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Vaccines manufacture and enhancement for preventing infection and promoting quality of life are of great concern worldwide. For vaccine enhancement, to date, only limited adjuvants have been approved globally. One of them is alum, which presents several side effects and limitations. Related to vaccine administration, mucosal vaccination is a promising method since it can induce both mucosal and systemic immunity since oral mucosa is the most exposed site of the body to various microbes, pathogens, and environmental particles. Consequently, an escalated specific local immunity is required in which stability and integrity of an encapsulated antigen is expected to result in a stable mucosal vaccine to protect the antigens from degradative chemical reactions occurring in the oral cavity and act as immunomodulator. Carbonate apatite (CHA) has been one of the most innovative materials as a newly developed vaccine adjuvant since it can adequately enhance drug and protein stability and delivery in various disease therapies. However, CHA fabrication that meets the parameters for adjuvants and immunomodulators remains challenging. In the form of nanoparticles, CHA is reported to enable targeted delivery of dendritic cells (DC), enhance uptakes, cross presentation, and biodistribution, as well as immune responses. Therefore, the development of nano-CHA-encapsulated vaccine antigens is required to enhance oral mucosal vaccinations and their effectiveness to prevent diseases. This study focuses on factors and strategies that affect the designing, fabrication, and testing of CHA nanoparticles for oral mucosal vaccines, especially in the aspect of physicochemical, immunological, cellular, surface chemistry, and biofunctionalization of the nanoparticle.

Nowadays, antibiotic resistance poses a threat to public health worldwide. For this reason, non-traditional antibacterial products, such as silver nanoparticles (AgNPs), offer an opportunity to address this issue. Although AgNPs have been proven to be effective antimicrobial agents, we studied the antibacterial and antibiofilm effects of two novel AgNPs (AgNP-Aloe-1 and AgNP-Aloe-2) obtained by green synthesis, their cytotoxicity on a cell line derived from human keratinocytes, and their skin penetration. These AgNPs were obtained here for the first time from an Aloe maculata aqueous extract as a reducing and capping agent of Ag(I), with varying the initial silver concentrations (5 and 9 mM of AgNO₃ for AgNP-Aloe-1 and AgNP-Aloe-2, respectively). For all the assessments, these were compared with AgNPs obtained from a traditional chemical method employing hydroxylamine hydrochloride as a reducing agent and AgNO₃ (AgNP–NH₂OH·HCl). The AgNPs were characterized physicochemically by TEM, DLS, Zeta potential, UV–vis, fluorescence, and Raman spectroscopy. Additionally, the concentration of silver forming AgNPs and the reaction yield were determined. Both green-synthesized AgNPs showed an improvement in the inhibition of bacterial growth after 24 h of incubation for E. coli and S. aureus. AgNP-Aloe-1 presented a MIC 4 times lower for both bacteria compared to AgNP–NH₂OH·HCl, while AgNP-Aloe-2 presented a MIC 32 and 8 time lower for E. coli and S. aureus, respectively. Moreover, they produced a decrease in the biofilm biomass formation from P. aeruginosa at lower concentrations (6.25 μg/ml for AgNP-Aloe-1 and 1.56 μg/ml for AgNP-Aloe-2) than AgNP-NH₂OH·HCl which only showed a reduction of 30% at the maximum concentration tested. However, AgNP-Aloe-1 and AgNP-Aloe-2 were less efficient in eradicating pre-formed biofilm. Even though AgNP-Aloe-2 showed a lower reaction yield (31.7%) compared to AgNP-Aloe-1 (68.5%), they showed the best antibacterial activity. On the other hand, green-synthesized AgNPs were mainly retained in the stratum corneum of intact skin and reached lower concentrations in the viable epidermis than AgNP–NH₂OH·HCl. Moreover, AgNP-Aloe-1 and AgNP-Aloe-2 did not show cytotoxic effects on human keratinocytes at the antibacterial concentrations. Their improved performance and lower skin penetration could be attributed to their physicochemical properties, such as size (10–25 nm), charge (around −10 mV), and shape (tendency towards a spherical shape), but mainly to the presence of phytocompounds from the extract that remained attached to the AgNPs, as observed by Raman spectroscopy and UV–vis. For the reasons mentioned above, these novel AgNPs obtained by a more environmentally friendly method have the potential to be used as antibacterial agents, particularly for topical applications.

Long acting injectables (LAIs) using polymeric microspheres has been developed to increase patient compliance and reduce side effects. Among many methods for manufacturing polymeric microspheres, microfluidics technology is known to have excellent characteristics in that the produced polymeric microspheres have perfect spherical shape without surface defect and uniform size, and thus have outstanding efficacy without initial burst. However, the mass production of polymeric microspheres was not realized by the inherent limitation that microfluidics is suitable for small quantity manufacturing. Overcoming such limitations, we could show mass production of finasteride-loaded polymeric microspheres (PLGA 7525) for LAIs using our microfluidic manufacturing platform technology, IVL-DrugFluidic®. The microfluidic channels used in manufacturing were optimized through computational fluid dynamics (CFD) simulation to minimize the flow variation between microchannels and eliminated disturbance outside of microchannels by resistance channels. In addition, the solvent removal was improved by applying the baffle and foam breaker system. Therefore, microspheres were mass-produced in the GMP manufacturing environment in perfect spherical shape, smooth surface, and even size distribution. The encapsulation efficiency was almost 100% and the residual solvent was under the Standard of regulation. In the clinical trial using microspheres mass-produced by IVL-DrugFluidic®, we confirmed that the drug release was stably maintained for a month, the target period without initial burst. It was also confirmed that the drug release by dose of microspheres was uniformly proportional. In conclusion, the microsphere manufacturing platform technology, IVL-DrugFluidic® has been proven to be an appropriate system for mass production of polymeric microspheres optimized for LAIs through physicochemical characteristics and clinical trial.

A major driving factor for antimicrobial resistance which leads to treatment failure for microbial infections ascribed to C. albicans are the formation of biofilms. 3D printing is a rapidly evolving innovation which could revolutionize drug delivery based on its unprecedented opportunity for targeted and improved delivery. Herein, we designed and 3D printed microbots via two photon-polymerisation. Subsequently, characterisation was performed and the activity of microbots independently and in combination with allicin against C. albicans biofilms were investigated. The microbots independently did not affect C. albicans biofilm formation and adhesion nor was there any significant synergistic interaction between microbots and allicin combination. However, this study has pioneered the utilisation of microbots for microbiological applications such as in combination with an antimicrobial to target biofilms. These prototype microbots will act as a guide for the next generation of microbots which will be functionalised to disrupt biofilms magnetically, enhancing allicin delivery and activity.

Dunaliella salina, a green microalga, is among the main sources of bioactive β-carotene and zeaxanthin. Hence, it will be investigated for its antioxidant effectiveness in wound healing. The current study's objective is to create new chitosan nanoparticle loaded D. salina hexane: ethyl acetate extract (HEAE-CNPs) and methanol extract (ME-CNPs) to be used in accelerating wound healing in-vivo. Double emulsion technique was utilized to prepare the nanoparticles. The prepared HEAE-CNPs and ME-CNPs were examined for in-vitro release and in-vivo wound healing efficacy in Wistar rats. Results confirmed that D. salina hexane:ethyl acetate extract (HEAE) contains 19.167 mg/g β-carotene and 16.196 mg/g zeaxanthin, whereas the extract of methanol (ME) contains only small amounts of zeaxanthin 0.313 mg/g as quantified by HPLC. The D. salina loaded chitosan gel greatly slowed the total carotenoids release, according to the in-vitro release assays, in comparison with D. salina nanoparticle dispersion with a particle size in the nanorange. By decreasing factor alpha (TNF-α) of tumor necrosis and increasing vascular endothelial growth factor (VEGF) and collagen skin contents, both HEAE-CNPs and ME-CNPs demonstrated wound healing and regeneration.

Gold nanoparticles (AuNPs) are one of the most extensively studied nanomaterials and their distinct physicochemical properties make them suitable for versatile applications. Herein, we synthesized concave cube-shaped gold nanoparticles (CCAuNPs) and functionalized them with lactoferrin (Lf), a natural antimicrobial protein, through electrostatic interaction as well as weak covalent formation. The functionalization of CCAuNPs was confirmed through UV–Visible (i.e., bathochromic shift of the surface plasmon resonance peak by 7 nm), Fourier transform infrared (FTIR) and X-ray diffraction (XRD) spectroscopy, and their surface zeta potential. The concave cusp of CCAuNPs was confirmed through atomic force microscopy (AFM). The Lf-functionalized CCAuNPs (Lf-CCAuNPs) exhibited superior antibacterial propensity against a series of bacteria when compared to that of CCAuNPs. However, they didn't demonstrate any antibacterial activity against Lactobacillus plantarum, one of the key beneficial gut bacteria. The lipid peroxidation (LPO) assay confirmed the oxidation of fatty acids in the bacterial membrane upon interaction with AuNPs, which made the bacterial membrane porous. The resultant membrane-impaired dead bacteria were visualized through CellTox™ Green assay as well as the Trypan Blue dye exclusion assay. Both the nanoparticles demonstrated excellent hemocompatibility as well as biocompatibility (both in vitro and in vivo) as confirmed by MTT assay and the levels of important functional biomarkers of liver (e.g., ALT, AST, and ALP) and kidney (e.g., creatinine, uric acid, and BUN) in the serum. Overall, Lf-CCAuNPs with excellent hemocompatibility, and biocompatibility can be deployed as potential antibacterial agents to tackle the menace of pathogenic bacteria.

Oral cancer has a poor survival rate despite comprehensive therapy. Current treatments result in acute side effects and fail to eliminate an aggressive group of cells overexpressing CD44. Such cells are capable of tumour initiating, self-renewal, invasion and metastasis, resulting in tumour relapse and resistance. This study aims to synthesise and characterise hyaluronic acid/chitosan-coated poly (lactic-co-glycolic acid) nanoparticles and assess their effectiveness in delivering Paclitaxel and Temozolomide to human tongue squamous cell carcinoma cell line that expresses high CD44 levels, in terms of cell cytotoxicity and apoptosis. This study also assesses the coordinated administration of Paclitaxel and Temozolomide and whether they exhibit significant synergistic cell inhibition effects with reduced introduced drug concentration if co-delivered simultaneously. Nanoparticles were synthesised with solvent evaporation method and characterised to assess their size, homogeneity, and zeta potential. Cell viability assay and real-time cell analysis were performed to examine the cell inhibitory effect of the drug-loaded nanoparticles. Cell apoptosis and cell cycle alteration were detected, and reactive oxygen species induction, mitochondrial membrane potential, and expressed genes associated with cell inhibition and death were evaluated. The synthesised nanoparticles had a nano-sized diameter of 260.40±11.54 nm, a positive zeta potential of +14.31±1.37 mV and a low polydispersity index value of 0.15±0.03. Paclitaxel, Temozolomide, and their combination have inhibited cell proliferation with half maximal inhibitory concentrations of 4 nM, 1000 μM and 2nM:300 μM, respectively. Compared to free drugs, the single-loaded and co-loaded drugs induced more cytotoxicity. Paclitaxel and Temozolomide showed a considerable synergistic inhibitory effect which was discovered to be more significant when the drugs were loaded in the nanoparticles. Drug-loaded nanoparticles were verified to induce higher cell apoptosis rates, cell proportion arrested at the S-phase of the cell cycle, reactive oxygen species generation, mitochondrial collapse and expression of genes associated with cellular inhibition and death than free drugs. These results demonstrate that the established nanoparticles could be a potential candidate for oral cancer therapy since they could deliver and improve the efficacy of single and dual drugs against oral cancer cells.

Lipid Nanoparticles (LNPs) are promising drug delivery systems for various RNAs such as small interfering (siRNA) and messenger RNA (mRNA). Microfluidic mixing is a common technique to encapsulate RNA in LNPs. However, high flow rates and lipid concentrations are used for LNP formation to control LNP size as well as RNA encapsulation efficiency. We investigated the feasibility of downscaling siRNA and mRNA LNP manufacturing to save materials and enable a broader access to this technology. To optimize such a down-scaled procedure, we evaluated physicochemical nanoparticle characteristics including hydrodynamic diameter, zeta potential, particle concentration, encapsulation efficiency, and recovery for LNPs produced with three different microfluidic methods. We observed differences in nanoparticle characteristics and in vitro performance regarding cellular uptake, gene silencing, and mRNA expression. We determined the gene knockdown ability of the best siRNA LNPs formulation ex vivo using precision-cut lung slices to highlight the translational character of LNPs for inhalation and observed comparable efficacy as with a commercially available transfection reagent.

Due to the fact that bacterial contamination of wounds is the cause of increased morbidity and mortality today, various antimicrobial wound dressings are developing to prevent wound contamination. In addition, an ideal wound dressing should have proper mechanical and hemostatic properties to maintain wound healing conditions. Here, a double-layer sponge hydrogel nanocomposite wound dressing was designed and manufactured by combining zinc oxide nanoparticles (ZnONPs) with different concentrations in the hydrogel layer and green carbon dots in the sponge layer. The surface morphology of two layers was investigated using a scanning electron microscope. X-ray diffraction proved the presence of ZnONPs. Physical tests showed a decrease in water absorption and water vapor transmission rate and an increase in blood absorption in the presence of ZnO. The sponge layer showed suitable absorption support in the presence of carbon dots. By combining nanoparticles in both layers, the mechanical properties were greatly enhanced. The sponge hydrogel with the highest concentration of ZnO showed excellent inhibition of 41 mm against Pseudomonas aeruginosa bacteria and 25 mm of inhibition against Staphylococcus aureus. Finally, in vitro blood clotting and animal tests confirmed the increase in the hemostatic power of the sponge hydrogel with the maximum concentration of ZnO.