Name: synthesis of strong adhesive hydrogel, gelatin o-nitrosobenzaldehyde
Uploaded: 2022-11-11T14:50:00
Description: Watch this Scientific Journal Video about Synthesis of Strong Adhesive Hydrogel, Gelatin O-Nitrosobenzaldehyde at JoVE.com

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...

<ol>
	<li>Tam, R. Y., Smith, L. J., Shoichet, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineering+cellular+microenvironments+with+photo-+and+enzymatically+responsive+hydrogels:+toward+biomimetic+3D+cell+culture+models.">Engineering cellular microenvironments with photo- and enzymatically responsive hydrogels: toward biomimetic 3D cell culture models.</a> Accounts of Chemical Research. 50 (4), 703-713 (2017).</li><li>Xu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioadhesive+hydrogels+demonstrating+pH-independent+and+ultrafast+gelation+promote+gastric+ulcer+healing+in+pigs.">Bioadhesive hydrogels demonstrating pH-independent and ultrafast gelation promote gastric ulcer healing in pigs.</a> Science Translational Medicine. 12 (558), (2020).</li><li>Zheng, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Directed+self-assembly+of+herbal+small+molecules+into+sustained+release+hydrogels+for+treating+neural+inflammation.">Directed self-assembly of herbal small molecules into sustained release hydrogels for treating neural inflammation.</a> Nature Communications. 10 (1), 1604 (2019).</li><li>Seif-Naraghi, S. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+and+efficacy+of+an+injectable+extracellular+matrix+hydrogel+for+treating+myocardial+infarction.">Safety and efficacy of an injectable extracellular matrix hydrogel for treating myocardial infarction.</a> Science Translational Medicine. 5 (173), (2013).</li><li>Mao, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GelNB+molecular+coating+as+a+biophysical+barrier+to+isolate+intestinal+irritating+metabolites+and+regulate+intestinal+microbial+homeostasis+in+the+treatment+of+inflammatory+bowel+disease.">GelNB molecular coating as a biophysical barrier to isolate intestinal irritating metabolites and regulate intestinal microbial homeostasis in the treatment of inflammatory bowel disease.</a> Bioactive Materials. 19, 251-267 (2022).</li><li>Nan, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+highly+elastic+and+fatigue-resistant+natural+protein-reinforced+hydrogel+electrolyte+for+reversible-compressible+quasi-solid-state+supercapacitors.">A highly elastic and fatigue-resistant natural protein-reinforced hydrogel electrolyte for reversible-compressible quasi-solid-state supercapacitors.</a> Advanced Science. 7 (14), 2000587 (2020).</li><li>Matsumoto, K., Sakikawa, N., Miyata, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermo-responsive+gels+that+absorb+moisture+and+ooze+water.">Thermo-responsive gels that absorb moisture and ooze water.</a> Nature Communications. 9 (1), 2315 (2018).</li><li>Liu, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=resilient,+adhesive,+and+anti-freezing+hydrogels+cross-linked+with+a+macromolecular+cross-linker+for+wearable+strain+sensors.">resilient, adhesive, and anti-freezing hydrogels cross-linked with a macromolecular cross-linker for wearable strain sensors.</a> ACS Applied Materials &amp; Interfaces. 13 (35), 42052-42062 (2021).</li><li>Hong, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+strongly+adhesive+hemostatic+hydrogel+for+the+repair+of+arterial+and+heart+bleeds.">A strongly adhesive hemostatic hydrogel for the repair of arterial and heart bleeds.</a> Nature Communications. 10 (1), 2060 (2019).</li><li>Yang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue-integratable+and+biocompatible+photogelation+by+the+imine+crosslinking+reaction.">Tissue-integratable and biocompatible photogelation by the imine crosslinking reaction.</a> Advanced Materials. 28 (14), 2724-2730 (2016).</li><li>Ofner, C. M., Bubnis, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+and+swelling+evaluations+of+amino+group+crosslinking+in+gelatin+and+modified+gelatin+matrices.">Chemical and swelling evaluations of amino group crosslinking in gelatin and modified gelatin matrices.</a> Pharmaceutical Research. 13 (12), 1821-1827 (1996).</li><li>Zhang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+long-term+retaining+molecular+coating+for+corneal+regeneration.">A long-term retaining molecular coating for corneal regeneration.</a> Bioactive Materials. 6 (12), 4447-4454 (2021).</li><li>Liang, Y., Li, Z., Huang, Y., Yu, R., Guo, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dual-dynamic-bond+cross-linked+antibacterial+adhesive+hydrogel+sealants+with+on-demand+removability+for+post-wound-closure+and+infected+wound+healing.">Dual-dynamic-bond cross-linked antibacterial adhesive hydrogel sealants with on-demand removability for post-wound-closure and infected wound healing.</a> ACS Nano. 15 (4), 7078-7093 (2021).</li></ol>

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...

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. Traditional hydrogels include gellan gum, gelatin, hyaluronic acid, polyethylene glycol (PEG), layered, hydrophobic/hydrophilic, alginate/polyacrylamide, double network, and polyamphoteric hydrogels6, all of which have good histocompatibility and mechanical properties. However, these traditional hydrogels are vulnerable to moisture and air in the environment. If they are exposed to air for a long time, they will lose water and dry; if they are immersed in the water for a long time, they will absorb water and expand7, thus reducing their flexibility and mechanical function. In addition, maintaining the tissue adhesion of conventional hydrogels is a major challenge8.
Based on this, we designed and synthesized a nanoscale hydrogel gelatin-NB, which is a novel hydrogel formed by modifying biological gelatin with NB (Figure 1). NB has a strong adhesion ability to -NH2 on the tissue, which can form a large number of C = N bonds, thus increasing the adhesiveness of the hydrogel-tissue interface. This strong adhesion can make the hydrogel firmly adhere to the tissue surface, thus forming a nano-level molecular coating. In the team&#39;s previous studies, it has been confirmed that this kind of modified hydrogel coating has improved tissue adhesion9; it can stably adhere to corneal and intestinal organs and tissues and play anti-inflammation, barrier isolation, and regeneration promotion roles. The goal is to introduce the specific synthesis process of gelatin-NB in detail here, so that gelatin-NB can be applied in more scenarios of damage repair. Moreover, we encourage other researchers to further strengthen and expand the nature of this material to suit more application scenarios.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1-(3Dimethylaminopropyl)-3-ethylcarbodimide hydrochloride (EDC)</td>
			<td>Aladdin</td>
			<td>L287553</td>
			<td></td>
		</tr>
		<tr>
			<td>4-Hydroxy-3-(methoxy-D3) benzaldehyde</td>
			<td>Shanghai Acmec Biochemical Co., Ltd</td>
			<td>H946072</td>
			<td></td>
		</tr>
		<tr>
			<td>DCM</td>
			<td>Aladdin</td>
			<td>D154840</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichloromethane</td>
			<td>Sigma-Aldrich</td>
			<td>270997</td>
			<td></td>
		</tr>
		<tr>
			<td>Dimethyl sulfoxide (DMSO)</td>
			<td>Sigma-Aldrich</td>
			<td>20-139</td>
			<td></td>
		</tr>
		<tr>
			<td>dimethylformamide (DMF)</td>
			<td>Sigma-Aldrich</td>
			<td>PHR1553</td>
			<td></td>
		</tr>
		<tr>
			<td>gelatin</td>
			<td>Sigma-Aldrich</td>
			<td>1288485</td>
			<td></td>
		</tr>
		<tr>
			<td>magnesium sulfate</td>
			<td>Sigma-Aldrich</td>
			<td>M7506</td>
			<td></td>
		</tr>
		<tr>
			<td>MeOH</td>
			<td>Sigma-Aldrich</td>
			<td>1424109</td>
			<td></td>
		</tr>
		<tr>
			<td>methyl 4-(4-formyl-2-methoxyphenoxy methoxyphenyl) butanoic acid methyl ester</td>
			<td>chemsrc</td>
			<td>141333-27-9</td>
			<td></td>
		</tr>
		<tr>
			<td>methyl 4-bromobutyrate</td>
			<td>Aladdin</td>
			<td>M158832</td>
			<td></td>
		</tr>
		<tr>
			<td>NaBH4</td>
			<td>Sigma-Aldrich</td>
			<td>215511</td>
			<td></td>
		</tr>
		<tr>
			<td>N-hydroxysuccinimide (NHS)</td>
			<td>Aladdin</td>
			<td>D342712</td>
			<td></td>
		</tr>
		<tr>
			<td>nitric acid</td>
			<td>Sigma-Aldrich</td>
			<td>225711</td>
			<td></td>
		</tr>
		<tr>
			<td>potassium carbonate</td>
			<td>Sigma-Aldrich</td>
			<td>209619</td>
			<td></td>
		</tr>
		<tr>
			<td>SEM (Nova Nano 450)</td>
			<td>Thermo FEI</td>
			<td>17024560</td>
			<td></td>
		</tr>
		<tr>
			<td>THF/EtOH</td>
			<td>Aladdin</td>
			<td>D380010</td>
			<td></td>
		</tr>
		<tr>
			<td>trifluoroacetic acid (TFA)</td>
			<td>Sigma-Aldrich</td>
			<td>8.0826</td>
			<td></td>
		</tr>
	</tbody>
</table>

synthesis of strong adhesive hydrogel, gelatin o-nitrosobenzaldehyde

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.

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...

Watch this Scientific Journal Video about Synthesis of Strong Adhesive Hydrogel, Gelatin O-Nitrosobenzaldehyde at JoVE.com

Synthesis of Strong Adhesive Hydrogel, Gelatin O-Nitrosobenzaldehyde

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.

Adhesive materials have become popular biomaterials in the field of biomedical and tissue engineering. In our previous work, we presented a new material - ...

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