title: Non-invasive delivery of nanoparticles to hair follicles: a perspective for transcutaneous immunization.
abstract: Transfollicular vaccination aims to reach the peri-follicular antigen presenting cells without impairing the stratum corneum (SC) barrier. This would be an optimal vaccination strategy under critical hygienic conditions. Nanoparticles (NPs) are the ideal vehicles for transfollicular delivery of vaccines as they are able to (i) penetrate deeper into the hair follicles than molecules in solution, (ii) can help to stabilize protein based antigen and (iii) improve and modulate the immune response. This study investigates the potential of transfollicular delivery of polymeric NPs using ovalbumin (OVA) as a model antigen. NPs were prepared by a double emulsion method from pharmaceutically well characterized biocompatible and biodegradable polymers poly(lactide-co-glycolide) (PLGA) or chitosan-coated PLGA (Chit-PLGA) using polyvinyl alcohol as stabilizer. The NP formulations are available as freeze dried product which can be re-constituted with water or cell culture medium before use to yield any desired OVA/NP concentration. OVA was protected from cleavage or aggregation inside the NPs and retained its biological activity to 74% (PLGA) and 64% (Chit-PLGA). Thus, when applying a typical dose of 8.5 μl/cm(2) NP formulation (50mg NPs/ml, 54.3±0.047 and 66.5±0.044 μg OVA/mg NPs for PLGA and Chit-PLGA NPs, respectively) an effective dose of 17 μg/cm(2) (PLGA) or 18 μg/cm(2) (Chit-PLGA) of active OVA is administered. In a cell culture assay encapsulated OVA stimulated the proliferation of CD4+ (PLGA and Chit-PLGA) and CD8+ T-cells (only Chit-PLGA) to a larger extent than OVA in solution. An adoptive transfer experiment demonstrated that the model antigen OVA can be delivered via the transfollicular route. This preliminary experiment is a proof of concept that by this transfollicular immunization approach it is possible to deliver antigens, thereby stimulating antigen-specific T cells. Both NP formulations improved the delivery efficiency of OVA into the hair follicles on excised pig ears by a factor of 2-3 compared to OVA solution. This delivery efficiency could further be increased by increasing the number of NPs applied per skin area by a factor of ≈2-2.4. Consequently formulation of OVA into PLGA and Chit-PLGA NPs may offer to reduce the dose which needs to be applied for transfollicular immunization.
authors.affiliation: Saarland University, Biopharmaceutics and Pharmaceutical Technology, Saarbruecken, Germany.
authors.forename: Ankit
authors.initials: A
authors.lastname: Mittal
authors.forename: Anne S
authors.initials: AS
authors.lastname: Raber
authors.forename: Ulrich F
authors.initials: UF
authors.lastname: Schaefer
authors.forename: Sebastian
authors.initials: S
authors.lastname: Weissmann
authors.forename: Thomas
authors.initials: T
authors.lastname: Ebensen
authors.forename: Kai
authors.initials: K
authors.lastname: Schulze
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Guzmán
authors.forename: Claus-Michael
authors.initials: CM
authors.lastname: Lehr
authors.forename: Steffi
authors.initials: S
authors.lastname: Hansen
mesh_terms: D000279:Administration, Cutaneous; D000818:Animals; D000941:Antigens / Q000008:administration & dosage / Q000276:immunology; D049109:Cell Proliferation; D048271:Chitosan / Q000008:administration & dosage / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology; D005260:Female; D018859:Hair Follicle / Q000276:immunology* / Q000378:metabolism; D007114:Immunization / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D010047:Ovalbumin / Q000008:administration & dosage / Q000276:immunology; D011098:Polyglactin 910 / Q000737:chemistry; D013552:Swine; D013601:T-Lymphocytes / Q000276:immunology; D014612:Vaccines / Q000276:immunology
chemical_list: D000941:Antigens; D014612:Vaccines; D011098:Polyglactin 910; D010047:Ovalbumin; D048271:Chitosan
keywords: CD4+; CD8+; Chitosan; Drug delivery; Nanoparticle; Ovalbumin; PLGA; Transcutaneous immunization; Transfollicular immunization