Fabrication and characterization of matrix metalloproteinase-responsive G4 PAMAM-IBU/GelMA hydrogel

doi:10.3969/j.issn.1674-8115.2021.02.003

Abstract

Abstract: Objective

·To fabricate matrix metalloproteinase (MMP)-responsive gelatin (Gel) hydrogel loaded with generation 4 polyamindoamine (G4 PAMAM)/ibuprofen (IBU) polyplexes (G4 PAMAM-IBU), and investigate its characteristics in vitro.

Methods

·The G4 PAMAM was mixed with IBU powder to form G4 PAMAM-IBU polyplexes under ultrasound vortex. The concentration of IBU in the saturated G4 PAMAM-IBU solution was determined by ultraviolet spectrophotometer. The solubilization ability of G4 PAMAM for IBU and the release of IBU in G4 PAMAM-IBU were assessed. And the ability of IBU and G4 PAMAM-IBU to inhibit the proliferation of rat fibroblasts was verified by CCK-8 assay. Meanwhile, to achieve the on-demand release of G4 PAMAM-IBU, Gel and methacrylic anhydride were used to synthesize methacrylate modified gelatin (GelMA) through an addition reaction, MMP-responsive hydrogel was formed under 365 nm light, and the G4 PAMAM-IBU polyplexes were embedded in the hydrogel. The microscopic morphology of the hydrogel was observed by scanning electron microscope (SEM) and the amount of released IBU was measured by ultraviolet spectrophotometer. Then, drug-loaded GelMA hydrogel was co-cultured with fibroblasts, and the effect of hydrogel on proliferation of fibroblasts was evaluated by CCK-8 assay and live/dead cell staining. The differences in the quantitative data among the groups were analyzed by using one-way ANOVA.

Results

·G4 PAMAM-IBU was formed successfully, the solubility of IBU in water increased obviously, and G4 PAMAM-IBU could gradually dissociate within 12 h and thereby release IBU as well. Compared with the blank control group, the concentration of IBU alone needed to reach 300 μg/mL to inhibit the proliferation of fibroblasts (P=0.023), while the concentration of IBU in G4 PAMAM-IBU was 100 μg/mL to significantly inhibit the proliferation of fibroblasts (P=0.000). The SEM images and released IBU detection results showed that the internal porosity of the prepared G4 PAMAM-IBU/GelMA hydrogel increased and the release of the drug accelerated. Besides, the in vitro results showed that compared with the IBU/GelMA hydrogel, there were less live cells and more dead cells in the G4 PAMAM-IBU/GelMA hydrogel-treated fibroblasts. G4 PAMAM-IBU/GelMA hydrogel had a greater proliferation inhibitory effect than the IBU/GelMA hydrogel. And the addition of MMP further enhanced the inhibitory effect on fibroblasts.

Conclusion

·G4 PAMAM can significantly increase the solubility of IBU and improve its drug effect; the G4 PAMAM-IBU/GelMA hydrogel has a sustained drug release behavior responding to MMP and can inhibit the proliferation of fibroblasts sustainably.

Key words: hydrogel, generation 4 polyamindoamine (G4 PAMAM), ibuprofen (IBU), peritendinous adhesion

CLC Number:

R686.1

Chuan-dong CAI, Fei WANG, Wen-guo CUI, Cun-yi FAN, Shen LIU. Fabrication and characterization of matrix metalloproteinase-responsive G4 PAMAM-IBU/GelMA hydrogel[J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(2): 140-146.

Figures/Tables 4

Fig 1 Characterization of combination and dissociation of G4 PAMAM-IBU

Fig 2 MMP-induced GelMA hydrogel degradation and MMP-responsive IBU release from G4 PAMAM-IBU/GelMA hydrogel

Fig 3 Inhibitory effects of G4 PAMAM and G4 PAMAM-IBU on cell proliferation

Fig 4 Inhibitory effect of MMP-responsive G4 PAMAM-IBU/GelMA hydrogel on cell proliferation

TrendMD

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