Supercritical Carbon Dioxide Treatment of Porous Silicon Increases Biocompatibility with Cardiomyocytes.

Porous silicon is of current interest for cardiac tissue engineering applications. While porous silicon is considered to be a biocompatible material, it is important to assess whether post-etching surface treatments can further improve biocompatibility and perhaps modify cellular behavior in desirable ways. In this work, porous silicon was formed by electrochemically etching with hydrofluoric acid, and was then treated with oxygen plasma or supercritical carbon dioxide (scCO2). These processes yielded porous silicon with a thickness of around 4 µm. The different post-etch treatments gave surfaces that differed greatly in hydrophilicity: oxygen plasma-treated porous silicon had a highly hydrophilic surface, while scCO2 gave a more hydrophobic surface. The viabilities of H9c2 cardiomyocytes grown on etched surfaces with and without these two post-etch treatments was examined; viability was found to be highest on porous silicon treated with scCO2. Most significantly, the expression of some key genes in the angiogenesis pathway was strongly elevated in cells grown on the scCO2-treated porous silicon, compared to cells grown on the untreated or plasma-treated porous silicon. In addition, the expression of several apoptosis genes were suppressed, relative to the untreated or plasma-treated surfaces.

Slow skeletal muscle troponin T acts as a potential prognostic biomarker and therapeutic target for hepatocellular carcinoma.

Slow skeletal muscle troponin T (TNNT1) as a poor prognostic indicator is upregulated in colon and breast cancers. However, the role of TNNT1 in the disease prognosis and biological functions of hepatocellular carcinoma (HCC) is still unclear. The Cancer Genome Atlas (TCGA), real-time quantitative RT-PCR (qRT-PCR), immunoblot, and immunohistochemical analyses were applied to evaluate the TNNT1 expression of human HCC. The impact of TNNT1 levels on disease progression and survival outcome was studied using TCGA analysis. Moreover, the bioinformatics analysis and HCC cell culture were used to investigate the biological functions of TNNT1. Besides, the immunoblot analysis and enzyme-linked immunosorbent assay (ELISA) were used to detect the extracellular TNNT1 of HCC cells and circulating TNNT1 of HCC patients, respectively. The effect of TNNT1 neutralization on oncogenic behaviors and signaling was further validated in the cultured hepatoma cells. In this study, tumoral and blood TNNT1 was upregulated in HCC patients based on the analyses using bioinformatics, fresh tissues, paraffin sections, and serum. From the multiple bioinformatics tools, the TNNT1 overexpression was associated with advanced stage, high grade, metastasis, vascular invasion, recurrence, and poor survival outcome in HCC patients. By the cell culture and TCGA analyses, TNNT1 expression and release were positively correlated with epithelial-mesenchymal transition (EMT) processes in HCC tissues and cells. Moreover, TNNT1 neutralization suppressed oncogenic behaviors and EMT in hepatoma cells. In conclusion, TNNT1 may serve as a non-invasive biomarker and drug target for HCC management. This research finding may provide a new insight for HCC diagnosis and treatment.

Doping of MXenes enhances the electrochemical response of peptide-imprinted conductive polymers for the recognition of C-Reactive protein.

The level of C-reactive protein (CRP) in serum is frequently used to evaluate risk of coronary heart disease, and its concentration is related to cardiovascular disease, fibrosis and inflammation, cancer, and viral infections. In this work, three novel peptides, never previously used as imprinted templates, were selected, synthesized, and employed for epitope imprinting. Various imprinting concentrations of the template and various ratios of aniline (AN) to m-aminobenzenesulfonic acid (MSAN) were used in electropolymerization to form molecularly imprinted polymers (MIPs). The imprinting template and functional monomer concentrations were optimized to maximize the electrochemical response to target peptides. The surface morphologies of peptide- and non-imprinting poly(AN-co-MSAN) were observed using a scanning electron microscope (SEM) and an atomic force microscope (AFM). Moreover, the effect of doping of MIPs with a very small percentage of an MXene (e.g. Ti2C at 0.1 wt% in the preparation solution) on the electrochemical response was also studied. Ti2C doping dramatically increased sensing range from 0.1 to 100 fg/mL to 10000 fg/mL, and electrochemical responses were amplified by a factor of approximately 1.3 within the sensing range. Finally, commercially available serum was diluted and then measured using the MXene-doped PIP-coated electrodes to estimate the accuracy compared with ELISA results.

Synthesis of ginsenoside Rb1-imprinted magnetic polymer nanoparticles for the extraction and cellular delivery of therapeutic ginsenosides.

Background: Panax ginseng (ginseng) is a traditional medicine that is reported to have cardioprotective effects; ginsenosides are the major bioactive compounds in the ginseng root. Methods: Magnetic molecularly imprinted polymer (MMIP) nanoparticles might be useful for both the extraction of the targeted (imprinted) molecules, and for the delivery of those molecules to cells. In this work, plant growth regulators were used to enhance the adventitious rooting of ginseng root callus; imprinted polymeric particles were synthesized for the extraction of ginsenoside Rb1 from root extracts, and then employed for subsequent particle-mediated delivery to cardiomyocytes to mitigate hypoxia/reoxygenation injury. Results: These synthesized composite nanoparticles were first characterized by their specific surface area, adsorption capacity, and magnetization, and then used for the extraction of ginsenoside Rb1 from a crude extract of ginseng roots. The ginsenoside-loaded MMIPs were then shown to have protective effects on mitochondrial membrane potential and cellular viability for H9c2 cells treated with CoCl2 to mimic hypoxia injury. The protective effect of the ginsenosides was assessed by staining with JC-1 dye to monitor the mitochondrial membrane potential. Conclusion: MMIPs can play a dual role in both the extraction and cellular delivery of therapeutic ginsenosides.

Sensing of C-Reactive Protein Using an Extended-Gate Field-Effect Transistor with a Tungsten Disulfide-Doped Peptide-Imprinted Conductive Polymer Coating.

C-reactive protein (CRP) is a non-specific biomarker of inflammation and may be associated with cardiovascular disease. In recent studies, systemic inflammatory responses have also been observed in cases of coronavirus disease 2019 (COVID-19). Molecularly imprinted polymers (MIPs) have been developed to replace natural antibodies with polymeric materials that have low cost and high stability and could thus be suitable for use in a home-care system. In this work, a MIP-based electrochemical sensing system for measuring CRP was developed. Such a system can be integrated with microfluidics and electronics for lab-on-a-chip technology. MIP composition was optimized using various imprinting template (CRP peptide) concentrations. Tungsten disulfide (WS2) was doped into the MIPs. Doping not only enhances the electrochemical response accompanying the recognition of the template molecules but also raises the top of the sensing range from 1.0 pg/mL to 1.0 ng/mL of the imprinted peptide. The calibration curve of the WS2-doped peptide-imprinted polymer-coated electrodes in the extended-gate field-effect transistor platform was obtained and used for the measurement of CRP concentration in real human serum.

Self-assembly Synthesis of Molecularly Imprinted Polymers for the Ultrasensitive Electrochemical Determination of Testosterone.

Molecularly imprinted polymers (MIPs) can often bind target molecules with high selectivity and specificity. When used as MIPs, conductive polymers may have unique binding capabilities; they often contain aromatic rings and functional groups, which can undergo π－π and hydrogen bonding interactions with similarly structured target (or template) molecules. In this work, an electrochemical method was used to optimize the synthetic self-assembly of poly(aniline-co-metanilic acid) and testosterone, forming testosterone-imprinted electronically conductive polymers (TIECPs) on sensing electrodes. The linear sensing range for testosterone was from 0.1 to 100 pg/mL, and the limit of detection was as low as ~pM. Random urine samples were collected and diluted 1000-fold to measure testosterone concentration using the above TIECP sensors; results were compared with a commercial ARCHITECT ci 8200 system. The testosterone concentrations in the tested samples were in the range of 0.33 ± 0.09 to 9.13 ± 1.33 ng/mL. The mean accuracy of the TIECP-coated sensors was 90.3 ± 7.0%.

Immunotherapy of Hepatocellular Carcinoma with Magnetic PD-1 Peptide-Imprinted Polymer Nanocomposite and Natural Killer Cells.

Programmed cell death protein 1 (PD-1) is a biomarker on the surface of cells with a role in promoting self-tolerance by suppressing the inflammatory activity of T cells. In this work, one peptide of PD-1 was used as the template for molecular imprinting to form magnetic peptide-imprinted poly(ethylene-co-vinyl alcohol) composite nanoparticles (MPIP NPs). The nanoparticles were characterized by dynamic light scattering (DLS), high-performance liquid chromatography (HPLC), Brunauer-Emmett-Teller (BET) analysis, and superconducting quantum interference device (SQUID) analysis. Natural killer 92 (NK-92) cells were added to these composite nanoparticles and then incubated with human hepatoma (HepG2) cells. The viability and the apoptosis pathway of HepG2 were then studied using cell counting kit-8 (CCK8) and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. These nanoparticles were found to significantly enhance the activity of natural killer cells toward HepG2 cells by increasing the expression of nuclear factor kappa B (NF-κB), caspase 8, and especially caspase 3.