AAPS PharmSciTech

, Volume 19, Issue 7, pp 3009–3018 | Cite as

Morphological, Stability, and Hypoglycemic Effects of New Gliclazide-Bile Acid Microcapsules for Type 1 Diabetes Treatment: the Microencapsulation of Anti-diabetics Using a Microcapsule-Stabilizing Bile Acid

  • Sangeetha Mathavan
  • Nigel Chen-Tan
  • Frank Arfuso
  • Hani Al-SalamiEmail author
Research Article


When we administered orally a mixture of the anti-diabetic drug, gliclazide (G) and a primary bile acid, they exerted a hypoglycemic effect in a rat model of type 1 diabetes (T1D), but stability of mixture was limited. We aimed to develop and characterize microcapsules incorporating G with a microcapsule-stabilizing bile acid, ursodeoxycholic acid (UDCA). Sodium alginate (SA)-based microcapsules were prepared with either G or G with UDCA and analyzed in terms of morphological, physico-chemical, and electro-chemical characteristics at different pH and temperatures. The microcapsules’ effects on viability on muscle cell line (C2C12) and on diabetic rats’ blood glucose levels and inflammatory profiles were also examined. Bile acid-based microcapsules maintained their morphology, showed good stability, and compatibility profiles, and the incorporation of UDCA resulted in less G content per microcapsule (p < 0.01) and production of stronger microcapsules that were more resistant to mechanical pressure (p < 0.01). G-UDCA-SA microcapsules enhanced muscle cell viability at higher glucose concentrations compared with control (G-SA and UDCA-SA), and they had strong anti-inflammatory effects on diabetic rats. In addition, the incorporation of UDCA into G microcapsules enhanced the physical characteristics of the microcapsules and optimized G delivery after oral administration.


drug stability ursodeoxycholic acid diabetes microencapsulation cell viability 



The authors acknowledge the Curtin Health Innovation Research Institute for provision of laboratory space and technology platforms utilized in this study. The authors would also like to acknowledge Professor Deidre Coombe from the School of Biomedical Sciences at Curtin University for the generous donation of the C2C12 cell line. HAS is partially supported by the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 690876.

Funding Information

The authors acknowledge the Curtin CUPS Scholarship for their support and also acknowledge the use of equipment, scientific, and technical assistance of the Curtin University Electron Microscope Facility, which has been partially funded by the University, State and Commonwealth Governments.

Compliance with Ethical Standards


The authors declare that they have no conflict of interest.


  1. 1.
  2. 2.
    Gaglia JL, Guimaraes AR, Harisinghani M, Turvey SE, Jackson R, Benoist C, et al. Noninvasive imaging of pancreatic islet inflammation in type 1A diabetes patients. J Clin Invest. 2011;121(1):442–5.CrossRefGoogle Scholar
  3. 3.
    Rabkin Z, Israel O, Keidar Z. Do hyperglycemia and diabetes affect the incidence of false-negative 18F-FDG PET/CT studies in patients evaluated for infection or inflammation and cancer? A comparative analysis. J Nucl Med. 2010;51(7):1015–20.CrossRefGoogle Scholar
  4. 4.
    Whelehan M, Marison IW. Microencapsulation using vibrating technology. J Microencapsul. 2011;28(8):669–88.CrossRefGoogle Scholar
  5. 5.
    Mooranian A, Negrulj R, Mathavan S, Martinez J, Sciarretta J, Chen-Tan N, et al. An advanced microencapsulated system: a platform for optimized oral delivery of antidiabetic drug-bile acid formulations. Pharm Dev Technol. 2015;20(6):702–9.CrossRefGoogle Scholar
  6. 6.
    Rakel A, Renier G, Roussin A, Buithieu J, Mamputu JC, Serri O. Beneficial effects of gliclazide modified release compared with glibenclamide on endothelial activation and low-grade inflammation in patients with type 2 diabetes. Diabetes Obes Metab. 2007;9(1):127–9.CrossRefGoogle Scholar
  7. 7.
    Mathavan S, Chen-Tan N, Arfuso F, Al-Salami H. A comprehensive study of novel microcapsules incorporating gliclazide and a permeation enhancing bile acid: hypoglycemic effect in an animal model of Type-1 diabetes. Drug Deliv. 2016;23(8):2869–80.CrossRefGoogle Scholar
  8. 8.
    Mathavan S, Chen-Tan N, Arfuso F, Al-Salami H. The role of the bile acid chenodeoxycholic acid in the targeted oral delivery of the anti-diabetic drug gliclazide, and its applications in type 1 diabetes. Artif Cells Nanomed Biotechnol. 2016;44(6):1508–19.CrossRefGoogle Scholar
  9. 9.
    Al-Salami H, Butt G, Tucker I, Mikov M. Influence of the semisynthetic bile acid (MKC) on the ileal permeation of gliclazide in healthy and diabetic rats. Pharmacol Rep. 2008;60(4):532–41.PubMedGoogle Scholar
  10. 10.
    Lazaridis KN, Gores GJ, Lindor KD. Ursodeoxycholic acid ‘mechanisms of action and clinical use in hepatobiliary disorders’. J Hepatol. 2001;35(1):134–46.CrossRefGoogle Scholar
  11. 11.
    Paumgartner G, Beuers U. Ursodeoxycholic acid in cholestatic liver disease: mechanisms of action and therapeutic use revisited. Hepatology. 2002;36(3):525–31.CrossRefGoogle Scholar
  12. 12.
    Ikegami T, Matsuzaki Y. Ursodeoxycholic acid: mechanism of action and novel clinical applications. Hepatol Res. 2008;38(2):123–31.PubMedGoogle Scholar
  13. 13.
    Kumar D, Tandon RK. Use of ursodeoxycholic acid in liver diseases. J Gastroenterol Hepatol. 2001;16(1):3–14.CrossRefGoogle Scholar
  14. 14.
    Engin F, Yermalovich A, Nguyen T, Hummasti S, Fu W, Eizirik DL, et al. Restoration of the unfolded protein response in pancreatic β cells protects mice against type 1 diabetes. Sci Transl Med. 2013;5(211):211ra156.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Mooranian A, Negrulj R, Chen-Tan N, Fakhoury M, Arfuso F, Jones F, et al. Advanced bile acid-based multi-compartmental microencapsulated pancreatic β-cells integrating a polyelectrolyte-bile acid formulation, for diabetes treatment. Artif Cells Nanomed Biotechnol. 2016;44(2):588–95.CrossRefGoogle Scholar
  16. 16.
    Mooranian A, Negrulj R, Mathavan S, Martinez J, Sciarretta J, Chen-Tan N, et al. Stability and release kinetics of an advanced gliclazide-cholic acid formulation: the use of artificial-cell microencapsulation in slow release targeted oral delivery of antidiabetics. J Pharm Innov. 2014;9(2):150–7.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Mooranian A, Negrulj R, Chen-Tan N, Watts GF, Arfuso F, Al-Salami H. An optimized probucol microencapsulated formulation integrating a secondary bile acid (deoxycholic acid) as a permeation enhancer. Drug Des Devel Ther. 2014;8:1673–83.PubMedPubMedCentralGoogle Scholar
  18. 18.
    Barakat NS, Shazly GA, Almedany AH. Influence of polymer blends on the characterization of gliclazide—encapsulated into poly (epsilon-caprolactone) microparticles. Drug Dev Ind Pharm. 2013;39(2):352–62.CrossRefGoogle Scholar
  19. 19.
    Wang YJ. Development of new polycations for cell encapsulation with alginate. Mater Sci Eng C. 2000;13(1):59–63.CrossRefGoogle Scholar
  20. 20.
    Stetinova V, Kvetina J, Pastera J, Polaskova A, Prazakova M. Gliclazide: pharmacokinetic-pharmacodynamic relationships in rats. Biopharm Drug Dispos. 2007;28(5):241–8.CrossRefGoogle Scholar
  21. 21.
    Korec R. Treatment of alloxan and streptozotocin diabetes in rats by intrafamiliar homo (allo) transplantation of neonatal pancreases. Endocrinol Exp. 1980;14(3):191–8.PubMedGoogle Scholar
  22. 22.
    Carvalho EN, Carvalho NA, Ferreira LM. Experimental model of induction of diabetes mellitus in rats. Acta Cirurgica Brasileira. 2003;18(SPE):60–4.CrossRefGoogle Scholar
  23. 23.
    Al-Salami H, Butt G, Tucker I, Golocorbin-Kon S, Mikov M. Probiotics decreased the bioavailability of the bile acid analog, monoketocholic acid, when coadministered with gliclazide, in healthy but not diabetic rats. Eur J Drug Metab Pharmacokinet. 2012;37(2):99–108.CrossRefGoogle Scholar
  24. 24.
    Alam MJ, Rahman MA. Changes in the saccharoid fraction in rats with alloxan-induced diabetes or injected with epinephrine. Clin Chem. 1971;17(9):915–20.PubMedGoogle Scholar
  25. 25.
    Greenwood R. Review of the measurement of zeta potentials in concentrated aqueous suspensions using electroacoustics. Adv Colloid Interf Sci. 2003;106(1–3):55–81.CrossRefGoogle Scholar
  26. 26.
    Mooranian A, Negrulj R, Al-Salami H. Viability and topographical analysis of microencapsulated β-cells exposed to a biotransformed tertiary bile acid: an ex vivo study. Int J Nano Biomaterials. 2016;6:74.CrossRefGoogle Scholar
  27. 27.
    Mooranian A, Negrulj R, Al-Salami H. The impact of allylamine-bile acid combinations on cell delivery microcapsules in diabetes. J Microencapsul. 2016;33(6):569–74.CrossRefGoogle Scholar
  28. 28.
    Mooranian A, Negrulj R, Al-Salami H. Flow vibration-doubled concentric system coupled with low ratio amine to produce bile acid-macrocapsules of beta-cells. Ther Deliv. 2016;7(3):171–8.CrossRefGoogle Scholar
  29. 29.
    Gill P, Moghadam TT, Ranjbar B. Differential scanning calorimetry techniques: applications in biology and nanoscience. J Biomol Tech. 2010;21(4):167–93.PubMedPubMedCentralGoogle Scholar
  30. 30.
    Sarmento B, Ferreira D, Veiga F, Ribeiro A. Characterization of insulin-loaded alginate nanoparticles produced by ionotropic pre-gelation through DSC and FTIR studies. Carbohydr Polym. 2006;66(1):1–7.CrossRefGoogle Scholar
  31. 31.
    Kazarian SG, Kong KWT, Bajomo M, Van Der Weerd J, Chan KLA. Spectroscopic imaging applied to drug release. Food Bioprod Process. 2005;83(2):127–35.CrossRefGoogle Scholar
  32. 32.
    Barakat NS, Almurshedi AS. Design and development of gliclazide-loaded chitosan microparticles for oral sustained drug delivery: in-vitro/in-vivo evaluation. J Pharm Pharmacol. 2011;63(2):169–78.CrossRefGoogle Scholar
  33. 33.
    Prakash K, Raju P, Shanta K, Lakshmi M. Preparation and characterization of lamivudine microcapsules using various cellulose polymers. Trop J Pharm Res. 2007;6(4):841–7.CrossRefGoogle Scholar
  34. 34.
    Sartori C, Finch DS, Ralph B, Gilding K. Determination of the cation content of alginate thin films by FTi.R. Spectroscopy. Polymer. 1997;38(1):43–51.CrossRefGoogle Scholar
  35. 35.
    Yang L, Xu Y, Su Y, Wu J, Zhao K, Chen J, et al. FTIR spectroscopic study on the variations of molecular structures of some carboxyl acids induced by free electron laser. Spectrochim Acta A Mol Biomol Spectrosc. 2005;62(4–5):1209–15.CrossRefGoogle Scholar
  36. 36.
    Prajapati SK, Tripathi P, Ubaidulla U, Anand V. Design and development of gliclazide mucoadhesive microcapsules: in vitro and in vivo evaluation. AAPS PharmSciTech. 2008;9(1):224–30.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Prajapati VD, Mashru KH, Solanki HK, Jani GK. Development of modified release gliclazide biological macromolecules using natural biodegradable polymers. Int J Biol Macromol. 2013;55:6–14.CrossRefGoogle Scholar
  38. 38.
    Pasparakis G, Bouropoulos N. Swelling studies and in vitro release of verapamil from calcium alginate and calcium alginate-chitosan beads. Int J Pharm. 2006;323(1–2):34–42.CrossRefGoogle Scholar
  39. 39.
    Mooranian A, Negrulj R, Al-Salami H. The effects of ionic gelation-vibrational jet flow technique in fabrication of microcapsules incorporating β-cell: applications in diabetes. Curr Diabetes Rev. 2017;13(1):91–6.CrossRefGoogle Scholar
  40. 40.
    Mooranian A, Negrulj R, Al-Salami H. Primary bile acid chenodeoxycholic acid-based microcapsules to examine beta-cell survival and the inflammatory response. Bionanoscience. 2016;6(2):103–9.CrossRefGoogle Scholar
  41. 41.
    Mooranian A, Negrulj R, Al-Salami H. The influence of stabilized deconjugated ursodeoxycholic acid on polymer-hydrogel system of transplantable NIT-1 cells. Pharm Res. 2016;33(5):1182–90.CrossRefGoogle Scholar
  42. 42.
    Yang J, Gao C. Fabrication of diverse microcapsule arrays of high density and good stability. Macromol Rapid Commun. 2010;31(12):1065–70.CrossRefGoogle Scholar
  43. 43.
    Zheng G, Liu X, Wang X, Chen L, Xie H, Wang F, et al. Improving stability and biocompatibility of alginate/chitosan microcapsule by fabricating bi-functional membrane. Macromol Biosci. 2014;14(5):655–66.CrossRefGoogle Scholar
  44. 44.
    De AK, Sana S, Datta S, Mukherjee A. Protective efficacy of ursodeoxycholic acid nanoparticles in animal model of inflammatory bowel disease. J Microencapsul. 2014;31(8):725–37.CrossRefGoogle Scholar
  45. 45.
    Al-Salami H, Butt G, Tucker I, Fawcett PJ, Golocorbin-Kon S, Mikov I, et al. Gliclazide reduces MKC intestinal transport in healthy but not diabetic rats. Eur J Drug Metab Pharmacokinet. 2009;34(1):43–50.CrossRefGoogle Scholar
  46. 46.
    Rodrigues CM, Fan G, Ma X, Kren BT, Steer CJ. A novel role for ursodeoxycholic acid in inhibiting apoptosis by modulating mitochondrial membrane perturbation. J Clin Invest. 1998;101(12):2790–9.CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Rodrigues C, Fan G, Wong PY, Kren BT, Steer CJ. Ursodeoxycholic acid may inhibit deoxycholic acid-induced apoptosis by modulating mitochondrial transmembrane potential and reactive oxygen species production. Mol Med. 1998;4(3):165–78.CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Mikov M, Boni N, Al-Salami H, Kuhajda K, Kevresan S, Golocorbin-Kon S, et al. Bioavailability and hypoglycemic activity of the semisynthetic bile acid salt, sodium 3α, 7α-dihydroxy-12-0X0-5β-cholanate, in healthy and diabetic rats. Eur J Drug Metab Pharmacokinet. 2007;32(1):7–12.CrossRefGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2018

Authors and Affiliations

  • Sangeetha Mathavan
    • 1
  • Nigel Chen-Tan
    • 2
  • Frank Arfuso
    • 3
  • Hani Al-Salami
    • 4
    Email author
  1. 1.Biotechnology and Drug Development Research Laboratory, School of Pharmacy and Biomedical Sciences, Curtin Health Innovation Research InstituteCurtin UniversityPerthAustralia
  2. 2.Faculty of Science and EngineeringCurtin UniversityPerthAustralia
  3. 3.Stem Cell and Cancer Biology Laboratory, School of Biomedical Sciences, Curtin Health Innovation Research InstituteCurtin UniversityPerthAustralia
  4. 4.School of PharmacyCurtin UniversityPerthAustralia

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