Progress 05/01/11 to 04/30/16

Outputs
Target Audience:Academia, food and pharmaceutical industries. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?A research associate, a post-doctoral researcher, a research scientist-part timeand severalundergraduate students were hired on the project. They were all trained on nanoparticle synthesis and characterization, tissue analysis and new methodologies for nanoparticle detection in various cells and tissues. How have the results been disseminated to communities of interest?The results of this work have been presented at numerous national and international conferences, as follows: National Academy of Engineering, American Chemical Society, Food and Drug Administration -CFSAN, Institute of food Technologists, Romanian Diaspora Conference,Gordon Conference on Environmental Nanotechnology, and Gordon Conference on Nanoscale Science & Engineering for Agriculture & Food Systems among others. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Nano-particle research and potential applications are increasing in scale and broadening in scope. Available data on nanoparticles documents the increased efficacy of bioactives delivered with polymeric nanoparticles when compared to emulsions or other delivery systems. The limited data on safety of the nanomaterials to both animals and the environment is of concern to scientists, governments, non-governmental organizations, members of the public and stake holders. However, special promising physiochemical properties of nanoparticles can result in very important applications food and agriculture. Louisiana State University AgCenter teamed up with Tufts University, Mississippi State University and the Biochemistry Institute, Romania to address safety and efficacy of polymeric nanoparticles. The team focused on determining the in-vivo biodistribution of two types of polymeric nanoparticles, their in-vitro and in-vivo safety, and the impact of nanoparticle biodistribution on the pharmacokinetics of nanodelivered bioactive. It was found that nanoparticles packed mostly in the intestine and liver, where they exhibited a minimum toxicity in rats orally exposed to nanoparticles daily for 21 days, but no detrimental effect was found in other organs sampled. Bioavailability of nodelivered lutein was significantly improved in rat plasma and selected tissues. In conclusion, nanodelivery with polymeric nanoparticles was identified as a viable approach to improve bioavailability of entrapped bioactive without major safety concerns to the consumer.

Publications

• Type: Journal Articles Status: Published Year Published: 2016 Citation: Simon, L. C., C. M. Sabliov, R. W. Stout. 2016. Bioavailability of orally delivered alpha-tocopherol by poly(lactic-co-glycolic) acid (PLGA) nanoparticles and chitosan covered PLGA nanoparticles in F344 rats. Nanobiomedicine. NBM.2016.0007R1.
• Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Chuacharoen, T. and C. M. Sabliov. 2016. Stability and Controlled Release of Lutein Loaded in Zein Nanoparticles with and without Lecithin and Pluronic F127 Surfactants. Colloids and Surfaces A: Physicochemical and Engineering Aspects. COLSUA-D-16-00327. In press.
• Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Chuacharoen, T. and C. M. Sabliov. 2016. The Potential of Zein Nanoparticles to Protect Entrapped ?-Carotene in the Presence of Milk under Simulated Gastrointestinal (GI) Conditions. LWT. LWT-D-16-00474. In press.
• Type: Journal Articles Status: Accepted Year Published: 2016 Citation: M. Navarro, Timothy Morgan, Carlos E. Astete, Rhett Stout, Diana Coulon, Pete Mottram & Cristina M. Sabliov. 2016. Biodistribution of PLGA, PLGA/Chitosan nanoparticles in rats following daily oral exposure for 7, 14, and 21 days. Nanomedicine. In press.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Kamil, A., D. E. Smith, J. B. Blumberg, C. Astete, C. M. Sabliov. 2016. Bioavailability and biodistribution of nanodelivered lutein. Journal of Food Chemistry. 192:915-23. doi: 10.1016/j.foodchem.2015.07.106. Epub 2015 Jul 23.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Trif, M, P. E. Florian, A. Roseanu, M. Moisei, O. Craciunescu, C. E. Astete and C. M. Sabliov. 2015. Cytotoxicity and Intracellular Fate of PLGA and Chitosan-coated PLGA Nanoparticles in Madin-Darby Bovine Kidney (MDBK) and Human Colorectal Adenocarcinoma (Colo 205) Cells. Journal of Biomedical Materials Research Part A. 103(1):3599-3611. DOI: 10/1002/jbm.a.35498.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Alqahtani, S., L. Simon, C. E. Astete, A. Alayoubi, P. W. Sylvester, S. Nazzal, Y. Shen, Z. Xu, A. Kaddoumi, C. M. Sabliov. 2015. Cellular uptake, antioxidant and antiproliferative activity of entrapped ?-tocopherol and ?-tocotrienol in poly (lactic-co-glycolic) acid (PLGA) and chitosan covered PLGA nanoparticles (PLGA-Chi). Journal of Colloid and Interface Science. 445:243-251. http://dx.doi.org/10.1016/j.jcis.2014.12.083.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Navarro, S., C. Darensbourg, L. Cross, R. Stout, D. Coulon, C. E. Astete, T. Morgan, C M. Sabliov. 2014. Biodistribution of poly(lactic-co-glycolic) acid (PLGA) and PLGA/chitosan nanoparticles after repeat-dose oral delivery in F344 rats for seven days. Therapeutic Delivery. 5(11), 11911201.
• Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Yada, R. Y., N. Buck, R. Canady, C. DeMerlis, T. Duncan, G. Janer, L. Juneja, M. Lin, J. McClements, G. Noonan, J. Oxley, C. M. Sabliov, L. Tsytsikova, S. V�zquez-Campos, J. Yourick, Q. Zhong, S. Thurmond. 2014. Engineered Nanoscale Food Ingredients: Evaluation of Current Knowledge on Material Characteristics Relevant to Uptake from the Gastrointestinal Tract. Comprehensive Reviews in Food Science and Food Safety. 13(4):730-744.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Borel, T. and C. M. Sabliov. 2014. Nanodelivery of Bioactive Components for Food Applications: Types of Delivery Systems, Properties, and their Effect on ADME Profiles and Toxicity of Nanoparticles. Annu. Rev. Food Sci. Technol. 2014. 5:12.112.17. DOI: 10.1146/annurev-food-030713-092354.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Simon, L. and C. M. Sabliov. 2014. The effect of nanoparticle properties, detection method, delivery route and animal model on poly(lactic-co-glycolic) acid nanoparticles biodistribution in mice and rats. Drug Metab Rev. 46(2):128-41. doi: 10.3109/03602532.2013.864664. Epub 2013 Dec 5.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Simon, L. and C. M. Sabliov. 2013. Time analysis of poly(lactic-co-glycolic) acid nanoparticle uptake by major organs following acute intravenous and oral administration in mice and rats. Invited paper. Journal of Industrial Biotechnology. 9(1): 19-23.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Li, J. and C. M. Sabliov. 2013. PLA/PLGA nanoparticles for delivery of drugs across the blood-brain barrier. Nanotechnology Reviews. 2(3): 241-257.
• Type: Other Status: Published Year Published: 2015 Citation: Nanoparticles could slash use of pesticides in agriculture. June 2015. Controlled Environments. http://www.cemag.us/news/2015/07/nanoparticles-could-slash-use-pesticides-agriculture.
• Type: Other Status: Published Year Published: 2015 Citation: Undergraduate learns research ropes through nanotechnology. The Daily Reveille. January 29, 2015. http://www.lsureveille.com/daily/undergraduate-learns-research-ropes-through-nanotechnology/article_179834f8-a754-11e4-bd64-5b19a16aa0fa.html
• Type: Other Status: Published Year Published: 2013 Citation: Comprehensive Reviews in Food Science and Food Safety. 2013 IFT International Food Nanoscience Conference: Proceedings. Rosetta Newsome.
• Type: Other Status: Published Year Published: 2013 Citation: Researchers study use of nanoparticles in ag chemicals. AgProfessional. http://www.agprofessional.com/news/Researchers-study-use-of-nanoparticles-in-ag-chemicals-211450961.html. 2013.
• Type: Other Status: Published Year Published: 2013 Citation: Material Characteristics Relevant to the Uptake and Bioavailability of Engineered Nanomaterials Delivered via Human Diet- A White Paper from the NanoRelease Food Additive Project. ILSI, Canada. http://www.ilsi.org/ResearchFoundation/RSIA/Pages/NRFA_TaskGroup1.aspx2013. 2013.
• Type: Other Status: Published Year Published: 2013 Citation: Neil H. Mermelstein (with material provided by C. M. Sabliov). 2013. Testing Nanomaterial Safety. Food Technology.
• Type: Other Status: Published Year Published: 2013 Citation: Scientist Utilizing Nanotechnology to Improve the Food Safety and Nutrition. IFT website. http://www.ift.org/newsroom/news-releases/2011/june/14/scientist-utilizing-nanotechnology-to-improve-the-food-safety-and-nutrition.aspx. 2011.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2016 Citation: Bodoki, E. and C. M. Sabliov. November 2016. Sustainable Nanotechnology Meeting. Zein nanoparticles as pesticide delivery systems. Orlando, FL.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2016 Citation: Ye, F. and C. M. Sabliov. September 2016. Entrapment and delivery of ?-tocopherol by a self-assembled, alginic acid-conjugated, prodrug nanostructure for effective inhibition of lipid oxidation Conference of Food Engineering. Columbus, OH.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Sabliov, C. M. December 2015. Polymeric nanoparticles for oral delivery of drugsefficacy and safety assessment. Nanotechnology for Health Care. A Winthrop Rockefeller Conference. Morrilton, AR.
• Type: Other Status: Published Year Published: 2015 Citation: Sabliov, C. M. September 2015. Nanodelivery systems, efficacy and safety assessment. LSU Board of Supervisors Meeting. Baton Rouge, LA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Whaley, M., T. Borel, B. Novak, C. E. Astete, D. Moldovan, C. M. Sabliov. April 2015. Nanoparticle cellular uptake- effect on bioactive functionality. LA-Sigma meeting. Baton Rouge, LA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Sabliov, C. M. June 2015. Data on Polymeric-Based Nanodelivery Systems  Efficacy and Safety Assessment. Gordon Conference. Nanoscale Science & Engineering for Agriculture & Food Systems. Waltham, MA.
• Type: Other Status: Published Year Published: 2012 Citation: Sabliov, C. M. March 2012. Polymeric nanoparticles as delivery systems for disease prevention and treatment. Division of Human Nutrition and Food. LSU, Baton Rouge, LA.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2015 Citation: Chuacharoen, T. and C. M. Sabliov. August 2015. Influence of combined lecithin and pluronic F127 surfactant on stability of lutein-loaded zein nanoparticles as a function of time and temperature. 250th American Chemical Society National Meeting and Exposition. Boston, MA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Chuacharoen, T. and C. M. Sabliov. July 2015. The potential of zein nanoparticles to protect ?-carotene in the presence of milk under simulated gastrointestinal (GI) environments. Institute of Food Technologists Annual meeting. Chicago, IL.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Sabliov, C. M. March 2015. Nano- and colloidal delivery systems for Food, Agriculture, and Health applications. PBRC meeting. Baton Rouge, LA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Alqahtani, S., L. Simon, C. E. Astete, A. Alayoubi, P. W. Sylvester, S. Nazzal, Y. Shen, Z. Xu, A. Kaddoumi, C. M. Sabliov. March 2015. Cellular uptake, antioxidant and antiproliferative activity of entrapped alpha-tocopherol and gamma-tocotrienol in poly (lactic-co-glycoliC) acid (PLGA) and chitosan covered PLGA (PLGA-Chi) nanoparticles. LA-CRI program. Monroe, LA.
• Type: Other Status: Published Year Published: 2015 Citation: Sabliov, C. M. March 2015. Nano- and colloidal delivery systems for Food, Agriculture, and Health applications. Cultivian Meeting. Baton Rouge, LA.
• Type: Other Status: Published Year Published: 2014 Citation: Sabliov C. M. November 2014. Polymeric nanoparticles as delivery systems in foods. BAEN Seminar. College Station, TX.
• Type: Other Status: Published Year Published: 2013 Citation: Sabliov, C. M. February 2013. Research briefs. LSU AgCenter Internal Leadership Program. Baton Rouge, LA.
• Type: Other Status: Published Year Published: 2012 Citation: Sabliov, C. M. June 2012. Polymeric nanoparticles to improve human health. Summer undergraduate research program. Baton Rouge, LA.
• Type: Other Status: Published Year Published: 2012 Citation: Sabliov, C. M. April 2012. BAE Advisory Council. Nano- and colloidal delivery systems for improved food quality, food safety, and human health. Baton Rouge, LA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Sabliov, C. M. August, 2014. An overview of nanoscale systems to enhance delivery efficacy of bioactives and micronutrients in food for healthy outcomes. IUFOST. 17th World Congress of Food Science and Technology & Expo. Montreal, Canada.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Borel, T., M. Whaley, C. M Sabliov, Z. Xu, Y. Shen. March 2014. Poly (DL-lactide Co-glycolide) acid and PLGA/chitosan nanoparticles with entrapped lutein: Stability and antioxidant activity. 247th ACS National Meeting and Exposition. Dallas, TX.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Simon, L. and C. M. Sabliov. July 2013. Bioavailability of alpha-tocopherol delivered with polymeric nanoparticles. Institute of Food Technologists International Food Nanoscience Conference. Chicago, IL.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sabliov, C. M. June 2013. Biodistribution and safety of PLGA nanoparticles. Gordon Conference on Environmental Nanotechnology. Stoweflake Resort and Conference Center. Stowe, VT.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sabliov, C. M., S. Navaro, C. Darensbourg, L. Cross, Diana Coulon, R. Stout, T. Morgan, C. Astete. June 2013. Biodistribution of poly (lactic-co-glycolic) acid (PLGA) and PLGA/Chitosan nanoparticles in F344 rats orally exposed to nanoparticles for seven days. TAPPI International Conference on Nanotechnology for Renewable Materials. Stockholm, Sweden.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Sabliov, C. M. October 2012. Polymeric nanoparticles as delivery systems: applications to food, health, and agriculture. Japan-America Frontiers of Engineering. National Academy of Engineering. Irvine, CA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Sabliov, C. M. September 2012. Targeted nanoparticles for cancer prevention and treatment. Diaspora Conference. Bucharest, Romania.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Sabliov, C. M. and T. Morgan. June 2012. Biodistribution of polymeric nanoparticles used for enhanced vitamin delivery. IFT, Las Vegas, NV.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Sabliov, C. M. February 2012. Materials and properties- food packaging and food. Basic nanotechnology science course at FDA-CFSAN. Washington, DC.

Progress 05/01/13 to 04/30/14

Outputs
Target Audience: Academia, food and pharmaceutical industries. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? A post-doctoral researcher was hired to work on the cell-studies due to her prior expertise with confocal microscopy. She was trained on FACS analysis and new methodologies for nanoparticle detection in various cells. Four undergraduate students continued to be actively involved in the study, performing tasks ranging from nanoparticle synthesis and characterization to processing tissue and measuring fluorescence. How have the results been disseminated to communities of interest? The results of this work have been presented at numerous national and international conferences, as follows: American Chemical Society National Meeting, Institute of Food Technologists International Nanoscience Conference, and Gordon Conference on Environmental Nanotechnology. What do you plan to do during the next reporting period to accomplish the goals? A new study will be initiated on nanoparticle safety at a higher dose. Lutein loaded nanoparticles will be tested in a pharmacokinetic study conducted in F344 rats at Tufts University Two papers on biodistribution and histology and a paper on cell studies will be submitted for publication in 2014.

Impacts
What was accomplished under these goals? A review of poly(lactic-co-glycolic) acid (PLGA) nanoparticle (NP) biodistribution was conducted with the intent of identifying particle behavior for drug delivery applications. The papers included in the review were limited to those that report biodistribution data in terms of % dose particles/g tissue in the liver, kidney, spleen, lung, heart and brain. Noted trends involved particle behavior based on individual organ, particle size, animal model, type of indicator (entrapped versus covalently linked) and method of delivery (oral or i.v.). The liver showed the highest uptake of particles in mice, and the lung showed the highest uptake in rats. Minimal amounts of particles were detected in both the heart and brain of rats and mice. In rats, the concentration of particles approached 0% dose/g or decreased significantly over 24 h after administration of a single dose of particles. Higher concentrations of smaller particles were evident in the liver, kidney and spleen. Orally delivered drugs showed little to no uptake within the 24h analysis when compared with i.v. delivered NPs. Differences in particle concentrations between rats and mice were also observed as expected when expressed as % dose/g organ. Particles with covalently linked indicators showed lower concentrations in tissues than particles with physically entrapped indicators. Further research on oral delivery of PLGA NPs as well as distribution beyond 24 h is needed to fully understand particle behavior in vivo for successful application of NPs in drug delivery. Nanoparticle (NP) interaction with mammalian cells was assessed by cytotoxicity measurements, fluorescence microscopy investigations and flow cytometry quantitations. Results showed that NPs did not induce cell toxicity up to a concentration of 2500µg/ml. At higher concentration, MDBK (Madin-Darby) bovine kidney cells showed decrease in proliferation, but Colo 205 (human intestinal epithelial cells) showed a good viability even at high concentrations of nanoparticles (5000 µg/ml). Kinetic studies on nanoparticle internalization by MDBK cells revealed that NPs were internalized as soon as 30 minutes after treatment. With all NPs tested an intracellular accumulation was observed after 24h, except with PLGA, which seemed to have peaked at 3h. Colo205 cells showed a slow internalization of NPs. After 24h, 85% of the cells internalized the NPs without chitosan, compared to only 65% of cells in the case of NPs with chitosan. It appears that the presence of chitosan inhibit nanoparticles internalization or delay the endocytic process. Regarding ER compartment co-localization in MDBK cells, it was shown that the NPs without chitosan partially overlay with ER compartment marker after 24h. Comparatively, the NPs with chitosan did not co-localize with the ER, despite their perinuclear localization. Colo205 cells revealed a diffuse pattern of NP internalization after 24h for both types of NPs studied. Interestingly, the NPs did not seem to target the ER compartment of Colo 205. Moreover, NPs agglomeration at plasma membrane was detected, indicating that the endocytosis of the NPs in Colo205 cells was a very slow and highly specific process.

Publications

• Type: Journal Articles Status: Published Year Published: 2014 Citation: Borel, T. and C. M. Sabliov. Nanodelivery of Bioactive Components for Food Applications: Types of Delivery Systems, Properties, and their Effect on ADME Profiles and Toxicity of Nanoparticles. Annu. Rev. Food Sci. Technol. 2014. 5:12.112.17. DOI: 10.1146/annurev-food-030713-092354.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Simon, L. and C. M . Sabliov. The effect of nanoparticle properties, detection method, delivery route and animal model on poly(lactic-co-glycolic) acid nanoparticles biodistribution in mice and rats. 2013 Dec 5. [Epub ahead of print]
• Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: Borel, T., M. Whaley, C. M Sabliov, Z. Xu, Y. Shen. March 2014. Poly (DL-lactide Co-glycolide) acid and PLGA/chitosan nanoparticles with entrapped lutein: Stability and antioxidant activity. 247th ACS National Meeting and Exposition. Dallas, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Simon, L. and C. M. Sabliov. July 2013. Bioavailability of alpha-tocopherol delivered with polymeric nanoparticles. Institute of Food Technologists International Food Nanoscience Conference. Chicago, IL.
• Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Sabliov, C. M. June 2013. Biodistribution and safety of PLGA nanoparticles. Gordon Conference on Environmental Nanotechnology. Stoweflake Resort and Conference Center. Stowe, VT.
• Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Sabliov, C. M., S. Navaro, C. Darensbourg, L. Cross, Diana Coulon, R. Stout, T. Morgan, C. Astete. June 2013. Biodistribution of poly (lactic-co-glycolic) acid (PLGA) and PLGA/Chitosan nanoparticles in F344 rats orally exposed to nanoparticles for seven days. TAPPI International Conference on Nanotechnology for Renewable Materials. Stockholm, Sweden.

Progress 05/01/12 to 04/30/13

Outputs
Target Audience: Academia, food and pharmaceutical industries. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? A research associate was hired on the project to conduct the animal studies. She was trained on tissue analysis and new methodologies for nanoparticle detection in various tissues. Four undergraduate students were actively involved in the study, performing tasks ranging from nanoparticle synthesis and characterization to processing tissue and measuring fluorescence. How have the results been disseminated to communities of interest? The results of this work have been presented at numerous national and international conferences, as follows: National Academy of Engineering, Institute of Food Technologists, Food and Drug Administration –CFSAN, Institute of food Technologists and Gordon Conference on Environmental Nanotechnology. In addition, presentations were made to academic institutions including Louisiana State University, and Carol Davila University of Medicine, Romania. What do you plan to do during the next reporting period to accomplish the goals? Nanoparticle interaction with mammalian cells will be assessed by cytotoxicity measurements, fluorescence microscopy investigations and flow cytometry quantitations. In-vivo histological evaluations of various tissues derived from treated F344 rats in the repeat-dose study will be completed. Lutein loaded nanoparticles will be made, characterized, and tested in a pharmacokinetic study conducted in F344 rats.

Impacts
What was accomplished under these goals? A repeat-dose study was performed by our group to decipher how daily exposure to nanoparticles (3 mg/dose) for 7, 14, and 21 days may affect biodistribution and toxicity of the PLGA and PLGA/Chitosan in a F344 rat model. PLGA and PLGA/Chi particles showed that the amount of particles detected stabilized in tissues by 7 days, suggesting that the particles were not bio-accumulating at the dosage used. The results are not surprising, as the polymer is eliminated from the body by excretion or degradation and no long-term accumulation is expected for biodegradable polymers. The percentage of daily dose of nanoparticles recovered in all the tissues analyzed was 10.9% daily dose/g for PLGA and 12.3% daily dose/g for PLGA/Chi nanoparticles, showing a difference of 1.4% daily/g in favor of PLGA/Chi relative to PLGA nanoparticles. Statistically, the difference was not significant. The relative NP biodistribution in various organs indicated that the highest amount of PLGA nanoparticles concentrated in the spleen (3.9% daily dose/g), followed by kidney (2.1% daily dose/g), intestine (1.98% daily dose/g), lung (1.18% daily dose/g), liver (0.98% daily dose/g or 0.14 % total dose/g), brain (0.59% daily dose/g) and heart (0.18% daily dose/g). Liver and spleen are known to be two major organs for systemic distribution of engineered nanoparticles. The significant presence of the particles in the kidney may indicate that particles, or parts of the particles that contained the fluorophore, were cleared by the kidney into the urine. Interestingly, fluorescence was present in the heart; however, in these studies, samples included heart blood, and it is likely that much of the fluorescence seen in the heart was actually due to particles in the heart blood. Histological studies indicated that PLGA and PLGA/Chi ENPs made with poly(vinyl alcohol) (PVA) surfactant, measuring 120 nm did not show evidence of toxicity or increased inflammation in the organs evaluated (liver, kidney, spleen, intestine), in F344 rats exposed to EPNs daily for as long as 21 days. As biodegradable nanoparticles are believed to be removed by macrophages of the RES, further more in-depth studies are needed to fully comprehend the extent of possible ENP immunogenicity.

Publications

• Type: Journal Articles Status: Published Year Published: 2013 Citation: Simon, L. and C. M. Sabliov. 2013. Time analysis of poly(lactic-co-glycolic) acid nanoparticle uptake by major organs following acute intravenous and oral administration in mice and rats: a review. Invited paper. Journal of Industrial Biotechnology. 9(1): 19-23.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Li, J. and C. M. Sabliov. 2013. PLA/PLGA nanoparticles for delivery of drugs across the blood-brain barrier. Nanotechnology Reviews. Volume 0, Issue 0, Pages 118, ISSN (Online) 2191-9097, ISSN (Print) 2191-9089, DOI: 10.1515/ntrev-2012-0084.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2012 Citation: 3. Sabliov, C. M. October 2012. Polymeric nanoparticles as delivery systems: applications to food, health, and agriculture. Japan-America Frontiers of Engineering. National Academy of Engineering. Irvine, CA. 4. Sabliov, C. M. September 2012. Targeted nanoparticles for cancer prevention and treatment. Diaspora Conference. Bucharest, Romania. 5. Sabliov, C. M. and T. Morgan. June 2012. Biodistribution of polymeric nanoparticles used for enhanced vitamin delivery. IFT, Las Vegas, NV. 6. Sabliov, C. M. March 2012. Polymeric nanoparticles as delivery systems for disease prevention and treatment. Division of Human Nutrition and Food. LSU, Baton Rouge, LA. 7. Sabliov, C. M. February 2012. Materials and properties- food packaging and food. Basic nanotechnology science course at CFSAN. Washington, DC. 8. Sabliov, C. M. November 2012. Polymeric nanoparticles for disease prevention and treatment. FDSC 7050 Food Protein Biotechnology. Baton Rouge, LA.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2013 Citation: 1. Simon, L. and C. M. Sabliov. July 2013. Bioavailability of alpha-tocopherol delivered with polymeric nanoparticles. Institute of Food Technologists Nanotechnology Meeting. Chicago, IL. 2. Sabliov, C. M. June 2013. Biodistribution and safety of PLGA nanoparticles. Gordon Conference on Environmental Nanotechnology. Stoweflake Resort and Conference Center. Stowe, Vermont. 3. Sabliov, C. M. February 2013. Research briefs. LSU AgCenter Internal Leadership Program. Baton Rouge, LA.

Progress 05/01/11 to 04/30/12

Outputs
OUTPUTS: The results of this work have been presented at national and international conferences, and were broadcasted as follows: 1. Biodistribution of polymeric nanoparticles used for enhanced vitamin delivery. IFT, Las Vegas, NV (June 2012). 2. Materials and properties food packaging and food. Basic nanotechnology science course at CFSAN. Washington, DC, (February 2012). 3. Polymeric nanoparticles as delivery systems for disease prevention and treatment. Joint NIH and USDA Using nanotechnology to improve nutrition through enhanced bioavailability and efficacy workshop. Washington, DC (November 2011). 4. Study results from Louisiana State University. VerticalNews Agriculture, Agricultural and Food Chemistry (April 2011). PARTICIPANTS: Cristina Sabliov (PI), Sara Navarro, Diana Coulon, Carlos Astete, Meocha Whaley, Lacey Simon, Linda Cross, Caleb Darensbourg and Mike Stout, LSU AgCenter; Tim Morgan, MSU; Jeffrey Blumberg and Alison Kamil, Tufts; Mihaela Trif, Romania; Jenny Pepping, DLAM, LSU Vet School. TARGET AUDIENCES: Academia, food and pharmaceutical industries PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
The goal of the project was to assess the bioreactivity (biotoxicity, inflammatory effects and immune-reactivity) of polymeric nanoparticles (NP) specifically designed for delivery of biologically active compounds via food, with a focus on quantifying the bioreactivity as a function of NP properties and dose, type of entrapped active ingredient, and characteristics of the meal with which it was administered. Red-fluorescent poly(lactic-co-glycolic) acid (PLGA) was synthesized following a carbodiimide method. NMR confirmed successful tagging of PLGA with red fluorescent tetramethyl rhodamine isothiocyanate (TRITC). PLGA-TRITC was used to make fluorescent nanoparticles for the biodistribution study. Polymeric NP made of empty or lutein-loaded PLGA, or PLGA/TMC (modified chitosan) NP (150-200 nm, of negative charge (PLGA)), and positive charge ((PLGA/TMC), at pH=5.8) were synthesized by emulsion evaporation. Lutein and EGCG were loaded in the PLGA NPs at different entrapment efficiencies, and the stability of the NP systems was studied over a pH range 1.5-8.0 pH for oral delivery. EGCG NPs were shipped to Tufts University, where the pharmacokinetic profile of the entrapped EGCG will be compared to that of the free bioactive in two feeding studies (normal and high-fat diet). An acute and a chronic animal feeding (seven days) study was carried out at the LSU AgCenter. For the acute study, F344 rats were gavaged 3 mg fluorescent PLGA-TRITC and PLGA/Chi-TRITC NP (150-200 nm). Blood and tissue were collected 3, 12, 24, 48, 96 hrs post-administration. The chronic study consisted of feeding animals 3 mg NP/dose daily for seven days. Histopathology analysis of the tissues indicated that brain, heart, lung, and liver samples from treated animals were within normal limits for the acute and seven day chronic studies for both PLGA and PLGA/Chi NP systems. Cellular uptake studies performed by Mihaela Trif (Romania) showed that positively charged PLGA/Chi NP had a higher tendency to be endocytosed by B16F10 cells. Biodistribution studies, are important in understanding the relative distribution of the NPs in different organs as a function of their properties (size and charge).

Publications

• No publications reported this period