Surface-modified carboxylated nanometre-sized diamond (cND) has been applied for the conjugation of biological molecules such as DNA and protein. In this study, we evaluated the biocompatibility and detection of cNDs and carbon nanotubes on human lung A549 epithelial cells and HFL-1 normal fibroblasts. Treatment with 5 or 100 nm cND particles, 0.1-100 μg ml −1 , did not reduce the cell viability and alter the protein expression profile in lung cells; however, carbon nanotubes induced cytotoxicity in these cells. The cNDs particles were accumulated in A549 cells, which were observed by atomic force microscopy and laser scanning confocal microscopy. Both 5 and 100 nm cNDs particles exhibited the green fluorescence and were ingested into cells. Moreover, the fluorescence intensities were increased in cells via a concentration-dependent manner after treatment with 5 and 100 nm cNDs, which can be detected by flow cytometer analysis. The fluorescence intensities of 5 nm cNDs were relative higher than 100 nm cNDs in cells at equal concentration treatment. The observation demonstrated that cND-interacting with cell is detectable by a confocal microscope, flow cytometer and atomic force microscope. These nanoparticles may be useful for further biomedical applications based on the properties of uptake ability, detectability and little cytotoxicity in human cells.

A novel method is proposed using nanometer-sized diamond particles as detection probes for biolabeling. The advantages of nanodiamond's unique properties were demonstrated in its biocompatibility, nontoxicity, easily detected Raman signal, and intrinsic fluorescence from its natural defects without complicated pretreatments. Carboxylated nanodiamond's (cND's) penetration ability, noncytotoxicity, and visualization of cND-cell interactions are demonstrated on A549 human lung epithelial cells. Protein-targeted cell interaction visualization was demonstrated with cND-lysozyme complex interaction with bacteria Escherichia coli. It is shown that the developed biomolecule-cND complex preserves the original functions of the test protein. The easily detected natural fluorescent and Raman intrinsic signals, penetration ability, and low cytotoxicity of cNDs render them promising agents in multiple medical applications.

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Theranostic nanomedicine is capable of diagnosis, therapy, and monitoring the delivery and distribution of drug molecules and has received growing interest. Herein, a self-monitored and self-delivered photosensitizer-doped FRET nanoparticle (NP) drug delivery system (DDS) is designed for this purpose. During preparation, a donor/acceptor pair of perylene and 5,10,15,20-tetro (4-pyridyl) porphyrin (H2TPyP) is co-doped into a chemotherapeutic anticancer drug curcumin (Cur) matrix. In the system, Cur works as a chemotherapeutic agent. In the meantime, the green fluorescence of Cur molecules is quenched (OFF) in the form of NPs and can be subsequently recovered (ON) upon release in tumor cells, which enables additional imaging and real-time self-monitoring capabilities. H2TPyP is employed as a photodynamic therapeutic drug, but it also emits efficient NIR fluorescence for diagnosis via FRET from perylene. By exploiting the emission characteristics of these two emitters, the combinatorial drugs provide a real-time dual-fluorescent imaging/tracking system in vitro and in vivo, and this has not been reported before in self-delivered DDS which simultaneously shows a high drug loading capacity (77.6%Cur). Overall, our carrier-free DDS is able to achieve chemotherapy (Cur), photodynamic therapy (H2TPyP), and real-time self-monitoring of the release and distribution of the nanomedicine (Cur and H2TPyP). More importantly, the as-prepared NPs show high cancer therapeutic efficiency both in vitro and in vivo. We expect that the present real-time self-monitored and self-delivered DDS with multiple-therapeutic and multiple-fluorescent ability will have broad applications in future cancer therapy.

Sepsis is a serious and life-threatening syndrome that often occurs in intensive care unit (ICU) patients. During sepsis, inflammatory cytokines and nitric oxide (NO) can be overproduced, causing tissue and cell injury. Propofol is an intravenous agent used for sedation of ICU patients. Our previous study showed that propofol has immunosuppressive effects on macrophage functions. This study was designed to evaluate the anti-inflammatory and antioxidative effects of propofol on the biosyntheses of tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta), IL-6, and NO in lipopolysaccharide (LPS)- activated macrophages. Exposure to a therapeutic concentration of propofol (50 microM), LPS (1 ng/mL), or a combination of these two drugs for 1, 6, and 24 h was not cytotoxic to the macrophages. ELISA revealed that LPS increased macrophage TNF-alpha, IL-1beta, and IL-6 protein levels in a time-dependent manner, whereas propofol significantly reduced the levels of LPS-enhanced TNF-alpha, IL-1beta, and IL-6 proteins. Data from RT-PCR showed that LPS induced TNF-alpha, IL-1beta, and IL-6 mRNA, but propofol inhibited these effects. LPS also increased NO production and inducible nitric oxide synthase (iNOS) expression in macrophages. Exposure of macrophages to propofol significantly inhibited the LPS-induced NO biosynthesis. The present study shows that propofol, at a therapeutic concentration, has anti-inflammatory and antioxidative effects on the biosyntheses of TNF-alpha, IL-1beta, IL-6, and NO in LPS-activated macro-phages and that the suppressive effects are exerted at the pretranslational level.

Hepatic assembly of triacylglycerol (TAG)-rich very low density lipoproteins (VLDL) is achieved through recruitment of bulk TAG (presumably in the form of lipid droplets within the microsomal lumen) into VLDL precursor containing apolipoprotein (apo)BHepatic assembly of very low density lipoproteins (VLDL) rich in triacylglycerol (TAG) 5 is initiated during and after translation and translocation of apolipoprotein (apo) B-100 across the endoplasmic reticulum membrane. The nascent VLDL particle is further enlarged in TAG content through a "second step" lipidation process, where a bulk TAG (presumably present within the microsomal lumen in the form of lipid droplet) is incorporated (1). Although much has been learned about the co-and post-translational lipidation of apoB-100, little is known about the nature or the dynamics of these lumenal lipid droplets (LLD) that are utilized as lipid precursor for VLDL assembly. Early studies with the abetalipoproteinemia gene MTTP suggested that the microsomal triglyceride transfer protein (MTP) is required for the partitioning of VLDL lipid precursor, mainly TAG, into the microsomal lumen in mouse liver (2), cultured murine primary hepatocytes (3), and the rat hepatoma McA-RH7777 cells (4). Thus, inactivation of MTP in the liver cells was associated with lack of LLD and invariably resulted in diminished assembly and secretion of VLDL. Recent proteomic and lipid characterization of LLD isolated from murine liver microsomes has shown that these lipid-rich entities are devoid of apoB but contain proteins such as TAG hydrolase, carboxylesterase 1, MTP, and apoE (5). Working with McA-RH7777 cells stably expressing recombinant human apoC-III, we have obtained evidence for the presence of a metabolically active TAG-rich entity within the microsomal lumen, which has a buoyant density resembling that of intermediate density lipoproteins (IDL) and low density lipoproteins (LDL)

This letter presents direct observation of growth hormone receptor in one single cancer cell using nanodiamond-growth hormone complex as a specific probe. The interaction of surface growth hormone receptor of A549 human lung epithelial cells with growth hormone was observed using nanodiamond’s unique spectroscopic signal via confocal Raman mapping. The growth hormone molecules were covalent conjugated to 100nm diameter carboxylated nanodiamonds, which can be recognized specifically by the growth hormone receptors of A549 cell. The Raman spectroscopic signal of diamond provides direct and in vitro observation of growth hormone receptors in physiology condition in a single cell level.

SummaryX-linked juvenile retinoschisis (XLRS), linked to mutations in the RS1 gene, is a degenerative retinopathy with a retinal splitting phenotype. We generated human induced pluripotent stem cells (hiPSCs) from patients to study XLRS in a 3D retinal organoid in vitro differentiation system. This model recapitulates key features of XLRS including retinal splitting, defective retinoschisin production, outer-segment defects, abnormal paxillin turnover, and impaired ER-Golgi transportation. RS1 mutation also affects the development of photoreceptor sensory cilia and results in altered expression of other retinopathy-associated genes. CRISPR/Cas9 correction of the disease-associated C625T mutation normalizes the splitting phenotype, outer-segment defects, paxillin dynamics, ciliary marker expression, and transcriptome profiles. Likewise, mutating RS1 in control hiPSCs produces the disease-associated phenotypes. Finally, we show that the C625T mutation can be repaired precisely and efficiently using a base-editing approach. Taken together, our data establish 3D organoids as a valid disease model.