Drug Delivery and Translational Research最新文献

Exosomes are small extracellular vesicles naturally secreted by cells into body fluids, enriched with bioactive molecules such as RNAs, proteins, and lipids. These nanosized vesicles play a crucial role in physiological and pathological processes by facilitating intercellular communication and modulating cellular responses, particularly within the central nervous system (CNS). Their ability to cross the blood-brain barrier and reflect the characteristics of their parent cells makes exosomal cargo a promising candidate for biomarkers in the early diagnosis and clinical assessment of neurological conditions. This review offers a comprehensive overview of current knowledge on the characterization of mammalian-derived exosomes, their application as drug delivery systems for neurological disorders, and ongoing clinical trials involving exosome-loaded cargo. Despite their promising attributes, a significant challenge remains the lack of standardized isolation methods, as current techniques are often complex, costly, and require sophisticated equipment, affecting the scalability and affordability of exosome-based therapies. The review highlights the engineering potential of exosomes, emphasizing their ability to be customized for targeted therapeutic delivery through surface modification or conjugation. Future advancements in addressing these challenges and leveraging the unique properties of exosomes could lead to innovative and effective therapeutic strategies in neurology.

Antibiotic resistance is a cause of serious illness and death, originating often from insufficient permeability into gram-negative bacteria. Nanoparticles (NP) can increase antibiotic delivery in bacterial cells, however, may as well increase internalization in mammalian cells and toxicity. In this work, NP in liposome (NP-Lip) formulations were used to enhance the selectivity of the antibiotics (3C and tobramycin) and quorum sensing inhibitor (HIPS-1635) towards Pseudomonas aeruginosa by fusing with bacterial outer membranes and reducing uptake in mammalian cells due to their larger size. Poly (lactic-co-glycolic) acid NPs were prepared using emulsion solvent evaporation and incorporated in larger liposomes. Cytotoxicity and uptake studies were conducted on two lung cell lines, Calu-3 and H460. NP-Lip showed lower toxicity and uptake in both cell lines. Then formulations were investigated for suitability for oral inhalation. The deposition of NP and NP-Lip in the lungs was assessed by next generation impactor and corresponded to 75% and 45% deposition in the terminal bronchi and the alveoli respectively. Colloidal stability and mucus-interaction studies were conducted. NP-Lip showed higher diffusion through mucus compared to NPs with the use of nanoparticle tracking analyzer. Moreover, the permeation of delivery systems across a liquid-liquid interface epithelial barrier model of Calu-3 cells indicated that NP-Lip could cause less systemic toxicity upon in-vivo like administration by aerosol deposition. Monoculture and Pseudomonas aeruginosa biofilm with Calu-3 cells co-culture experiments were conducted, NP-Lip achieved highest toxicity towards bacterial biofilms and least toxicity % of the Calu-3 cells. Therefore, the NP- liposomal platform offers a promising approach for enhancing antibiotic selectivity and treating pulmonary infections.

The delivery of drugs directly from the nose to the brain has been explored for the treatment of neurological diseases, such as glioblastoma, by overcoming the blood-brain barrier. Nanocarriers have demonstrated outstanding ability to enhance drug bioavailability in the brain, following intranasal administration. However, the performance of these nanosystems may be hindered by inadequate interactions with the nasal mucosa, limiting their effectiveness in reaching the olfactory region, and consequently, the translocation of particles to the brain. Here, we designed hybrid lipid-polymer nanoparticles (LPNP), containing the cationic lipid DOTAP and the triblock copolymer Pluronic^® F127 to combine the mucoadhesiveness and mucus-penetrating properties. Perillyl alcohol (POH), a molecule currently under clinical trials against glioblastoma, via intranasal route, was entrapped in the nanoparticles. LPNP-POH exhibited a balanced profile of mucus adhesion and penetration, suggesting that the formulation may enhance mucosal retention while maintaining effective mucus diffusivity. In vivo evaluations displayed higher translocation of LPNP-POH from the nasal cavity to the brain. LPNP-POH resulted in a 2.5-fold increase in the concentration of perillyl acid (a primary metabolite of POH) in the cerebral tissue compared to the free drug. In vitro assays demonstrated that LPNP-POH increased the cytotoxicity and reduced the tumor growth of U87MG glioma cells. These results highlighted that the engineered formulation, with optimized mucoadhesiveness and mucus penetration properties, improved nose-to-brain delivery of POH, offering a promising potential for glioblastoma therapy.

Wound healing is a complex process which is crucial for recovery. Delayed wound healing which is caused by the presence of pathogens has posed significant clinical implications affecting millions of patients globally. Wounds infection caused by Pseudomonas aeruginosa present significant challenges due to their resistance to multiple antimicrobial drugs. The Gram-negative bacteria secretes endotoxin lipopolysaccharide (LPS), which impede wound healing and may lead to severe complications, including life-threatening sepsis. Previously, our laboratory has successfully developed a new hydrogel containing a synthetic antimicrobial peptide as an alternative therapy to conventional antibiotics. This hydrogel contains LL37 microspheres embedded into activated carbon-chitosan hydrogel (LL37-AC-CS). LL37-AC-CS has shown desirable physicochemical properties as well as promising antimicrobial and antitoxin activities in vitro. This current study has two main objectives. The first is to evaluate the in vivo antimicrobial efficacy of LL37-AC-CS hydrogel in full-thickness rat wounds infected with P. aeruginosa. The second objective is to investigate the antitoxin efficacy on the rat wound models treated with E. coli endotoxins LPS. The wound healing efficacy was assessed in terms of the macroscopic appearance, wound contraction rate, histology, and wound tissue biochemical markers. As a result, the LL37-AC-CS hydrogel exhibited remarkable antimicrobial and antitoxin efficacy as compared to the controls. The wound healing efficacy was evident in increased wound closure rate and decrease in bacterial bioburden, and favourable changes in wound healing biomarkers namely the myeloperoxidase, interleukin-6 and tumour necrosis factor α. The elevation of hydroxyproline levels in the LPS-treated wound model indicates there was collagen synthesis. In conclusion, the results presented in this study have significantly enhanced our comprehension of the LL37-AC-CS hydrogel's potential in wound healing. Specifically, the research highlights its effectiveness in eliminating endotoxins and preventing bacterial growth.

This study aims to create glyco-based nanoparticles (NPs) with high drug-loading capability for targeted cancer treatment, specifically against MDA-MB-231 breast cancer cells. Traditional NPs have faced limitations due to low drug-loading capacities, leading to suboptimal therapeutic effectiveness and significant side effects. To overcome these limitations, DOX@pB-pM NP were synthesized using a self-assembly combination method of two poly(ε-caprolactone) (PCL) based polymers, mannoside-b-PCL (pM) and phenylboronic acid (PBA)-mPEG-t-PCL (pB). The pM polymer synthesis includes a Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAc) reaction. DOX@pB-pM NP's mannose moiety is specifically engineered to target MDA-MB-231 cells, while the core of the NPs is made of hydrophobic, biodegradable polyester PCL. The functions of mPEG and PBA in the pB tri-block copolymer are to enhance biocompatibility and drug-loading efficiency, respectively. Additionally, mPEG can reduce nonspecific interactions. The PBA on the pB introduces a hydrophobic segment to the copolymer, which can improve the interaction with water-insoluble drugs, doxorubicin (DOX). The PBA moiety can also provide additional functionality, such as pH-responsive and H₂O₂-responsive drug release, which is particularly useful in targeting the tumor's acidic and oxidative microenvironment. The PBA groups convert them to boronic acid and 4-(hydroxymethyl) phenol, which destroys the NP core and causes DOX release, resulting in cell death. The in vitro release profile of DOX from the DOX@pB-pM NPs was evaluated under various conditions, including different pH levels and the presence or absence of H₂O₂, to simulate the acidic tumor microenvironment. The cytotoxicity of the DOX@pB-pM NPs was assessed using the MTT assay, which demonstrated significant inhibition of MDA-MB-231 breast cancer cell growth by DOX@pB-pM NPs. By combining mannose for the targeting of MDA-MB-231 breast cancer cells and fine-tuning the ratio of pM and pB polymers, the NPs showed good therapeutic efficacy. Importantly, pB-pM NPs displayed good biocompatibility, with no significant effect on cell survival even at high concentrations, indicating their potential as safe drug carriers. These data show that DOX@pB-pM NPs can potentially improve cancer therapeutic efficacy and safety.

This study evaluates the in vivo mucoadhesive properties of thiolated cyclodextrins (CDs) with varying S-protection using polyethylene glycol (PEG) of different chain lengths. Free thiol groups of thiolated β-CDs (CD-SH) were S-protected with 1 kDa and 2 kDa PEG bearing a terminal thiol group, leading to third-generation of thiolated CDs (CD-SS-PEG). The structure of these thiolated CDs was confirmed and characterized by FT-IR, 1 H NMR, and colorimetric assays. Thiolated and S-protected CDs were evaluated regarding viscosity, cellular uptake and, in vitro and in vivo mucoadhesion. The viscosity of CD-SH, CD-SS-PEG 1 kDa, and CD-SS-PEG 2 kDa mixtures with mucus increased 9-, 7-, and 5.5-fold, respectively, compared to unmodified CD within 3 h. Cellular uptake on Caco-2 cells was 1.75 times higher for highly thiolated CDs than for unmodified CD. In vitro residence time on porcine intestine was prolonged 7-, 8.4-, and 7.9-fold for CD-SH, CD-SS-PEG 1 kDa, and CD-SS-PEG 2 kDa, respectively. In vivo results indicated CD-SS-PEG 1 kDa had the highest potential. Our comprehensive in vitro, ex vivo, and in vivo ffindings demonstrate that CD-SS-PEG 1 kDa is a highly promising candidate for mucoadhesive drug delivery systems.

Intracerebral hemorrhage (ICH) is a serious cerebrovascular disease with high morbidity, mortality, and disability rates, largely due to neuroinflammation. Diosmetin, a natural flavonoid, has known neuroprotective effects in cerebral ischemia/reperfusion models but has been less studied in ICH. Our previous study developed diosmetin-loaded lactoferrin-modified long-circulating liposomes (Lf-Dios-Lcl), which penetrate the BBB and improve diosmetin bioavailability and brain distribution. In this study, we found that diosmetin reduced the levels of proinflammatory cytokines (IL-1β and TNF-α) and increased the level of the anti-inflammatory cytokine IL-10 in LPS-induced BV2 cells, promoting microglial polarization toward the anti-inflammatory M2 phenotype. In ICH model rats, Lf-Dios-Lcl (1 mg/kg) effectively reduced neuroinflammation, decreased IL-1β and TNF-α levels, increased IL-10 levels, and increased the proportion of CD206-positive microglia in brain tissues. Moreover, Lf-Dios-Lcl significantly downregulated p-p38 expression, suggesting that p38 signaling activation was inhibited. Overall, Lf-Dios-Lcl demonstrated brain-targeting properties and antineuroinflammatory effects by modulating microglial polarization via the p38 pathway.

Oleanolic acid (OA) ischaracterized by its low water solubility, poor permeability and majorly metabolized by cytochrome P450 (CYP) isozymes in the intestinal tract, particularly CYP3A, which contribute to the low oral bioavailability. OA has multiple pharmacological actions including hepatoprotective, anti-inflammatory, antidiabetic and antiviral effects. OA classified as a BCS IV drug which have restricted its potential clinical application. In this study D-α-Tocopheryl polyethylene glycol succinate (TPGS) and Pluronics F68 based stabilized OA loaded mixed micellar system (OA-MMs) developed to improve the solubility and permeability. Mixed micelles were characterized by dynamic light scattering studies as a function of temperature, salt addition, and OA solubilisation followed byXRD, FE-SEM and IR analysis confirmed the formation of stabilized OA-MMs with the least size and PDI (10.041 ± 1.35 nm, 0.313 ± 0.012). Scattering studies results demonstrates the formation of stable micelles with no significant alterations insize upon salt addition (up to 150mM NaCl), OA incorporation (up to 150 mM) and temperature rise till 40 °C.Solubility of the pure OA and OA-MMs was found to be 0.042 mg/ml and 1.98 mg/ml. The % cumulative release of drug from alone OA, OA + TPGS and OA-MMs was found to be 4.363 ± 0.025%, 57.18 ± 0.034% and 92.269 ± 0.017% respectively up to 24 h. Single-pass intestinal perfusion studies (SPIP) showed that K_a and P_effective of OA-MMs was improved30 fold as compared with that of pure OA and this was mainly due to the improved permeability and inhibitory effect of Pluronic F68 on CYP3A. The in vivo Pharmacokinetic study showed that C_max increased markedly from 12.76 to 20.49 and 39.17 µg/ml in case of OA alone, OA + TPGS and OA-MMs. Parallel to the C_max there was an increase in the AUC_0-24133.68 to 164.56 and 296.50 respectively. All of the produced OA-MMs formulation's results demonstrated a notable increase in OA's bioavailability through increased permeability and solubility along with metabolic inhibition OA.

Topically applied therapies must not only be effective at the molecular level but also efficiently access the target site which can be on milli/centimetre-scales. This bottleneck is particularly inhibitory for peptide and nucleic acid macromolecule drug delivery strategies, especially when aiming to target wounded, infected, and poorly perfused tissues of significant volume and geometry. Methods to drive fluid-flow or to enhance physical distribution of such formulations after local administration in accessible tissues (skin, eye, intestine) would be transformative in realizing the potential of such therapeutics. We previously developed a technology termed Glycosaminoglycan (GAG)-binding enhanced transduction (GET) to efficiently deliver a variety of cargoes intracellularly, using GAG-binding peptides and cell penetrating peptides (CPPs) in the form of nanoparticles. Herein, we demonstrate that the most simplistic GET formulation is relatively poor in diffusing into tissue matrix (tested in collagen scaffolds). Changing nanoparticle physicochemical properties can enhance penetration, however the use of a pressure differential, generating fluid-flow significantly enhances effective gene delivery over milli/centimetre scales. We adapted clinically used pressure systems to administer both negative (Negative pressure (NP) wound therapy; NPWT) and positive pressures (PP; Insufflator). Pressure differences generated enhanced distribution, and we were able to show for the first-time localized gene transfer in vitro in cell scaffolds and enhanced transfection of ex vivo skin explants. The ability to simply control intra-tissue localization of gene delivery on milli/centimetre scales using pressure application will facilitate new drug delivery strategies for accessible tissues. Importantly site-specific enhancement of penetration and activity of novel nanotechnologies and gene therapeutics could be transformative for future regenerative medicine strategies.

Owing to faulty DNA damage repair system, triple negative breast cancer (TNBC) exhibits high susceptibility towards DNA damaging drugs such as platinum compounds e.g., oxaliplatin. Nevertheless, the clinical utility of oxaliplatin (OXA) has been constrained due to chemoresistance and chronic toxicities. Hence, to confer systemic inertness, tumor specific delivery, and multifaceted action, a octahedral OXA-CBL prodrug was synthesized using chlorambucil (CBL) as an axial ligand. The combination of OXA and CBL exhibited synergistic anti-cancer action in TNBC cell lines. Further, to potentiate the cellular internalization, targeting efficiency, and in-vivo performance, the synthesized prodrug was loaded into bovine serum albumin nanoparticles (OXA-CBL/BSA-NPs). The prepared nanoparticles had optimal particle size < 200 nm and high drug loading (∼ 5.863 ± 0.16%). As relative to free conjugate, the nanoparticles exhibited amplified cellular internalization and reduced the IC₅₀ in 4T1 (∼ 1.38-fold) and MDA-MB-231 (∼ 1.43-fold) cell line. The anti-cancer study in 4T1-based TNBC model in BALB/c mice demonstrated significantly higher tumor inhibition rate, and reduced tumor burden in OXA-CBL/BSA-NPs treated group. Toxicity assessment revealed no signs of hepato- and/or renal toxicity. Also, nanoparticles exhibited sufficient compatibility with erythrocytes. Overall, delivery of OXA-CBL via virtue of albumin nanoparticles presents safer and efficacious approach to combat TNBC.