Stimulated Cell Proliferation Research Articles

Optimizing mass transfer performance in triply periodic minimal surface porous scaffolds through isosurface offset

Efficient bone tissue regeneration remains a critical challenge in orthopedic medicine, with scaffold mass transfer capabilities playing a pivotal role. Triply periodic minimal surface (TPMS) scaffolds have emerged as promising candidates due to their unique structure characterized by smooth, continuous surfaces with zero mean curvature and high specific surface area. However, optimizing their mass transfer performance to meet the diverse needs of bone tissues at different anatomical sites has been a persistent challenge. This study addresses this gap by investigating the effects of isosurface offset on mass transfer performance in three TPMS scaffolds (Fisher-Koch S, Gyroid, and Split-P) using computational fluid dynamics (CFD). The results showed that isosurface offset significantly increased the effective scaffold permeability range (by 116.8%, 5.3%, and 64.3% for F, G, and S scaffolds, respectively) and improved the wall shear stress (WSS) distribution, enhancing the area that effectively stimulates cell proliferation (by 25.2%, 8.7%, and 14.3% increase, respectively). Additionally, it was found that porosity, specific surface area, the ratio of maximum pore size to tortuosity, and curvature significantly influenced the permeability and WSS distribution of the scaffolds. Finally, permeation experiments using porous scaffolds fabricated by laser powder bed fusion (LPBF) technology were performed to validate the simulation results. This study provides new insights into the design of TPMS porous scaffolds and customized bone implants, enhancing their application prospects in bone tissue engineering.

Aloe vera and the Proliferative Phase of Cutaneous Wound Healing: Status Quo Report on Active Principles, Mechanisms, and Applications.

Aloe vera is commonly used as traditional medicine for cutaneous wound healing. Nonetheless, the wound healing mechanisms of Aloe vera remain unclear. This review aims to provide insight into the molecular mechanisms of Aloe vera in promoting cutaneous wound healing, with particular emphasis on the mechanisms that stimulate cell proliferation and migration. Aloe vera has been shown to upregulate growth factors such as keratinocyte growth factor-1 (KGF-1), transforming growth factor-β (TGF-β), cyclin D1, insulin-like growth factor 1 (IGF-1), vascular endothelial growth factor (VEGF), basic fibroblastic growth factor (bFGF), and microfibril-associated glycoprotein 4 (MFAP4), as well as collagen, fibrillin, elastin, α-smooth muscle actin (α-SMA), integrins, and platelet endothelial cell adhesion molecule 1 (PECAM-1, also known as CD31), while downregulating the expression of matrix metalloproteinases (MMPs). In addition, Aloe vera was also found to upregulate PI3K/Akt and MAPK pathways, as well as the TGF-β signalling pathway via Smad proteins. Furthermore, molecular docking studies revealed that certain chemical constituents of Aloe vera bind to some of the forementioned growth factors or signalling molecules. With regards to current applications, although human clinical trials have reported positive results from using Aloe vera in healing open wounds and burns and alleviating some inflammatory skin diseases, the current commercial uses of Aloe vera remain largely focused on cosmetic products. Thus, greater advances are required to promote the use of Aloe vera products in clinical settings.

Tailoring cell therapies for diabetic metabolic phenotypes: a comparative study on the efficacy of various umbilical cord-derived cell regimens.

Given the high heterogeneity of type 2 diabetes mellitus (T2DM), it is imperative to develop personalized stem cell infusion regimen for targeted metabolic phenotype in order to ensure optimal therapeutic efficacy. In this study, we conducted a comparative analysis of 4 infusion regimens involving single and repeated infusions of human umbilical cord Wharton's jelly-derived MSCs (hucMSCs), single infusions of umbilical cord blood mononuclear cells (UCB), and sequential infusions of hucMSCs and UCB in T2DM rats. Results showed all 4 infusion regimens exhibited comparable efficacy in lowering fasting blood glucose levels and suppressing glucagon secretion. Single and double infusions of hucMSCs exhibited a tendency to migrate to the liver, thereby better at ameliorating hepatic glucose metabolism by enhancing glycogen synthesis and storage, promoting glycolysis, inhibiting gluconeogenesis, and improving insulin signal transduction. The sequential infusion of hucMSCs and UCB demonstrated specific cell tropism toward the pancreas, leading to prolonged glucose-lowering effects following a glucose tolerance test, restoration of early-phase insulin secretion, stimulation of islet beta cell proliferation and improvement in the beta/alpha ratio. Multiple injections, regardless of cell type, reduced the expression of systemic chronic inflammatory markers such as IL-1β, IL-6, IL-17, IL-22, and IFN-γ. Finally, a single dose of UCB exhibited a greater tendency to target visceral fat and enhanced effectiveness in regulating levels of total cholesterol and triglycerides. In conclusion, our study provided personalized stem cell regimens for diverse T2DM metabolic phenotypes, thereby offering improved treatment alternatives for future clinical trials and applications.

Circ_0124346 facilitates cell proliferation of pancreatic adenocarcinoma cells by regulating lipid metabolism via miR-223-3p/ACSL3 axis

BackgroundBoth lipid metabolism and cyclic RNAs (circRNAs) have been found to be involved in pancreatic adenocarcinoma (PAAD) progression, but the relationship between lipid metabolism and circRNAs remains unclear.MethodsThe expression levels of miR-223-3p, circ_0124346, and acyl-CoA synthetase long chain family member 3 (ACSL3) were determined through qRT-PCR and Western blot analysis. Cell proliferation was evaluated using the CCK-8 and EdU incorporation assays. Cholesterol (CH) and triglyceride (TG) levels were quantified using relevant kits. The relationships between miR-223-3p and circ_0124346 or ACSL3 mRNA were examined by bioinformatics analysis, luciferase reporter, RNA-RNA pull-down, and RIP assays.ResultsWe observed a significant elevation in circ_0124346 expression in both pancreatic adenocarcinoma (PAAD) tissues and cell lines, and its expression level was shown to be correlated with tumor size. Circ_0124346 stimulated cell proliferation and facilitated lipid synthesis in PAAD cells. Additionally, we found that circ_0124346 functioned as a sponge for miR-223-3p, preventing miR-223-3p’s binding to the 3'-UTR of ACSL3 mRNA, which subsequently led to an elevation in ACSL3 expression and promoted lipid synthesis. Accordingly, circ_0124346 knockdown resulted in a significant decrease in lipid synthesis and cell proliferation in PAAD cells, with partial reversal of these effects achieved via inhibiting miR-223-3p or overexpressing ACSL3.ConclusionOur study demonstrated that circ_0124346 regulates lipid metabolism in PAAD cells via the miR-223-3p/ACSL3 axis, suggesting that targeting circ_0124346 may serve as a potential strategy for treating PAAD and assisting in its diagnosis.

The Effectivity of Acemannan Sponge and Calcium Phosphate Cement-Calcium Sulfate Hemihydrate (CPC-CSH)-Diode Laser-Assisted Method on the Direct Pulp Capping

Dental caries, the decay of tooth, affects approximately 2.3 billion people worldwide and reaches 88.8% of the total Indonesians. Caries reaching the pulp requires direct pulp capping (DPC) therapy to preserve pulp vitality. Ca(OH)2 is the golden standard DPC material, but its adhesion and mechanical properties are poor. Therefore, an alternative therapy is needed to improve the success of DPC using acemannan sponge and calcium phosphate cement-calcium sulfate hemihydrate (CPC-CSH)-diode laser. Acemannan, polymannose extracted from aloe vera gel, plays a role in reparative dentin formation that is immunomodulatory, antimicrobial, biocompatible, and accelerates wound healing. Calcium Phosphate Cement (CPC), bioactive material used in tissue engineering. Calcium sulfate hemihydrate (CSH), inorganic component and biocompatible. The combination of CPC-CSH can shorten the setting time, maintain compressive strength, and has good handling properties. Low level diode laser (LLDL) plays a role in reparative dentin formation by altering growth factors expression to stimulate cell proliferation and fibroblast development. LLDL is applied before pulp capping material application between the pulp-dentin so that aggregated collagen fibrils are collected and stimulate the formation of odontoblasts. This paper aims to describe The Effectiveness of Acemannan Sponge and Calcium Phosphate Cement-Calcium Sulfate Hemihydrate (CPC-CSH)-Diode Laser Assisted Method in Direct Pulp Capping Therapy. Article searching uses the Preferred Reporting Items for Systematic Reviews and Meta-analyses with keywords (MeSH): "DPC and Acemannan", "DPC and CPC-CSH", and "DPC and Diode Laser" from Pubmed, ScienceDirect, and Google Scholar databases in 2019-2024. Acemannan Sponge and Calcium Phosphate Cement-Calcium Sulfate Hemihydrate (CPC-CSH)-Diode Laser Assisted Method are effective as Alternative Direct Pulp Capping Therapy.

Broad-Spectrum Bactericidal Multifunctional Tiny Silicon-Based Nanoparticles Modified with Tannic Acid for Healing Infected Diabetic Wounds.

Infected chronic wounds, in particular, diabetic wounds, are hard to heal, posing a global health concern with high morbidity and mortality rates. Diabetic full-thickness wounds infected with E. coli belong to the most difficult to heal chronic infected wounds. Here, we introduced tannic acid-modified silicon-based nanoparticles (TA-SiNPs) with broad-spectrum bactericidal activity that bacteria develop minimal resistance to, and they can effectively treat full-thickness wounds in diabetic mice infected with E. coli. Our findings indicate that these TA-SiNPs could achieve 100% antibacterial efficiency against S. aureus and 99.83% against E. coli, underlied by a positive surface charge and tannic acid groups facilitating bacterial membrane chemical composition depletion and depolarization of the membrane. In addition, we showed that spraying TA-SiNPs onto the skin wound of diabetic mice infected with E. coli resulted in wound healing with 98% closure after 12 days, in stark contrast to 49% of the control (PBS) and 68% of the one treated with Ofloxacin. Along with infection inhibition and ROS scavenging, we identified cell proliferation stimulation, inflammatory cytokine downregulation, and healing cytokine upregulation in the lesion, favoring the healing process. This study not only demonstrates the feasibility of employing silicon-based nanoparticles in diabetic wound healing for the first time, but also reports the first broad-spectrum bactericidal silicon nanodots. Furthermore, this provides novel insights into the mechanism of tannin-based nanoparticles disrupting bacterial membranes by depleting their chemical constituents. Our results highlighted that the developed TA-SiNPs are an effective nanomaterial for treating the infected chronic wounds.

N-acetylglucosaminyltransferase V drives colorectal cancer metastasis by facilitating ZO-1 ubiquitination and degradation

Increasing evidence supports the crucial role of Epithelial-Mesenchymal Transition (EMT) in cancer invasion and metastasis. N-acetylglucosaminyltransferase V (MGAT5), which is associated with multiantenna glycosylation, can contribute to tumorigenesis, yet its specific role in promoting colorectal cancer (CRC) metastasis remains unclear. Bioinformatics analysis of CRC datasets revealed that elevated MGAT5 expression was associated with EMT and a poor prognosis. In vitro experiments confirmed the pivotal role of MGAT5 as an EMT regulator in CRC cells. MGAT5 overexpression stimulated cell proliferation and migration, while MGAT5 knockdown had the opposite effect. Mechanistically, MGAT5 promoted EMT through multiantenna glycosylation of ZO-1, promoting its ubiquitination and reducing its expression. Clinically, MGAT5 upregulation in the CRC TMA correlated negatively with ZO-1 expression, which is indicative of malignancy and a poor prognosis. This study revealed that MGAT5 promotes EMT in CRC via interactions between multiple antenna glycosylation products and ZO-1 ubiquitination/degradation, indicating that MGAT5 could serve as a promising therapeutic target for CRC.Graphical

Mitochondrial double-stranded RNA homeostasis depends on cell-cycle progression.

Mitochondrial gene expression is a compartmentalised process essential for metabolic function. The replication and transcription of mitochondrial DNA (mtDNA) take place at nucleoids, whereas the subsequent processing and maturation of mitochondrial RNA (mtRNA) and mitoribosome assembly are localised to mitochondrial RNA granules. The bidirectional transcription of circular mtDNA can lead to the hybridisation of polycistronic transcripts and the formation of immunogenic mitochondrial double-stranded RNA (mt-dsRNA). However, the mechanisms that regulate mt-dsRNA localisation and homeostasis are largely unknown. With super-resolution microscopy, we show that mt-dsRNA overlaps with the RNA core and associated proteins of mitochondrial RNA granules but not nucleoids. Mt-dsRNA foci accumulate upon the stimulation of cell proliferation and their abundance depends on mitochondrial ribonucleotide supply by the nucleoside diphosphate kinase, NME6. Consequently, mt-dsRNA foci are profuse in cultured cancer cells and malignant cells of human tumour biopsies. Our results establish a new link between cell proliferation and mitochondrial nucleic acid homeostasis.

Hydroxyapatite/Chondroitin Sulphate Nanocomposite Infused with Black Seed Oil for Bone Tissue Applications: Physicochemical Characterizations and In Vitro Screening

AbstractMorbidities associated with natural bone have highlighted the need of advanced bioactive implants for the replacement of damaged bone. Known for their beneficial bioactivity and biocompatibility, Nigella sativa seeds (kalonji/black seed) are traditionally used in many medications. This research focuses on the positive impact of black seed extract on the chondroitin sulfate‐based hydroxyapatite nanocomposite for bone tissue regeneration applications. Novel nanocomposites with varied concentrations of black seed extract (0, 1, 2, and 3 wt%) were fabricated through a coprecipitation approach and analyzed for physicochemical and biological applications. The results of X‐ray diffraction and FTIR spectroscopy revealed that the HCSK3 nanocomposite was successfully developed with appreciably strong molecular interactions among their components. HCSK3 nanocomposite exhibits higher rough surface morphology (Ra −1.13 µm and Rz −4.8 µm), uniform particle distribution (average particle size −9.9 nm), and good thermal stability with total weight loss of 13%. It also provides better nucleation sites for apatite formation and protein adsorption stimulating cell attachment, proliferation, and differentiation. Moreover, HCSK3 nanocomposite showed excellent cell viability, higher ARS/ALP activities, better antibacterial property, and nonhemolytic property (below 3%). Thus, it can be concluded that HCSK3 nanocomposite manifested the highest biocompatibility and negligible cell toxicity. Therefore, it can be employed as an ideal bone implant in the bone tissue restoration phenomenon.

The role of glycolysis in tumorigenesis: From biological aspects to therapeutic opportunities

Glycolytic metabolism generates energy and intermediates for biomass production. Tumor-associated glycolysis is upregulated compared to normal tissues in response to tumor cell-autonomous or non-autonomous stimuli. The consequences of this upregulation are twofold. First, the metabolic effects of glycolysis become predominant over those mediated by oxidative metabolism. Second, overexpressed components of the glycolytic pathway (i.e. enzymes or metabolites) acquire new functions unrelated to their metabolic effects and which are referred to as “moonlighting” functions. These functions include induction of mutations and other tumor-initiating events, effects on cancer stem cells, induction of increased expression and/or activity of oncoproteins, epigenetic and transcriptional modifications, bypassing of senescence and induction of proliferation, promotion of DNA damage repair and prevention of DNA damage, antiapoptotic effects, inhibition of drug influx or increase of drug efflux. Upregulated metabolic functions and acquisition of new, non-metabolic functions lead to biological effects that support tumorigenesis: promotion of tumor initiation, stimulation of tumor cell proliferation and primary tumor growth, induction of epithelial-mesenchymal transition, autophagy and metastasis, immunosuppressive effects, induction of drug resistance and effects on tumor accessory cells. These effects have negative consequences on the prognosis of tumor patients. On these grounds, it does not come to surprise that tumor-associated glycolysis has become a target of interest in antitumor drug discovery. So far, however, clinical results with glycolysis inhibitors have fallen short of expectations. In this review we propose approaches that may allow to bypass some of the difficulties that have been encountered so far with the therapeutic use of glycolysis inhibitors.

Boron in wound healing: a comprehensive investigation of its diverse mechanisms.

Chronic wounds present a significant clinical challenge due to their prolonged healing time and susceptibility to infection. Boron, a trace element with diverse biological functions, has emerged as a promising therapeutic agent in wound healing. This review article comprehensively investigates the mechanisms underlying the beneficial effects of boron compounds in wound healing. Boron exerts its healing properties through multiple pathways, including anti-inflammatory, antimicrobial, antioxidant, and pro-proliferative effects. Inflammation is a crucial component of the wound-healing process, and boron has been shown to modulate inflammatory responses by inhibiting pro-inflammatory cytokines and promoting the resolution of inflammation. Furthermore, boron exhibits antimicrobial activity against a wide range of pathogens commonly associated with chronic wounds, thereby reducing the risk of infection and promoting wound closure. The antioxidant properties of boron help protect cells from oxidative stress, a common feature of chronic wounds that can impair healing. Additionally, boron stimulates cell proliferation and migration, as well as essential tissue regeneration and wound closure processes. Overall, this review highlights the potential of boron as a novel therapeutic approach for treating chronic wounds, offering insights into its diverse mechanisms of action and clinical implications.

MicroRNA-145-5p restricts cardiac scar resolution during cardiac regeneration

Abstract Background Human hearts have a very limited capacity to renew cardiomyocytes following injury. The lost myocardium is permanently replaced with noncontractile fibrotic tissue, consequently leading to heart failure. In contrast, zebrafish retain cardiac regenerative capacity during their lifetime. After injury, zebrafish hearts undergo transient fibrotic scarring that subsequently resolves and is replaced with new cardiomyocytes over time. Thus, understanding the molecular mechanisms governing cardiac scar resolution in zebrafish could offer insights into enhancing the regenerative capacities of human hearts. Objective We aimed to decipher the molecular pathways associated with zebrafish cardiac scar resolution. Methods We investigated the mRNA and miRNA responses to injury at 14 days post-injury by high-throughput transcriptome profiling of RNAs isolated from healthy or cryoinjured hearts. The prediction of miRNA-mRNA interactions was performed to identify the most differentially expressed miRNAs and their target genes. These miRNAs and genes were validated in zebrafish regenerating hearts and in the left ventricles of ischemic cardiomyopathy or dilated cardiomyopathy patients. The potential role of identified miRNAs in cardiac scar resolution was investigated in human cardiac fibroblasts. Results Principal component analyses identify two distinct clusters in both the mRNAome and miRNAome corresponding to healthy and injured hearts, indicating significant changes in transcriptome profiles during zebrafish cardiac regeneration. Gene set enrichment analyses indicate that molecular pathways involved in extracellular matrix organization and inflammatory response are markedly activated whereas genes associated with mitochondrial organization and oxidative phosphorylation are downregulated in injured hearts. We observed the downregulation of miR-145-5p and the upregulation of procollagen V in the injured hearts. In contrast to zebrafish, in the left ventricles of ischemic cardiomyopathy patients, miR-145-5p is significantly upregulated and procollagen V α1 is downregulated. Theoretical prediction of miRNA-mRNA interactions indicates procollagen V α1 is negatively correlated with miR-145-5p. Overexpression of miR-145-5p in human cardiac fibroblasts downregulates collagen V α1 and alpha smooth muscle actin, upregulates procollagen I and matrix metalloproteinase 9, and inhibits cell proliferation and migration. In contrast, inhibition of miR-145-5p upregulates collagen V α1 and alpha smooth muscle actin, downregulates procollagen I and matrix metalloproteinase 9, and stimulates cell proliferation and migration. Conclusion Our study suggests that miR-145-5p may play a role in the suppression of myofibroblast transdifferentiation via collagen V α1, which restricts scar resolution and cardiac repair.

Platelet Rich Plasma in the Management of Recurrent Lumbar Disc Surgery; A Study of Ten Cases

Low back pain (LBP) is a very common entity, now regarded as the first cause of disability worldwide. Intervertebral disc (IVD) degeneration leads to posterior rupture annulus fibrosus (AF) with extrusion of Central Nucleus pulposus (NP) through the injured area which has little potential to repair as it has no blood supply and causes mechanical pressure and inflammatory changes in the nerve root causing LBP. Autologous platelet rich plasma (PRP) contain concentrate blood that contain a natural concentration of autologous growth factors and cytokines and currently widely used in the clinical setting for tissue regeneration and repair. PRP had great potential to stimulate cell proliferation and metabolic activity of IVD effectively restoring structural change, improving matrix integrity, IVD regeneration, reducing discogenic back pain. Our Study of 10 cases, where repeat or re-exploration surgery (after primary PLID Surgery, fenestration and discectomy) was done, where removal of recurrent disc 5, release of epidural fibrosis and perineural scarring 4 and surgical debridement for discitis 1 was done. In addition, PRP was given in the disc space & around the nerve root which provides lots of growth factor, cytokines which helps in disc regeneration and prevent adhesion & fibrosis. These dual actions enhance symptomatic recovery of all cases. Large scale studies may be required to confirm the clinical evidence of PRP for the treatment of discogenic LBP Bang. J Neurosurgery 2024; 13(2): 70-74

Phytochemical Characterization and Assessment of the Wound Healing Properties of Three Eurasian Propolis.

The objective of this study is to evaluate the wound healing potential of Eurasian propolis by analyzing the phytochemical profile and the biological effects of three representative propolis samples. Specific colorimetric assays were used to estimate the total phenolic and flavonoid contents and the triterpenoids content. Some of the main components of Eurasian propolis (pinocembrin, pinobanksin, CAPE, chrysin and galangin) were analyzed using HPLC-DAD. Scavenging activity and total antioxidant capacity were assessed through DPPH and ORAC assays, respectively. Human keratinocyte, fibroblast, and monocytic cell lines were used for the biological in vitro analyses. The direct wound healing properties were tested through scratching assays and ELISA kits for the assessment of the production of growth factors (FGF-7, Latency Associated Peptide-LAP), while the indirect effects were evaluated through the estimation of the levels of MMP9, IL-1β, IL-8, and TNF-α using ELISA kits together with a cell-free test on the inhibition capacity on collagenases. Network Pharmacology analysis was employed to further explore possible mechanisms of the action of propolis on the healing process. The analyses confirmed the high phenolic content of Eurasian propolis (142.50-211.30 mg GAE/g), dominated by flavonoids (95.50-196.80 mg Galangin Equivalents/g), and terpenes (431.50-650.00 mg β-sitosterol Equivalents/g), while also verifying the significant antioxidant (4.9-8.9 mM/g Trolox Equivalents) and antiradical (DPPH IC50 26.1-54.4 μg/mL) activities. The samples showed indirect wound healing properties by mitigating inflammation and remodeling (reduced IL-1β and MMP9) and potentially modulating the immune response (upregulated IL-8). In vitro studies confirmed these effects, demonstrating decreased MMP9 production and collagenase inhibition when cells were co-treated with propolis and a stressor. Propolis also suppressed IL-1β release in fibroblasts, although its impact on TNF-α was inconclusive. Notably, co-treatment upregulated IL-8 in monocytes, suggesting a potential immunomodulatory role. Eurasian propolis may not directly stimulate cell proliferation during wound healing. Its anti-inflammatory and immunomodulatory properties could indicate an indirect contribution in helping the process.

Anti-fibrotic Potential of WJ-MSC Exosomes in Liver Fibrosis: Mechanistic Insights and Dose-Response Efficacy

Background: Hepatic fibrosis is a biological response characterized by the accumulation of extracellular matrix (ECM) during the wound healing process. Hepatic stellate cells (HSCs) play a pivotal role in fibrogenesis, transitioning from quiescent to myofibroblast cell types and leading to excessive ECM production. Platelet-derived growth factor (PDGF), a potent mitogen, is produced by activated HSCs, stimulating cell proliferation and migration. Objectives: This study aims to analyze the impact of Wharton's jelly mesenchymal stem cell (WJ-MSC)-derived exosomes on HSC activation induced by PDGF during liver fibrosis. Methods: Hepatic stellate cells-T6 cells were treated with PDGF-BB for 24 hours to induce activation, followed by treatment with varying concentrations of WJ-MSC-derived exosomes (0, 25, and 50 µg/mL) for another 24 hours. The effects of exosome treatment on HSC activation were evaluated through flow cytometry, differentiation assays, dynamic light scattering (DLS), transmission electron microscopy (TEM), RT-PCR, and western blot analysis. Results: Our study yields promising results, highlighting the potential therapeutic effects of WJ-MSC-derived exosomes on liver fibrosis. The dose-dependent decrease in fibrotic markers such as α-SMA, COLA1, and phosphorylated AKT protein in PDGF-BB-treated HSC-T6 cells suggests that WJ-MSC exosomes exert an anti-fibrotic effect by inhibiting HSC activation. Conclusions: These findings suggest that exosomes derived from WJ-MSCs hold therapeutic promise for liver fibrosis treatment by targeting key pathways involved in HSC activation and fibrogenesis. Further investigation into the underlying mechanisms of this anti-fibrotic effect and the potential clinical applications of WJ-MSC-derived exosomes in liver fibrosis management is warranted.

The immunomodulatory effects of vitamins in cancer.

Nutrition may affect animal health due to the strong link between them. Also, diets improve the healing process in various disease states. Cancer is a disease, where the harmful consequences of tumors severely impair the body. The information regarding the evolution of this disease is extrapolated from human to animal because there are few specific studies regarding nutritional needs in animals with cancer. Thus, this paper aims to review the literature regarding the immunomodulatory effects of vitamins in mammal cancer. An adequate understanding of the metabolism and requirements of nutrients for mammals is essential to ensuring their optimal growth, development, and health, regardless of their food sources. According to these: 1) Some species are highly dependent on vitamin D from food, so special attention must be paid to this aspect. Calcitriol/VDR signaling can activate pro-apoptotic proteins and suppress anti-apoptotic ones. 2) Nitric oxide (NO) production is modulated by vitamin E through inhibiting transcription nuclear factor kappa B (NF-κB) activation. 3) Thiamine supplementation could be responsible for the stimulation of tumor cell proliferation, survival, and resistance to chemotherapy. 4) Also, it was found that the treatment with NO-Cbl in dogs is a viable anti-cancer therapy that capitalizes on the tumor-specific properties of the vitamin B12 receptor. Therefore, diets should contain the appropriate class of compounds in adequate proportions. Also, the limitations of this paper are that some vitamins are intensively studied and at the same time regarding others, there is a lack of information, especially in animals. Therefore, some subsections are longer and more heavily debated than others.

6376 Wnt/beta-catenin Pathway is Associated with Cell Proliferation in Growth Hormone-producing Pituitary Tumors

Abstract Disclosure: S. Kuwabara: None. H. Kameda: None. H. Nomoto: None. K. Cho: None. A. Nakamura: None. Y. Ishi: None. H. Motegi: None. M. Fujimura: None. T. Atsumi: None. Objective: It has been well recognized that the Wnt/β-catenin pathway regulates cell proliferation, and that translocation of β-catenin from the cytoplasm to the nucleus promotes cell proliferation. There have been, however, limited reports on the Wnt/β-catenin pathway in growth hormone-producing pituitary tumors (GHomas), and we aimed to investigate the impact of the Wnt/β-catenin pathway on cell proliferation in GHomas. Methods: We performed three types of experiments with patient pathological specimens, disease model cell lines, and primary culture of GHoma cells. 1.The intracellular localization of β-catenin was evaluated using immunofluorescence staining of pathological specimens from GHoma patients and subsequently its association with clinical data was analyzed. 2.The Wnt/β-catenin inhibitor iCRT14 was administered to rat GHoma cells GH1, using the WST-1 assay to measure cell viability and real-time PCR to assess GH and cell cycle genes expression. 3. The Wnt/β-catenin inhibitor iCRT14 was added to cultured cells obtained from GHoma patients to evaluate cell viability and gene expression. Results: 1. Samples from 26 cases (42%) exhibited strong dot-like expression of β-catenin in the nucleus (dot pattern (DP) group), and samples from 36 cases (58%) showed β-catenin expression in the cytoplasm or cell membrane (non-DP group). Tumors in the DP group had a larger size than non-DP group (diameters 18.3±9.2 vs. 13.4±7.4 mm, p<0.05), and the postoperative remission rate was lower (23 vs. 50%, p<0.05). 2. Viability evaluation using WST-1 assay showed a maximum decrease of 25%. Gene expression showed concentration-dependent reduction in GH gene expression by up to 40% (p<0.05) and cyclin D1 gene up to 50% (p<0.05) following iCRT14 administration. 3. Among 6 cases, 3 showed a tendency toward cell growth inhibition, and 4 showed a tendency toward decreased cyclin D1 gene expression. Conclusion: It is suggested that the Wnt/β-catenin pathway is activated in the DP group because of the nucleus accumulation of β-catenin, and given the larger tumor size and lower postoperative remission rate in DP group, the Wnt/β-catenin pathway may stimulate cell proliferation in GHomas. In addition, cyclin D1-mediated effects were considered. Wnt/β-catenin pathway could play a role in GHomas growth, therefore this pathway would be a potential therapeutic target for treating affected patients. Presentation: 6/3/2024

8636 Characterization Of The Role Of Neutrophils In Promoting Progression Of Lymphangioleiomyomatosis

Abstract Disclosure: T.R. Henson: None. B. Minor: None. S.R. Hammes: None. Lymphangioleiomyomatosis (LAM) is a rare, multisystemic disease that is associated with cystic lung destruction. LAM is caused by small smooth muscle cell-like tumors throughout the lung that contain deleterious mutations in the tuberous sclerosis complex genes TSC1 or TSC2. This attenuates the activity of the mammalian target of rapamycin complex 1 (mTORC1) causing increased proliferation of the smooth muscle cells. LAM is a sexually dimorphic disease that primarily affects genetic females. Clinical observations implicate the role of estrogen in disease progression, as it is exacerbated during pregnancy and by use of exogenous estrogens in birth control. LAM tumor-derived cells lines have shown that direct estrogen stimulation of cell proliferation is modest compared to the dramatic in vivo effects of estradiol (E2) stimulation in a uterine-specific TSC2-null mouse model. Creation of a monoclonal cell line from these mice resulted in the generation of the LTM3 cell line, which also demonstrates low response to direct estrogen stimulation in vitro. This indicates that direct E2 stimulation may act upon other cell types in the tumor microenvironment. Immunophenotyping of uterine-specific TSC2-null mice has shown that the tumor microenvironment is enriched with myeloid cells specifically neutrophils. Functional analysis of neutrophil suppression in vivo has shown a reduction in myometrial growth in the uterine-specific TSC2-null mice, indicating a key role for neutrophil-mediated LAM tumor growth. Altered transcriptional regulation of neutrophils further highlights the importance of neutrophils in LAM progression. Bulk messenger RNA sequencing revealed E2 dependent increase in the neutrophil derived serine protease neutrophil elastase (NE). NE in vitro has been shown to increase migration, invasion, and proliferation of LAM cell lines, while inhibition in vivo has shown a reduction to the myometrial growth. Previously published data by the lab has shown that E2 increases tumor induced expansion of neutrophils in the bone marrow. Isolation and purification of Neutrophils from mouse bone marrow stimulated with tumor conditioned media from the LTM3 cells has shown signs of activation via upregulated gene expression. The focus of the current research has been the characterization of bone marrow derived neutrophils. Currently working with an in vitro system to study the proliferation of LTM3 and the rat leiomyoma (ELT3) cell lines when they are co-cultured with neutrophils. Understanding the mechanism by which neutrophils are activated and interact with LAM cells can give understanding to disease progression may provide insight into future treatment options. Presentation: 6/3/2024

Nitrogen-doped carbon quantum dot regulates cell proliferation and differentiation by endoplasmic reticulum stress

ABSTRACT Quantum dots have diverse biomedical applications, from constructing biological infrastructures like medical imaging to advancing pharmaceutical research. However, concerns about human health arise due to the toxic potential of quantum dots based on heavy metals. Therefore, research on quantum dots has predominantly focused on oxidative stress, cell death, and other broader bodily toxicities. This study investigated the toxicity and cellular responses of mouse embryonic stem cells (mESCs) and mouse adult stem cells (mASCs) to nitrogen-doped carbon quantum dots (NCQDs) made of non-metallic materials. Cells were exposed to NCQDs, and we utilized a fluorescent ubiquitination-based cell system to verify whether NCQDs induce cytotoxicity. Furthermore, we validated the differentiation-inducing impact of NCQDs by utilizing embryonic stem cells equipped with the Oct4 enhancer-GFP reporter system. By analyzing gene expression including Crebzf, Chop, and ATF6, we also observed that NCQDs robustly elicited endoplasmic reticulum (ER) stress. We confirmed that NCQDs induced cytotoxicity and abnormal differentiation. Interestingly, we also confirmed that low concentrations of NCQDs stimulated cell proliferation in both mESCs and mASCs. In conclusion, NCQDs modulate cell death, proliferation, and differentiation in a concentration-dependent manner. Indiscriminate biological applications of NCQDs have the potential to cause cancer development by affecting normal cell division or to fail to induce normal differentiation by affecting embryonic development during pregnancy. Therefore, we propose that future biomedical applications of NCQDs necessitate comprehensive and diverse biological studies.

CAPE derivatives: Multifaceted agents for chronic wound healing.

Chronic wounds significantly impact the patients' quality of life, creating an urgent interdisciplinary clinical challenge. The development of novel agents capable of accelerating the healing process is essential. Caffeic acid phenethyl ester (CAPE) has demonstrated positive effects on skin regeneration. However, its susceptibility to degradation limits its pharmaceutical application. Chemical modification of the structure improves the pharmacokinetics of this bioactive phenol. Hence, two novel series of CAPE hybrids were designed, synthesized, and investigated as potential skin regenerative agents. To enhance the stability and therapeutic efficacy, a caffeic acid frame was combined with quinolines or isoquinolines by an ester (1a-f) or an amide linkage (2a-f). The effects on cell viability of human gingival fibroblasts (HGFs) and HaCaT cells were evaluated at different concentrations; they are not cytotoxic, and some proved to stimulate cell proliferation. The most promising compounds underwent a wound-healing assay in HGFs and HaCaT at the lowest concentrations. Antimicrobial antioxidant properties were also explored. The chemical and thermal stabilities of the best compounds were assessed. In silico predictions were employed to anticipate skin penetration capabilities. Our findings highlight the therapeutic potential of caffeic acid phenethyl ester (CAPE) derivatives 1a and 1d as skin regenerative agents, being able to stimulate cell proliferation, control bacterial growth, regulate ROS levels, and being thermally and chemically stable. An interesting structure-activity relationship was discussed to suggest a promising multitargeted approach for enhanced wound healing.