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Insulin-Loaded Nanoparticles Based on N-Trimethyl Chitosan: In Vitro (Caco-2 Model) and Ex Vivo (Excised Rat Jejunum, Duodenum, and Ileum) Evaluation of Penetration Enhancement Properties

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Abstract

The aim of this paper was to evaluate the penetration enhancement properties of nanoparticles (NP) based on N-trimethyl chitosan (TMC 35% quaternization degree) loaded with insulin. The permeation performances of TMC NP were compared with those of chitosan (CS) NP and also with TMC and CS solutions. To estimate the mechanism of penetration enhancement, two different approaches have been taken into account: an in vitro study (Caco-2 cells) and an ex vivo study (excised rat duodenum, jejunum, and ileum). Insulin-loaded CS and TMC NP had dimensions of about 250 nm and had high yield and high encapsulation efficiency. The in vitro study evidenced that TMC and CS were able to enhance insulin permeation to the same extent. Penetration enhancement properties of TMC NP seem to be prevalently related to endocytosis while the widening of tight junctions appeared more important as mechanism in the case of CS NP. The ex vivo study put in evidence the role of mucus layer and of its microclimate pH. In duodenum (pH 5–5.5), CS and TMC solutions were more effective than NP while TMC NP were more efficient towards jejunum tissue (pH 6–6.5) for their high mucoadhesive potential. Confocal laser scanning microscopy study supported the hypothesis that penetration enhancement due to TMC NP was mainly due to internalization/endocytosis into duodenum and jejunum epithelial cells. The good penetration enhancement properties (permeation and penetration/internalization) make TMC NP suitable carriers for oral administration of insulin.

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References

  1. Aboubakar M, Couvreur P, Pinto-Alphandary H, Gouritin B, Lacur B, Farinotti R, et al. Insulin-loaded nanocapsules for oral administration: in vitro and in vivo investigation. Drug Dev Res. 2000;49:109–17.

    Article  CAS  Google Scholar 

  2. Sadeghi AM, Dorkoosh FA, Avadi MR, Weinhold M, Bayat A, Delie F, et al. Permeation enhancer effect of chitosan and chitosan derivatives: comparison of formulations as soluble polymers and nanoparticulate systems on insulin absorption in Caco-2 cells. Eur J Pharm Biopharm. 2008;70:270–8.

    Article  CAS  PubMed  Google Scholar 

  3. Ponchel G, Montisci M-J, Dembri A, Durrer C, Duchene D. Mucoadhesion of colloidal particulate systems in the gastrointestinal tract. Eur J Pharm Biopharm. 1997;44:25–31.

    Article  CAS  Google Scholar 

  4. Hans ML, Lowman AM. Biodegradable nanoparticles for drug delivery and targeting. Curr Opin Solid State Mater Sci. 2002;6:319–27.

    Article  CAS  Google Scholar 

  5. Janes KA, Calvo P, Alonso MJ. Polysaccharide colloidal particles as delivery systems for macromolecules. Adv Drug Del Rev. 2001;47:83–57.

    Article  CAS  Google Scholar 

  6. Goycoolea FM, Lollo G, Remuñán-López C, Quaglia F, Alonso MJ. Chitosan-alginate blended nanoparticles as carriers for the transmucosal delivery of macromolecules. Biomacromolecules. 2009;10:1736–43.

    Article  CAS  Google Scholar 

  7. Kotzé AF, de Leeuw BJ, Lueßen HL, de Boer AG, Verhoef JC, Junginger HE. Chitosans for enhanced delivery of therapeutic peptides across intestinal epithelia: in vitro evaluation in Caco-2 cell monolayers. Int J Pharm. 1997;159:131–6.

    Article  Google Scholar 

  8. Thanou M, Verhoef JC, Roemeijn SG, Nagelkerke JF, Merkus WH, Junginger HE. Effects of N-trimethyl chitosan chloride, a novel absorption enhancer, on Caco-2 intestinal epithelia and the ciliary beat frequency of chicken embryo trachea. Int J Pharm. 1999;185:73–8.

    Article  CAS  PubMed  Google Scholar 

  9. Sandri G, Bonferoni MC, Rossi S, Ferrari F, Gibin S, Zambito Y, et al. Nanoparticles based on N-trimethyl chitosan: evaluation of absorption properties using in vitro (Caco-2 cells) and ex vivo (excised rat jejunum) models. Eur J Pharm Biopharm. 2007;65:78–67.

    Article  Google Scholar 

  10. Sieval AB, Thanou M, Kotzè AF, Verhoef JC, Brussee J, Junginger HE. Preparation and NMR characterization of highly substituted N-trimethyl chitosan chloride. Carbohyd Res. 1998;36:175–65.

    Google Scholar 

  11. Di Colo G, Burgalassi S, Zambito Y, Monti D, Chetoni P. Effects of different N-trimethyl chitosans on in vitro/in vivo ofloxacin transcorneal permeation. J Pharm Sci. 2004;93:2851–62.

    Article  PubMed  Google Scholar 

  12. Calvo P, Remunan-Lopez C, Vila-Jato JL, Alonso MJ. Novel hydrophilic chitosan–polyethylene oxide nanoparticles as protein carriers. J Appl Polym Sci. 1997;63:125–32.

    Article  CAS  Google Scholar 

  13. Dorkoosh FA, Verhoef JC, Ambagts MHC, Rafiee-Tehrani M, Borchard G, Junginger HE. Peroral delivery systems based on superporous hydrogel polymers: release characteristics for the peptide drugs buserelin, octreotide and insulin. Eur J Pharm Sci. 2002;15:433–9.

    Article  CAS  PubMed  Google Scholar 

  14. Dodane V, Amin Khan A, Mervin JR. Effect of chitosan on epithelial permeability and structure. Int J Pharm. 1999;182:21–32.

    Article  CAS  PubMed  Google Scholar 

  15. Smith J, Wood E, Dornish M. Effect of chitosan on epithelial cell tight junctions. Pharm Res. 2004;21:43–9.

    Article  CAS  PubMed  Google Scholar 

  16. Ma Z, Lim L-Y. Uptake of chitosan associated insulin in Caco-2 cell monolayers: a comparison between chitosan molecules and chitosan nanoparticles. Pharm Res. 2003;20:1812–9.

    Article  CAS  PubMed  Google Scholar 

  17. Bayat A, Dorkoosh FA, Dehpour AR, Moezi L, Larijani B, Junginger HE, et al. Nanoparticles of quaternized chitosan derivatives as a carrier for colon delivery of insulin: ex vivo and in vitro study. Int J Pharm. 2008;356:259–66.

    Article  CAS  PubMed  Google Scholar 

  18. Yin L, Ding J, He C, Cui L, Tang C, Yin C. Drug permeability and mucoadhesion properties of thiolated trimethyl chitosan nanoparticles in oral insulin delivery. Biomaterials. 2009;30:5691–700.

    Article  CAS  PubMed  Google Scholar 

  19. Jonker C, Hamman JH, Kotze AF. Intestinal paracellular permeation enhancement with quaternized chitosan: in situ and in vitro evaluation. Int J Pharm. 2002;238:205–13.

    Article  CAS  PubMed  Google Scholar 

  20. Lee K-J, Johnson N, Castelo J, Sinko PJ, Grass G, Holme K, et al. Effect of experimental pH on the in vitro permeability in intact rabbit intestines and Caco-2 monolayer. Eur J Pharm Sci. 2005;25:193–200.

    CAS  PubMed  Google Scholar 

  21. Horter D, Dressman JB. Influence of physicochemical properties on dissolution of drugs in the gastrointestinal tract. Adv Drug Deliv Rev. 1997;25:3–14.

    Article  Google Scholar 

  22. Schillin RJ, Mitra AK. Intestinal mucosal transport of insulin. Int J Pharm. 1990;62:53–64.

    Article  Google Scholar 

  23. Aoki Y, Morishita M, Asai K, Akikusa B, Hosoda S, Takayama K. Region-dependent role of the mucous/glycocalyx layers in insulin permeation across rat small intestinal membrane. Pharm Res. 2005;22:1854–62.

    Article  CAS  PubMed  Google Scholar 

  24. Aoki Y, Morishita M, Takayama K. Role of the mucous/glycocalyx layers in insulin permeation across the rat ileal membrane. Int J Pharm. 2005;297:98–109.

    CAS  PubMed  Google Scholar 

  25. Morishita M, Aoki Y, Sakagami M, Nagai T, Takayama K. In situ ileal absorption of insulin in rats: effects of hyaluronidase pretreatment diminishing the mucous/glycocalyx layers. Pharm Res. 2004;21:309–16.

    Article  CAS  PubMed  Google Scholar 

  26. Sarmento B, Ribeiro A, Veiga F, Ferreira D, Neufeld R. Oral bioavailability of insulin contained in polysaccharide nanoparticles. Biomacromolecules. 2007;8:3054–60.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Pharmaceutical Chemistry, School of Pharmacy, University of Pavia, viale Taramelli 12, 27100, Pavia, Italy

    Giuseppina Sandri, Maria Cristina Bonferoni, Silvia Rossi, Franca Ferrari & Carla Caramella

  2. Department of Applied and Experimental Pharmacology, University of Pavia, viale Taramelli 14, 27100, Pavia, Italy

    Cinzia Boselli

Authors
  1. Giuseppina Sandri
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  2. Maria Cristina Bonferoni
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  3. Silvia Rossi
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  4. Franca Ferrari
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  5. Cinzia Boselli
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  6. Carla Caramella
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Correspondence to Carla Caramella.

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Sandri, G., Bonferoni, M.C., Rossi, S. et al. Insulin-Loaded Nanoparticles Based on N-Trimethyl Chitosan: In Vitro (Caco-2 Model) and Ex Vivo (Excised Rat Jejunum, Duodenum, and Ileum) Evaluation of Penetration Enhancement Properties. AAPS PharmSciTech 11, 362–371 (2010). https://doi.org/10.1208/s12249-010-9390-3

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