Additional file 2: of Mathematical modelling of interacting mechanisms for hypoxia mediated cell cycle commitment for mesenchymal stromal cells

The effect of initial conditions on steady state E2F concentration. (DOCX 42 kb

Additional file 3: of Mathematical modelling of interacting mechanisms for hypoxia mediated cell cycle commitment for mesenchymal stromal cells

Compiled hypoxic MSCs growth data. (DOCX 23 kb

Additional file 1: of Mathematical modelling of interacting mechanisms for hypoxia mediated cell cycle commitment for mesenchymal stromal cells

Parameter values and initial conditions. (DOCX 23 kb

Screening study on significant Chinese herb for anti-idiopathic pulmonary fibrosis by combining clinical experience prescriptions and molecular dynamics simulation technologies

Various techniques such as data mining, network pharmacology, molecular docking and molecular dynamics simulation were used in this study to screen and validate effective herbal medicines for the treatment of idiopathic pulmonary fibrosis (IPF) and to reveal their mechanisms of action at the molecular level. The use of this approach will provide new tools and ideas for future drug screening, especially for the application of herbal medicines in the treatment of complex diseases. Among them, the five identified core targets, including IL6, TP53, AKT1, VEGFA, and TNF, as well as a series of major active compounds, will be important references for future anti-IPF drug development. This information will accelerate the discovery and development of relevant drugs. Meanwhile, this study further confirmed the potential value of four Chinese herbal medicines, including Gancao, Danshen, Huangqin, and Sanqi, in the treatment of IPF. This will promote more clinical trials and practices to confirm and optimise the application of these herbs. Finally, this study is an important theoretical guide to enhance the advantages of Chinese herbal medicines in the prevention and treatment of major and difficult diseases, as well as to understand and utilise the potential efficacy of Chinese herbal medicines. This will further promote the scientific research and clinical application of herbal medicines and provide more possibilities for future disease treatment Communicated by Ramaswamy H. Sarma</p

Morphological analysis of human umbilical vein endothelial cells co-cultured with ovarian cancer cells in 3D: An oncogenic angiogenesis assay

<div><p>Antiangiogenic therapy for cancer is a strategy targeted at tumour vasculature, often in combination with conventional cytotoxicity treatments. Animal testing is still the most common method used for evaluating the efficacy of new drugs but tissue-engineered <i>in vitro</i> models are becoming more acceptable for replacing and reducing the use of animals in anti-cancer drug screening. In this study, a 3D co-culture model of human endothelial cells and ovarian cancer cells was developed. This model has the potential to mimic the interactions between endothelial cells and ovarian cancer cells. The feasibility of applying this model in drug testing was explored here. The complex morphology of the co-culture system, which features development of both endothelial tubule-like structures and tumour structures, was analysed quantitatively by an image analysis method. The co-culture morphology integrity was maintained for 10 days and the potential of the model for anti-cancer drug testing was evaluated using Paclitaxel and Cisplatin, two common anti-tumour drugs with different mechanisms of action. Both traditional cell viability assays and quantitative morphological analyses were applied in the drug testing. Cisplatin proved a good example showing the advantages of morphological analysis of the co-culture model when compared with mono-culture of endothelial cells, which did not reveal an inhibitory effect of Cisplatin on the tubule-like endothelial structures. Thus, the tubule areas of the co-culture reflected the anti-angiogenesis potential of Cisplatin. In summary, <i>in vitro</i> cancer models can be developed using a tissue engineering approach to more closely mimic the characteristics of tumours <i>in vivo</i>. Combined with the image analysis technique, this developed 3D co-culture angiogenesis model will provide more reproducible and reliably quantified results and reveal further information of the drug’s effects on both tumour cell growth and tumour angiogenesis.</p></div

Image processing procedure used for bright field images.

<p>(A) The original bright field images of 3D mono-culture of OVCAR8 cells, on-top co-culture of OVCAR8 cells and HUVECs, and 3D co-culture of OVCAR8 cells and HUVECs at day 10. (B) Background corrected images of each original image. (C) Different contour lines detected in each background corrected image. (D) Segmentation of spheroids (in black colour) and tubules (in grey colour) in each image.</p

Apoptosis assay comparing co-culture of OVCAR8 with HUVECs and mono-culture of OVCAR8 at day 10.

<p>(A) Fluorescence images of co-culture. It is noticeable that nearly no apoptotic signal was observed with thinner tubule structures, while more apoptosis was detected with the structures with larger dimensions. (B) Fluorescent images of mono-culture. Cleaved caspase 3 (magenta); DAPI (blue), HUVECs (red), and OVCAR8 (green). Scale bar: 100μm. (C) Zen localisation coefficient comparison between co-culture and mono-culture. No significant difference was observed.</p

Morphology characterisation of different <i>in vitro</i> models used in the study.

<p>(A) Representative pictures of (A) HUVECs in mono-culture, (B) OVCAR8 in mono-culture, (C) 3D co-culture of both cell types in the Matrigel sandwich, and (D) on-top co-culture of both cell types on top of the Matrigel at day 1 (24 hours after seeding on day 0), day 3, day 5, day 7 and day 10. It is noticeable that the tubule structures of HUVECs mono-culture started to degrade at day 3.</p

Schematic procedure for 3D co-culture of HUVECs and OVCAR8 in Matrigel sandwich.

<p>(A) HUVECs seeded on polymerised Matrigel; (B) After 4 hours, HUVECs started to form tubules; (C) OVCAR8 cell suspension in medium containing 10% Matrigel was added; (D) 24 hours later, Matrigel sandwich structure formed and co-culture stabilised, ready for longer term culture or further drug testing.</p

Semi-automated image analysis comparing morphologies of different <i>in vitro</i> models.

<p>(A) Comparison of 3D on-top and 3D sandwich co-culture of OVCAR8 and HUVECs with regard to: (i) Total spheroid areas in μm² and (ii) Mean tubule areas. (B) Comparison of 3D mono-culture of OVCAR8 and 3D co-culture of HUVECs and OVCAR8 in terms of: (i) Total spheroid areas in μm² and (ii) Mean spheroid areas in μm². (C) Angiogenesis parameter comparisons for 3D on-top and 3D sandwich co-culture of OVCAR8 and HUVECs over 10 days for: (i) Total tubule areas and (ii) Branching points. Results are presented as (mean ± SD), and are considered significantly different when p < 0.05 based on Student’s t-test (marked as ‘*’ in the figures showing significant difference between the compared models on the same day). Note that since HUVECs mono-culture lost their tubule network at day 2 and eventually lost signs of living cells over longer term culture (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0180296#pone.0180296.g002" target="_blank">Fig 2(A))</a>, only mono-culture of cancer cell OVCAR8 and co-culture of OVCAR8 and HUVECs were compared here over 10 days of culture.</p