Microfluidic hydrodynamic trapping for single cell analysis: mechanisms, methods and applications

The development of hydrodynamic-based microfluidic biochips has been increasing over the years. In this technique, the cells or particles are trapped in a particular region for single cell analysis (SCA) usually without any application of external force fields such as optical, electrical, magnetic or acoustic. There is a need to explore the insights of SCA in the cell's natural state and development of these techniques is highly essential for that study. Researchers have highlighted the vast potential field that needs to be explored to develop biochip devices to suit market/researcher demands. Hydrodynamic microfluidics facilitates the development of passive lab-on-chip applications. This review gives an account of the recent advances in this field, along with their mechanisms, methods and applications

In vitro evaluation of porcupine bezoar extracts as anticancer agent on A549 -A preliminary study

Porcupine bezoar (PB) was reported to possess medicinal properties in old medical manuscript. However, its potential as anticancer agent on human lung cancer cells (A549) is not yet studied. In present study, porcupine bezoar was tested to observe its ability in inhibiting cell growth of cancer cell (A549) and its cytotoxicity on Normal Human Gingival Fibroblast Cell (HGF-1). A549 cells morphology was observed after treated with bezoars for 72 hours. The ability of bezoars to induce DNA damage and apoptosis was analyzed by staining cells with Hoechst 33428(nucleus) and Rhodamine Phalloidin (f-actin). The A549 IC50 is 13.6±1.58μg/ml A549 growths was inhibited in dose-dependent pattern, but no inhibition found on normal HGF-1 cells. Treated A549 morphology shows sign of apoptosis such as DNA fragmentation, cytoplasm shrunk and vacuolation. The finding in this study suggests PB extracts able to inhibit cell growth, induce DNA damage and apoptosis, further analysis need to be done to verify the mechanism

Direct cell imprint lithography in superconductive carbon black polymer composites: process optimization, characterization and in vitro toxicity analysis

Cell imprint lithography (CIL) or cell replication plays a vital role in fields like biomimetic smart culture substrates, bone tissue engineering, cell guiding, cell adhesion, tissue engineering, cell microenvironments, tissue microenvironments, cell research, drug delivery, diagnostics, therapeutics and many other applications. Herein we report a new formulation of superconductive carbon black photopolymer composite and its characterization towards a CIL process technique. In this article, we demonstrated an approach of using a carbon nanoparticle-polymer composite (CPC) for patterning cells. It is observed that a 0.3 wt % load of carbon nanoparticles (CNPs) in a carbon polymer mixture (CPM) was optimal for cell-imprint replica fabrication. The electrical resistance of the 3-CPC (0.3 wt %) was reduced by 68% when compared to N-CPC (0 wt %). This method successfully replicated the single cell with sub-organelle scale. The shape of microvesicles, grooves, pores, blebs or microvilli on the cellular surface was patterned clearly. This technique delivers a free-standing cell feature substrate. In vitro evaluation of the polymer demonstrated it as an ideal candidate for biomimetic biomaterial applications. This approach also finds its application in study based on morphology, especially for drug delivery applications and for investigations based on molecular pathways

Identification and quantification of quercetin, a major constituent of Artocarpus altilis by targeting related genes of apoptosis and cell cycle: in vitro cytotoxic activity against human lung carcinoma cell lines

Nine phenolic compounds were identified and quantified in Artocarpus altilia fruit. One of the main compounds was quercetin, which is the major class of flavonoids has been identified and quantified in pulp part of A. altilis fruit of methanol extract. The aim of this study was to evaluate in vitro cytotoxic assay. Inhibitory concentration 50% concentration was determined using trypan blue exclusion assay. Apoptosis induction and cell cycle regulation were studied by flow cytometric analysis. The expression of apoptosis and cell cycle-related regulatory genes were assessed by RT-qPCR study of the methanol extract of pulp part on human lung carcinoma (A549) cell line. A significant increase of cells at G2/M phases was detected (P<0.05). Furthermore, the pulp of the fruit downregulated the expression of antiapoptosis gene BCL-2 and upregulated the expression of pro-apoptosis gene BAX. CASPASE3 was also activated by the fruit, which started a CASPASE-3-depended mitochondrial pathway to induce apoptosis. As the results, the pulp was the most active in terms of all tests, due to high amount of quercetin in pulp part, 78% of total flavonoids. Taken together, these findings suggested that A. altilis induces apoptosis in a mitochondrial-dependent pathway by releasing and upregulating CYTOCHROME C expression and regulates the expression of downstream apoptotic components, including BCL-2 and BAX

Alpha-glucosidase inhibitory effect of psychotria malayana jack leaf: A rapid analysis using infrared fingerprinting

The plant Psychotria malayana Jack belongs to the Rubiaceae family and is known in Malaysia as “meroyan sakat/salung”. A rapid analytical technique to facilitate the evaluation of the P. malayana leaves’ quality has not been well-established yet. This work aimed therefore to develop a validated analytical technique in order to predict the alpha-glucosidase inhibitory action (AGI) of P. malayana leaves, applying a Fourier Transform Infrared Spectroscopy (FTIR) fingerprint and utilizing an orthogonal partial least square (OPLS). The dried leaf extracts were prepared by sonication of different ratios of methanol-water solvent (0, 25, 50, 75, and 100% v/v) prior to the assessment of alpha-glucosidase inhibition (AGI) and the following infrared spectroscopy. The correlation between the biological activity and the spectral data was evaluated using multivariate data analysis (MVDA). The 100% methanol extract possessed the highest inhibitory activity against the alpha-glucosidase (IC50 2.83 ± 0.32 µg/mL). Different bioactive functional groups, including hydroxyl (O-H), alkenyl (C=C), methylene (C-H), carbonyl (C=O), and secondary amine (N-H) groups, were detected by the multivariate analysis. These functional groups actively induced the alpha-glucosidase inhibition effect. This finding demonstrated the spectrum profile of the FTIR for the natural herb P. malayana Jack, further confirming its medicinal value. The developed validated model can be used to predict the AGI of P. malayana, which will be useful as a tool in the plant’s quality control

Potential anticancer agents identification of Hystrix Brachyura Bezoar through gas chromatography-mass spectrometry-based metabolomics and protein-ligand interaction with molecular docking analyses

Background: Bezoar (PB) is a rare, solidified form of undigested food commonly found in the gastrointestinal tract of porcupine (Hystrix brachyura). It is believed to be traditionally used to treat various diseases including different kinds of cancers in Malaysia. However, its active principles have not been found out yet. The purpose of this study was to investigate the anticancer property of PB extract as well as to identify the metabolites responsible for its anticancer effect through a widely acclaimed metabolomics approach. Methods: Initially, 25 PB extracts using various solvent ratios of methanol–water (100, 75, 50, 25, 0% v/v) were prepared in regard to metabolomics approach and subsequently the cytotoxicity of each extract was determined against (melanoma) A375 cell line. The metabolites profiling of the most potent extract was conducted using gas chromatography mass spectrometry (GC-MS) and in silico investigation was performed on Bcl-2 and cyclin/CDK1 complex protein. Results: The correlation of the bioactivity with GC-MS data produced an orthogonal partial least square (OPLS) model which pinpointed four putative active compounds namely (1) cholest-7-en-3-beta-ol,4,4-dimethyl-,acetate; (2) 4-androsten-4-ol-3,17-dione; (3) isolongifolol and (4) gallic acid. The in silico data suggested the binding score and binding mode of active metabolites with the amino acid residues of protein via hydrophobic interactions. Conclusion: This study is the first to report the identified anticancer compounds from PB extract and evaluate them using molecular docking. This further confirms and justifies its traditional usage as an alternative medicine for the treatment of cancer in Malaysia

Porcupine bezoar exhibit as anti-cancer activities on human breast adenocarcinoma cells (MCF-7)

Purpose: Currently there were many treatments available to treat cancer patients, for instance chemotherapy, radiation therapy, immune therapy, and many more. Unfortunately, advancement in cancer therapy has resulted in increasing numbers of survivors which left to deal with side effects of their treatments. Therefore, discovery of alternative treatment with minimal side effect is crucial. Thus, study on bezoar take place as there were no scientific studies about porcupine bezoar to this date in Malaysia. This lead to the premise that porcupine bezoar could have greater potential for the chemoprevention activities. Methods: MCF-7 cells were treated with bezoar for cell proliferation, cell cycle, apoptosis and qPCR analyses. Cell viability of treated and untreated MCF-7 was examined by Trypan Blue Exclusion method. The cellular morphology of MCF-7 cells was observed by phase contrast microscopy. Flow cytometry analyzed cell apoptosis with annexin V/ propidium iodide (PI) and cell cycle with PI. qPCR was run in detecting gene expression involved in the apoptosis and cell cycle arrest. Result(s): Based on the conducted study and experiments, it is evident that the bezoar treatment in human periodontal ligament cells didn’t show cytotoxicity effect. Upon treated with optimized concentration of bezoar for 24, 48 and 72 hours, it shows inhibition of proliferation by 45.5, 31.25 and 37.1% respectively, significantly apoptosis and cell cycle arrest occurs. Bezoar treatment also induced apoptotic-liked in morphological changes. Upon qPCR analysis, its indicated mcf7 undergo intrinsic pathway of apoptosis and arrest at G1/G0 as gene expression high in cdk2, cyclin D and p21. Conclusion(s): Porcupine bezoar can be alternative treatment for cancer with minimal side effects. Yet, furthermore insights need to be studied

Characterization of alpha-glucosidase inhibitory activity of Tetracera scandens leaves by Fourier transform infrared spectroscopy-based metabolomics

Tetracera scandens is a medicinal shrub that belongs to Dilleniaceae. The leaves of the plant have been traditionally used in the treatment of diabetes mellitus in Malaysia. The conventional quality control analysis of medicinal plants that relies on the quantification of few major metabolites is considered time-consuming since it requires extensive sample preparation and neglects the possible impacts that the minor metabolites could have on the activity. This study was aimed to investigate the α-glucosidase inhibitory (AGI) potential of different hydromethanolic extracts of T. scandens leaves and to establish a predictive multivariate model that could be used for the quality evaluation of T. scandens leaf based on the Fourier transform infrared (FT-IR) spectra of its extracts. Different solvent ratios (0%, 20%, 40%, 60%, 80% and 100% methanol in water) were used to prepare a total of 36 extracts. The AGI potential and the FT-IR fingerprint spectrum were acquired for each extract. A four components orthogonal partial least squares (OPLS) model (1 + 3 + 0) with R2Y of 0.951 and Q2Y of 0.916 was established to describe the correlation between the fingerprint FT-IR spectra of different T. scandens extracts and their corresponding AGI activities. The carbon-halide, carbon–oxygen single bonds, as well as the hydroxyl and carbonyl groups were identified to be positively correlated with the AGI activity. To sum up, an OPLS model was successfully developed as a rapid quality evaluation method to predict the AGI activity of T. scanden

Skin cancer detection using non-invasive techniques

Skin cancer is the most common form of cancer and is globally rising. Historically, the diagnosis of skin cancers has depended on various conventional techniques which are of an invasive manner. A variety of commercial diagnostic tools and auxiliary techniques are available to detect skin cancer. This article explains in detail the principles and approaches involved for non-invasive skin cancer diagnostic methods such as photography, dermoscopy, sonography, confocal microscopy, Raman spectroscopy, fluorescence spectroscopy, terahertz spectroscopy, optical coherence tomography, the multispectral imaging technique, thermography, electrical bio-impedance, tape stripping and computer-aided analysis. The characteristics of an ideal screening test are outlined, and the authors pose several points for clinicians and scientists to consider in the evaluation of current and future studies of skin cancer detection and diagnosis. This comprehensive review critically analyses the literature associated with the field and summarises the recent updates along with their merits and demerits

Direct cell imprint lithography in superconductive carbon black polymer composites: Process optimization, characterization and in vitro toxicity analysis

Cell imprint lithography (CIL) or cell replication plays a vital role in fields like biomimetic smart culture substrates, bone tissue engineering, cell guiding, cell adhesion, tissue engineering, cell microenvironments, tissue microenvironments, cell research, drug delivery, diagnostics, therapeutics and many other applications. Herein we report a new formulation of superconductive carbon black photopolymer composite and its characterization towards a CIL process technique. In this article, we demonstrated an approach of using a carbon nanoparticle-polymer composite (CPC) for patterning cells. It is observed that a 0.3 wt % load of carbon nanoparticles (CNPs) in a carbon polymer mixture (CPM) was optimal for cell-imprint replica fabrication. The electrical resistance of the 3-CPC (0.3 wt %) was reduced by 68% when compared to N-CPC (0 wt %). This method successfully replicated the single cell with sub-organelle scale. The shape of microvesicles, grooves, pores, blebs or microvilli on the cellular surface was patterned clearly. This technique delivers a free-standing cell feature substrate. In vitro evaluation of the polymer demonstrated it as an ideal candidate for biomimetic biomaterial applications. This approach also finds its application in study based on morphology, especially for drug delivery applications and for investigations based on molecular pathways