Tumor-targeted drug delivery systems are promising for their advantages in enhanced tumor accumulation and reduced toxicity towards normal organs. However, few nanomedicines have been successfully ...translated into clinical application. One reason is the gap between current pre-clinical and clinical studies. The prevalent in vitro models utilized in pre-clinical phase are mainly based on the two-dimensional (2D) cell culture and are limited by the difficulty of simulating three-dimensional physiological conditions in human body, such as three-dimensional (3D) architecture, cell heterogeneity, nutrient gradients and the interaction between cells and the extracellular matrix (ECM). In addition, traditional animal models have drawbacks such as high-cost, long periods and physiological differences between animal and human. On the other hand, the employment of 3D tumor cell culture models, especially multicellular tumor spheroids (MCTS), has increased significantly in recent decades. These models have been shown to simulate 3D structures of tumors in vitro with relatively low cost and simple protocols. Currently, MCTS have also been widely exploited in drug delivery system research for comprehensive study of drug efficacy, drug penetration, receptor targeting, and cell recruitment abilities. This review summarizes the delivery barriers for nano-carriers presented in tumor microenvironment, the characteristics and formation methods for applicable multicellular tumor spheroid culture models and recent studies related to their applications in tumor-targeted drug delivery system research.
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Neural stem cells (NSCs), capable of ischemia‐homing, regeneration, and differentiation, exert strong therapeutic potentials in treating ischemic stroke, but the curative effect is limited in the ...harsh microenvironment of ischemic regions rich in reactive oxygen species (ROS). Gene transfection to make NSCs overexpress brain‐derived neurotrophic factor (BDNF) can enhance their therapeutic efficacy; however, viral vectors must be used because current nonviral vectors are unable to efficiently transfect NSCs. The first polymeric vector, ROS‐responsive charge‐reversal poly(2‐acryloyl)ethyl(p‐boronic acid benzyl)diethylammonium bromide (B‐PDEA), is shown here, that mediates efficient gene transfection of NSCs and greatly enhances their therapeutics in ischemic stroke treatment. The cationic B‐PDEA/DNA polyplexes can effectively transfect NSCs; in the cytosol, the B‐PDEA is oxidized by intracellular ROS into negatively charged polyacrylic acid, quickly releasing the BDNF plasmids for efficient transcription and secreting a high level of BDNF. After i.v. injection in ischemic stroke mice, the transfected NSCs (BDNF‐NSCs) can home to ischemic regions as efficiently as the pristine NSCs but more efficiently produce BDNF, leading to significantly augmented BDNF levels, which in turn enhances the mouse survival rate to 60%, from 0% (nontreated mice) or ≈20% (NSC‐treated mice), and enables more rapid and superior functional reconstruction.
The first nonviral gene carrier, reactive‐oxygen‐species‐responsive charge‐reversal poly(2‐acryloyl)‐ethyl(p‐boronic acid benzyl)diethylammonium bromide (B‐PDEA), is shown to mediate efficient gene transfection to neural stem cells (NSCs). When BDNF gene plasmids are used, the transfected NSCs homing to the ischemic regions increase animal survival and reconstruct functions.
Skin wound therapy aims not only to restore skin protection but also to recover excitation functions through nerve regeneration. During the restoration of skin nerves, the recruitment of endogenous ...stem cells and promotion of neuronal regeneration on site work stepwise are foundations of in situ regeneration. However, current therapeutic systems usually execute each process separately, leading to limited regeneration and recovery of excitation functions. Herein, a novel self‐adaptive all‐in‐one delivery chip (G:P:Al‐Chip) is constructed that combines therapeutic protein release, gene delivery, and electrical conduction in a single microfluidic chip by 3D coaxial printing. G:P:Al‐Chip consists of an outer conductive hydrogel shell anchored with chemokine and an inner microchannel filled with enzyme‐initiated vector/plasmid DNAs microcomplexes. G:P:Al‐Chip delivers chemokine, functional plasmid DNAs, and promotes electrical conduction with a self‐adaptive program that significantly enhances the recruitment of endogenous mesenchymal stem cells and promotes neuronal regeneration. G:P:Al‐Chip is shown to enhance nerve regeneration with excitation functions within 23 days. G:P:Al‐Chip organizes recruitment and neuronal regeneration cues along with bioelectrical signal in one degradable chip for accelerated skin nerve regeneration.
An all‐in‐one delivery chip that combines protein release, gene delivery, and electrical conduction in a single microfluidic chip is developed via 3D coaxial printing. This chip organizes recruitment and neuronal regeneration cues for endogenous mesenchymal stem cells for in situ neuronal regeneration, providing a bottom‐up approach to regenerative biology.
Nanoparticles are used in many fields and in everyday products. Silver nanoparticles are the most frequently used nanoparticles; for example, in food-related products, owing to their antibacterial ...activity. However, it has been pointed out that they might have unexpected biological effects, and evaluation of their effects is underway. Although there is a growing body of evidence that nanoparticles can also induce epigenetic changes, there is still little information on the underlying mechanisms. Here, we evaluated changes in DNA methylation induced by silver nanoparticles and attempted to elucidate the induction mechanism. Immunofluorescence staining analysis revealed that silver nanoparticles with a diameter of 10, 50, or 100 nm (nAg10, nAg50, and nAg100, respectively) decreased the content of methylated DNA in A549 alveolar epithelial cells. The level of DNA methyltransferase 1 (Dnmt1) protein, which is involved in maintaining methylation during DNA replication, was significantly decreased, whereas that of Dnmt3b, which is responsible for de novo DNA methylation, was significantly increased by nAg10 treatment. Co-treatment with nAg10 and cycloheximide, which inhibits translation by inhibiting the translocation step of protein synthesis, decreased the level of Dnmt1 in comparison with nAg10-treated A549 cells, indicating a post-translational effect of nAg10. Furthermore, pretreatment with the proteasome inhibitor lactacystin restored the levels of Dnmt1 protein and DNA methylation in nAg10-treated cells. Collectively, these results suggest that nAg10 induced DNA hypomethylation through a proteasome-mediated degradation of Dnmt1.
Despite more effective chemotherapy combined with limb-salvage surgery for the osteosarcoma treatment, survival rates for osteosarcoma patients have stagnated over the past three decades due to the ...poor prognosis. Osteosarcoma cancer stem cells (OSCs) are responsible for the growth and metastasis of osteosarcoma. The existence of OSCs offers a theoretical explanation for therapeutic failures including tumor recurrence, metastasis, and drug resistance. Understanding the pathways that regulate properties of OSCs may shed light on mechanisms that lead to osteosarcoma and suggest better modes of treatment. In this study, we showed that the expression level of Kruppel-like factor 4 (KLF4) is highly associated with human osteosarcoma cancer stemness. KLF4-overexpressed osteosarcoma cells displayed characteristics of OSCs: increased sphere-forming potential, enhanced levels of stemness-associated genes, great chemoresistance to adriamycin and CDDP, as well as more metastasis potential. Inversely, KLF4 knockdown could reduce colony formation in vitro and inhibit tumorigenesis in vivo, supporting an oncogenic role for KLF4 in osteosarcoma pathogenesis. Furthermore, KLF4 was shown to activate the p38 MAPK signaling pathway to promote cancer stemness. Altogether, our studies uncover an essential role for KLF4 in regulation of OSCs and identify KLF4-p38 MAPK axis as a potential therapeutic target for osteosarcoma treatment.
The toxicity evaluation of inorganic nanoparticles has been reported by an increasing number of studies, but toxicity studies concerned with biodegradable nanoparticles, especially the neurotoxicity ...evaluation, are still limited. For example, the potential neurotoxicity of Polysorbate 80-modified chitosan nanoparticles (Tween 80-modified chitosan nanoparticles, TmCS-NPs), one of the most widely used brain targeting vehicles, remains unknown. In the present study, TmCS-NPs with a particle size of 240 nm were firstly prepared by ionic cross-linking of chitosan with tripolyphosphate. Then, these TmCS-NPs were demonstrated to be entered into the brain and specially deposited in the frontal cortex and cerebellum after systemic injection. Moreover, the concentration of TmCS-NPs in these two regions was found to decrease over time. Although no obvious changes were observed for oxidative stress in the in vivo rat model, the body weight was found to remarkably decreased in a dose-dependent manner after exposure to TmCS-NPs for seven days. Besides, apoptosis and necrosis of neurons, slight inflammatory response in the frontal cortex, and decrease of GFAP expression in the cerebellum were also detected in mouse injected with TmCS-NPs. This study is the first report on the sub-brain biodistribution and neurotoxicity studies of TmCS-NPs. Our results provide new insights into the toxicity evaluation of nanoparticles and our findings would help contribute to a better understanding of the neurotoxicity of biodegradable nanomaterials used in pharmaceutics.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Purposes
Liquid protein-based biopharmaceutical formulations have been reported to form aggregation and protein sub-visible particles (SbVPs) during dropping (Randolph
et al
., J Pharm Sci 2015, 104, ...602). However, effects of secondary package on liquid biopharmaceutical formulation stability during dropping are overlooked and have not been reported so far. This study reports the first real-world evaluation on effects of secondary package on liquid biopharmaceutical formulation stability during dropping, using two monoclonal antibodies (mAb-1 and mAb-2) and one fusion protein (FP-1) as model biopharmaceuticals.
Methods
The potential protective effects of secondary package and formulation composition on liquid biopharmaceutical formulations during dropping were evaluated with micro-flow imaging (MFI) and dynamic light scattering (DLS).
Results
The dropping-induced degradation could be detected with the two sensitive particle analyzing techniques MFI and DLS. Formulation compositions have dramatic impact on biopharmaceutical stability during dropping. Surprisingly, unlike the primary packages that have been reported to impact liquid biopharmaceutical stability, the secondary packaging system as described in our current preliminary design has little or no protective effect during dropping.
Conclusions
Our study is the first real-world data showing that the secondary package system has little to no effect on the liquid biopharmaceutical formulation quality during dropping. On the contrary, the stability of liquid biopharmaceutical formulations during dropping is more relevant to formulation compositions and primary packages.
Owing to the diversity and ease of preparation of nanomaterials, the rational nanocarriers with a rational design have become increasingly popular in medical researches. Although nanoparticle-based ...drug delivery exhibits great potential, there are some challenges facing like rapid plasma clearance, triggering or aggravation of immune response, etc. Herein, cell-based targeted drug delivery systems have drawn more and more attention owing to low immunogenicity and intrinsic mutation rate, and innate ability to allow targeted delivery. Mesenchymal stem cells (MSCs) have been used in gene and drug delivery. The use of MSCs is a promising approach for the development of gene transfer systems and drug loading strategies because of their intrinsic properties, including homing ability and tumor tropism. By combining the inherent cell properties and merits of synthetic nanoparticles (NPs), cell membrane coated NPs emerge as the time requires. Overall, we provide a comprehensive overview of the utility of MSCs in drug and gene delivery as well as MSC membrane coated nanoparticles for therapy and drug delivery, aiming to figure out the significant room for development and highlight the potential future directions.
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Abstract α-Mangostin (α-M) is a polyphenolic xanthone that protects and improves the survival of cerebral cortical neurons against Aβ oligomer-induced toxicity in rats. α-M is a potential candidate ...as a treatment for Alzheimer's disease (AD). However, the efficacy was limited by the poor penetration of the drug through the blood–brain barrier (BBB). In this study, we modified the α-M liposome with transferrin (Tf) and investigated the intracellular distribution of liposomes in bEnd3 cells. In addition, the transport of α-M across the BBB in the Tf(α-M) liposome group was examined. In vitro studies demonstrated that the Tf(α-M) liposome could cross the BBB in the form of an integrated liposome. Results of the in vivo studies on the α-M distribution in the brain demonstrated that the Tf(α-M) liposome improved the brain delivery of α-M. These results indicated that the Tf liposome is a potential carrier of α-M against AD.
Spinal cord injury (SCI) has seriously affected the lives of patients and brought economic and medical burdens to society. Biopharmaceuticals, including transplanted cells, growth factors, enzyme ...drugs, and microRNA, present multidimensional therapeutic effects in tissue repair and regeneration, and thus are widely utilized for the treatment of SCI. Nevertheless, they still encounter great challenges. The turbulent microenvironment accompanied by various destructive components following SCI has led to poor stability, low local accumulation, and reduced therapeutic efficacy of biopharmaceuticals. To this end, biomaterial‐based defenders (BBDs) that are capable of resisting the destructive components have shown great promise in improving the survival of transplanted cells or retaining the activity of the protein and nucleic acid drugs for the treatment of SCI. In this review, various types of biopharmaceuticals applied in SCI and their limitations are introduced. The destructive components that threaten the activities of biopharmaceuticals in the rigorous microenvironment are then summarized. In particular, the current research progress about BBDs boosting the efficacy of biopharmaceuticals for the treatment of SCI is illustrated with specific examples. The existing challenges and future perspectives are also provided.
Although various types of biopharmaceuticals have presented multidimensional therapeutic effects in spinal cord injury (SCI), the rigorous microenvironment after SCI impedes their capacity. Biomaterial‐based defenders (BBDs) are designed to protect biopharmaceuticals passively or actively to reduce their inactivation and loss. Herein, this review summarizes the latest advances in BBDs boosting the therapeutic efficacy of biopharmaceuticals for SCI treatment.
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