Aerosolized Non-viral Nucleic Acid Delivery in the Vaginal Tract of Pigs

Abstract

As aerosolization was an easy and effective method to deliver complexes to the vaginal tract of pigs, we believe this application technique has future potential for both vaginal and perhaps nasal vaccination using non-viral gene delivery vectors.

Plasmid‐Templated Control of DNA–Cyclodextrin Nanoparticle Morphology through Molecular Vector Design for Effective Gene Delivery

“…For practical reasons, we have chosen cyclomaltoheptaose (βCD) as the basic core to test the gemini‐type preorganization strategy for molecular vector elaboration; nevertheless, the synthetic methods are general and can be easily extended to other native CD representatives. Approaches allowing the installation of polycationic clusters at either the primary or the secondary rim of the CD core have been previously reported . The first arrangement, involving selective functionalization of the sterically more accessible primary hydroxyl groups, bears advantages in terms of simplicity and has been preferred in this work.…”

“…Approachesa llowing the installation of polycationic clusters at either the primary or the secondary rim of the CD core have been previously reported. [23][24][25][26][27][28][29][30][71][72][73] The first arrangement, involvings elective functionalization of the sterically more accessible primary hydroxyl groups,b ears advantages in terms of simplicity andh as been preferred in this work. Dimerizationc an then be effected through single-point differentiation at aprimary or asecondary position.I na ny event, the axial C 7 symmetry of the cyclooligosaccharide is lost.…”

“…[9][10][11][12][13] Further developmentso fa ccurates ynthetic manipulation [14] and (bio)orthogonal click-chemistry methodologies [15][16][17] have enabled the access to single-isomer,n anometric-sized molecules having ap ersistent topology and ap re-oriented pattern of functional groups, generally termed molecular nanoparticles, with surgical precision. [18][19][20] Ab attery of single-isomer3 Dd isplays tailored towards nucleic acid complexation, targeting, and delivery has thus been produced, [21,22] examples record on covering cyclodextrin (CD), [23][24][25][26][27][28][29][30] calixarene, [31][32][33][34][35][36] macrocyclic peptides, [37][38][39][40] fullerene, [41][42][43][44] pillarene, [45][46][47] or cyclotrehalanp latforms. [48,49] These studies have focused on the influence of the molecular architecture of the vectoro nt he self-assembling and the nucleic acid delivery capabilities; however, none of them has pursued shape control of the vector-nucleic acid nanoparticles for the systematic study of its role fort he transfection efficiency.…”

High‐Pressure Nebulization as Application Route for the Peritoneal Administration of siRNA Complexes

“…If some siRNA is released during nebulization, however, it can be expected that this effect is larger for the lower amounts of siRNA (i.e., 1 and 2 pmol) as the threshold for a sufficient siRNA concentration might no longer be reached. The decrease in transfection efficiency may also be attributed to the change in structure of some siRNA molecules due to the pressure applied, as suggested for pDNA in literature . Again, however, it is a possibility that when higher volumes (i.e., more complexes) are added on the cells, the amount of siRNA entering the cell and able to exert its function is still sufficient which can lead to nonsignificant decrease in transfection efficiency.…”

“…To evaluate the effect of aerosolization on the physicochemical properties and transfection efficiency of the complexes, 300 µL of complexes (containing 0.5 pmol µL −1 siRNA) were aerosolized using a Microsprayer Aerosolizer in combination with a high‐pressure syringe (Penn‐Century, Wyndmoor, USA) as described before . Aerosolized complexes were collected in Eppendorf vials (typically >90% recovery).…”