution to obtain hydrogel precursor solution; (2) And adding freeze-dried CGF powder into the hydrogel precursor liquid, stirring and uniformly mixing, and performing photo-curing treatment to obtain the GelMA/HA/CGF composite hydrogel.
  2. 2. The method according to claim 1, wherein in the step (1), the hydrogel precursor liquid is obtained by stirring uniformly in a water bath at 37 ℃.
  3. 3. The preparation method according to claim 1 or 2, wherein in the step (1), the mass ratio of the CHO-HA to the GelMA solution is 1:9-11, the feed liquid ratio of the CHO-HA to the PBS solution is 5 mg/1 mL, the PBS solution contains 0.25% (w/v) LAP, and the mass-volume ratio of the GelMA solution is 4.5-5.5%.
  4. 4. A process according to any one of claims 1 to 3, wherein in step (1), the process comprises The preparation method of the methacryloylated gelatin (GelMA) comprises heating LAP photoinitiator solution in water bath, periodically oscillating during the heating, adding GelMA, heating for reaction, and periodically stirring to ensure uniformity of the solution.
  5. 5. The preparation method according to claim 4, wherein the mass-to-volume ratio of the LAP photoinitiator solution is 0.25%, the mass ratio of the LAP to the GelMA is 1:1, the water bath heating is performed in a 45 ℃ water bath for 15min, and the heating reaction is performed in a light-shielding condition for 30min at 60 ℃.
  6. 6. The method according to any one of claims 1 to 3, wherein in the step (1), the oxidized hyaluronic acid (CHO-HA) is prepared by adding 0.565 g sodium periodate (NaIO 4 ) to the hyaluronic acid solution, stirring and reacting for 2 hours, and finally adding 1 mL ethylene glycol, and stirring for 1 hour. And (3) putting the obtained solution into a dialysis bag with the molecular weight cut-off of 8000-14000, dialyzing for 4 days by using deionized water, and freeze-drying after the dialysis is finished to obtain a final product.
  7. 7. The preparation method according to claim 6, wherein the concentration of the hyaluronic acid solution is 5mg/mL, the mass ratio of the hyaluronic acid to the sodium periodate (NaIO 4 ) is 1:1.13, the stirring reaction time is 2h, the feed liquid ratio of the hyaluronic acid to the glycol is 0.5 g/1 mL, and the stirring continuing time is 1h.
  8. 8. The method according to any one of claims 1 to 7, wherein in the step (2), the mass fraction of the lyophilized CGF powder in the hydrogel precursor liquid is 1 to 3%, the temperature of stirring is 37 ℃, and the conditions of the photo-curing treatment are that the photo-curing treatment is carried out at 395 to 405 nm wavelength for 30 to 120s.
  9. 9. A GelMA/HA/CGF composite hydrogel prepared by the method of any one of claims 1-8.
  10. 10. Use of a GelMA/HA/CGF composite hydrogel according to claim 9 in the fields of rapid repair of skin wounds, cartilage defect regeneration, bone regeneration, periodontal regeneration.

Description

GelMA-HA-CGF composite hydrogel and preparation method and application thereof Technical Field The invention belongs to the technical field of biological materials, relates to injectable hydrogel for tissue engineering and regenerative medicine, and in particular relates to GelMA-HA-CGF composite hydrogel for the fields of cell culture, tissue repair, regeneration and the like, and a preparation method and application thereof. Background At present, in the field of tissue engineering repair, injectable hydrogels have become a research hotspot because of the advantages of minimally invasive implantation, good tissue compatibility and capability of filling irregular defect parts. Among them, methacryloylated gelatin (GelMA) based hydrogels are a class of materials that are widely used. Hyaluronic Acid (HA) is often used for hydrogel modification due to the excellent moisture retention, anti-inflammatory and cell adhesion promotion characteristics, for example, hybrid hydrogels based on GelMA and hyaluronic acid for tissue engineering applications discloses a technology for preparing hydrogel by physically blending unmodified HA and GelMA, and the preparation method comprises the steps of directly dissolving HA powder in GelMA solution, uniformly stirring and then photo-crosslinking to obtain the gel material by introducing HA to improve the moisture retention and biocompatibility of the hydrogel. However, the gel material is only combined through weak intermolecular interaction, HA is easy to dissolve out rapidly in the initial stage of hydrogel degradation (1-4 days), the dissolution rate can reach more than 60%, the moisturizing and anti-inflammatory effects cannot be continuously exerted, and a large amount of dissolved HA can possibly cause local tissue osmotic pressure imbalance, increase wound surface edema risk and influence the repair process. The GelMA-based hydrogel prepared by the prior art has single crosslinking density regulation means, is realized only by adjusting the illumination time or the photoinitiator concentration, is degraded too slowly when the mechanical strength is too high and is not matched with the repair cycle of tissues such as skin, cartilage and the like, has insufficient mechanical strength when the degradation speed is matched, cannot support the contradiction of early stress of cartilage defect parts, and limits the application of the GelMA-based hydrogel in repairing load-bearing tissues. Disclosure of Invention Aiming at the problems of poor combination stability of HA and GelMA, insufficient biological activity, weak action persistence, poor suitability of mechanical property and degradation property and the like in the GelMA-HA-CGF composite hydrogel in the prior art, the invention provides a preparation method of the GelMA-HA-CGF composite hydrogel. The hydrogel prepared by the method can be used as a scaffold material for cell growth in tissue engineering, can assist in repairing and functional recovery of damaged tissues in regenerative medicine, can be applied to the scenes of bone defect filling, chronic inflammatory tissue repairing such as periodontitis and the like, and provides a three-dimensional repairing microenvironment with bioactivity for the damaged tissues. The invention also provides the GelMA-HA-CGF composite hydrogel prepared by the preparation method. The invention also aims to provide application of the GelMA-HA-CGF composite hydrogel in the fields of rapid repair of skin wounds, regeneration of cartilage defects, bone regeneration and periodontal regeneration. The technical scheme adopted by the invention for achieving the purpose is as follows: The invention provides a preparation method of GelMA-HA-CGF composite hydrogel, which comprises the following steps: (1) Dissolving oxidized hyaluronic acid (CHO-HA) and methacryloylated gelatin (GelMA) in PBS solution to obtain hydrogel precursor solution; (2) And adding freeze-dried CGF powder into the hydrogel precursor liquid, stirring and uniformly mixing, and performing photo-curing treatment to obtain the GelMA/HA/CGF composite hydrogel. Preferably, in the step (1), the hydrogel precursor liquid is obtained by uniformly stirring under the condition of 37 ℃ water bath. Preferably, in the step (1), the mass ratio of the CHO-HA to the GelMA solution is 1:9-11, the feed liquid ratio of the CHO-HA to the PBS solution is 5mg to 1mL, the PBS solution contains 0.25% (w/v) LAP, and the mass volume ratio of the GelMA solution is 4.5-5.5%. Preferably, in the step (1), the methacryloylated gelatin (GelMA) is prepared by heating the LAP photoinitiator solution in a water bath, periodically oscillating during the heating, then adding GelMA, heating for reaction, and periodically stirring to ensure the uniformity of the solution, thereby preparing the GelMA solution. Preferably, the mass volume ratio of the LAP photoinitiator solution is 0.25%, the mass ratio of the LAP to the GelMA is 1:1, the water bath heating