Identification and characterization of intermediate states in mammalian neural crest cell epithelial to mesenchymal transition and delamination.

Epithelial to mesenchymal transition (EMT) is a cellular process that converts epithelial cells to mesenchymal cells with migratory potential in both developmental and pathological processes. Although originally considered a binary event, EMT in cancer progression involves intermediate states between a fully epithelial and a fully mesenchymal phenotype, which are characterized by distinct combinations of epithelial and mesenchymal markers. This phenomenon has been termed epithelial to mesenchymal plasticity (EMP), however, the intermediate states remain poorly described and it's unclear whether they exist during developmental EMT. Neural crest cells (NCC) are an embryonic progenitor cell population that gives rise to numerous cell types and tissues in vertebrates, and their formation is a classic example of developmental EMT. An important feature of NCC development is their delamination from the neuroepithelium via EMT, following which NCC migrate throughout the embryo and undergo differentiation. NCC delamination shares similar changes in cellular state and structure with cancer cell invasion. However, whether intermediate states also exist during NCC EMT and delamination remains unknown. Through single cell RNA sequencing, we identified intermediate NCC states based on their transcriptional signature and then spatially defined their locations in situ in the dorsolateral neuroepithelium. Our results illustrate the progressive transcriptional and spatial transitions from premigratory to migratory cranial NCC during EMT and delamination. Of note gene expression and trajectory analysis indicate that distinct intermediate populations of NCC delaminate in either S phase or G2/M phase of the cell cycle, and the importance of cell cycle regulation in facilitating mammalian cranial NCC delamination was confirmed through cell cycle inhibition studies. Additionally, transcriptional knockdown revealed a functional role for the intermediate stage marker Dlc1 in regulating NCC delamination and migration. Overall, our work identifying and characterizing the intermediate cellular states, processes, and molecular signals that regulate mammalian NCC EMT and delamination furthers our understanding of developmental EMP and may provide new insights into mechanisms regulating pathological EMP.

Natural deep eutectic solvent supported targeted solid-liquid polymer carrier for breast cancer therapy.

Solid-liquid nanocarriers (SLNs) are at the front of the rapidly emerging field of medicinal applications with a potential role in the delivery of bioactive agents. Here, we report a new SLN of natural deep eutectic solvent (NADES) and biotin-conjugated lysine-polyethylene glycol copolymer. The SLN system was analyzed for its functional groups, thermal stability, crystalline nature, particle size, and surface morphology through the instrumental analysis of FT-IR, TGA, XRD, DLS, SEM, and TEM. Encapsulation of PTX (paclitaxel) and 7-HC (7-hydroxycoumarin) with the SLN was carried out by dialysis, and UV-visible spectra evidenced the drug loading capacity and higher encapsulation efficiency obtained. The enhanced anticancer potential of PTX- and 7-HC-loaded SLN was assessed in vitro, and the system reduces the cell viability of MDA-MB-231 cells. The PTX- and 7-HC-loaded SLN system was investigated in a breast cancer-induced rat model via in vivo studies. It shows decreased lysosomal enzymes and increased levels of caspase to cure breast tumors. It very well may be reasoned that the designed PTX- and 7-HC-loaded SLN system has strong anticancer properties and exhibits potential for delivery of drug molecules in cancer treatment.

Development of biotin molecule targeted cancer cell drug delivery of doxorubicin loaded κ-carrageenan grafted graphene oxide nanocarrier.

Cervical cancer is one of the most occurring cancers and the fourth leading occurrence of cancer in women, worldwide. In this study, we planned to synthesis κ-Carrageenan grafted graphene oxide nanocarrier conjugated with biotin (GO-κ-Car-biotin) for targeted cervical cancer. Doxorubicin (DOX) is a well-known anticancer drug for any type of cancer and it is used to entrap over on the graphene oxide surface via π-π stacking interaction. The chemical function and crystalline nature of the synthesized nanocarrier was characterized by Fourier Transformed Infrared Spectroscopy (FT-IR) and X-ray diffraction Analysis (XRD). The surface morphological study was carried out through Scanning Electron Microscopy (SEM), Transmission electron microscopy (TEM) and Atomic force microscopy (AFM). The in-vitro drug release profile of DOX was carried out by UV-Vis spectrometer at the λmax value of 480 nm. The entrapment of DOX on GO-κ-car-biotin has been observed at 94%. The hydrophilic DOX drug has excellent pH-sensitive drug released in an in-vitro study. The anticancer efficiency of the synthesized GO-based nanocarrier was examined using HeLa cell line in-vitro. Cell viability, proliferation, cytotoxicity, and nuclear chromatin condensation was studied by trypan blue assay, triphosphate assay (ATP), lactate dehydrogenase assay (LDH) and Hoechst staining respectively. Finally, biotin leading GO-κ-Car carrier demonstrated is a promising drug delivery system for cervical cancer treatment.

Folate receptor targeted delivery of paclitaxel to breast cancer cells via folic acid conjugated graphene oxide grafted methyl acrylate nanocarrier.

The adaptability, joint with a large surface area, electronic flexibility, high intrinsic mobility, high mechanical strength and supreme thermal conductivity have condensed graphene family materials attractive as technological tools of the drug delivery system. In this present study, investigate a modified graphene oxide-methyl acrylate (GO-g-MA) nanocarrier for targeted anti-cancer drug delivery in breast cancer cells and the GO-g-MA fascinated with folic acidas a targeting ligand to target the cancer cells. Paclitaxel (PTX) was assembled through π-π stacking, hydrophophic interaction on the surface of the GO-g-MA/FA carrier. Structural modification of GO-g-MA, functionalization of targeting ligands GO-g-MA/FA and drug loaded GO-g-MA/FA-PTX was characterized and confirmed through FTIR, XRD, SEM,TEM and AFM analysis. The in-vitro drug release pattern of PTX from the GO-g-MA/FA was examined in different pH ranges. An MTT assay was performed to evaluate the cytotoxicity behaviour of the carrier and PTX loaded nanocarrier in the human breast cancer cell line (MDA-MB-231). GO-g-MA/FA-PTX carrier showed that 39% of cytotoxic effect. Furthermore, the in-vivo (DMBA induced breast cancer rats) studies were carried out and treatment with PTX- loaded GO-g-MA/FA nanocarrier attenuates the levels of mitochondrial citric acids enzymes to near normal.

Deep Eutectic Solvent-Mediated FA-g-ß-Alanine-co-PCL Drug Carrier for Sustainable and Site-Specific Drug Delivery.

An amphiphilic polymer that consisted of a deep eutectic solvent (DES)-mediated drug carrier was designed, where the DES influenced the formation of folic acid (FA)-tagged g-ß-alanine-co-PCL polymer (DES@FA-g-ß-alanine-co-PCL); the nature of the carrier was investigated through emission analysis and pyrene used as a model probe (CMC = 0.4 mg/mL). The amphiphilic polymer was self-assembled into a sphere (≈204 nm diameter) with a surface charge of -3 ± 0.5 mV. The doxorubicin was incorporated and the structural changes were analyzed by UV-visible spectroscopy, FT-IR, XRD, Raman, and TGA analysis, while size and morphological analysis was performed by DLS, AFM, SEM, and TEM. The controlled release of drug from the carrier was observed at different pH levels. The enhanced anticancer potential of DOX-loaded polymeric micelle was studied both in vitro and in vivo breast cancer model. The treatment of DOX-loaded polymeric micelle reduces the viability and proliferation of MDA-MB-231 cells. From the results of the current investigation it concludes that the DOX-loaded polymeric micelle has enhance anticancer effect and it exhibits its potential effect at the dosage of 5 mg/kg body weight in mammary carcinoma-bearing rats. From the observed results, synthesized DOX-loaded polymeric micelle holds strong anticancer properties compared with free DOX and can be used as a potential carrier in the pharmaceutical industry.