Microplastics and Nanoplastics Impair the Biophysical Function of Pulmonary Surfactant by Forming Heteroaggregates at the Alveolar-Capillary Interface.

Microplastics (MPs) are ubiquitous environmental pollutants produced through the degradation of plastic products. Nanoplastics (NPs), commonly coexisting with MPs in the environment, are submicrometer debris incidentally produced from fragmentation of MPs. We studied the biophysical impacts of MPs/NPs derived from commonly used commercial plastic products on a natural pulmonary surfactant extracted from calf lung lavage. It was found that in comparison to MPs/NPs derived from lunch boxes made of polypropylene or from drinking water bottles made of poly(ethylene terephthalate), the MP/NP derived from foam packaging boxes made of polystyrene showed the highest adverse impact on the biophysical function of the pulmonary surfactant. Accordingly, intranasal exposure of MP/NP derived from the foam boxes also induced the most serious proinflammatory responses and lung injury in mice. Atomic force microscopy revealed that NP particles were adsorbed on the air-water surface and heteroaggregated with the pulmonary surfactant film. These results indicate that although the incidentally formed NPs only make up a small mass fraction, they likely play a predominant role in determining the nano-bio interactions and the lung toxicity of MPs/NPs by forming heteroaggregates at the alveolar-capillary interface. These findings may provide novel insights into understanding the health impact of MPs and NPs on the respiratory system.

Polystyrene nanobeads exacerbate chronic colitis in mice involving in oxidative stress and hepatic lipid metabolism.

BACKGROUND: Nanoplastics (NPs) are omnipresent in our lives as a new type of pollution with a tiny size. It can enter organisms from the environment, accumulate in the body, and be passed down the food chain. Inflammatory bowel disease (IBD) is a nonspecific intestinal inflammatory disease that is recurrent and prevalent in the population. Given that the intestinal features of colitis may affect the behavior and toxicity of NPs, it is imperative to clarify the risk and toxicity mechanisms of NPs in colitis models. METHODS AND RESULTS: In this study, mice were subjected to three cycles of 5-day dextran sulfate sodium (DSS) exposures, with a break of 7 to 11 days between each cycle. After the first cycle of DSS exposure, the mice were fed gavagely with water containing 100 nm polystyrene nanobeads (PS-NPs, at concentrations of 1 mg/kg·BW, 5 mg/kg·BW and 25 mg/kg·BW, respectively) for 28 consecutive days. The results demonstrated that cyclic administration of DSS induced chronic inflammation in mice, while the standard drug "5-aminosalicylic acid (5-ASA)" treatment partially improved colitis manifestations. PS-NPs exacerbated intestinal inflammation in mice with chronic colitis by activating the MAPK signaling pathway. Furthermore, PS-NPs aggravated inflammation, oxidative stress, as well as hepatic lipid metabolism disturbance in the liver of mice with chronic colitis. CONCLUSION: PS-NPs exacerbate intestinal inflammation and injury in mice with chronic colitis. This finding highlights chronically ill populations' susceptibility to environmental hazards, which urgent more research and risk assessment studies.

First Report of Fusarium denticulatum Causing Chlorotic Leaf Distortion of Sweetpotato in China.

Sweetpotato (Ipomoea batatas) is one of the most important crops in China. To gain a clearer picture of the occurrence of diseases on sweetpotato, 50 fields (100 plants/field) were randomly surveyed in prominent sweetpotato growing areas of Lulong county, Hebei Province in the years 2021-2022. Plants showing chlorotic leaf distortion with mildly twisted young leaves and stunted vines were observed frequently. It was similar to the symptoms of chlorotic leaf distortion of sweetpotato (Clark et al. 2013). The incidence of disease with patch pattern ranged from 15% to 30%. Ten symptomatic leaves were excised, surface disinfested with 2% sodium hypochlorite for 1 min, rinsed three times in sterilized ddH2O, and cultured on potato dextrose agar (PDA) at 25°C. Nine fungal isolates were obtained. A pure culture of representative isolate FD10 obtained after serial hyphal tip transfer was examined for morphological and genetic characters. Colonies of isolate FD10 on PDA at 25°C were slow growing (4±0.1mm/day) with aerial, white-to-pink mycelium. Lobed colonies had greyish-orange pigmentation in reverse and conidia aggegated in false heads. Conidiophores were prostrate and short. Phialides were mostly monophialidic but occasionally polyphialidic. Polyphialidic openings often denticulate in a rectangular arrangement. Microconidia were abundant, long, oval to allantoid, mostly none or one septate, and 4.79 to 9.53 × 2.08 to 3.22 µm (n = 20). Macroconidia were fusiform to falcate with a beaked apical cell and a footlike basal cell, 3 to 5 septate, and 25.03 to 52.92 × 2.56 to 4.49 µm. Chlamydospores were absent. All in agreement with the morphology of Fusarium denticulatum (Nirenberg and O'Donnell 1998). Genomic DNA of isolate FD10 was extracted. The EF-1α and ß-tubulin genes were amplified and sequenced (O'Donnell and Cigelnik 1997; O'Donnell et al. 1998). The obtained sequences were deposited in GenBank (accession nos. OQ555191 and OQ555192). BLASTn revealed that those sequences showed 99.86% (EF-1α) and 99.93% (ß-tubulin) homology with the relative sequences of F. denticulatum type strain CBS407.97 (accession nos. MT011002.1 and MT011060.1), respectively. Furthermore, a neighbor-joining phylogenetic tree based on the EF-1α and ß-tubulin sequences revealed that the isolate FD10 clustered with F. denticulatum. Based on morphological characteristics and sequence analysis, the isolate FD10 associated with chlorotic leaf distortion of sweetpotato was identified as F. denticulatum. Pathogenicity tests were performed by immersing ten 25-cm-long vine-tip cuttings of cultivar Jifen 1 origin from tissue culture in a conidial suspension (1 × 106 conidia per ml) of isolate FD10. Vines immersed in sterile distilled water used as a control. All inoculated plants in 25-cm plastic pots were incubated in a climate chamber at 28℃ and 80% RH for two and half months and the control were incubated in a separate climate chamber. Nine inoculated plants developed chlorotic terminals, moderate interveinal chlorosis and slight leaf distortion. No symptoms were observed on the control plants. The pathogen was reisolated from inoculated leaves and matched the morphological and molecular characteristics of the original isolates, thus fulfilling Koch's postulates. To our knowledge, this is the first report of F. denticulatum causing chlorotic leaf distortion of sweetpotato in China. Its identification will promote the management of this disease in China.

The ex vivo and in vivo biological performances of graphene oxide and the impact of surfactant on graphene oxide's biocompatibility.

Graphene oxide (GO) displays promising properties for biomedical applications including drug delivery and cancer therapeutics. However, GO exposure also raises safety concerns such as potential side effects on health. Here, the biological effects of GO suspended in phosphate buffered saline (PBS) with or without 1% nonionic surfactant Tween 80 were investigated. Based on the ex vivo experiments, Tween 80 significantly affected the interaction between GO and peripheral blood from mice. GO suspension in PBS tended to provoke the aggregation of diluted blood cells, which could be prevented by the addition of Tween 80. After intravenous administration, GO suspension with or without 1% Tween 80 was quickly eliminated by the mononuclear phagocyte system. Nevertheless, GO suspension without Tween 80 showed greater accumulation in lungs than that containing 1% Tween 80. In contrast, less GO was found in livers for GO suspension compared to Tween 80 assisted GO suspension. Organs including hearts, livers, lungs, spleens, kidneys, brains, and testes did not reveal histological alterations. The indexes of peripheral blood showed no change upon GO exposure. Our results together demonstrated that Tween 80 could greatly alter GO's biological performance and determine the pattern of its biodistribution in mice.

Enhanced hepatic cytotoxicity of chemically transformed polystyrene microplastics by simulated gastric fluid.

Microplastics pollution has emerged as one of the top-ranked global environmental issues, receiving worldwide attention in recent years. However, knowledge about the detrimental effects of microplastics on human health is still limited. In real-world settings, the physicochemical characteristics of microplastics were modified by environmental and biological transformation, largely changing their ultimate toxicity. Nonetheless, the toxicity change related to transformation of microplastics has not been considered in most published studies thus far. In the current study, we investigated the cytotoxicity of transformed polystyrene microplastics in hepatocytes. Our results revealed that 500 nm polystyrene microplastics, which were chemically transformed by simulated gastricfluid, exacerbated their adverse effects on SMMC-7721 cells at 20 µg/mL for 24 h treatment, including morphological alteration, membrane damage and increased cell apoptosis via oxidative stress. This exacerbated cytotoxicity could be at least partially explained by the degradation, changed surface charge and altered surface chemistry of these polystyrene microplastics after transformation. In conclusion, our study demonstrates that the hepatic cytotoxicity of polystyrene microplastics is enhanced after transformation.

Bone morphogenetic protein 7 promotes odontogenic differentiation of dental pulp stem cells in vitro.

AIMS: in vitro effects of bone morphogenetic protein 7 (BMP-7) on proliferation and differentiation of dental pulp stem cells (DPSCs) have not been investigated, nor has an appropriate dose been established. MAIN METHODS: Human DPSCs obtained from healthy volunteers were cultured with BMP-7 at 25, 50, and 100 ng/ml. Cell viability was measured by Cell Counting Kit-8 assay. Expression profiles of selected odontogenic differentiation-related markers in DPSCs were evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunocytochemistry, and western blot analysis. Mineralization of DPSCs was evaluated by alizarin red staining. The Smad5 signaling pathway was examined by qRT-PCR and western blot analysis. KEY FINDINGS: Diminished cell viability was found in DPSCs induced with 25, 50, and 100 ng/ml of BMP-7 for 7 days, showing a dose-response effect (P-trend = 0.03). DSPP, OCN, DMP-1, and RUNX2 were upregulated by BMP-7 induction after 7 and 14 days, especially at 50 and 100 ng/ml (P < 0.05). Immunocytochemical staining revealed strong expression of DSPP, DMP-1 and ALP in DPSCs induced by BMP-7, whereas null or weak expression in untreated cells. Western blot analysis confirmed over-expression of DSPP in cells induced by BMP-7. Alizarin red staining confirmed formation of mineralized nodules 4 weeks after BMP-7 induction. BMP-7 treated cells showed dose-dependently increased expression of BMPR1A, Smad5, and p-Smad5. SIGNIFICANCE: Our data indicated that BMP-7 at 50 ng/ml and 100 ng/ml was capable to induce DPSCs toward odontogenic differentiation through the Smad5 signaling pathway and not dramatically halt cell proliferation in vitro.

Imperatorin suppresses proliferation and angiogenesis of human colon cancer cell by targeting HIF-1α via the mTOR/p70S6K/4E-BP1 and MAPK pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Angelica dahurica is a commonly used traditional Chinese medicine to treat migraine headache, toothache and cancer. Imperatorin is an active natural furocoumarin component originating from Angelica dahurica and has been shown to exhibit multiple bioeffector functions, including anti-cancer activity. However, the mechanism by which imperatorin inhibits tumor growth is not fully understood. AIM OF THE STUDY: The aim of this study was to investigate the effectiveness of imperatorin as a treatment of cancer and to identify the underlying mechanisms of its anticancer activity. MATERIALS AND METHODS: HCT116, HeLa, and Hep3B cells were used in this study. Major assays were promoter-reporter gene assay, MTT, western blot analysis, immunofluorescence assay, reverse transcription-PCR (RT-PCR), flow cytometric analysis, clonogenic assay, EdU labeling and immunofluorescence, xenografted assay, and VEGF ELISA. RESULTS: We here demonstrated the effect of imperatorin on hypoxia-inducible factor-1 (HIF-1) activation. Imperatorin showed a potent inhibitory activity against HIF-1 activation induced by hypoxia in various human cancer cell lines. This compound markedly decreased the hypoxia-induced accumulation of HIF-1α protein dose-dependently, whereas it did not affect the expressions of HIF-1ß and topoisomerase-I (Topo-I). Further analysis revealed that imperatorin inhibited HIF-1α protein synthesis, without affecting the expression level of HIF-1α mRNA or degradation of HIF-1α protein. Moreover, the phosphorylation levels of mammalian target of rapamycin (mTOR), ribosomal protein S6 kinase (p70S6K), eIF4E binding protein-1 (4E-BP1), eukaryotic initiation factor 4E (eIF4E), extracellular signal-regulated kinase-1/2 (ERK1/2), SAPK/JNK and p38 were significantly suppressed by imperatorin. Furthermore, imperatorin prevented hypoxia-induced expression of HIF-1 target genes and flow cytometric analysis indicated that imperatorin induced G1 phase arrest in human colon cancer cell (HCT116). We found that imperatorin administration inhibits tumor growth and blocks tumor angiogenesis in a xenograft tumor model. CONCLUSIONS: These results show that imperatorin inhibited HIF-1α protein synthesis by downregulating the mTOR/p70S6K/4E-BP1 and MAPK pathways. These conclusions suggest that imperatorin is an effective inhibitor of HIF-1 and provide new perspectives into the mechanism of its anticancer activity.

In vitro cell proliferation evaluation of porous nano-zirconia scaffolds with different porosity for bone tissue engineering.

The selection of scaffold materials and the optimization of scaffold morphological and mechanical properties are critical for successful bone tissue engineering. We fabricated porous scaffolds of nano-sized zirconia using a replication technique. The study aimed to explore the relationship between porosity, pore size, mechanical strength, cell adhesion, and cell proliferation in the zirconia scaffolds. Macro- and micro-structures and compressive strength were comparatively tested. Beagle bone marrow stromal cells were seeded onto the scaffolds to evaluate cell seeding efficiency and cell proliferation profile over 14 d of incubation. The zirconia scaffolds presented a complex porous structure with good interconnectivity of pores. By increasing the sinter cycles, the porosity and pore size of the scaffolds decreased, with mean values ranging from 92.7-68.0% and 830-577 µm, respectively, accompanied by increased compressive strengths of 0.6-4.4 MPa. Cell seeding efficiency and cell proliferation over the first 7 d of incubation increased when the porosity decreased, with cell viability highest in the scaffold with a porosity of 75.2%. After 7 d of incubation, the cell proliferation increased when the porosity increased, highest in the scaffolds with a porosity of 92.7%. These results showed that the zirconia scaffold with a porosity of 75.2% possesses favorable mechanical and biological properties for future applications in bone tissue engineering.

DNA nanosensor based on biocompatible graphene quantum dots and carbon nanotubes.

An ultrasensitive nanosensor based on fluorescence resonance energy transfer (FRET) between biocompatible graphene quantum dots and carbon nanotubes for DNA detection was reported. We take advantage of good biocompatibility and strong fluorescence of graphene quantum dots, base pairing specificity of DNA and unique fluorescence resonance energy transfer between graphene quantum dots and carbon nanotubes to achieve the analysis of low concentrations of DNA. Graphene quantum dots with high quantum yield up to 0.20 were prepared and served as the fluorophore of DNA probe. FRET process between graphene quantum dots-labeled probe and oxidized carbon nanotubes is easily achieved due to their efficient self-assembly through specific π-π interaction. This nanosensor can distinguish complementary and mismatched nucleic acid sequences with high sensitivity and good reproducibility. The detection method based on this nanosensor possesses a broad linear span of up to 133.0 nM and ultralow detection limit of 0.4 nM. The constructed nanosensor is expected to be highly biocompatible because of all its components with excellent biocompatibility.