Development and Characterization of Chitosan Microparticles-in-Films for Buccal Delivery of Bioactive Peptides

Nowadays, bioactive peptides are used for therapeutic applications and the selection of a carrier to deliver them is very important to increase the efficiency, absorption, release, bioavailability and consumer acceptance. The aim of this study was to develop and characterize chitosan-based films loaded with chitosan microparticles containing a bioactive peptide (sequence: KGYGGVSLPEW) with antihypertensive properties. Films were prepared by the solvent casting method, while the microparticles were prepared by ionic gelation. The final optimized chitosan microparticles exhibited a mean diameter of 2.5 µm, a polydispersity index of 0.46, a zeta potential of +61 mV and a peptide association efficiency of 76%. Chitosan films were optimized achieving the final formulation of 0.79% (w/v) of chitosan, 6.74% (w/v) of sorbitol and 0.82% (w/v) of citric acid. These thin (±0.100 mm) and transparent films demonstrated good performance in terms of mechanical and biological properties. The oral films developed were flexible, elastic, easy to handle and exhibited rapid disintegration (30 s) and an erosion behavior of 20% when they came into contact with saliva solution. The cell viability (75–99%) was proved by methylthiazolydiphenyl-tetrazolium bromide (MTT) assay with TR146 cells. The chitosan mucoadhesive films loaded with peptide–chitosan microparticles resulted in an innovative approach to perform administration across the buccal mucosa, because these films present a larger surface area, leading to the rapid disintegration and release of the antihypertensive peptide under controlled conditions in the buccal cavity, thus promoting bioavailability.

[1]  Bruno Sarmento,et al.  Combination of PLGA nanoparticles with mucoadhesive guar‐gum films for buccal delivery of antihypertensive peptide , 2018, International journal of pharmaceutics.

[2]  B. Sarmento,et al.  Recent insights in the use of nanocarriers for the oral delivery of bioactive proteins and peptides , 2018, Peptides.

[3]  Priscilla A T Pereira,et al.  Microparticles prepared with 50-190kDa chitosan as promising non-toxic carriers for pulmonary delivery of isoniazid. , 2017, Carbohydrate polymers.

[4]  Saad M Ahsan,et al.  Chitosan as biomaterial in drug delivery and tissue engineering. , 2017, International journal of biological macromolecules.

[5]  R. Lourenço,et al.  Properties of gelatin-based films incorporated with chitosan-coated microparticles charged with rutin. , 2017, International journal of biological macromolecules.

[6]  B. Sarmento,et al.  Optimization of two biopolymer-based oral films for the delivery of bioactive molecules. , 2017, Materials science & engineering. C, Materials for biological applications.

[7]  D. Mcclements,et al.  Preparation, characterization, and properties of chitosan films with cinnamaldehyde nanoemulsions , 2016 .

[8]  Lili He,et al.  Enhanced and Extended Anti-Hypertensive Effect of VP5 Nanoparticles , 2016, International journal of molecular sciences.

[9]  Omid C Farokhzad,et al.  Nanotechnology for protein delivery: Overview and perspectives. , 2016, Journal of controlled release : official journal of the Controlled Release Society.

[10]  H. García,et al.  Insightful understanding of the role of clay topology on the stability of biomimetic hybrid chitosan-clay thin films and CO2-dried porous aerogel microspheres. , 2016, Carbohydrate polymers.

[11]  Ankita D. Chonkar,et al.  Development of fast dissolving oral films containing lercanidipine HCl nanoparticles in semicrystalline polymeric matrix for enhanced dissolution and ex vivo permeation. , 2016, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[12]  Lídia M D Gonçalves,et al.  Effect of Experimental Parameters on Alginate/Chitosan Microparticles for BCG Encapsulation , 2016, Marine drugs.

[13]  Gajanand Sharma,et al.  Fabrication, characterization and cytotoxicity studies of ionically cross-linked docetaxel loaded chitosan nanoparticles. , 2016, Carbohydrate polymers.

[14]  Tarek A. Ahmed,et al.  Preparation, characterization, and potential application of chitosan, chitosan derivatives, and chitosan metal nanoparticles in pharmaceutical drug delivery , 2016, Drug design, development and therapy.

[15]  H. Byun,et al.  Biological effects of chitosan and its derivatives , 2015 .

[16]  P. Doshi,et al.  Fabrication of chitosan microspheres using vanillin/TPP dual crosslinkers for protein antigens encapsulation. , 2015, Carbohydrate polymers.

[17]  B. Sarmento,et al.  Oral films as breakthrough tools for oral delivery of proteins/peptides. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[18]  A. Brandelli,et al.  Whey as a source of peptides with remarkable biological activities , 2015 .

[19]  H. E. Flores,et al.  Protein adsorption onto alginate-pectin microparticles and films produced by ionic gelation , 2015 .

[20]  M. Pintado,et al.  Development of Oral Strips Containing Chitosan as Active Ingredient: A Product for Buccal Health , 2015 .

[21]  M. Qadir,et al.  Orally disintegrating films: A modern expansion in drug delivery system , 2015, Saudi pharmaceutical journal : SPJ : the official publication of the Saudi Pharmaceutical Society.

[22]  L. G. L. Torre,et al.  Scalable production of highly concentrated chitosan/TPP nanoparticles in different pHs and evaluation of the in vitro transfection efficiency , 2015 .

[23]  Bruno Sarmento,et al.  The impact of nanoparticles on the mucosal translocation and transport of GLP-1 across the intestinal epithelium. , 2014, Biomaterials.

[24]  Xinyi He,et al.  Development and characterization of GRGDSPC-modified poly(lactide-co-glycolide acid) porous microspheres incorporated with protein-loaded chitosan microspheres for bone tissue engineering. , 2014, Colloids and Surfaces B: Biointerfaces.

[25]  Chuan Tang,et al.  Preparation of ibuprofen-loaded chitosan films for oral mucosal drug delivery using supercritical solution impregnation. , 2014, International journal of pharmaceutics.

[26]  A. Mitra,et al.  Recent developments in protein and peptide parenteral delivery approaches. , 2014, Therapeutic delivery.

[27]  K. Sawant,et al.  Improvement in antihypertensive and antianginal effects of felodipine by enhanced absorption from PLGA nanoparticles optimized by factorial design. , 2014, Materials science & engineering. C, Materials for biological applications.

[28]  J. Boateng,et al.  An integrated buccal delivery system combining chitosan films impregnated with peptide loaded PEG-b-PLA nanoparticles. , 2013, Colloids and surfaces. B, Biointerfaces.

[29]  J. Boateng,et al.  Development and characterisation of chitosan films impregnated with insulin loaded PEG-b-PLA nanoparticles (NPs): a potential approach for buccal delivery of macromolecules. , 2012, International journal of pharmaceutics.

[30]  V. Balcão,et al.  Nanocarrier possibilities for functional targeting of bioactive peptides and proteins: state-of-the-art , 2012, Journal of drug targeting.

[31]  B. Sarmento,et al.  Chitosan formulations as carriers for therapeutic proteins. , 2011, Current drug discovery technologies.

[32]  F. Malcata,et al.  Novel whey-derived peptides with inhibitory effect against angiotensin-converting enzyme: In vitro effect and stability to gastrointestinal enzymes , 2011, Peptides.

[33]  Jun Jie Wang,et al.  Recent advances of chitosan nanoparticles as drug carriers , 2011, International journal of nanomedicine.

[34]  M. Rezaei,et al.  Development and evaluation of a novel biodegradable film made from chitosan and cinnamon essential oil with low affinity toward water , 2010 .

[35]  Liqun Zhang,et al.  In vitro degradation of starch/PVA films and biocompatibility evaluation , 2010 .

[36]  S. Saha,et al.  A review on mouth dissolving films. , 2009, Current drug delivery.

[37]  J. Boateng,et al.  Development and mechanical characterization of solvent-cast polymeric films as potential drug delivery systems to mucosal surfaces , 2009, Drug development and industrial pharmacy.

[38]  J. Shaji,et al.  Protein and Peptide Drug Delivery: Oral Approaches , 2008, Indian journal of pharmaceutical sciences.

[39]  N. Peppas,et al.  Is the oral route possible for peptide and protein drug delivery? , 2006, Drug discovery today.

[40]  S. Rossi,et al.  Buccal drug delivery: A challenge already won? , 2005, Drug discovery today. Technologies.

[41]  Yanyun Zhao,et al.  Incorporation of a high concentration of mineral or vitamin into chitosan-based films. , 2004, Journal of agricultural and food chemistry.

[42]  Duane D. Miller,et al.  Emerging trends in oral delivery of peptide and protein drugs. , 2003, Critical reviews in therapeutic drug carrier systems.

[43]  H. M. Nielsen,et al.  The Potential of Chitosan in Enhancing Peptide and Protein Absorption Across the TR146 Cell Culture Model—An in Vitro Model of the Buccal Epithelium , 2002, Pharmaceutical Research.

[44]  R. A. Jain,et al.  The manufacturing techniques of various drug loaded biodegradable poly(lactide-co-glycolide) (PLGA) devices. , 2000, Biomaterials.

[45]  H. M. Nielsen,et al.  TR146 cells grown on filters as a model of human buccal epithelium: IV. Permeability of water, mannitol, testosterone and beta-adrenoceptor antagonists. Comparison to human, monkey and porcine buccal mucosa. , 2000, International journal of pharmaceutics.

[46]  M. Alonso,et al.  Novel hydrophilic chitosan‐polyethylene oxide nanoparticles as protein carriers , 1997 .

[47]  P. Sharma,et al.  Buccal Film: An Advance Technology for Oral Drug Delivery , 2014 .