A chronic inability to maintain blood glucose homeostasis leads to diabetes, which can damage multiple organs. The pancreatic islets regulate blood glucose levels through the coordinated action of islet cell-secreted hormones, with the insulin released by β-cells playing a crucial role in this process. Diabetes is caused by insufficient insulin secretion due to β-cell loss, or a pancreatic dysfunction. The restoration of a functional β-cell mass might, therefore, offer a cure. To this end, major efforts are underway to generate human β-cells de novo, in vitro, or in vivo. The efficient generation of functional β-cells requires a comprehensive knowledge of pancreas development, including the mechanisms driving cell fate decisions or endocrine cell maturation. Rapid progress in single-cell RNA sequencing (scRNA-Seq) technologies has brought a new dimension to pancreas development research. These methods can capture the transcriptomes of thousands of individual cells, including rare cell types, subtypes, and transient states. With such massive datasets, it is possible to infer the developmental trajectories of cell transitions and gene regulatory pathways. Here, we summarize recent advances in our understanding of endocrine pancreas development and function from scRNA-Seq studies on developing and adult pancreas and human endocrine differentiation models. We also discuss recent scRNA-Seq findings for the pathological pancreas in diabetes, and their implications for better treatment.

Articular cartilage and meniscus injuries are prevalent disorders with insufficient regeneration responses offered by available treatment methods. In this regard, 3D bioprinting has emerged as one of the most promising new technologies, offering novel treatment options. Additionally, the latest achievements from the fields of biomaterials and tissue engineering research identified constituents facilitating the creation of biocompatible scaffolds. In this study, we looked closer at hyaluronic acid and multi-walled carbon nanotubes as bioink additives. Firstly, we assessed the minimal concentrations that stimulate cell viability, and decrease reactive oxygen species and apoptosis levels in 2D cell cultures of normal human knee articular chondrocytes (NHAC) and human adipose-derived mesenchymal stem cells (hMSC-AT). In this regard, 0.25 mg/ml of hyaluronic acid and 0.0625 mg/ml of carbon nanotubes were selected as the most optimal concentrations. In addition, we investigated the protective influence of 2-phospho-L-ascorbic acid in samples with carbon nanotubes. Tests conducted on 3D bioprinted constructs revealed that only a combination of components positively impacted cell viability throughout the whole experiment. Gene expression analysis of COL1A1, COL6A1, HIF1A, COMP, RUNX2, and POU5F1 showed significant changes in the expression of all analyzed genes with a progressive overall loss of transcriptional activity in most of them.

Carbon nanotubes (CNTs) are one of the most promising nanomaterials synthesized to date. Thanks to their unique mechanical, electronic, and optical properties, they have found a wide application in electronics in the production of biosensors and nanocomposites. The functionalization of multiwalled carbon nanotubes (MWCNTs) is aimed at making them biocompatible by adding hydrophilic groups on their surface, increasing their solubility and thus rendering them applicable in the regenerative medicine. So far, there is conflicting information about carbon nanotubes in biological systems. This paper investigates the effect of functionalized, oxidized, multiwalled carbon nanotubes (MWCNT-Ox) on the cytotoxicity of normal human articular chondrocytes (NHAC-kn cell line). Since absorbance-based and fluorescence-based assays were shown to interfere with carbon nanotubes, luminescence-based tests were carried out, as they work on a different method of detection and provide advantages over the mentioned ones. Cell viability and reactive oxygen species (ROS) tests were carried out. The cell viability assay showed that with the increasing MWCNTs concentration, the number of viable chondrocytes was significantly decreasing. Exposure to MWCNT-Ox indicated oxidative stress in the lowest investigated concentration with a decreased amount of ROS with higher concentrations. However, control experiments with adenosine triphosphate (ATP) and H2O2—molecules that are detected by the assays—showed that carbon nanotubes interfere directly with measurement, thus rendering the results unreliable. To understand the exact interference mechanisms, further studies must be taken. In conclusion, this study shows that luminescence-based tests yield erroneous results, confirming that in vitro experiments in the literature concerning carbon nanotubes should be analyzed with caution.