S1. Materials and Methods S1.1. Materials Carboxymethyl cellulose sodium salt (M w ~ 90 kDa), tyramine hydrochloride (TYR), N-(3dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), Tween 20, Triton-X100, deuterium oxide (D 2 O), dialysis tubing (MWCO 124000), and bovine serum albumin (BSA) were obtained from Sigma-Aldrich Pty. Ltd. (Castle Hill, NSW, Australia). Gelatin (M w = 80-140 kDa, pI = 5) and horseradish peroxidase (HRP; 100 units/mg) were obtained from Wako Pure Chemical Industries (Novachem Pty. Ltd., Collingwood, VIC, Australia). Fetal bovine serum (FBS), LIVE/DEAD  Cell Viability Assays, calcein AM, 4',6-diamidino-2-phenylindole (DAPI), Dulbecco's Modified Eagle's Medium (DMEM), penicillin-streptomycin, trypsin-EDTA, and phosphate buffered saline (PBS) were acquired from Invitrogen (Life Technologies Pty. Ltd., Mulgrave, VIC, Australia). Hydrogen peroxide (H 2 O 2) was obtained from Merck Pty. Ltd.

Background: Breast cancer is most serious reasons of women death around worldwide result in increasing its morbidity and mortality. MicroRNAs are considered as significant regulators of cancer biological processes. The main aim of this study is restoration of miR-126 could lead to modulate breast cell line and impairs their proliferation by targeting vascular endothelial growth factor gene (VEGF-A). Methods: Breast cancer cell line (MCF7) was transfected by miR-126 lipofectamine and negative miR control for 24 hr. Cytotoxic effects of miR-126 lipofectamine were determined by cell viability assay. Cell proliferation and cell cycle were quantitatively measured using PicoGreen assay and DAPI stain-flow cytometer analysis. For further investigation, Taq-Man real time PCR assay was performed to detect relative VEGF-A and miRNA-126 level. Results: MiR-126 was overexpressed in treated breast cancer cell (MCF7) compared with control cells. miR-126 expression has been associated-with a decrease in cell proliferation and arrested MCF7 cells at G1 phase. The study found that vascular endothelial growth factor is regulated by miR-126. Hence, VEGF-A is considered as functional vital and direct target to miR-126 in breast cancer cell line (MCF7). Conclusions: This study provided that manipulated miR-126 level may suggest a novel therapeutic approach in breast cancer treatment. However, an animal models study is needed to address and prove predictive ability of miR-126 on breast cancer controlling.

Despite the promise of stem cell therapy for lung therapeutics and repair, there are few viable means for directly delivering stem cells to locally target the respiratory airways via inhalation. This is not surprising given the significant challenges in aerosolising stem cells, particularly given their susceptibility to damage under the large stresses involved in the nebulisation process. Here, we present promising results using a microfluidic acoustic nebulisation platform that is not only low cost and portable, but also its high MHz order frequencies are effective for preserving the structural and functional integrity of mesenchymal stem cells (MSCs) during the nebulisation process. This is verified through an assessment of the viability, structure, metabolic activity, proliferation ability and genetic makeup of the nebulised MSCs using a variety of assays, including cell viability staining, flow cytometry, reverse transcription and quantitative polymerase chain reaction, and immunophenotyping, thus demonstrating the platform as a promising method for efficient pulmonary stem cell delivery.

The ability to spatially organise the microenvironment of tissue scaffolds unlocks the potential of many scaffold-based tissue engineering applications. An example application is to aid the regeneration process of peripheral nerve injuries. Herein, we present a promising approach for three-dimensional (3D) micropatterning of nerve cells in tissue scaffolds for peripheral nerve repair. In particular, we demonstrate the 3D micropatterning of PC12 cells in a gelatin-hydroxyphenylpropionic acid (Gtn-HPA) hydrogel using ultrasound standing waves (USWs). PC12 cells were first aligned in 3D along nodal planes by the USWs in Gtn-HPA hydrogel precursor solution. The precursor was then crosslinked using horseradish peroxidase (HRP) and diluted hydrogen peroxide (H2O2), thus immobilising the aligned cells within 90–120 s. This micropatterning process is cost effective and can be replicated easily without the need for complex and expensive specialised equipment. USW-aligned PC12 cells showed no adverse effect in terms of viability or ability to proliferate. To our best knowledge, this is the first report on the effect of USW alignment on neural cell differentiation. Differentiated and USW-aligned PC12 cells showed directional uniformity after 20 d, making this technique a promising alternative approach to guide the nerve regeneration process.