In Vitro and in Vivo characterization of alginate-chitosan-alginate artificial microcapsules for therapeutic oral delivery of live bacterial cells.

Oral administration of artificial cell microcapsules entrapping live bacterial cells is a promising approach in disease therapy. However, the current technology of microcapsules limits this approach. In this study, alginate-chitosan-alginate (ACA) microcapsules entrapping live bacterial cells were prepared with the purpose of oral delivery for therapy, and their in vitro and in vivo properties were investigated. Genetically engineered Escherichia coli DH5 were used as the model bacterial strain. ACA microcapsules remained intact and stable in simulated gastrointestinal fluid and the entrapped bacteria cells survived and grew normally. Moreover, ACA microcapsules were more stable than alginate-polylysine-alginate microcapsules in the rat gastrointestinal tract, which was attributed to the enhanced resistance of the ACA microcapsules to enzymatic digestion. Therefore, these results reinforce the potential of ACA microcapsules for the therapeutic oral delivery of live bacterial cells.

[1]  T. Chang,et al.  Artificial cells for bioencapsulation of cells and genetically engineered E. coli. For cell therapy, gene therapy, and removal of urea and ammonia. , 1997, Methods in molecular biology.

[2]  H. Rautelin,et al.  In Vitro Adhesion and Platelet Aggregation Properties of Bacteremia-Associated Lactobacilli , 1999, Infection and Immunity.

[3]  M. Playne,et al.  A probiotic strain of L. acidophilus reduces DMH-induced large intestinal tumors in male Sprague-Dawley rats. , 1999, Nutrition and cancer.

[4]  R. Muzzarelli,et al.  Biological activity of chitosan: ultrastructural study. , 1988, Biomaterials.

[5]  I. Barchia,et al.  Immune response to orally consumed antigens and probiotic bacteria , 2000, Immunology and cell biology.

[6]  G. Skjåk-Bræk,et al.  Microcapsules of alginate-chitosan. II. A study of capsule stability and permeability. , 1999, Biomaterials.

[7]  G. Skjåk-Bræk,et al.  Long-term reversal of diabetes by the injection of immunoprotected islets. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[8]  P. Chang Encapsulation for Somatic Gene Therapy , 1999, Annals of the New York Academy of Sciences.

[9]  S. Salminen,et al.  Safety of probiotic bacteria. , 1996, Asia Pacific journal of clinical nutrition.

[10]  X. Li The use of chitosan to increase the stability of calcium alginate beads with entrapped yeast cells , 1996, Biotechnology and applied biochemistry.

[11]  R. Neufeld,et al.  Stability of chitosan and poly-L-lysine membranes coating DNA-alginate beads when exposed to hydrolytic enzymes. , 1999, Journal of microencapsulation.

[12]  W. D. de Vos,et al.  Demonstration of safety of probiotics -- a review. , 1998, International journal of food microbiology.

[13]  T. Chang,et al.  Growth and survival of renal failure rats that received oral microencapsulated genetically engineered E. coli DH5 cells for urea removal. , 1998, Artificial cells, blood substitutes, and immobilization biotechnology.

[14]  T. Chang,et al.  Semipermeable Microcapsules , 1964, Science.

[15]  M. Moo-young,et al.  Immobilization of hybridoma cells in chitosan alginate beads , 1991 .

[16]  R. Hausinger,et al.  Microbial ureases: significance, regulation, and molecular characterization. , 1989, Microbiological reviews.

[17]  T. Chandy,et al.  Chitosan/calcium alginate microcapsules for intestinal delivery of nitrofurantoin. , 1996, Journal of microencapsulation.

[18]  W. Fiers,et al.  Treatment of murine colitis by Lactococcus lactis secreting interleukin-10. , 2000, Science.

[19]  Wei Ouyang,et al.  Microencapsulated Genetically Engineered Lactobacillus plantarum 80 (pCBH1) for Bile Acid Deconjugation and Its Implication in Lowering Cholesterol , 2004, Journal of biomedicine & biotechnology.

[20]  M. Sefton,et al.  Making microencapsulation work: conformal coating, immobilization gels and in vivo performance. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[21]  J. Sell,et al.  Effect of orally administered Eubacterium coprostanoligenes ATCC 51222 on plasma cholesterol concentration in laying hens. , 1996, Poultry Science.

[22]  G. Reuter [Probiotics--possibilities and limitations of their application in food, animal feed, and in pharmaceutical preparations for men and animals]. , 2001, Berliner und Munchener tierarztliche Wochenschrift.

[23]  M. Goosen,et al.  Alginate‐Polylysine Microcapsules of Controlled Membrane Molecular Weight Cutoff for Mammalian Cell Culture Engineering , 1987 .

[24]  S. Prakash,et al.  Procedures for microencapsulation of enzymes, cells and genetically engineered microorganisms , 2001, Molecular biotechnology.

[25]  T. Chang Artificial cells with emphasis on cell encapsulation of genetically engineered cells. , 1998, Artificial organs.

[26]  F. Lim,et al.  Microencapsulated islets as bioartificial endocrine pancreas. , 1980, Science.

[27]  D. Laux,et al.  Colonization of the streptomycin-treated mouse large intestine by a human fecal Escherichia coli strain: role of adhesion to mucosal receptors , 1988, Infection and immunity.

[28]  R. Hausinger,et al.  Regulation of gene expression and cellular localization of cloned Klebsiella aerogenes (K. pneumoniae) urease. , 1989, Journal of general microbiology.

[29]  S. Prakash,et al.  Artificial Cell Therapy: New Strategies for the Therapeutic Delivery of Live Bacteria. , 2005, Journal of biomedicine & biotechnology.

[30]  S. Prakash,et al.  Artificial cell microcapsule for oral delivery of live bacterial cells for therapy: design, preparation, and in-vitro characterization. , 2004, Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, Societe canadienne des sciences pharmaceutiques.

[31]  A. Sun,et al.  Generation of alginate-poly-l-lysine-alginate (APA) biomicrocapsules: the relationship between the membrane strength and the reaction conditions. , 1994, Artificial cells, blood substitutes, and immobilization biotechnology.

[32]  T. Chang,et al.  Therapeutic uses of microencapsulated genetically engineered cells. , 1998, Molecular medicine today.

[33]  D. Ecker,et al.  Xenotransplantation of porcine and bovine islets without immunosuppression using uncoated alginate microspheres. , 1995, Transplantation.

[34]  T. Chang,et al.  Microencapsulated genetically engineered live E. coli DH5 cells administered orally to maintain normal plasma urea level in uremic rats , 1996, Nature Medicine.

[35]  A. King,et al.  The effect of host factors and capsule composition on the cellular overgrowth on implanted alginate capsules. , 2001, Journal of biomedical materials research.