An Injectable Hyaluronic Acid-Based Composite Hydrogel by DA Click Chemistry With pH Sensitive Nanoparticle for Biomedical Application.

Xiaohong Hu,Huiming Wang,Juan Pang,Huaping Tan,Xincheng Mao,Ziyu Gao

doi:10.3389/fchem.2019.00477

Abstract

Hydrogels with multifunctional properties attracted intensively attention in the field of tissue engineering because of their excellent performance. Also, object-oriented design had been supposed to an effective and efficient method for material design as cell scaffold in the field of tissue engineering. Therefore, a scaffold-oriented injectable composite hydrogel was constructed by two components. One was pH-sensitive bifunctional nanoparticles for growth factor delivery to improve biofunctionability of hydrogel scaffold. The other was Diels-alder click crosslinked hyaluronic acid hydrogel as matrix. pH dependent release behavior of nanoparticle component was confirmed by results. And, its bioactivity was verified by in vitro cell culture evaluation. In consideration of high-efficiency and effectiveness, low toxicity, controllability and reversibility, dynamic covalent and reversible Diels-alder click chemistry was used to design a HA hydrogel with two kinds of crosslinking points. The properties of hydrogel like gelation time and swelling ratio were influenced by pH value and polymer concentration. Composite hydrogel was formed by in situ polymerization, which exhibited acceptable mechanical property as a scaffold for biomedical field. Lastly, in vitro evaluation from results of viability, DNA content and cell morphology confirmed that hydrogels could maintain cell activity and support cell growth. Compared with pure hydrogel, composite hydrogel possessed better properties.

Highlights

Hydrogel, a water-swollen polymer network, is widely used in fields of biomedical fields due to their physiological-like aqueous environment (Bai et al, 2017; Celie et al, 2019; Massaro et al, 2019; Song et al, 2019; Zhao et al, 2019)
The absorbance of 200 μL MTT/Dimethyl sulfoxide (DMSO) solution at 560 nm was recorded by a microplate reader (Infinite M200 PRO).For DNA assay, detached cells of each well (12well culture plate) was digested by 500 μL 10 mg/mL papain solution at 65◦C overnight, which was qualified by QuantiTTM PicoGreen R dsDNA kit
Adhesive structure was found in the TEM image, which might be due to slight crosslinking of nanogel during the process of in situ polymerization and conglutination in drying process of HEP nanogel. These results indicated that nanogel had been successfully synthesized during the preparation of nanoparticle

Summary

INTRODUCTION

A water-swollen polymer network, is widely used in fields of biomedical fields due to their physiological-like aqueous environment (Bai et al, 2017; Celie et al, 2019; Massaro et al, 2019; Song et al, 2019; Zhao et al, 2019). HA-furan-CHO was obtained after the solution was dialyzed and freeze-dried according to above-mentioned conditions. These derivatives were characterized by 1H nuclear magnetic resonance (1H NMR, Bruker, AV500) using D2O as solvent. The absorbance of 200 μL MTT/DMSO solution at 560 nm was recorded by a microplate reader (Infinite M200 PRO).For DNA assay, detached cells of each well (12well culture plate) was digested by 500 μL 10 mg/mL papain solution at 65◦C overnight, which was qualified by QuantiTTM PicoGreen R dsDNA kit. SCHEME 3 | Scheme illustration to show the formation of composite hydrogel

AND DISCUSSION

Findings

CONCLUSION