TY - JOUR
T1 - Chitosan-zein nano-in-microparticles capable of mediating in vivo transgene expression following oral delivery
AU - Farris, Eric
AU - Brown, Deborah M.
AU - Ramer-Tait, Amanda E.
AU - Pannier, Angela K.
N1 - Funding Information:
Nebraska Research Initiative, UNL IANR ARD, National Science Foundation (CBET-1254415), Center for Nanohybrid Functional Materials (NSF EPS-1004094), American Heart Association (#10SDG2640217), the University of Nebraska Foundation (Layman Funds), National Institute of General Medical Sciences of the National Institutes of Health (P20GM104320), the Crohn's and Colitis Foundation of America (#3578), the Nebraska Corn Board, UNL Research Council Interdisciplinary Seed Grant, UNL Research Council-Tobacco Settlement Funds Biomedical Seed Grant and USDA CSREES-Nebraska [NEB-21-146 and NEB-26-211] are acknowledged for funding. We wish to thank Dr. Abby Geis, Hatem Kittana, and Carlos Gomes Neto from Dr. Amanda Ramer-Tait's laboratory for their assistance with oral gavage. We also want to thank Anna Lampe from Dr. Deborah Brown's laboratory for her assistance with oral gavage and antibody analysis. The authors have no conflict of interest to declare.
PY - 2017/3/10
Y1 - 2017/3/10
N2 - The oral route is an attractive delivery route for the administration of DNA-based therapeutics, specifically for applications in gene therapy and DNA vaccination. However, oral DNA delivery is complicated by the harsh and variable conditions encountered throughout gastrointestinal (GI) transit, leading to degradation of the delivery vector and DNA cargo, and subsequent inefficient delivery to target cells. In this work, we demonstrate the development and optimization of a hybrid-dual particulate delivery system consisting of two natural biomaterials, zein (ZN) and chitosan (CS), to mediate oral DNA delivery. Chitosan-Zein Nano-in-Microparticles (CS-ZN-NIMs), consisting of core Chitosan/DNA nanoparticles (CS/DNA NPs) prepared by ionic gelation with sodium tripolyphosphate (TPP), further encapsulated in ZN microparticles, were formulated using a water-in-oil emulsion (W/O). The resulting particles exhibited high CS/DNA NP loading and encapsulation within ZN microparticles. DNA release profiles in simulated gastric fluid (SGF) were improved compared to un-encapsulated CS/DNA NPs. Further, site-specific degradation of the outer ZN matrix and release of transfection competent CS/DNA NPs occurred in simulated intestinal conditions with CS/DNA NP cores successfully mediating transfection in vitro. Finally, CS-ZN-NIMs encoding GFP delivered by oral gavage in vivo induced the production of anti-GFP IgA antibodies, demonstrating in vivo transfection and expression. Together, these results demonstrate the successful formulation of CS-ZN-NIMs and their potential to improve oral gene delivery through improved protection and controlled release of DNA cargo.
AB - The oral route is an attractive delivery route for the administration of DNA-based therapeutics, specifically for applications in gene therapy and DNA vaccination. However, oral DNA delivery is complicated by the harsh and variable conditions encountered throughout gastrointestinal (GI) transit, leading to degradation of the delivery vector and DNA cargo, and subsequent inefficient delivery to target cells. In this work, we demonstrate the development and optimization of a hybrid-dual particulate delivery system consisting of two natural biomaterials, zein (ZN) and chitosan (CS), to mediate oral DNA delivery. Chitosan-Zein Nano-in-Microparticles (CS-ZN-NIMs), consisting of core Chitosan/DNA nanoparticles (CS/DNA NPs) prepared by ionic gelation with sodium tripolyphosphate (TPP), further encapsulated in ZN microparticles, were formulated using a water-in-oil emulsion (W/O). The resulting particles exhibited high CS/DNA NP loading and encapsulation within ZN microparticles. DNA release profiles in simulated gastric fluid (SGF) were improved compared to un-encapsulated CS/DNA NPs. Further, site-specific degradation of the outer ZN matrix and release of transfection competent CS/DNA NPs occurred in simulated intestinal conditions with CS/DNA NP cores successfully mediating transfection in vitro. Finally, CS-ZN-NIMs encoding GFP delivered by oral gavage in vivo induced the production of anti-GFP IgA antibodies, demonstrating in vivo transfection and expression. Together, these results demonstrate the successful formulation of CS-ZN-NIMs and their potential to improve oral gene delivery through improved protection and controlled release of DNA cargo.
KW - Chitosan
KW - DNA delivery
KW - DNA vaccination
KW - Gene therapy
KW - Oral delivery
KW - Zein
UR - http://www.scopus.com/inward/record.url?scp=85011807558&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85011807558&partnerID=8YFLogxK
U2 - 10.1016/j.jconrel.2017.01.035
DO - 10.1016/j.jconrel.2017.01.035
M3 - Article
C2 - 28153762
AN - SCOPUS:85011807558
VL - 249
SP - 150
EP - 161
JO - Journal of Controlled Release
JF - Journal of Controlled Release
SN - 0168-3659
ER -