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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

The protocol presented here shows the synthesis of a strong adhesive hydrogel gelatin o-nitrosobenzaldehyde (gelatin-NB). Gelatin-NB has rapid and efficient tissue adhesion ability, which can form a strong physical barrier to protect wound surfaces, so it is expected to be applied to the field of injury repair biotechnology.

Abstract

Adhesive materials have become popular biomaterials in the field of biomedical and tissue engineering. In our previous work, we presented a new material - gelatin o-nitrosobenzaldehyde (gelatin-NB) - which is mainly used for tissue regeneration and has been validated in animal models of corneal injury and inflammatory bowel disease. This is a novel hydrogel formed by modifying biological gelatin with o-nitrosobenzaldehyde (NB). Gelatin-NB was synthesized by activating the carboxyl group of NB-COOH and reacting with gelatin through 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). The obtained compound was purified to generate the final product, which can be stably stored for at least 18 months. NB has a strong adhesion to -NH2 on the tissue, which can form many C = N bonds, thus increasing the adhesion of gelatin-NB to the tissue interface. The preparation process comprises steps for the synthesis of the NB-COOH group, modification of the group, synthesis of gelatin-NB, and purification of the compound. The goal is to describe the specific synthesis process of gelatin-NB in detail and to demonstrate the application of gelatin-NB to damage repair. Moreover, the protocol is presented to further strengthen and expand the nature of the material produced by the scientific community for more applicable scenarios.

Introduction

Hydrogel is a type of three-dimensional polymer formed by water swelling. In particular, hydrogel derived from an extracellular matrix is widely used in the field of biosynthesis and regenerative medicine because of its excellent biocompatibility and therapeutic effectiveness1. Hydrogels have been reported for the treatment of gastric ulcers, neuritis, myocardial infarction2,3,4, and other diseases. Further, it has been proved that gelatin-NB can promote the outcome of inflammation ininflammatory bowel disease (IBD)5. Traditiona....

Protocol

The C57BL/6 mice were purchased from Zhejiang University School of Medicine Sir Run Run Shaw Hospital. The New Zealand rabbits were purchased from Zhejiang University. The animals were maintained in natural light-dark cycle conditions and given food and drinking water freely. All experimental procedures were approved ethically by the institutional guidelines of the Zhejiang University Ethics Committee standard guidelines (ZJU20200156) and Zhejiang University School of Medicine Sir Run Run Shaw Hospital Animal Care and Us.......

Representative Results

Figure 2A shows a schematic of the main chemical reactions involved in the synthesis of gelatin-NB, which promotes tissue integration by grafting NB groups onto gelatin. Figure 2B shows that the O-nitrobenzene of the gelatin-NB hydrogel converts to an NB group immediately after UV irradiation, and then the active aldehyde group can be crosslinked with an amino group to form a Schiff base. Figure 2C indicates that different ratios of.......

Discussion

Adhesive materials are a new class of material. More and more researchers are committed to the synthesis of various types of adhesive materials, and are trying to find their applications in biotechnology, tissue engineering, regenerative medicine, and other fields, which has led to vigorous development in recent years. In addition to focusing on the strong adhesion of adhesive materials, researchers are also paying more attention to other properties, such as injectability, self-healing, hemostatic, antibacterial, control.......

Acknowledgements

None.

....

Materials

NameCompanyCatalog NumberComments
1-(3Dimethylaminopropyl)-3-ethylcarbodimide hydrochloride (EDC)AladdinL287553
4-Hydroxy-3-(methoxy-D3) benzaldehydeShanghai Acmec Biochemical Co., LtdH946072
DCMAladdinD154840
DichloromethaneSigma-Aldrich270997
Dimethyl sulfoxide (DMSO)Sigma-Aldrich20-139
dimethylformamide (DMF)Sigma-AldrichPHR1553
gelatinSigma-Aldrich1288485
magnesium sulfateSigma-AldrichM7506
MeOHSigma-Aldrich1424109
methyl 4-(4-formyl-2-methoxyphenoxy methoxyphenyl) butanoic acid methyl esterchemsrc141333-27-9
methyl 4-bromobutyrateAladdinM158832
NaBH4Sigma-Aldrich215511
N-hydroxysuccinimide (NHS)AladdinD342712
nitric acidSigma-Aldrich225711
potassium carbonateSigma-Aldrich209619
SEM (Nova Nano 450)Thermo FEI17024560
THF/EtOHAladdinD380010
trifluoroacetic acid (TFA)Sigma-Aldrich8.0826

References

  1. Tam, R. Y., Smith, L. J., Shoichet, M. S. Engineering cellular microenvironments with photo- and enzymatically responsive hydrogels: toward biomimetic 3D cell culture models. Accounts of Chemical Research. 50 (4), 703-713 (2017).
  2. Xu, X., et al.

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