Nanomechanical Atomic Force Microscopy to Probe Cellular Microplastics Uptake and Distribution

The concerns regarding microplastics and nanoplastics pollution stimulate studies on the uptake and biodistribution of these emerging pollutants in vitro. Atomic force microscopy in nanomechanical PeakForce Tapping mode was used here to visualise the uptake and distribution of polystyrene spherical microplastics in human skin fibroblast. Particles down to 500 nm were imaged in whole fixed cells, the nanomechanical characterization allowed for differentiation between internalized and surface attached plastics. This study opens new avenues in microplastics toxicity research.

Co-Transplantation of Marginal Mass Allogeneic Islets with 3D-Culture-Derived Adult Human Skin Cells Improves Glycemia in Diabetic Mice

Background. Pancreatic islets transplantation represents a promising therapeutic option for severe type 1 diabetes (T1D). Maintenance of long-term viability of transplanted islets still requires improvement. Stem cell use represents an option to repair and replace damaged islets or, alternatively, b cells in T1D. Mesenchymal stem cells (MSC) have been proposed as adjuvants for islet transplantation, facilitating grafting and improving their functionality. Aggregation of stem cells has gained interest in providing physiological interactions between cells and enhancing the in situ concentration of modulators of inflammation and immunity.Methods. We established a hanging-drop culture by the spontaneous aggregation of adult human skin fibroblast-like cells as spheroids. Adult skin spheroid-derived cells (SphCs) were characterized in vitro and in vivo. We assessed the potential benefit of SphCs as adjuvants to improve islet functionality by cotransplantation with a marginal mass of allogeneic islets in an experimental diabetic mouse model. We characterized the secretome of SphCs by mass spectrometry-based proteomics.Results. SphCs were characterized as multipotent progenitors by their surface expression of markers analyzed by flow cytometry and multilineage germ differentiation capacity. Coculture of SphCs with anti-CD3-stimulated mouse splenocytes diminished the proliferation of T-CD4+ lymphocytes and biased splenocyte cytokine secretion through an increase in the Th2/Th1 ratio. SphCs conditioned media attenuated apoptosis of islets induced by inflammatory cytokine challenge in vitro. Administration (i.t.) of SphCs showed the absence of tumorigenicity in immune-deficient mice.SphCs improved glycemic control when cotransplanted with a marginal mass of allogeneic islets in an experimental diabetic mouse model without pharmacological immunosuppression. SphCs’ protein secretome differed from its paired skin fibroblast-like counterpart in containing 70% of up- and downregulated proteins and biological processes that overall positively influence islets such as cytoprotection, cellular stress, metabolism, and survival.Conclusions. Hanging-drop cell culture methodology might contribute to the development of an efficient way to improve transplantation outcome by reducing undesirable consequences of pharmacological immunosuppression as well as the number of allogeneic islets required to achieve normoglycemia in T1D transplanted patients. Further studies might determine whether the identified proteins sustain immunomodulation and/or cytoprotective effects in transplanted allogeneic islets.

Synthesis and Evaluation of Novel 1,2,6-Thiadiazinone Kinase Inhibitors as Potent Inhibitors of Solid Tumors

A focused series of substituted 4H-1,2,6-thiadiazin-4-ones was designed and synthesized to probe the anti-cancer properties of this scaffold. Insights from previous kinase inhibitor programs were used to carefully select several different substitution patterns. Compounds were tested on bladder, prostate, pancreatic, breast, chordoma, and lung cancer cell lines with an additional skin fibroblast cell line as a toxicity control. This resulted in the identification of several low single digit micro molar compounds with promising therapeutic windows, particularly for bladder and prostate cancer. A number of key structural features of the 4H-1,2,6-thiadiazin-4-one scaffold are discussed that show promising scope for future improvement.

Niosome Encapsulated Bromelain Reduced IL-6 and TNF-α in LPS Induced Human Skin Fibroblast Cell Line

The topical delivery of bromelain as an anti-inflammatory solution for skin inflammation has attracted the attention of researchers. Due to the skin barrier issue, a new method was designed for the effective delivery of specific doses of bromelain to the desired action sites. A niosome was selected as a novel and practical transdermal vehicle for the delivery of bromelain to inflamed sites. In this regard, a lipopolysaccharide (LPS)-induced human skin fibroblast (HSF1184) cell line was assembled in-vitro as a simulated model. The levels of interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF-α), the two immune-modulatory regulators of cell responses to inflammation, were measured to determine the response towards the niosome-encapsulated bromelain treatment. The results showed that the niosome-encapsulated bromelain significantly reduced the levels of IL-6 and TNF-α compared to the non-encapsulated bromelain, the vehicle (niosome) and the control.

Anti-inflammatory and antiaging properties of chlorogenic acid on UV-induced fibroblast cell

Background Skin aging is the most common dermatological problem caused by intrinsic and extrinsic factor, such as exposure to (ultraviolet) UV rays. Chlorogenic acid (CA) is a phenolic compound which is known for its antioxidant properties against oxidative stress. Objective This study investigates the antiaging and anti-inflammatory properties of CA on UV-induced skin fibroblast cells. Methods Anti-inflammatory properties of CA were assessed by measuring inflammatory-related proteins IL-1β and TNF-α, while antiaging properties of CA were assessed by measuring reactive oxygen species (ROS), apoptosis, live and necrotic cells, and COL-3 gene expression level. Results Treating UV-induced skin fibroblast cells with CA decreased the level of ROS, IL-1β, TNF-α, apoptotic cells, and necrotic cells and increased live cells and COL-3 gene expression. Conclusion CA has the potential as the protective compound against inflammation and aging by decreasing the level ROS, pro-inflammatory cytokines IL-1β and TNF-α, apoptotic cells, and necrotic cells and by increasing live cells and COL-3 gene expression.

Effect of Cerebrospinal Fluid on Fibroblasts Concerning Epidural Fibrosis: An In Vitro Study

One of the most common treatments for lumbar disc herniation and other lumbar disorders is lumbar laminectomy. There may be some unwanted and serious complications with this procedure such as the “failed back surgery syndrome (FBSS)”. Epidural fibrosis (EF), mainly due to fibroblast proliferation, emerges as the main cause of failed back surgery syndrome. According to the current literature and practice techniques, different agents are being used to prevent EF formation. To date there is no single agreed upon treatment method of EF. In this study, dilutional effect of CSF, together with low potassium levels, on primary skin fibroblast cultures was studied as a possible material for EF prevention. CSF at different concentrations (0-100%) were tested to see its effect on Skin fibroblast proliferation. A wound healing assay was also performed to see the effect of CSF on wound healing. The cell proliferation goes up from 24h to 72hr in all CSF percentages from 0-75% but the proliferation was inhibited at 100% CSF. The “wound” is closed successfully in all CSF percentages between 0-75. The 100% CSF fails to completely close the wound. Adverse effects of low concentrations of potassium levels and dilutional effect of CSF may be a promising solution in the prevention of EF. Further in vivo and in vitro experiments are required to characterize its use.

Response to Comment on “Circadian rhythms in the absence of the clock gene Bmal1”

Ness-Cohn et al. claim that our observations of transcriptional circadian rhythms in the absence of the core clock gene Bmal1 in mouse skin fibroblast cells are supported by inadequate evidence. They claim that they were unable to reproduce some of the original findings with their reanalysis. We disagree with their analyses and outlook.

Chromatic intervention and biocompatibility assay for biosurfactant derived from Balanites aegyptiaca (L.) Del

Extraction of biosurfactants from plants is advantageous than from microbes. The properties and robustness of biosurfactant derived from the mesocarp of Balanites aegyptiaca have been reported. However, the dark brown property of biosurfactant and lack of knowledge of its biocompatibility limits its scope. In the present work, the decolorization protocol for this biosurfactant was optimized using hydrogen peroxide. The hemolytic potential and biocompatibility based on cell toxicity and proliferation were also investigated. This study is the first report on the decolorization and toxicity assay of this biosurfactant. For decolorization of biosurfactant, 34 full factorial design was used, and the data were subjected to ANOVA. Results indicate that 1.5% of hydrogen peroxide can decolorize the biosurfactant most efficiently at 40 °C in 70 min at pH 7. Mitochondrial reductase (MTT) and reactive oxygen species (ROS) assays on M5S mouse skin fibroblast cells revealed that decolorized biosurfactant up to 50 µg/mL for 6 h had no significant toxic effect. Hemolysis assay showed ~ 2.5% hemolysis of human RBCs, indicating the nontoxic effect of this biosurfactant. The present work established a decolorization protocol making the biosurfactant chromatically acceptable. Biocompatibility assays confirm its safer use as observed by experiments on M5S skin fibroblast cells under in vitro conditions.