Advanced pharmaceutical bulletin最新文献

Purpose: Studies have shown the potential of exosomes as therapeutic agents with anti-inflammatory properties. However, the clinical application of mammalian-derived exosomes is hindered by mass production challenges and strict regulations. Plant-derived exosome-like nanoparticles (PELNs) are a more economical alternative possessing a similar therapeutic potential. Ginger is a readily available plant with components that are clinically proven to inhibit inflammation. Therefore, it is interesting to investigate the potential of red ginger and emprit ginger, cultivated varieties in Indonesia possessing the most potent anti-inflammatory activities, as a PELN source for anti-inflammatory therapy.

Methods: In this work, PELNs from the rhizomes of red ginger (RG-ELN) and emprit ginger (EG-ELN) were obtained through differential centrifugation and polymer precipitation using PEG6000. The PELNs were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), and bicinchoninic acid assay. Their internalization and effect on RAW 246.7 cell viability were also assessed. The anti-inflammatory potential of PELNs was investigated by assessing interleukin 6 (IL-6) expression of lipopolysaccharide (LPS)-stimulated macrophages treated with RG-ELN and EG-ELN.

Results: Both RG-ELN and EG-ELN exhibited cup-shaped morphologies with average sizes of 195.83±1.35 and 194.40±8.40 nm, respectively. Both PELNs can be internalized within 2 h and did not significantly affect RAW 264.7 cell viability after 24 h. The reverse transcription quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay results indicated a significantly lower expression and secretion of IL-6 in the macrophage cells pre-treated with RG-ELN and EG-ELN.

Conclusion: The RG-ELN and EG-ELN samples were successfully obtained through the polymer precipitation method, as confirmed by the TEM and DLS results which aligned with typical PELN characteristics. The pre-treatment of RG-ELN and EG-ELN to activated RAW 264.7 cells decreased the pro-inflammatory cytokine IL-6 expression relative to activated controls.

Purpose: SARS-CoV-2 infection may lead to a worse prognosis in COVID-19 patients by inducing syncytia formation which implies intercellular transmission and immune evasion. Hesperidin (HSD) and hesperetin (HST) are two citrus flavonoids that demonstrate the potential to interfere with spike/human angiotensin-converting enzyme-2 (hACE2) binding and show an inhibitory effect in the SARS-CoV-2 pseudovirus internalization model. Here, we determined the effects of HSD and HST to inhibit syncytia formation using in vitro cell models.

Methods: We confirmed spike, hACE2, and transmembrane protease, serine 2 (TMPRSS2) ectopic expressions by immunofluorescence staining (IF) after transfection using polyethylene imine (PEI) in 293T cells. Then, the cells were transfected with a set of plasmids encoding spike/hACE2/TMPRSS2 or spike/hACE2 to induce syncytia formation. Cell treatment with HSD/HST was performed 4-5 h after transfection and then incubated for another 16-18 h. Syncytia were observed using an inverted microscope or a high content screening (HCS) platform. The data obtained from syncytia formation assays were statistically analyzed using ANOVA (Bonferroni).

Results: We successfully observed spike, hACE2, and TMPRSS2 expression in 293T cells by IF staining. Furthermore, we showed that HSD 10 and 100 µM significantly inhibited the formation of small-to-medium-sized syncytia compared to the control cells by manual syncytia observation. In the HCS assay, 10 µM HSD showed an inhibitory effect of syncytia induced by spike WT. In contrast, 100 µM HSD, 10 and 100 µM HST, and 10 µg/mL citrus peel extract containing HSD prepared by the hydrodynamic cavitation method (HCV) inhibited syncytia formation induced by spike Omicron.

Conclusion: HSD and HST show the potential inhibitory activity of SARS-CoV-2 intercellular transmission. Further study is needed to confirm the mechanism of action of the antiviral activity.

Purpose: Cervical cancer (CxCa) is primarily caused by high-risk human papillomaviruses (hrHPV), which disrupt p53 and pRb regulation, leading to uncontrolled growth and progression. Co-infection with polyomaviruses like MCPyV in some HPV-positive cases suggests a potential combined effect on tumor development. Cisplatin is commonly used for advanced CxCa, but resistance remains a challenge. This study examines whether MCPyV sT oncoprotein and HPV-18 oncoproteins affect key gene transcription, influencing proliferation and cisplatin resistance in CxCa.

Methods: The sT gene was cloned into the pCMV6 vector, and HeLa cells were transfected with pCMV6-sT using Lipofectamine 3000. Transfection efficiency was assessed via fluorescence microscopy and flow cytometry. Protein expression was analyzed using SDS-PAGE and Western blotting. Cytotoxicity was measured with the MTT assay, gene expression was analyzed by RT-qPCR, Ki-67 staining was performed on cell blocks, and cisplatin-induced effects on proliferation and apoptosis were examined.

Results: Cytotoxicity assays showed a significant increase in cell viability at 0.2 μg of sT plasmid after 72 hours (13.76%, P<0.05). MCPyV sT expression significantly upregulated E1 (4.22-fold), E6/E7 (3.80-fold), and MMP1 (6-fold) mRNA levels (P<0.001). Increased Ki-67 positivity indicated enhanced proliferation. Additionally, sT expression reduced cisplatin-induced apoptosis, with fewer apoptotic cells observed in the sT+cisplatin group than in the cisplatin-only group (25.9% vs. 38.3%, P<0.05).

Conclusion: The presence of MCPyV sT and HPV oncoproteins together enhances resistance to cisplatin-induced apoptosis in CxCa cells, highlighting the need for further investigation into viral oncoprotein interactions to overcome therapeutic resistance.

Purpose: we investigated the synergistic influence of amnion and keratinocytes on dermal regeneration in mice.

Methods: A scaffold derived from amnion and gelatin via electrospinning was used to synthesize a polyurethane-based scaffold. polyurethane/gelatin/amnion (PU/G/A) scaffold was characterized by scanning electron microscopy (SEM), FTIR, and tensile test. biocompatibility of the corresponding scaffold was investigated using the MTT method in the culture of keratinocytes.

Results: The SEM images showed sufficient cell adhesion on the PU/G/A scaffold. The tensile test results indicated that the scaffold containing PU/G/A with the lowest Young's modulus (12 MPa±2.1) displayed higher elasticity than the scaffold without amnion. Furthermore, the MTT assay revealed that the amniotic scaffold contributed to 100% cell viability (P≤0.0001 compared to control) and proliferation. Moreover, an in vivo study conducted on mice showed that the PU/G/A/ keratinocytes scaffold results in increased granulation, tissue formation and wound closure (P<0.001 compared to control).

Conclusion: This innovative nanofiber device not only addresses the limitations of traditional dressings but also offers additional functionalities such as wound compatibility, gas exchange, promotion of angiogenesis in the injured area and a substrate that amplifies the biological functions of stem cells.

Purpose: Chitosan nanoparticles (CNs) have directed considerable research efforts towards developing biocompatible, biodegradable, inexpensive and efficient particulate drug delivery systems.

Methods: In the present investigation, we utilized green and safe inositol hexaphosphate (InsP6) as a physical cross-linker to obtain CNs (^InsP6CNs) and compared their size, zeta potential and cell uptake ability with the CNs cross-linked with tripolyphosphate (TPP) as a commonly used cross-linker (^TPPCNs). Methotrexate (MTX) as the model drug was physically incorporated within the both types of CNs (^InsP6CNs_MTX and ^TPPCNs_MTX) and their time-dependent anti-cancer behavior was evaluated on MCF-7 cell line.

Results: Compared to ^TPPCNs, ^InsP6CNs were bigger in hydrodynamic diameter and showed far different zeta potential value. The MTX encapsulation efficiency was much higher for ^InsP6CNs_MTX than that of ^TPPCNs_MTX. ^InsP6CNs and ^TPPCNs showed similar in vitro cell uptake behavior, examined on MCF-7 cell line. Furthermore, after 24 h, ^InsP6CNs_MTX had the most in vitro antitumor effect on the MCF-7 cells, compared to free MTX and ^TPPCNs_MTX.

Conclusion: Consequently, InsP6 can be presented as an accessible and cost-effective member of physical cross-linkers to prepare efficient CNs as drug delivery systems.

Cervical cancer ranks fourth in terms of diagnosis and cancer-related deaths in women worldwide. Despite the approval of prophylactic vaccines against cervical cancers, these vaccines are not able to eradicate the existing ones. Therefore, various platforms have been developed to design therapeutic vaccines against cervical cancers, including DNA/RNA-based, protein/peptide-based, vector-based, and cell-based platforms. Despite the advantages of each platform, therapeutic vaccines have displayed limited clinical benefit in patients with cervical cancer, which is partially associated with inefficient delivery of vaccine components. To address these issues, different nanoplatforms have been developed to carry cellular or molecular components of vaccines to target cells and lymphoid tissues, thus promoting the durability and potency of immune responses against tumor cells and antigens besides decreasing side effects. Moreover, nanoparticles (NPs), as adjuvants and/or carriers, provide other advantages, including sufficient antigen loading and uptake by antigen-presenting cells (APCs), adaptable antigen presentation, high immunogenicity, high stability, increased lymph node retention, and precise targeting. Thus, nanovaccines also lead us to design and develop personalized vaccines against cervical cancer. Here, we discuss platforms that have been used in clinical trials for the treatment of cervical cancer, their advantages and disadvantages, platforms for developing nanovaccines, and how they improve the therapeutic efficacy of vaccines.

The prevalence of nutrition-related non-communicable diseases like diabetes mellitus (DM) is exponentially increasing across the world. Particularly, type-2 diabetes mellitus (T2DM) is prevalent in sub-Saharan Africa (SSA) than in any other region of the world, with a significant effect on mortality and morbidity. T2DM is a disease known to be associated with elevated glucose levels in the blood, caused by numerous factors including dietary and lifestyle changes. Ensuring an adequate supply of a healthy diet through a transformed food system could be a potential strategy to mitigate T2DM in SSA. In plants, starch is the most common storage carbohydrate, and it is the major glucose-releasing carbohydrate in human diets. The rate of starch digestibility varies and is largely due to the proportion of its two polyglucan components, amylose and amylopectin. Although, no medication has been found to effectively treat T2DM, it could be managed through effective postprandial glycemia control. This article reviews the mechanism for slowing down the rate of starch digestion and absorption in the small intestine through direct alteration of amylose and amylopectin in starch crops. This strategy would ensure the supply of healthy diets for consumption and ultimately help to curb the increasing prevalence of T2DM.

Purpose: The present study aimed to determine the selective effects of BRAF V600E inhibitor on focal adhesion in melanoma cells with respect to their phenotypic reprogramming.

Methods: Flow cytometry was used to analyse the distribution of BRAFV600E and BRAFWT melanoma cells throughout the cell cycle post-vemurafenib treatment. Senescent cells were identified based on b-galactosidase activity and the mRNA expression of cell cycle proteins, CCND1 and RBL1. Centrifugal cell adhesion assay was used to determine the adhesive capacities of resting and proliferative BRAF mutant and BRAF wild-type melanoma cells under vemurafenib treatment. Fibronectin binding was evaluated by spectrophotometry and quantitative real-time PCR to measure the mRNA levels of integrins: ITGAV, ITGA5, ITGB1 and ITGB3.

Results: Vemurafenib increases the proportion of melanoma BRAFV600E-positive cells in the G0 phase of a cell cycle. Melanoma cells entering the G0 phase after vemurafenib treatment indicated an upregulation of senescence-associated markers. Non-proliferating melanoma cell number was elevated among vemurafenib-treated BRAFV600E cells with enhanced attachment. BRAFV600E-positive but not BRAFV600E-negative cells were characterised by upregulated ITGAV.

Conclusion: The current results demonstrated that vemurafenib induces the phenotypic switch in melanoma cells depending on their mutational status. It also strengthens the adhesive features of senescent cells, increasing their binding to fibronectin via ITGAV, which may be a part of the phenotypic mode of drug resistance or slow interaction of proliferating cancer cells with the extracellular matrix (ECM). Thus, targeting senescent cells by focal adhesion modulators may be a promising approach to control drug-resistant melanoma cells.