Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Jul 16:10:4565-80.
doi: 10.2147/IJN.S87816. eCollection 2015.

Enhanced therapeutic efficacy of budesonide in experimental colitis with enzyme/pH dual-sensitive polymeric nanoparticles

Muhammad Naeem  1 , Jiafu Cao  1 , Moonjeong Choi  1 , Woo Seong Kim  1 , Hyung Ryong Moon  1 , Bok Luel Lee  1 , Min-Soo Kim  1 , Yunjin Jung  1 , Jin-Wook Yoo  1
Affiliations

Enhanced therapeutic efficacy of budesonide in experimental colitis with enzyme/pH dual-sensitive polymeric nanoparticles

Muhammad Naeem et al. Int J Nanomedicine. .

Abstract

Current colon-targeted drug-delivery approaches for colitis therapy often utilize single pH-triggered systems, which are less reliable due to the variation of gut pH in individuals and in disease conditions. Herein, we prepared budesonide-loaded dual-sensitive nanoparticles using enzyme-sensitive azo-polyurethane and pH-sensitive methacrylate copolymer for the treatment of colitis. The therapeutic potential of the enzyme/pH dual-sensitive nanoparticles was evaluated using a rat colitis model and compared to single pH-triggered nanoparticles. Clinical activity scores, colon/body weight ratios, myeloperoxidase activity, and proinflammatory cytokine levels were markedly decreased by dual-sensitive nanoparticles compared to single pH-triggered nanoparticles and budesonide solution. Moreover, dual-sensitive nanoparticles accumulated selectively in inflamed segments of the colon. In addition, dual-sensitive nanoparticle plasma concentrations were lower than single pH-triggered nanoparticles, and no noticeable in vitro or in vivo toxicity was observed. Our results demonstrate that enzyme/pH dual-sensitive nanoparticles are an effective and safe colon-targeted delivery system for colitis therapy.

Keywords: azo-polyurethane; budesonide; colitis; colon-targeted nanoparticles; methacrylate copolymer.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Characterization of ES NPs and Azo-pu ES NPs. Notes: (A) SEM images; (B) size histograms by qNano; (C) DSC thermogram of budesonide, polymers, and NPs; (D) in vitro drug release from ES NPs and Azo-pu ES NPs at different pH values. Abbreviations: NPs, nanoparticles; ES, Eudragit S100; Azo-pu, azo-polyurethane; SEM, scanning electron microscopy; DSC, differential scanning calorimetry.
Figure 2
Figure 2
Clinical activity index during the whole experimental period and macroscopic evaluation of colitis treatment. Notes: (A) Clinical activity-score system for the colitis control group and budesonide-treated groups during the experimental period (error bars not shown for clarity reasons, n=5 animals/group, *P<0.05 compared with colitis control); (B) macroscopic evaluation of the colon; (C) colon length (***P<0.001); (D) open distal colon photographs; (E) colon/body-weight ratio (*P<0.05, ***P<0.001). Healthy group, rats not treated with TNBS; colitis control, rats treated with TNBS; budesonide solution group, colitis rats treated with budesonide in solution form; ES NPs group, colitis rats treated with budesonide-loaded ES NPs; Azo-pu ES NP group, colitis rats treated with budesonide loaded Azo-pu ES NPs administered by oral gavage. Abbreviations: NPs, nanoparticles; ES, Eudragit S100; Azo-pu, azo-polyurethane; TNBS, 2,4,6-trinitrobenzenesulfonic acid.
Figure 3
Figure 3
Histological evaluation of colon tissue from the colitis control and budesonide-treated groups. Notes: Representative microphotographs of healthy control, rats not treated with TNBS; colitis control, rats treated with TNBS; budesonide solution, colitis rats treated with budesonide in solution form, ES NPs, colitis rats treated with budesonide-loaded ES NPs; Azo-pu ES NPs, colitis rats treated with budesonide loaded Azo-pu ES NPs administered by oral gavage. (A) Hematoxylin and eosin staining for microscopic evaluation of the colon sections isolated from healthy control, colitis control, and budesonide-treated groups. Images of tissues are shown with 100× magnification. (B) Histological score of the colitis control and budesonide-treated groups. Data are presented as means ± standard deviation (n=3 animals/group). Azo-pu ES NPs showed statistically significant differences (*P<0.05, ***P<0.001) compared to the colitis control. Abbreviations: NPs, nanoparticles; ES, Eudragit S100; Azo-pu, azo-polyurethane; TNBS, 2,4,6-trinitrobenzenesulfonic acid.
Figure 4
Figure 4
Average MPO activity and cytokine expression in the healthy control group, colitis control group, and treated groups. Notes: (A) MPO assay; (B) IL-6; (C) TNF-α. Statistical comparisons were evaluated between the inflamed control group versus budesonide solution, ES, and Azo-pu ES NPs (*P<0.05, **P<0.01). Abbreviations: NPs, nanoparticles; ES, Eudragit S100; Azo-pu, azo-polyurethane; MPO, myeloperoxidase.
Figure 5
Figure 5
In vivo localization of ES NPs and Azo-pu ES NPs in healthy and inflamed colons. Notes: (A) Confocal images of C-6-loaded Azo-pu ES NPs in healthy control and inflamed control group colon cross sections prepared 24 hours after oral NP administration; (B) quantitative determination of C-6 in healthy and inflamed colon tissue 24 hours after oral administration of ES NPs and Azo-pu ES NPs. Data are presented as means ± standard deviation (n=3 animals/group, **P<0.01). Healthy colon, colon section from healthy rats; inflamed colon, colon section from 2,4,6-trinitrobenzenesulfonic acid-induced colitis rats. Abbreviations: NPs, nanoparticles; ES, Eudragit S100; Azo-pu, azo-polyurethane; C-6, coumarin 6.
Figure 6
Figure 6
Pharmacokinetic and blood glucose-level evaluations of colon-targeted delivery of ES NPs and Azo-pu ES NPs. Notes: (A) Plasma concentration of C-6 after oral administration of C-6 solution, ES NPs, and Azo-pu ES NPs. (B) Blood glucose levels in animals treated with budesonide solution, ES NPs, and Azo-pu ES NPs. Data are presented as means ± standard deviation (n=3 animals/group, *P<0.05, **P<0.01). Abbreviations: NPs, nanoparticles; ES, Eudragit S100; Azo-pu, azo-polyurethane; C-6, coumarin 6; Cmax, concentration maximum; Tmax, time to Cmax; AUC, area under curve.
Figure 7
Figure 7
In vivo and in vitro biocompatibility study of Azo-pu ES NPs. Notes: (A) Representative photomicrographs of the liver and kidney of colitis rats treated with Azo-pu ES NPs. (B) Small intestine and colon hematoxylin and eosin-stained sections (200× magnification) of healthy rats treated with blank Azo-pu ES NPs. Rats without any treatment were regarded as the control group (n=3 animals/group). (C) Changes in body weight of healthy rats treated with blank Azo-pu ES NPs. (D) In vitro cytotoxicity of blank Azo-pu ES NPs in the HT29 and HCT116 cell lines. Abbreviations: NPs, nanoparticles; ES, Eudragit S100; Azo-pu, azo-polyurethane.
Figure 8
Figure 8
Proposed mechanism of budesonide release from the single system (pH-sensitive NPs) and dual system (enzyme pH-sensitive NPs) under gastrointestinal tract-mimicking conditions. Abbreviations: NPs, nanoparticles; Azo-pu, azo-polyurethane.
See this image and copyright information in PMC

References

    1. Travis SP, Danese S, Kupcinskas L, et al. Once-daily budesonide MMX in active, mild-to-moderate ulcerative colitis: results from the randomised CORE II study. Gut. 2014;63(3):433–441. - PMC - PubMed
    1. Frank DN, St Amand AL, Feldman RA, Boedeker EC, Harpaz N, Pace NR. Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases. Proc Natl Acad Sci U S A. 2007;104(34):13780–13785. - PMC - PubMed
    1. Knutson CG, Mangerich A, Zeng Y, et al. Chemical and cytokine features of innate immunity characterize serum and tissue profiles in inflammatory bowel disease. Proc Natl Acad Sci U S A. 2013;110(26):E2332–E2341. - PMC - PubMed
    1. Seow CH, Benchimol EI, Griffiths AM, Otley AR, Steinhart AH. Budesonide for induction of remission in Crohn’s disease. Cochrane Database Syst Rev. 2008;3:CD000296. - PubMed
    1. Nakase H, Okazaki K, Tabata Y, et al. Development of an oral drug delivery system targeting immune-regulating cells in experimental inflammatory bowel disease: a new therapeutic strategy. J Pharmacol Exp Ther. 2000;292(1):15–21. - PubMed

Publication types