ACUTE TOXICITY OF CHITOSAN NANOPARTICLES CONTAINING MAHKOTA DEWA (PHALERIA MACROCARPA) LEAF EXTRACT AND ANTI-INFLAMMATORY EFFECTS IN A DEXTRAN SODIUM SULFATE-INDUCED MOUSE MODEL OF ULCERATIVE COLITIS

  • Ari Estuningtyas Department of Pharmacology, Faculty of Medicine, Universitas Indonesia, Jakarta 10430, Indonesia.
  • Santi Widiasari Student of Biomedical Science Magister Program Universitas Indonesia, Jakarta 10430, Indonesia
  • Kusmardi Kusmardi Department of Anatomic Pathology, Universitas Indonesia, Jakarta 10430, Indonesia.

Abstract

Objective: The plant mahkota dewa (Phaleria macrocarpa) is known to have anti-inflammatory effects. This study aimed to determine whether
chitosan nanoparticles containing mahkota dewa leaf extract would yield superior anti-inflammatory effects in the colon of a mouse model of dextran
sodium sulfate (DSS)-induced ulcerative colitis, compared with ethanol extract alone after testing the acute toxicities (lethal dose) of both preparations.
Methods: For acute toxicity testing, 10 Sprague-Dawley rats were administered 6000 mg/kg body weight (BW) of leaf extract alone or with nanoparticles.
Subsequently, mice were divided into the following six groups to determine the anti-inflammatory effects: Untreated, negative control (DSS 2% w/v), leaf
extract at 12.5 or 25 mg/kg BW, and leaf extract in chitosan nanoparticles at 6.25 or 12.5 mg/kg BW. To induce colitis, DSS (2% w/v) was administered
through drinking water for 6 weeks. The anti-inflammatory effect was observed histopathologically by imaging the inflammatory cells of the mice colon
with hematoxylin-eosin (HE) staining.
Results: For acute toxicity testing, 10 Sprague-Dawley rats were administered 6000 mg/kg BW of leaf extract alone or with nanoparticles. Subsequently,
mice were divided into the following six groups to determine the anti-inflammatory effects: Untreated, negative control (DSS 2% w/v), leaf extract at
12.5 or 25 mg/kg BW, and leaf extract in chitosan nanoparticles at 6.25 or 12.5 mg/kg BW. To induce colitis, DSS (1% w/v) was administered through
drinking water for 6 weeks. The anti-inflammatory effect was observed histopathologically by imaging the inflammatory cells of the mice colon with
HE staining.
Conclusion: Chitosan nanoparticles containing mahkota dewa leaf extract can be included in the practically non-toxic class of materials. However, an
ethanol extract of mahkota dewa leaf effectively inhibited DSS-induced inflammation in the mouse colon, regardless of delivery vehicle.

Keywords: Mahkota dewa leaf extract, Chitosan nanoparticles, Acute toxicity, Inflammation, Histopathology

References

1. FFakhoury M, Negrulj R, Mooranian A, Al-Salami H. Inflammatory
bowel disease: Clinical aspect and treatments. J Inflamm Res
2014;7:113-20.
2. Mulder DJ, Noble A, Justinich C, Duffin JM. A tile of two diseases: The
history of inflammatory bowel disease. J Crohn’s Colitis 2013;5:1-8.
3. Itzkowitz SH, Yio X. Inflammation and cancer IV. Colorectal cancer in
inflammatory bowel disease: The role of inflammation. Am J Physiol
Gastrointest Liver Physiol 2004;287:G7-17.
4. Lakatos PL, Lakatos L. Risk for colorectal cancer in ulcerative colitis:
Changes, causes and management strategies. World J Gastroenterol
2008;14:3937-47.
5. Carter MJ, Lobo AJ, Travis SP. Guidelines for the management of
inflammatory bowel disease in adults. Gut 2004;53:v1-v16.
6. Friis S, Riis AH, Erichsen R, Baron JA, Sorensen HT. Low-dose aspirin
or nonsteroidal anti-inflammatory drug use and colorectal cancer risk.
Ann Intern Med 2015;163:347-59.
7. Hendra R, Ahmad S, Sukari A, Shukor MY, Oskoueian E. Flavonoid
analyses and antimicrobial activity of various parts of Phaleria
macrocarpa (Scheff.) Boerl fruit. Int J Mol Sci 2011;12:3422-31.
8. De Padua LS, Bunyapraphatsara N, Lemmens RH. Plant Resources
of South East Asia. Medical and Poisonous Plants. Printed in Bogor.
Indonesia (PROSEA). Leiden, the Netherlands: Backhuys Publishers;
1999. p. 36.
9. Soeksmanto A, Hapsari Y, Simanjuntak P. Antioxidant content of parts of
Mahkota Dewa, Phaleria macrocarpa [Scheff] Boerl. (Thymelaceae).
Biodeversita 2007;8:92-5.
10. Suprapti T, Louisa M, Tedjo A, Fadilah K, Handjari DR, Yulhasri H.
Anti-inflammatory effect of Mahkota Dewa (Phaleria macrocarpa
(Scheff.) Boerl.) Leaves extract on colon carcinogenesis induced by
azoxymethane and dextran sodium sulphate: Focus on the iNOS,
β-catenin and COX-2. expressions. Asian J Appl Sci 2014;2014:511-27.
11. Nanoparticle Technology. Available from: http://www.gitam.edu. [Last
cited on 2016 Sep 05].
12. Ma Y. Controlled Delivery of Nanoparticles to the Colon for Tumour
Targeting. Australia: The University of Quesland; 2015.
13. Singh R, Lillard JW Jr. Nanoparticle-based targeted drug delivery. Exp
Mol Pathol 2009;86:215-23.
14. Muntiha M. Teknik Pembuatan Preparat Histopatologi Dari Jaringan
Hewandengan Pewarnaan Hematoksilin Dan Eosin (H and E) (in English:
Histopathology Preparation Technique Of Animal Network With
Staining Hematoxylin And Eosin (H and E)). Temu Teknis Fungsional
Non Peneliti (Non Technical Meeting Research Functional); 2001.
Fig. 4: Average inflammatory score
Fig. 5: Histopathologic analysis of colon sections from mice
(HE staining, ×400 magnification). (a) No treatment, (b) Negative
control (DSS), (c) Phaleria macrocarpa leaf extract, 25 mg/kg BW,
(d) P. macrocarpa leaf extract, 12.5 mg/kg BW, (e) P. macrocarpa leaf
extract in chitosan nanoparticle, 12.5 mg/kg BW, (f) P. macrocarpa
leaf extract in chitosan nanoparticle, 6.25 mg/kg BW
The 2nd Physics and Technologies in Medicine and Dentistry Symposium (PTMDS), Universitas Indonesia. Depok, Indonesia 10
Int J App Pharm, Vol 10, Special Issue 1, 2018
Estuningtyas et al.
15. Fischer AH, Jacobson KA, Rose J, Zeller R. Hematoxylin and eosin
staining of tissue and cell sections. CSH Protoc 2008;2008:pdb.prot4986.
16. Indonesian Agency of Drug and Food Control. Peraturan Kepala Badan
Pengawas Obat dan Makanan Nomor 7 Tahun 2014. Pedoman Uji
Toksisitas Nonklinik Secara in vivo (in English: Regulation of the Head
of the National Agency of Drug and Food Control No 7 Year 2014.
Non-clinical toxicity test guidelines in vivo) Jakarta; 2014.
17. Walum E. Acute oral toxicity. Environ Health Perspect 1998;106 Suppl
2:497-503.
18. Redbook 2000. Toxicolocigal Principals for The Safety of Food
Ingredients; Guideline for Reporting The Result of Toxicity Studies,
U.S. FDA; 2003
19. Organization for Economic Cooperation and Development. OECD
Guidelines for Testing of Chemical; 2001. p. 407-8.
20. Nijveldt RJ, van Nood E, van Hoorn DE, Boelens PG, van Norren K,
van Leeuwen PA, et al. Flavonoids: A review of probable mechanisms
of action and potential applications. Am J Clin Nutr 2001;74:418-25.
21. Zhang PC. The Effect of Cooked Common Beans on DSS-induced Colitis
in Mice. Thesis. Ontario, Canada: The University of Guelph; 2012.
22. Napsah R, Wahyuningsih I. Preparation of chitosan-Tpp/nanoparticles
ethanol extract of Mahkota Dewa fruit (Phaleria macrocarpa (Scheff)
Boerl) with ionic gelation method. J Pharm Sci Commun 2014;11:7-12.
23. Rogers R, Eastham-Anderson J, DeVoss J, Lesch J, Yan D. Image
analysis-based approaches for scoring mouse models of colitis. Vet
Pathol 2015;2015:1-11.
24. Perse M, Cerar A. Dextran sodium sulphate colitis mouse model: Traps
and tricks. J Biomed Biotech 2012;2012:1-13.
25. Kadioglu O, Nass J, Saeed ME, Schuler B, Efferth T. Kaempferol is
an anti-inflammatory compound with activity towards NF-κB pathway
proteins. Anticancer Res 2015;35:2645-50.
26. Tanaka T. Development of an inflammation-associated colorectal
cancer model and its application for research on carcinogenesis and
chemoprevention. Int J Inflam 2012;2012:658786.
27. Suzuki R, Kohno H, Sugie S, Nakagama H, Tanaka T. Strain differences
in the susceptibility to azoxymethane and dextran sodium sulfateinduced
colon carcinogenesis in mice. Carcinogenesis 2006;27:162-9.
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Estuningtyas, A., Widiasari, S., & Kusmardi, K. (2018). ACUTE TOXICITY OF CHITOSAN NANOPARTICLES CONTAINING MAHKOTA DEWA (PHALERIA MACROCARPA) LEAF EXTRACT AND ANTI-INFLAMMATORY EFFECTS IN A DEXTRAN SODIUM SULFATE-INDUCED MOUSE MODEL OF ULCERATIVE COLITIS. International Journal of Applied Pharmaceutics, 10(1), 6-10. https://doi.org/10.22159/ijap.2018.v10s1.02
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