Search Results (99)

Background: Three-dimensional (3D) cell models may bridge the gap between two-dimensional (2D) cell cultures and animal models. Technical advances have led to the development of 3D-bioprinted cell models, characterized by greater reproducibility and the ability to mimic in vivo conditions. Glioblastoma multiforme (GBM) is a highly aggressive brain tumor with poor clinical outcomes due to its heterogeneity, angiogenic activity, and invasiveness. Src family kinases (SFKs) play a crucial role in GBM progression, making them attractive targets for drug development. Here, we show results about the pharmacological profile of a new prodrug synthesized from a Src inhibitor, SI306. Methods: Three-dimensional-bioprinted GBM cell models were used in predicting the antitumor activity of the prodrug SI306-PD2 with respect to its precursor, SI306. Results: Since the prodrug releases the active inhibitor through the cleavage by specific enzymes, SI306-PD2 was analyzed for stability and release kinetics in various media, including fetal bovine serum (FBS), which is normally used in cell culture. In comparison to SI306, SI306-PD2 demonstrated higher solubility in water, higher permeability across gastrointestinal and blood–brain barrier membranes, and the ability to release the drug in the presence of FBS progressively. In the 2D GBM cell model, using U87 and U251 cell lines, both compounds similarly reduced tumor cell viability. In 3D-bioprinted cell models, in the presence of an FBS-free medium, SI306-PD2 exhibited a more effective antitumor activity compared to SI306, reducing the proliferation and diameter of U251 spheroids grown within the bioprinted scaffold in a statistically significant manner. The analysis of proteins extracted from 3D scaffolds confirmed that SI306-PD2 inhibited Src activation more efficiently than SI306. Conclusions: Our study suggests that, when tissue permeability represents a discriminating characteristic, bioprinted cell models can provide a valid alternative for studying the cytotoxicity of new antitumor compounds. This approach has permitted us to ascertain the potential of the prodrug SI306-PD2 as a therapeutic agent for GBM, demonstrating better tissue penetration and antiproliferative efficacy compared to the precursor compound SI306. Full article

(1) Background: Controlled skin perforations, such as ear tags, piercings, and microdermal implants, induce inflammation and stress in individuals undergoing these procedures. This localized trauma requires care to optimize healing, reduce inflammation, and prevent infections. (2) Methods: Two formulations were developed: an FB-suspension and an FB-gel. Their in vivo efficacy was evaluated, along with drug retention in porcine and human skin after 30 min of administration, chemical stability at different temperatures, cytotoxicity, histological changes induced via transdermal application, and irritative potential, assessed using the HET-CAM assay. (3) Results: Both formulations reduced inflammation when applied 30 min before perforation compared to the positive control. The FB-suspension demonstrated no cytotoxicity and exhibited greater efficacy than the free flurbiprofen solution, highlighting the advantages of using nanoparticle-mediated drug delivery. Moreover, the FB-gel maintained chemical stability for up to 3 months across a temperature range of 4 to 40 °C. Histologically, no significant changes in skin composition were observed. (4) Conclusions: The FB-suspension is viable for both pre- and post-perforation application, as it is a sterile formulation. In contrast, the FB-gel is a convenient and easy application, making it a practical alternative for use in both clinical and veterinary settings. Full article

Volatile compounds (VCs) from fungi can promote plant growth, but their application methods are limited. Edible mushroom fungi beds (FBs) provide a readily available alternative source of fungal VCs, although their biostimulatory functions remain unvalidated. In this study, a novel, non-contact exposure method for applying VCs emitted from FBs to rice seedlings was developed. This marks the first evaluation of mushroom FBs as a direct source of bioactive VCs for plant growth promotion. Volatiles from two different edible mushroom FBs promoted shoot growth and increased biomass for rice seedlings. VCs from shiitake FBs significantly increased biomass by 67.4% while VCs from enokitake FBs by 39.5% compared to the control. The biomass-increasing effects were influenced by the quantity of shiitake FBs applied, with significant increases at 15 g, 30 g and 60 g applications. The VCs effects remained significant even when the FBs were covered with two types of gas-permeable polymer film. Chemical analysis of VCs from FBs identified several organic compounds and subsequent bioassays using synthetic VCs determined key bioactive VCs contributing to biomass increase at specific concentrations. This study presents a utilization method of waste mushroom FBs as sustainable, scalable, and cost-effective agricultural biostimulants. Full article

Background/Objectives: DNA damage response (DDR) is a highly conserved and complex signal transduction network required for preserving genome integrity. DNA repair pathways downstream of DDR include the tyrosyl-DNA phosphodiesterase1 (TDP1) enzyme that hydrolyses the phosphodiester bond between the tyrosine residue of topoisomerase I (TopI) and 3′-phosphate end of DNA. A small TDP1 subfamily, composed of TDP1α and TDP1β, is present in plants. The aim of this work was to investigate the role of the two TDP1 genes in the DDR context. Methods: A series of Arabidopsis thaliana DDR single and double mutants defective in the sog1, e2fb, pol2A, atm, and atr genes, treated with the genotoxic agents camptothecin (CPT, inhibitor of TopI) and NSC120686 (NSC, inhibitor of TDP1), were used. These compounds were specifically used due to their known impact on the TDP1 function. The effect of the treatments was assessed via phenotypic analyses that included germination percentage, speed, and seedling growth. Subsequently, the expression of the TDP1α and TDP1β genes was monitored through qRT-PCR. Results: Overall, the gathered data indicate that the atm mutant was highly sensitive to NSC120686, both phenotypically and concerning the TDP1α gene expression profiles. Alternatively, the upregulation of TDP1β in e2fb, pol2a, and atr supports its implication in the replication stress response. Conclusions: The current study demonstrates that genotoxic stress induced by CPT and NSC has a genotype-dependent effect reflected by a differential expression of TDP1 genes and early phenotypic development. Full article

(1) Background: For the reconstruction of a human vagina, various surgical procedures are available that are often associated with complications due to their failure to mimic the physiology of the human vagina. We recently developed a vascularized, organ-specific matrix from healthy human vaginal wall tissue with suitable biomechanical properties. A superior graft would require further extensive colonization with autologous vaginal cells to reduce complications upon implantation. However, reports on isolation of vaginal cells from biopsies are scarce, and published protocols rarely contain sufficient details. In this study, we aimed to examine protocols for inconsistencies and identify (where possible) the optimal protocol in terms of reproducibility and efficiency for isolation of human vaginal fibroblasts (FBs), epithelial cells (VECs), and smooth muscle cells (SMCs). Overall, this study aims to guide other researchers and aid future tissue engineering solutions that rely on autologous cells. (2) Methods: A total of 41 isolation protocols were tested: four protocols specific to FBs, 13 protocols for VECs, and 24 protocols for SMCs. Protocols were derived from published reports on cell isolation by enzymes, with exclusion criteria including the need for specialized equipment, surgical separation of tissue layers, or missing protocol details. Enzymatic digestion with collagenase-I, collagenase-IV, and dispase-II was used for isolation of VECs, collagenase-IV for isolation of SMCs, and collagenase-IA for isolation of FBs. Fluorescent immunostaining was applied to identify VECs with cytokeratin, SMCs with desmin, endothelial cells with UEA-1, and FBs with vimentin. Protocols were assessed based on (>95%) homogeneity, duplicate consistency, cell viability, and time to first passage. (3) Results: A total of 9 out of the 41 protocols resulted in isolation and expansion of vaginal FBs. This involved 1 out of 13 VEC protocols, 6 out of 24 SMC protocols, and 2 out of 2 FB protocols. Isolation of vaginal SMCs or VECs was not achieved. The best results were obtained after digestion with 0.1% collagenase-IV, where pure FB colonies formed with high cell viability. (4) Conclusions: Today, vaginoplasty is considered the gold standard for surgically creating a neovagina, despite its considerable drawbacks and limitations. Tissue-engineered solutions carry great potential as an alternative, but cell seeding is desired to prevent complications upon implantation of grafts. In this study, we examined isolation of human vaginal FBs, SMCs, and VECs, and identified the most efficient and reliable protocol for FBs. We further identified inconsistencies and irreproducible methods for isolation of VECs and SMCs. These findings aid the clinical translation of cell-based tissue engineering for the reconstruction and support of vaginas, fulfilling unmet medic needs. Full article

Fetal bovine serum (FBS) has long been the standard supplement in cell culture media, providing essential growth factors and proteins that support cell growth and differentiation. However, ethical concerns and rising costs associated with FBS have driven researchers to explore alternatives, particularly human platelet lysate (HPL). Among these alternatives, fibrinogen-depleted HPL (FD-HPL) has gained attention due to its reduced thrombogenicity, which minimizes the risk of clot formation in cell cultures and enhances the safety of therapeutic applications. This study investigates two preparation methods for FD-HPL from human platelet concentrates: the calcium chloride method and a mechanical approach. The concentrations of critical growth factors, including vascular endothelial growth factor (VEGF), brain-derived neurotrophic factor (BDNF), insulin-like growth factor (IGF), and keratinocyte growth factor (KGF), were evaluated for both methods. Additionally, the impact of FD-HPL on the proliferation and morphology of umbilical cord-derived mesenchymal stem cells (UC-MSCs) was assessed. The findings revealed that the calcium chloride method produced significantly higher concentrations of all measured growth factors compared to the mechanical method. Moreover, UC-MSCs cultured in calcium chloride-prepared FD-HPL exhibited enhanced cellular characteristics, including increased cell size, elongation, and improved overall morphology compared to those cultured in mechanically processed FD-HPL. These results indicate that the preparation method significantly influences the biological properties of HPL and the effectiveness of UC-MSC culture. The calcium chloride method emerges as a superior technique for producing FD-HPL, offering a promising alternative to FBS in regenerative medicine applications. This study underscores the importance of preparation methods in optimizing HPL for cell culture and therapeutic uses. Full article

Background/Objectives: This paper will compare the outcomes—safety and efficacy—of three minimally invasive glaucoma surgeries (MIGSs),the Hydrus Microstent, iStent, and Gonioscopy-Assisted Transluminal Trabeculotomy (GATT), for intraocular pressure (IOP) reduction in patients with primary open-angle glaucoma (POAG). Methods: A literature search of Ovid Medline and Embase identified studies evaluating the Hydrus, iStent, and GATT. Data on IOP reduction, medication use, and complications were analyzed. Results: Studies show the Hydrus, iStent, and GATT reduce IOP and medication burden in POAG patients, with some complications. For the Hydrus, studies showed 37.09% (27.5 ± 4.4 to 17.3 ± 3.7 mmHg) and 25% (16.8 to 12.6 mmHg) IOP reduction. Meanwhile, medication burden decreased from 2.5 ± 0.7 to 1.0 and from 2.1 to 1.15. For the iStent, studies showed a 36.39% (21.1 to 13.4 mmHg) and 8.19% (17.1 to 15.7 mmHg) IOP drop. Medication burden decreased from 2.87 to 1.24 and from 1.7 to 0.26. For GATT, studies showed a 49.33% (27.70 ± 10.30 to 14.04 ± 3.75) and 39.09% (26.40 ± 6.37 to 16.08 ± 2.38) IOP drop. Medication burden reduced from 3.73 ± 0.98 to 1.82 ± 1.47 and from 3.12 ± 0.80 to 0.45 ± 0.96. Conclusions: The Hydrus, iStent, and GATT are effective alternatives to trabeculectomy for mild to moderate POAG. They reduce and control IOP and dependence on medications with manageable safety profiles. In all three options, there were some clinically significant complications based on the p-value. For the Hydrus, it was PAS. For the iStent, they were PAS, FB sensation, IOP spikes, and microhyphema. For GATT, it was IOP spikes. However, further long-term studies, especially randomized controlled trials, are needed to support these results. Full article

Cultured meat technology is a form of cellular agriculture where meat is produced from animal cells grown in a lab, instead of raising and slaughtering animals. This technology relies heavily on fetal bovine serum (FBS) in cell media; hence, production is costly and contributes significantly to ammonia and greenhouse gas emissions. Achieving the successful commercialization of cell-cultured food requires the critical resolution of manufacturing cost and safety concerns. Hence, our research efforts are focused on identifying commercially viable and ecologically sustainable alternatives to FBS. In this study, we evaluated the potential of twenty-six water-based algal and cyanobacterial extracts to stimulate cell growth for meat cultivation under 90% reduced serum conditions. The extracts were compared in viability, proliferation, and Trypan blue exclusion assays. In the first screening phase, the extracts were evaluated in a ZEM2S (zebrafish) cell culture in a 1% FBS regimen. Based on their ability to exhibit protein tolerance or promote cell proliferation, ten extracts were selected and further assayed in a QM7 cell culture. The QM7 cell line (myoblasts from Japanese quail) is highly relevant for meat cultivation because of its ability to differentiate into muscle fibers. Extracts derived from two microalgae species, Arthrospira platensis (Spirulina) and Dunaliella tertiolecta, demonstrated the highest tolerance in cell culture, above 10 μg/mL (expressed as total protein concentration). Tolerance at a 100 μg/mL concentration was demonstrated exclusively using an extract of blue spirulina (commercially purified Spirulina), which supported cell growth through multiple passages. Full article

Mobile-bearing (MB) total knee arthroplasty (TKA) implants were introduced as an alternative to fixed-bearing (FB) implants because of their theoretical advantages related to femorotibial rotational mismatch. The purpose of this study is to compare the clinical and radiologic outcomes of MB and FB TKA after approximately 13 years of follow-up. We compared the results of 88 patients with a mean age of 66 years who had received a rotating platform MB implant or a FB implant. The mean follow-up was 13.6 years. The patients were assessed clinically (VAS, ROM, KSS, WOMAC scores) and radiographically before and 13 years after operation. There were no statistically significant differences between the FB and MB groups in terms of clinical outcomes and radiological outcomes (p > 0.05 for all). Although the incidence of complications was higher in the MB group, the findings were not statistically significant compared to the FB group (p > 0.05 for all). Although there were no significant differences in the clinical and radiologic outcomes between the FB and MB groups, the possible higher risk of osteolysis or aseptic loosening in the MB group could be an important clinical implication when selecting the TKA implant. Full article

Background: fasting-based strategies (FBS) and continuous caloric restriction (CCR) are popular methods for weight loss and improving metabolic health. FBS alternates between eating and fasting periods, while CCR reduces daily calorie intake consistently. Both aim to create a calorie deficit, but it is still uncertain as to which is more effective for short- and long-term weight and metabolic outcomes. Objectives: this systematic review and meta-analysis aimed to compare the effectiveness of FBS and CCR on these parameters in obese adults. Methods: after screening 342 articles, 10 randomized controlled trials (RCTs) with 623 participants were included. Results: both interventions led to weight loss, with a reduction of 5.5 to 6.5 kg observed at the six-month mark. However, the results showed that FBS led to slightly greater short-term reductions in body weight (−0.94 kg, p = 0.004) and fat mass (−1.08 kg, p = 0.0001) compared to CCR, although these differences are not clinically significant. Both interventions had similar effects on lean mass, waist and hip circumference, blood pressure, lipid profiles, and glucose metabolism. However, FBS improved insulin sensitivity, with significant reductions in fasting insulin (−7.46 pmol/L, p = 0.02) and HOMA-IR (−0.14, p = 0.02). Conclusions: despite these short-term benefits, FBS did not show superior long-term outcomes compared to CCR. Both strategies are effective for weight management, but more research is needed to explore the long-term clinical relevance of FBS in obese populations. Full article

The removal of foreign bodies (FBs) from the airways of children is a critical procedure that can avert serious complications. While both flexible and rigid bronchoscopy techniques are employed for this purpose, their comparative efficacy and safety remain subjects of debate. Therefore, we conducted this investigation to compare between both procedures. Studies comparing flexible to rigid bronchoscopy (n = 14) were identified by searching PubMed, Scopus, Web of Science, Cochrane Library, and Google Scholar. We performed comparative meta-analyses of reported presentation characteristics and clinical outcomes, using fixed- and random-effects models. A diverse range of FB types and locations were identified. No difference was observed in the success rate of FB removal between flexible and rigid bronchoscopy (logOR = 0.27; 95%CI: −1.91:2.45). The rate of negative first bronchoscopy was higher in the flexible compared to the rigid group (logOR = 2.68; 95%CI: 1.68:3.67). Conversion rates to the alternative method were higher in the flexible bronchoscopy group. The overall complication rates were similar between both methods; however, the risk of desaturation was significantly lower with flexible bronchoscopy (logOR = −2.22; 95%CI: −3.36:−1.08). Flexible bronchoscopy was associated with a shorter length of hospital stay. The choice of bronchoscopy technique should be tailored to individual case characteristics. Full article

The development of serum-free media (SFM) is critical to advance cell culture techniques used in viral vaccine production and address the ethical concerns and contamination risks associated with fetal bovine serum (FBS). This study evaluated the effects of marine microalgal extracts and growth factor cocktails on the activity of Madin-Darby canine kidney (MDCK) and Vero cells. Five marine microalgal species were used: Spirulina platensis (SP), Dunaliella salina (DS), Haematococcus pluvialis (HP), Nannochloropsis salina (NS), and Tetraselmis sp. (TS). DS and SP extracts significantly increased the proliferation rate of both MDCK and Vero cells. DS had a proliferation rate of 149.56% and 195.50% in MDCK and Vero cells, respectively, compared with that in serum-free medium (SFM). Notably, DS and SP extracts significantly increased superoxide dismutase (SOD) activity, which was 118.61% in MDCK cells and 130.08% in Vero cells for DS, and 108.72% in MDCK cells and 125.63% in Vero cells for SP, indicating a reduction in intracellular oxidative stress. Marine microalgal extracts, especially DS and SP, are feasible alternatives to FBS in cell culture as they promote cell proliferation, ensure safety, and supply essential nutrients while reducing oxidative stress. Full article

The present study aimed at converting meat industry waste, particularly waste bones and chicken feathers, into biochar to recycle valuable nutrients present in it, which ultimately become part of the municipal waste. The bone biochar (BB) and feathers biochar (FB) were prepared at 550 °C, and their potential was evaluated as an organic amendment for the growth of sunflower. The ash content (AC) and fixed carbon (FC) improved significantly in prepared biochars as compared to raw feedstock. Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX) analyses signaled the occurrence of various functional groups viz. amide group and hydroxyapatite, porosity, and multiple nutrients. Application of BB and FB in potted soil alone as well as in composites (1:1, 1:2, 2:1) at 1%, 3%, and 5% (w/w) and synthetic fertilizer significantly increased soil pH, electrical conductivity (EC_e), organic matter (OM) and water holding capacity (WHC), while reducing the bulk density (BD). The growth of plants grown in soil treated with a 2:1 composite of feathers and bone biochar at 5% application rate showed significantly greater differences in plant height, total chlorophyll content, and plant dry weight than the control but was comparable to growth with chemical fertilizer, rendering it a potential alternative to chemical-based synthetic fertilizer. Full article

Germline variants in the NSD1 gene are responsible for Sotos syndrome, while somatic variants promote neoplastic cell transformation. Our previous studies revealed three alternative RNA isoforms of NSD1 present in fibroblast cell lines (FBs): the canonical full transcript and 2 alternative transcripts, termed AT2 (NSD1 Δ5Δ7) and AT3 (NSD1 Δ19–23 at the 5′ end). The precise molecular pathways affected by each specific isoform of NSD1 are uncharacterized to date. To elucidate the role of these isoforms, their expression was suppressed by siRNA knockdown in FBs and protein expression and transcriptome data was explored. We demonstrate that one gene target of NSD1 isoform AT2 is ARP3 actin-related protein 3 homolog B (ACTR3B). We show that loss of both canonical NSD1 and AT2 isoforms impaired the ability of fibroblasts to regulate the actin cytoskeleton, and we observed that this caused selective loss of stress fibers. Our findings provide novel insights into NSD1 function by distinguishing isoform function and demonstrating an essential role of NSD1 in regulating the actin cytoskeleton and stress fiber formation in fibroblasts. Full article

Currently, in cities in different regions of the world, forest bathing (FB) is considered a practice to promote public health in vulnerable groups, such as the elderly, who have a higher risk of hypertension. This practice has had limited development in Latin American countries, and therefore, the objective of this study was to evaluate the benefits of forest baths on blood pressure and heart rate in a group of older adults in Guadalajara, Mexico. A program of six sessions was designed, the first to welcome and recognize the natural environment of the urban forest and the other five dedicated to each of the senses (hearing, touch, smell, sight, and taste), using the methodology of the Forest Therapy Hub (FTHub). The type of study was observational (before and after) with a single group, where the participants’ blood pressure and heart rate were evaluated. The results obtained showed significant effects of FB on reduction in high systolic blood pressure and diastolic hypertension before and after FB, with no significance on heart rate. Forest baths can be considered as an alternative strategy to manage the risk of hypertension in older adults, due to their ability to induce relaxation and normalize blood pressure levels. Full article