مطالعه نقش حفاظتی دانکهای دوپوشینهای آلژینات کلسیم-کیتوزان- نانوذرات اودوراژیت S100 حاصل از ریزپوشانی باکتری لاکتوباسیلوس اسیدوفیلوس به عنوان فلور غالب روده انسان و حیوانات

نویسندگان

گروه بهداشت و کنترل کیفی مواد غذایی، دانشکده دامپزشکی دانشگاه تهران، تهران- ایران

چکیده

زمینه مطالعه: پروبیوتیک‌ها دارای اثرات سودمند زیادی می‌باشند و توانایی زنده‌مانی پایین آن‌ها در شرایط دشوار اسیدی صفراوی دستگاه‌گوارش و شرایط محصولات غذایی، محققین را همیشه به پیدا کردن راه‌های حل این مشکل تشویق کرده است. ریزپوشانی به عنوان یک روش کارا اثر قابل توجهی در این زمینه داشته است. هدف: این مطالعه با هدف بررسی نقش حفاظتی دانک‌های دوپوشینه‌ای آلژینات کلسیم-کیتوزان- نانوذرات اودوراژیت S100 حاصل از ریزپوشانی باکتری لاکتوباسیلوس اسیدوفیلوس بعنوان فلور غالب روده انسان و حیوانات انجام گرفت. روش کار: پس از فعال‌سازی باکتری استارتر ل. اسیدوفیلوس در محیط MRS broth، برای خالص‌سازی باکتری از سانتریفیوژ (سرعتrpm‌5000 به مدت min‌10) استفاده شد. ریزپوشانی باکتری پروبیوتیک به روش اکستروژن انجام گرفت. بررسی استحکام دانک‌ها در طول h 12 و بررسی میزان زنده‌مانی باکتری‌ها در طول min 120 در درون  اسیدهیدروکلریک، بافرفسفات و محلول پودر دایجستیو در دو حالت با و بدون تنش مکانیکی انجام گرفت. جهت کشت از محیط  MRS-Salicin-agar و روش کشت سطحی و گرمخانه‌گذاری در دمای oC‌37 به مدت  48 ساعت استفاده شد. داده‌ها بوسیله آزمون t-test مستقل مورد آنالیز قرار گرفتند. نتایج: شکل و اندازه دانک‌ها بوسیله میکروسکوپ نوری نشان داده شد. نتایج نشان دادند که میزان زنده‌مانی باکتری‌های ریزپوشانی-شده در شرایط فوق در دو حالت با و بدون تنش مکانیکی بطور معنی‌داری بیشتر از باکتری‌های آزاد می‌باشد (05/0>p). نتیجهگیری نهایی: ریزپوشانی با آلژینات کلسیم-کیتوزان-اودوراژیت S100 نقش مهمی در ارتقای میزان زنده‌مانی پروبیوتیک ل. اسیدوفیلوس ایفا می‌کند.

کلیدواژه‌ها


عنوان مقاله [English]

Study of protective role of double coated beads of calcium alginate-chitosan-eudragit s100 nanoparticles achieved from microencapsulation of Lactobacillus acidophilus as a predominant flora of human and animals gut

نویسندگان [English]

  • Hadi Pourjafar
  • negin noori
  • Hasan Gandomi Nasrabadi
  • Afshin Akhondzadeh Basti
Department of Food Hygiene, Faculty of Veterinary Medicine, University of Tehran, Tehran- Iran
چکیده [English]

BACKGROUND: Probiotics have more functional effects and less survival  under hard acidic-bile circumstances of digestive system, and foodstuff products situation has persuaded investigators to find techniques to resolve this problem. Microencapsulation as a useful method has a perceptible effect in this regard. Objectives: The aim of this study was to assess the protective role of double coated beads of calcium alginate-chitosan-eudragit S100 achieved from microencapsulation of Lactobacillus acidophilus as a predominant flora of human and animals gut. Methods: Following activation of starter culture of L.acidophilus in MRS-broth medium,  centrifuge (at aspeed of 5000 rpm for 10 minutes) was used to purify bacteria. Extrusion technique was used for Microencapsulation of probiotic bacterium. The survey of beads solidity was carried out for 12 hours and the study of survival of microencapsulated bacteria was done for 120 minutes inside hydrochloric acid, phosphate buffer and digestive powder solution.  MRS-Salicin-agar and pour plate method and incubation at 37oC for 48 h was done for cultivation. Data were analyzed by means of an independent t-test. Results: Shape and size of beads were shown by optical microscope. The consequences demonstrated that survivability of microencapsulated bacteria in the mentioned conditions, in both situation with and without mechanical tensions, is significantly more than free bacteria (p<0/05). Conclusions: Microencapsulation with calcium alginate- chitosan-eudragit S100 plays a significant role  in increasing the rate of L. acidophilus viability.

کلیدواژه‌ها [English]

  • eudragit S100
  • calcium alginate
  • Chitosan
  • lactobacillus acidophilus
  • Microencapsulation
Anal, A.K., Singh, H. (2007) Recent advances in microencapsulation of probiotics for industrial applications and targeted delivery. Trends Food Sci Tech.18: 240-251.

Arpita, D., Sohini, R., Utpal, R., Runu, C. (2014) Microencapsulation of probiotic bacteria and its potential application in food technology. Int J Agric Environ Biotechnol. 7: 47-53.

Badhana, S., Garud, N., Garud, A. (2013) Colon specific drug delivery of mesalamine using eudragit S100-coated chitosan microspheres for the treatment of ulcerative colitis. Int Curr Pharm J. 2: 42-48.

Chávarri, M., Marañón, I., Ares, R., Ibáñez, F.C., Marzo, F., Villarán, M. (2010) Microencapsulation of a probiotic and prebiotic in alginate-chitosan capsules improves survival in simulated gastro-intestinal conditions. Int J Food Microbiol. 142: 185-189.

Cui, J.H., Goh, J.S., Kim, P.H., Choi, S.H., Lee, B.J. (2000) Survival and stability of Bifidobacteria loaded in alginate poly-l-lysine microparticles. Int J Pharm. 210: 51-59.

Drisko, J.A., Giles, C.G., Bischoff, B.J. (2003) Probiotics in health maintenance and disease prevention. Alter Med Rev. 8: 143-215.

Gallardo, G., Guida, L., Martinez, V., Lopez, M.C., Bernhardt, D., Blasco, R., Pedroza-lslas, R., Hermida, L.G. (2013) Microencapsulation of linseed oil by spray drying for functional food application. Food Res Int. 52: 473-482.

Homayouni, A., Azizi, A., Ehsani, M.R, Yarmand, M.S., Razavi, S.H. (2008) Effect of microencapsulation and resistant starch on the probiotic survival and sensory properties of symbiotic ice cream. Food Chem. 111: 50-55.

Hu, D., Liu, L., Chen, W., Li, S., Zhao, Y. (2012) A novel preparation method for 5-Aminosalicylic acid loaded Eudragit S100 nanoparticles. Int J Mol Sci. 13: 6454-6468.

Kailasapathy, K., Harmstorf, I., Phillips, M. (2008) Survival of Lactobacillus acidophilus and Bifidobacterium animalis ssp. Lactis in stirred fruit yogurts. LWT-Food Sci Technol. 41: 1317-1322.

Kanmani, P., Satish Kumar, R., Yuvaraj, N., Paari, K.A., Pattukumar, V., Arul, V. (2011) Cryopreservation and microencapsulation of a probiotic in alginate-chitosan capsules improves survival in simulated gastrointestinal conditions. Biotechnol Bioprocess Eng. 16: 1106-1114.

Khaleghi, N. (2012) Nanoencapsulation of silibin with Eudragit RL for colon targeted delivery. Res Pharm Sci. 7: 222.

Klaenhammer, T.R. (2001) Food Microbiology: Fundamentals and Frontiers. (2nd ed.) ASM press. Washington D.C, USA.

Krasaekoopt, W., Bhandari, B., Deeth, H. (2004) The influence of coating materials on some properties of alginate beeds and survivability of microencapsulated probiotic bacteria. Int Dairy J. 14: 737-743.

Krasaekoopt, W., Bhandari, B., Deeth, H. (2005) Survival of probiotics encapsulated in chitosan-coated alginate beads in yoghurt from UHT- and conventionally treated milk during storage. LWT-Food Sci Technol. 14: 315-324.

Kristo, E., Bilianderis, C.G., Tzanetakis, N. (2003) Modelling of rheological, microbiological and acidification properties of Lactobacillus paracasei. Int Dairy J. 13: 517-528.

Lam, P.L., Gambari, R. (2014) Advanced progress of microencapsulation technologies: In vivo and in vitro models for studying oral and transdermal drug deliveries. J Control Release. 178: 25-45.

Lee, J.S., Cha, D.S., Park, H.J. (2004) Survival of freeze-dried Lactobacillus bulgaricus KFRI 673 in chitosan-coated calcium alginate microparticles. J Agric Food Chem. 52: 7300-7305.

Li, P., Dai, Y.N., Zhang, J.P., Wang, A.Q., Wei, Q. (2008) Chitosan-alginate nanoparticles as a novel drug delivery system for Nifedipine. Int J Biomed Sci. 4: 221-228.

Liserre, A.M., Ines Ré, M., Franco, B.D.G.M. (2007) Microencapsulation of Bifidobacterium animalis subsp. Lactis in Modified alginate-chitosan beads and evaluation of survival in simulated gastrointestinal conditions. Food  Biotechnol. 21: 1-16.

Madziva, H., Kailasapathy, K., Phillips, M. (2006) Evaluation of alginate-pectin capsules in cheddar cheese as a food carrier for the delivery of folic acid. Food Sci Technol-Lebenson Wiss Technol. 39: 146-151.

Mirzaei, H., pourjafar, H., Homayouni, A. (2011) The effect of microencapsulation with calcium alginate and resistant starch on the Lactobacillus acidophilus (LA5) survival rate in simulated gastrointestinal juice conditions. J Vet Res. 66: 337-342.

Mirzaei, H., Pourjafar, H., Homayouni, A. (2012) Effect of calcium alginate and resistant starch microencapsulation on the survival rate of Lactobacillus acidophilus La5 and sensory properties in Iranian white brined cheese. Food Chem. 132: 1966-1970.

Nikam, V.K., Kotade, K.B., Gaware, V.M. Dolas, R.T. Dhamak, K.B., Somwanshi, S.B., Khadse, A.N., Kashid, V.A. (2011) Eudragit A Versatile Polymer: A Review. Pharmacologyonline. 1: 152-164.

Özer B., Kirmacia H., Ebru S, Enel b, Atamer M.B., Hayaloglu A. (2009) Improving the viability of Bifidobacterium bifidum BB-12 and Lactobacillus acidophilus LA-5 in white-brined cheese by microencapsulation. Int Dairy J. 19: 22-29.

Picot, A., Lacroix, C. (2004) Encapsulation of Bifidobacteria in whey protein-based microcapsules and survival in simulated gastrointestinal conditions and in yoghurt. Int Dairy J. 14: 505-515.

Picout, D.R., Ross-Murphy S.B, Errington, N., Harding, S.E. (2001) Pressure cell assisted solution characterization of polysaccharides. 1. Guar gum. Biomacromolecules. 2: 1301-1309.

Porzio, M. (2008) Melt extrusion and melt injection. Perfumer Flavorist. 33: 48-53.

Pourjafar, H., Ghasemnezhad, R. (2010) Probiotics as a suitable replacement for common antibiotics against infectious disease. J IRIAF Health Administration. 13: 72-77.

Pourjafar, H., Ghasemnezhad, R. (2013) Role of probiotic bacteria in cancer prevention. Pejvad. 1: 129-134.

Pourjafar, H., Mirzaei, H., Ghasemnezhad, R., Homayouni, A. (2011) Study of morphological and protective characteristics of beads obtained from microencapsulation of Lactobacillus acidophilus probiotic as a predominant and natural flora in human gut. J Army Uni Med Sci. 9: 233-240.

Pourjafar, H., Mirzaei, H., Homayouni, A. (2010) Study of the survival rate of free and microencapsulated Lactobacillus acidophilus La5 in Iranian white cheese during manufacture and storage. Iranian Vet J. 7: 50-59.

Rathore, S., Desai, P.M., Liew, C.V., Chan, L.W., Sia Heng, P.W. (2013) Microencapsulation of microbial cells. J Food Eng. 116: 369-381.

Salminen, S., Bouley, C., Boutron-Ruault, M.C., Cummings, J.H., Franck, A., Gibson, G.R, Isolauri, E., Moreau, M.C., Roberforid, M., Rowland, I. (1998) Functional food science and gastrointestinal physiology and function. Br J Nutr. 1: 147-171.

Saxelin, M., Lassig, A., Karjalainen, H., Tynkkynen, S., Surakka, A., Vapaatalo, H., Järvenpää, S., Korpela, R., Mutanen, M., Hatakka, K. (2010) Persistence of probiotic strains in the gastrointestinal tract when administered as capsules, yoghurt, or cheese. Int J Food Microbiol. 144: 293-300.

Shah, N.P. (2000) Probiotic bacteria: Selective enumeration and survival in dairy foods. J Dairy Sci. 83: 894-907.

Shen, Q., Quek, S.Y. (2014) Microencapsulation of astaxanthin with blends of milk protein and fiber by spray drying. J Food Eng. 123: 165-171.

Shima, M., Morita, Y., Yamashita, M., Adachi, S. (2006) Protection of Lactobacillus acidophilus from the low pH of a model gastric juice by incorporation in a W/O/W emulsion. Food Hydrocolloid. 10: 1016-1027.

Simonoska Crcarevska, M., Glavas Dodov, M., Goracinova, K. (2008) Chitosan coated Ca-alginate microparticles loaded with budesonide for delivery to the inflamed colonic mucosa. Eur J Pharm Biopharm. 68: 565-578.

Sultana, K., Godward, G., Reynolds, N., Arumugaswamy, R., Peiris, P., KailasaPathy, K. (2000) Encapsulation of Probiotic bacteria with alginate-starch and Evaluation of Survival in Simulated Gastrointestinal Conditions and in Yoghurt. Int J Food Microbiol. 62: 47-55.

Tatar, F., Tugce, T., Dervisoglu, M., Cekmecelioglu, D., Kahyaoglu, T. (2014) Evaluation of hemicellulose as a coating material with gum Arabic for food microencapsulation. Food Res Int. 57: 168-175.

Vinderola, C.G., Reinheimer, J.A. (2000) Enumeration of Lactobacillus casei in the presence of L.acidophilus, Bifidobacteria and Lactic starter bacteria in fermented dairy products. Int Dairy J. 10: 271-275.

Yoo, J.W., Giri, N., Lee, C.H. (2011) pH-sensitive Eudragit nanoparticles for mucosal drug delivery. Int J Pharma. 403: 262-267.

Zarate, G., Nader-Macias, M.E. (2006) Viability and biological properties of probiotic vaginal Lactobacilli after lyophilization and refrigerated storage into gelatin capsules. Process Biochem. 41: 1779-1785.

Zuidam, N.J., Nedovic, V.A. (2010) Encapsulation Technologies for Active Food Ingredients and Food Processing. (1st ed.) St. Spring, Springe, New York, USA.