Effect of GelMA Hydrogel Properties on Long-Term Encapsulation and Myogenic Differentiation of C2C12 Spheroids.

Skeletal muscle regeneration and engineering hold great promise for the treatment of various muscle-related pathologies and injuries. This research explores the use of gelatin methacrylate (GelMA) hydrogels as a critical component for encapsulating cellular spheroids in the context of muscle tissue engineering and regenerative applications. The preparation of GelMA hydrogels at various concentrations, ranging from 5% to 15%, was characterized and correlated with their mechanical stiffness. The storage modulus was quantified and correlated with GelMA concentration: 6.01 ± 1.02 Pa (5% GelMA), 75.78 ± 6.67 Pa (10% GelMA), and 134.69 ± 7.93 Pa (15% GelMA). In particular, the mechanical properties and swelling capacity of GelMA hydrogels were identified as key determinants affecting cell sprouting and migration from C2C12 spheroids. The controlled balance between these factors was found to significantly enhance the differentiation and functionality of the encapsulated spheroids. Our results highlight the critical role of GelMA hydrogels in orchestrating cellular dynamics and processes within a 3D microenvironment. The study demonstrates that these hydrogels provide a promising scaffold for the long-term encapsulation of spheroids while maintaining high biocompatibility. This research provides valuable insights into the design and use of GelMA hydrogels for improved muscle tissue engineering and regenerative applications, paving the way for innovative approaches to muscle tissue repair and regeneration.

Effects of ß-glucan, probiotics, and synbiotics on obesity-associated colitis and hepatic manifestations in C57BL/6J mice.

PURPOSE: Probiotics and prebiotics are commonly used to improve the gut microbiota. Since prebiotics can support the growth of probiotics, co-administration of these is called synbiotics. It has been demonstrated that obesity-induced gut dysbiosis can worsen inflammatory bowel disease symptoms. This study evaluated how modulation of gut microbiota with Schizophyllum commune-derived ß-glucan (BG), probiotics (PRO), and synbiotics containing both BG and PRO (SYN) could improve the symptoms of obesity-associated colitis and hepatic manifestation. METHODS: Mice were fed a normal diet (ND), high-fat diet (HFD), and HFD with different additives (BG, PRO, and SYN) for 12 weeks, followed by 5 days of colitis induction. Mice were sacrificed before and after colitis induction. During the experiment, body weight, food and water consumption, and rectal bleeding were monitored. Proteins from the colon were subjected to western blotting, and serum biomarkers such as alanine transaminase, alkaline phosphatase, triglycerides, and total cholesterol were analyzed. Colon and liver samples were sectioned for histological analysis. The fecal microbiota was analyzed based on partial 16S rRNA gene sequences. RESULTS: Although BG and PRO secured intestinal tight junctions, these two treatments did not modulate inflammatory cell infiltration and inflammatory markers (i.e., IL-6 and TNF-α). In contrast, SYN demonstrated stronger and broader effects in reducing colonic inflammation. While BG treatment increased the abundance of indigenous Lactobacillus, PRO treatment decreased bacterial diversity by suppressing the growth of several species of bacteria. SYN treatment groups, however, supported the growth of both indigenous and supplemented bacteria while maintaining bacterial diversity. CONCLUSION: Obesity-associated colitis can be improved by modulating gut bacteria with ß-glucan and probiotics. The co-administration of both outperformed ß-glucan and probiotic treatment alone by fostering both indigenous and supplemented probiotic strains.

ß-glucan, "the knight of health sector": critical insights on physiochemical heterogeneities, action mechanisms and health implications.

ß-glucans, the class of biological response modifier has unceasing attention, not only for its immune stimulating but also for its role as prebiotics, modulator of physiological events etc. and is widely used in the treatment of cancer, diabetes, gastrointestinal disorders, cardiovascular diseases etc. However, ß-glucan with different physiochemical properties is found to have discrete clinical functions and thus careful selection of the types of ß-glucan plays pivotal role in providing significant and expected clinical outcome. Herein this review, we presented the factors responsible for diverse functional properties of ß-glucan, their distinct mode of actions in regulating human health etc. Further, clinical aspects of different ß-glucans toward the management of wound care, metabolic dysbiosis, fatty liver disorders and endurance training associated energy metabolism were compiled and exhibited in detail.

Redox Regulation of NOX Isoforms on FAK(Y397)/SRC(Y416) Phosphorylation Driven Epithelial-to-Mesenchymal Transition in Malignant Cervical Epithelial Cells.

Epithelial-to-mesenchymal transition (EMT) promulgates epithelial cell associated disease-defining characteristics in tumorigenesis and organ fibrosis. Growth factors such as epidermal growth factor and fibroblast growth factor in addition to cytokines such as transforming growth factor-ß1 (TGF-ß1) is said to play a prominent role in remodeling related pathological events of cancer progression such as invasion, metastasis, apoptosis, EMT, etc. through redox related cellular secondary messengers, in particular the reactive oxygen species (ROS). However, the signaling cascade underlying the redox mechanism and thereby the progression of EMT remains largely unknown. In this study, upon TGF-ß1 treatment, we observed an induction in NOX isoforms-NOX2 and NOX4-that have time (early and late) and cellular localization (nucleus and autophagosome co-localized) dependent effects in mediating EMT associated cell proliferation and migration through activation of the focal adhesion kinase (FAK)/SRC pathway in HeLa, human cervical cancer cells. Upon silencing NOX2/4 gene expression and using the SRC inhibitor (AZD0530), progression of TGF-ß1 induced EMT related cellular remodeling, extra cellular matrix (ECM) production, cell migration and invasion, got significantly reverted. Together, these results indicate that NOX2 and NOX4 play important, albeit distinct, roles in the activation of cytokine mediated EMT and its associated processes via tyrosine phosphorylation of the FAK/SRC pathway.

Dietary intervention using (1,3)/(1,6)-ß-glucan, a fungus-derived soluble prebiotic ameliorates high-fat diet-induced metabolic distress and alters beneficially the gut microbiota in mice model.

PURPOSE: Western diet, rich in carbohydrates and fat, is said to be a major factor underlying metabolic syndrome. Interventions with prebiotics, the key modulators of the gut microbiota, have paramount impact on host-associated metabolic disorders. Herein, we investigated the effect of fungus-derived (1,3)/(1,6)-ß-glucan, a highly soluble dietary fiber, on high-fat diet (HFD)-induced metabolic distress. METHODS: Male C57BL/6 J mice were fed with different diet groups (n = 11): control diet, HFD, 3 g/kg or 5 g/kg of ß-glucan-incorporated HFD. At the end of experimental study period (12th week), body weight, feces weight and fecal moisture content were observed. Further, colonic motility was measured using activated charcoal meal study. Proteins extracted from liver and intestine tissues were subjected to western blot technique. Paraffin-embedded intestinal tissues were sectioned for histochemical [Periodic acid-Schiff (PAS) and Alcian blue (AB) staining] analysis. Fecal microbiota analysis was performed using MOTHUR bioinformatic software. RESULTS: ß-glucan consumption exhibited anti-obesity property in mice groups fed with HFD. In addition, ß-glucan ameliorated HFD-induced hepatic stress, colonic motility and intestinal atrophy (reduction in colon length, goblet cells, and mucosal layer thickness). Further, ß-glucan incorporation shifted bacterial community by increasing butyrate-producing bacteria such as Anaerostipes, Coprobacillus, and Roseburia and decreasing reportedly obesity-associated bacteria such as Parabacteroides and Lactococcus. CONCLUSION: Altogether, the outcomes of this present pre-clinical animal study show ß-glucan to be a promising therapeutic candidate in the treatment of HFD-induced metabolic distress. Further comprehensive research has to be conducted to brace its clinical relevance, reproducibility and efficacy for aiding human health.

Cellular senescence and EMT crosstalk in bleomycin-induced pathogenesis of pulmonary fibrosis-an in vitro analysis.

With poor prognosis and aberrant lung remodeling, pulmonary fibrosis exhibits worldwide prevalence accompanied by an increase in burden in terms of hospitalization and death. Apart from genetic and non-genetic factors, fibrosis occurs as a side effect of bleomycin antineoplastic activity. Elucidating the cellular and molecular mechanism could help in the development of effective anti-fibrotic treatment strategies. In the present study, we investigated the underlying mechanism behind bleomycin-induced fibrosis using human alveolar epithelial cells (A549 cells). On the basis of the experimental observation, it was demonstrated that with transforming growth factor-ß (TGF-ß) as a central mediator of fibrosis progression, a cross-talk between epithelial-mesenchymal transition (EMT) and senescence upon bleomycin treatment occurs. This results in the advancement of this serious fibrotic condition. Fibrosis was initiated through integrin activation and imbalance in the redox state (NOX expression) of the cell. It progressed along the TGF-ß-mediated non-canonical pathway (via ERK phosphorylation) followed by the upregulation of α-smooth muscle actin and collagen synthesis. Additionally, in this process, the loss of the epithelial marker E-cadherin was observed. Furthermore, the expressions of senescence markers, such as p21 and p53, were upregulated upon bleomycin treatment, thereby intensifying the fibrotic condition. Accordingly, the molecular pathway mediating the bleomycin-induced fibrosis was explored in the current study.

ß-Glucan-Based Wet Dressing for Cutaneous Wound Healing.

Objective: Recognized as pathogen-associated molecular patterns (PAMPs), ß-glucans, a naturally occurring heterogeneous group of polysaccharides, were investigated for their ability to accelerate wound healing in the form of high water-retaining hydrogel dressing. Approach: Full-thickness wounds on the dorsal side of mice created using a 5-mm biopsy punch were treated with ß-glucan-based hydrogel for 2 weeks. Standardized photographs of the wound site were taken at regular time intervals to calculate the percentage of wound closure. Tissues isolated from the wound area were subjected to histological examination and immunoblot analysis. Results: ß-Glucan-based hydrogel significantly accelerated the duration of wound healing and enhanced the development of skin appendages in the regenerated skin tissue. Increased expression of transforming growth factor-ß3 in the skin tissue isolated from the healed wound site indicated that skin regeneration rather than skin repair occurred, thereby minimizing cutaneous scarring. The expression level of cytokeratin 10 and cytokeratin 14 in the isolated skin tissue revealed that the wounds treated with hydrogel showed proper differentiation and proliferation of keratinocytes in the epidermal layer. Innovation: Immunomodulating ß-glucan (responsible for fighting infections at the wound site, and enhancing the migration and proliferation of keratinocytes and fibroblasts) in the form of a three-dimensional hydrogel membrane that retains a high water content (responsible for cooling and soothing effect around the wound site, thereby reducing pain) was prepared and analyzed for its effects on the cutaneous wound healing mechanism. Conclusion: ß-Glucan-based hydrogels are promising as wet wound dressings in the health care industry.