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Advanced therapies are commonly administered via injection even when they act within the skin tissue, and this increases the chances of off-target effects. Here we report the use of a skin patch containing a hypobaric chamber that induces skin dome formation to enable needleless delivery of advanced therapies directly into porcine, rat, and mouse skin. Finite element method modeling showed that the hypobaric chamber in the patch opened the skin appendages by 32%, thinned the skin, and compressed the appendage wall epithelia. These changes allowed direct delivery of an H1N1 vaccine antigen and a diclofenac nanotherapeutic into the skin. Fluorescence imaging and infrared mapping of the skin showed needleless delivery via the appendages. The in vivo utility of the patch was demonstrated by a superior immunoglobulin G response to the vaccine antigen in mice compared to intramuscular injection and a 70% reduction in rat paw swelling in vivo over 5 h with diclofenac without skin histology changes.

There has been an increased interest in the use of protein therapeutics, especially antibodies, for the treatment of a variety of diseases due to their high specificity to tissues and biological pathways of interest. However, the use of antibodies can be hindered by physical aggregation, degradation, and diffusion when injected in vivo leading to the need for antibody-releasing depots for the controlled and localized delivery within tissues of interest. Here, we investigated photolabile hydrogel chemistries for creating on-demand and tunable antibody release profiles. Innovative, scalable synthetic procedures were established and applied for fabricating hydrogels with nitrobenzyl (NB) and coumarin (CMR) photolabile crosslinks that responded to clinically relevant doses of long-wavelength UV and short-wavelength visible light. This synthetic procedure includes a route to make a CMR linker possessing two functional handles at the same ring position with water-stable bonds. The photocleavage properties of NB and CMR crosslinked hydrogels were characterized, as well as their potential for translational studies by degradation through pig skin, a good human skin mimic. The mechanism of hydrogel degradation, bulk versus surface eroding, was determined to be dependent on the wavelength of light utilized and the molar absorptivity of the different photolabile linkers, providing a facile means for altering protein release upon hydrogel degradation. Further, the encapsulation and on-demand release of a model monoclonal antibody was demonstrated, highlighting the ability to control antibody release from these hydrogels through the application of light while retaining its bioactivity. In particular, the newly designed CMR hydrogels undergo surface erosion-based protein release using visible light, which is more commonly used clinically. Overall, this work establishes scalable syntheses and relevant pairings of formulation-irradiation conditions for designing on-demand and light-responsive material systems that provide controlled, tunable release of bioactive proteins toward addressing barriers to preclinical translation of light-based materials and ultimately improving therapeutic regimens.

Currently, most vaccines available on the market are for parental use; however, this may not be the best option on several occasions. Mucosal routes of administration such as intranasal, sublingual, and buccal generate great interest due to the benefits they offer. These range from increasing patient compliance to inducing a more effective immune response than that achieved through conventional routes. Due to the activation of the common mucosal immune system, it is possible to generate an effective systemic and local immune response, which is not achieved through parenteral administration. Protection against pathogens that use mucosal entry routes is provided by an effective induction of mucosal immunity. Mucosal delivery systems are being developed, such as films and microneedles, which have proven to be effective, safe, and easy to administer. These systems have multiple advantages over commonly used injections, which are simple to manufacture, stable at room temperature, painless for the patient since they do not require puncture. Therefore, these delivery systems do not require to be administered by medical personnel; in fact, they could be self-administered.

Sexually transmitted diseases (STDs) are a major global health issue. Approximately 250 million new cases of STDs occur each year globally. Currently, only three STDs (human papillomavirus (HPV), hepatitis A, and hepatitis B) are preventable by vaccines. Vaccines for other STDs, including gonorrhea, chlamydia, and syphilis, await successful development. Currently, all of these STDs are treated with antibiotics. However, the efficacy of antibiotics is facing growing challenge due to the emergence of bacterial resistance. Therefore, alternative therapeutic approaches, including the development of vaccines against these STDs, should be explored to tackle this important global public health issue. Mass vaccination could be more efficient in reducing the spread of these highly contagious diseases. Bacterial outer membrane vesicle (OMV) is a potential antigen used to prevent STDs. OMVs are released spontaneously during growth by many Gram-negative bacteria. They present a wide range of surface antigens in native conformation that possess interesting properties such as immunogenicity, adjuvant potential, and the ability to be taken up by immune cells, all of which make them an attractive target for application as vaccines against pathogenic bacteria. The major challenge associated with the use of OMVs is its fragile structure and stability. However, a particulate form of the vaccine could be a suitable delivery system that can protect the antigen from degradation by a harsh acidic or enzymatic environment. The particulate form of the vaccine can also act as an adjuvant by itself. This review will highlight some practical methods for formulating microparticulate OMV-based vaccines for STDs.

Breast cancer is the number one cause of cancer-related deaths among females. Current chemotherapy targets both tumor and normal cells, leading to pronounced side effects. Therefore, therapeutic vaccines acting against specific cancer cells would be the choice of treatment. We prepared microparticles entrapping the antigens obtained from a murine metastatic breast cancer cell line, 4 T1 using the spray drying technology. These microparticles were incorporated into microneedle patches to deliver to the animals for the efficacy study. An antineoplastic drug, cyclophosphamide, in a very low dose has been found to inhibit the immunosuppressive regulatory T cells (T

Transdermal vaccination route using biodegradable microneedles is a rapidly progressing field of research and applications. The fear of painful needles is one of the primary reasons most people avoid getting vaccinated. Therefore, developing an alternative pain-free method of vaccination using microneedles has been a significant research area. Microneedles comprise arrays of micron-sized needles that offer a pain-free method of delivering actives across the skin. Apart from being pain-free, microneedles provide various advantages over conventional vaccination routes such as intramuscular and subcutaneous. Microneedle vaccines induce a robust immune response as the needles ranging from 50 to 900 μm in length can efficiently deliver the vaccine to the epidermis and the dermis region, which contains many Langerhans and dendritic cells. The microneedle array looks like band-aid patches and offers the advantages of avoiding cold-chain storage and self-administration flexibility. The slow release of vaccine antigens is an important advantage of using microneedles. The vaccine antigens in the microneedles can be in solution or suspension form, encapsulated in nano or microparticles, and nucleic acid-based. The use of microneedles to deliver particle-based vaccines is gaining importance because of the combined advantages of particulate vaccine and pain-free immunization. The future of microneedle-based vaccines looks promising however, addressing some limitations such as dosing inadequacy, stability and sterility will lead to successful use of microneedles for vaccine delivery. This review illustrates the recent research in the field of microneedle-based vaccination.

With the need for safe and efficacious vaccines which could be administered via non-invasive procedure, alternatives to traditional injectables vaccines are sought after. The present study aimed to develop the microparticulate formulation of measles vaccine and explore the feasibility of transdermal delivery via ablative laser mediated skin microporation. Transdermal route offers several advantages including painless immunization and ease of administration. We propose to use P.L.E.A.S.E. ablative laser for transdermal immunization of the microparticulate measles vaccine. This laser emits energy at 2940 µm, enabling cold ablation. This creates the micropores of defined size for delivery of vaccines into the skin. We compared the efficacy of transdermal immunization using the particulate formulation of the vaccine to that of traditional subcutaneous immunization using soluble and particulate vaccine. The microparticles were formulated using the biocompatible and biodegradable bovine serum albumin (BSA)-based polymer matrix. These vaccine microparticles were non-cytotoxic to the antigen presenting cells (APCs) and could effectively stimulate the innate immune response, confirmed by release of nitric oxide (NO) from the Griess’s assay. The APCs when exposed to vaccine microparticles also showed a significantly higher expression of antigen-presenting molecules, MHC I and MHC II, and their co-stimulatory molecules, CD80 and CD40 as compared to the blank microparticles. The microparticulate measles vaccine was evaluated in vivo in the murine model. We compared the serum IgG and IgM levels in the mice receiving the vaccine subcutaneously and transdermally post-immunization. The results revealed that transdermal immunization with microparticulate vaccine is as efficient as the traditional subcutaneous administration.

Neisseria meningitidis is a gram-negative bacterium that causes a severe acute infection, called the meningococcal disease [...]

With the evolution of different challenging diseases, there is an urgent need of vaccine development against them to save millions of lives around the world. Particlulate delivery system plays an important role by acting as self-adjuvant in form of particles and thus assisting the immunogenicity of vaccines. Particulate vaccines have shown to have improved uptake by antigen presenting cells as compared to the soluble antigen. Traditional injectable vaccines are generally poor inducers of mucosal immunity and are therefore less effective against infections at the mucosal site. Mucosal vaccines have been reported to provide additional secretory antibody mediated protection at the mucosal site of entry of the pathogen. In this chapter, we discuss the benefits of particulate drug delivery systems for oral delivery, the role of immune system in the gut, and a case study ofa novel particulate vaccine formulated into oral dissolving film for immunization via the buccal route. Key formulation components, process parameters and their biophysical characterizations have been discussed as well.

Neisseria meningitidis is a leading cause of bacterial meningitis and sepsis and its capsular polysaccharides (CPS) are a major virulence factor in meningococcal infections and form the basis for serogroup designation and preventive vaccines. We have formulated a novel meningococcal nanoparticulate vaccine formulation that does not require chemical conjugation, but encapsulates meningococcal CPS polymers in a biodegradable material that slowly release antigens, thereby has antigen depot effect to enhance antigenicity. The novel vaccine formulation is inexpensive and can be stored as a dry powder with extended shelf life that does not require the cold-chain which facilitates storage and distribution. In order to enhance the antigenicity of meningococcal nanoparticulate vaccine, we screened various adjuvants formulated in nanoparticles, for their ability to potentiate antigen presentation by dendritic cells. Here, we report that MF59 and Alum are superior to TLR-based adjuvants in enhancing dendritic cell maturation and antigen presentation markers MHC I, MHC II, CD40, CD80 and CD86 in dendritic cells pulsed with meningococcal CPS nanoparticulate vaccine.

Neisseria gonorrhoeae is a strict human pathogen responsible for more than 100 million new sexually transmitted infections worldwide each year. Due to the global emergence of antibiotic resistance, the CDC recently listed N. gonorrhoeae as an urgent threat to public health. No vaccine is available in spite of the huge disease burden and the possibility of untreatable gonorrhea. The aim of this study is to investigate the immunogenicity of a novel whole-cell based inactivated gonococcal microparticle vaccine formulation loaded in dissolvable microneedles for transdermal administration. The nanotechnology-based vaccine formulation consists of inactivated whole-cell gonococci strain CDC-F62, spray dried and encapsulated into biodegradable cross-linked albumin matrix with sustained slow antigen release. The dry vaccine nanoparticles were then loaded in a dissolvable microneedle skin patch for transdermal delivery. The efficacy of the whole-cell microparticles vaccine formulation loaded in microneedles was assessed in vitro using dendritic ,cells and macrophages as well as in vivo mouse model. Antibody titers were measured using an ELISA and antigen-specific T lymphocytes were assessed in spleens and lymph nodes. Here we report that whole-cell based gonococcal microparticle vaccine loaded in dissolvable microneedles for transdermal administration induced significant increase in antigen-specific IgG antibody titers and antigen-specific CD4 and CD8 T lymphocytes in mice compared to gonococcal antigens in solution or empty microneedles. Significant increase in antigen-specific IgG antibody levels was observed at end of week 2 in groups that received the vaccine compared to the group receiving empty nanoparticles. The advantages of using formalin-fixed whole-cell gonococci that all immunogenic epitopes are covered and preserved from degradation. The spherical shaped micro and nanoparticles are biological mimics of gonococci, therefore present to the immune system as invaders but without the ability to suppress adaptive immunity. In conclusion, the transdermal delivery of microparticles vaccine via a microneedle patch was shown to be an effective system for vaccine delivery. The novel gonorrhea nanovaccine is cheap to produce in a stable dry powder and can be delivered in microneedle skin patch obviating the need for needle use or the cold chain.

This study provided a convenient approach for large scale production of hydrogenated soya phosphatidylcholine nano-liposome powders using beclometasone dipropionate as a model drug and sucrose as proliposome carrier. Fluid-bed coating was employed to manufacture proliposomes by coating sucrose with the phospholipid (5%, 10%, 15% and 20% weight gains), followed by hydration, size reduction using high pressure homogenization, and freeze-drying to yield stable nano-vesicles. High pressure homogenization was compared with probe-sonication in terms of liposome size, zeta potential and drug entrapment. Furthermore, the effect of freeze-drying on vesicle properties generated using both size reduction methods was evaluated. Results have shown that high-pressure homogenization followed by freeze-drying and rehydration tended to yield liposomes smaller than the corresponding vesicles downsized via probe-sonication, and all size measurements were in the range of 72.64-152.50 nm, indicating that freeze-drying was appropriate, regardless of the size reduction technique. The liposomes, regardless of size reduction technique and freeze drying had slightly negative zeta potential values or were almost neutral in surface charge. The entrapment efficiency of BDP in homogenized liposomes was found to increase following freeze-drying, hence the drug entrapment efficiency values in rehydrated liposomes were 64.9%, 57%, 69.5% and 64.5% for 5%, 10%, 15% and 20% weight gains respectively. In this study, we have reported a reliable production method of nano-liposomes based on widely applicable industrial technologies such as fluid-bed coating, high pressure homogenization and freeze-drying. Moreover, sucrose played a dual role as a carrier in the proliposome formulations and as a cryoprotectant during freeze-drying.

The aim of this study is to develop an orally disintegrating film (ODF) containing a microparticulate measles vaccine formulation for buccal delivery. The measles vaccine microparticles were made with biocompatible and biodegradable bovine serum albumin (BSA) and processed by spray drying. These vaccine microparticles were incorporated in the ODF, consisting of Lycoat RS720®, Neosorb P60W® and Tween 80. The yield of the microparticles was approximately 85–95%, w/w. The mean size of the vaccine microparticles was 3.65 ± 1.89 μm and had a slightly negative surface charge of 32.65 ± 2.4 mV. The vaccine particles were nontoxic to normal cells at high concentrations (500 μg/2.5 × 105 cells) of vaccine particles. There was a significant induction of innate immune response by vaccine microparticles which was observed in vitro when compared to blank microparticles (P

Recent years have witnessed the development and advancement of many nonparenteral biologics and vaccines for human use. This chapter discusses various nonparenteral routes of administration. The oral route of administration is the most preferred and patient compliant method of them all. Transdermal, buccal, and pulmonary routes are also discussed. We have developed novel technologies using nanoparticles and microparticles to deliver vaccines by the oral and transdermal route of administration. These new technologies enable the formulation of vaccine particles containing vaccine antigens, without loss of their biological activity during the formulation process. Also, multiple antigens, targeting ligands and adjuvants can all be encapsulated within the same particle. When administered orally, these particles are designed to withstand the acidic environment of the stomach and are targeted to the Peyer’s patches and the gut-associated mucosal immune system. Because these vaccines are particulate in nature, they are readily taken up by phagocytic antigen presenting cells (APCs), such as M cells, dendritic cells, and macrophages in the Peyer’s patches of the intestines, resulting in a strong immune response and antibody production. Of particular interest is the fact that the particles release the antigen in a slow and sustained manner over a prolonged time period, intracellularly into APCs, resulting in strong mucosal and systemic immunity after oral administration, without the need for added adjuvants that are typically present in current vaccine preparations. Because no needles are required for oral vaccines, this method of vaccine delivery is inexpensive and suitable for mass vaccination in the developing world as well as for the developed world. This chapter discusses studies conducted on a wide array of vaccines, including infectious disease vaccines and cancer vaccines. This method of vaccine delivery enables the delivery of a wide spectrum of vaccines for prophylactic and therapeutic use, including oral and transdermal vaccines for cancer such as human papillomavirus, melanoma, ovarian, breast, and prostate with encouraging results. With respect to cancer therapy, a comparison is made between the conventional cancer therapy and immunotherapy. With a wide range of nanocarriers available for delivery of biologics, vaccines, and cancer therapies, nanotechnology not only has gained the well-deserved limelight but has also attracted the attention of regulatory bodies, although it presents certain challenges that must be considered before marketing such nanocarriers.

Neisseria meningitidis is a leading cause of bacterial meningitis and sepsis, and its capsular polysaccharides (CPS) are a major virulence factor in meningococcal infections and form the basis for serogroup designation and protective vaccines. We formulated a novel nanovaccine containing meningococcal CPS as an antigen encapsulated in albumin-based nanoparticles (NPs) that does not require chemical conjugation to a protein carrier. These nanoparticles are taken up by antigen-presenting cells and act as antigen depot by slowly releasing the antigen. In this study, we determined the ability of CPS-loaded vaccine nanoparticles to induce co-stimulatory molecules, namely CD80, CD86, and CD95 that impact effective antigen presentation. Co-stimulatory molecule gene induction and surface expression on macrophages and dendritic cells pulsed with meningococcal CPS-loaded nanoparticles were investigated using gene array and flow cytometry methods. Meningococcal CPS-loaded NP significantly induced the surface protein expression of CD80 and CD86, markers of dendritic cell maturation, in human THP-1 macrophages and in murine dendritic cells DC2.4 in a dose-dependent manner. The massive upregulation was also observed at the gene expression. However, high dose of CPS-loaded NP, but not empty NP, induced the expression of death receptor CD95 (Fas) leading to reduced TNF-α release and reduction in cell viability. The data suggest that high expression of CD95 may lead to death of antigen-presenting cells and consequently suboptimal immune responses to vaccine. The CPS-loaded NP induces the expression of co-stimulatory molecules and acts as antigen depot and can spare antigen dose, highly desirable criteria for vaccine formulations.