Fluorescent Labeling Research Articles

Synthesis and Evaluations of Cleavable Triazene‐Modified Nucleotide for DNA Sequencing

AbstractA fluorescence‐labeled nucleotide with a cleavable triazene linker was designed and synthesized as a potential reversible terminator for DNA sequencing by synthesis (SBS). The key intermediate, a triazene‐modified nucleotide, was successfully synthesized through the triazenylation of an amino‐modified nucleotide, followed by fluorescence labeling to yield the desired product. The synthesized triazene‐modified nucleotide can effectively serve as a substrate for Klenow Fragment (exo−) DNA polymerase and be incorporated into DNA strands. Once the first modified nucleotide is incorporated, no further extension is possible, even with an unblocked 3′‐OH group. After the fluorescent label is completely removed under acidic conditions, the triazene reversible terminator can be incorporated into DNA strands again, thereby enabling the DNA sequencing cycles.

Automated production of batched unclonable micro-patterns anti-counterfeiting labels with strong robustness and rapid recognition speed

Anti-counterfeiting technologies are indeed crucial for information security and protecting product authenticity. Traditional anti-counterfeiting methods have their limitations due to their clonable nature. Exploring new technologies, particularly those based on pixel-level textures, is a promising avenue to address the clonable issue due to high encoding capacity. However, research in this field is still in its early stages. This work introduces a new fluorescent anti-counterfeiting label technology with four key characteristics: efficient laser etching, high-throughput fabrication and segmentation, robustness aided by data augmentation, and an exceptionally high recognition speed. Specifically, the etching achieves a speed of 1,200 labels/3s, the high-throughput procedures yield a rate of 2,400 labels/4 mins, and the total count is about 5.2 × 104 labels. The number of labels is further augmented to about 5.2 × 106 by implementing arbitrary rotation and brightness variation to enhance the robustness in the recognition procedure. We divide these labels into 44 categories based on differences in patterns. Utilizing machine learning methods, we have achieved a total recognition (including extraction and search process) time per label averaging 421.96 ms without classification, and 40.13 ms with classification. Specifically, the search process with classification is nearly fiftieth times shorter than the non-classification method, reaching 8.52 ms in average. The overall recognition time is much faster than previous works, and achieves an accuracy of over 98.7%. This work significantly increases the practical significance of pixel-level anti-counterfeiting labels.

Simultaneous dual-colour labelling of mitochondria and lysosomes: An indolium-based approach

Fluorescence is a valuable tool for understanding mitochondria-lysosome interactions, which is central for unveiling their roles in health and disease. Herein, we describe novel indolium-based molecules for labelling mitochondria and lysosomes simultaneously with organelle-specific fluorescence properties. These probes readily enable clear distinction of mitochondria and lysosomes by combining confocal and two-photon fluorescence microscopy. Our results show that this unique behaviour relies on aggregation. The functioning of these probes is not compromised by mitophagy, allowing the visualisation of these organelles in separate emission channels by using a single fluorescent marker under all scenarios. Their enhanced experimental simplicity compared to using multiple dyes makes these novel compounds well-suited for automated screening applications within living cells.

Green Synthesis and Characterization of Hydroxyapatite Nanorods with Enhanced Antibacterial and Anticancer Properties

This study reports the microwave synthesis of hydroxyapatite nanorods using Lantana camara L. flower extract (LCF-HA) and characterized by FTIR, XRD, TEM and SEM-EDX techniques. The extract coating on the hydroxyapatite nanorods improves the antibacterial efficacy against Gram-positive bacteria. The study also evaluates the mitochondrial membrane potential (MMP), generation of reactive oxygen species (ROS) and capacity to induce apoptosis of LCF-HA. It shows that LCF-HA efficiently inhibits the growth of osteosarcoma cells (MG-63) in a dose-dependent manner. The efficacy of LCF-HA in inducing apoptosis in osteosarcoma cells is established by dual acridine orange/ethidium bromide (AO/EB) fluorescence labelling. Additionally, LCF-HA induces an increase in reactive oxygen species (ROS) generation and a reduction in mitochondrial membrane potential (MMP) levels. The findings indicate that derived LCF-HA has significant anticancer efficacy by inducing apoptosis in osteosarcoma cells through enhanced ROS generation, suggesting that derived LCF-HA may serve as a promising therapeutic target for bone cancer cell lines. Furthermore, the study demostrates excellent compatibility with osteoblast cells, highlighting its potential for bone regeneration applications.

Long Noncoding RNAs Expressed in Mouse Pituitary Development and Mature Hormone-Producing Cells.

Mammalian genomes contain thousands of genes for long noncoding RNA (lncRNAs), some of which have been shown to affect protein coding gene expression through diverse mechanisms. The lncRNA transcripts are longer than 200 nucleotides and are often capped, spliced, and polyadenylated, but not translated into protein. Nuclear lncRNAs can modify chromatin structure and transcription in trans or cis by interacting with the DNA, forming R-loops, and recruiting regulatory proteins. Not much is known about the role of lncRNA in pituitary gland differentiation and function. We mined transcriptome data from mouse pituitary glands collected at embryonic days 12.5 and 14.5 and identified over 200 different lncRNA transcripts. To develop a research resource for the study of lncRNA, we used pituitary cre transgenes to tag pituitary cell types in adult mice with fluorescent markers, and enriched for thyrotropes, gonadotropes, and somatotropes using fluorescence-activated cell sorting. We determined the transcriptome of each cell population using RNA sequencing and mined the data for lncRNA. We detected hundreds of lncRNAs in adult pituitary cells; a few were located immediately nearby genes that encode pituitary hormones or lineage-specific transcription factors. The location of these lncRNAs suggests the possibility of a cis-acting regulatory role in pituitary development or function, and we observe coordinated expression of 2 of them with their putative target genes in transgenic mice. This research resource sets the foundation for examining the actions of lncRNAs on their putative target genes and determining whether they have roles during development and in response to physiological demand.

Targeted Magnetic Nanoparticles for Beta-Amyloid Detection

Background/Objectivities: The presence of beta-amyloid plaques is a part of the pathogenesis of Alzheimer’s disease, but there is currently no universally accepted method for magnetic resonance (MR) imaging of the disease. However, it is known that magnetic nanoparticles (MNPs) can improve the T2 contrast in MR images of various targets. Methods: We used cubic MNPs, which were produced by thermal decomposition and then it was covalently bonded to a modified fluorescently labeled tetrapeptide, HAEE-Cy5, for visualizing beta-amyloid plaques. The interaction of MNPs-HAEE-Cy5 and beta-amyloid was determinate by confocal microscopy using SH-SY5Y cell line. Results: MNPs exhibit relatively high relaxivity (approximately 200 mM−1s−1), which is crucial for enhancing target visibility in MR imaging. HAEE provides targeted delivery of MNPs by specifically interacting with beta-amyloid, while the fluorescent label Cy5 enables monitoring the efficacy of the interaction through confocal microscopy. Conclusions: The MNPs modified with HAEE-Cy5 demonstrated excellent binding to beta-amyloid plaques in vitro, as shown by experiments on the SH-SY5Y cell line. These results suggest that the proposed method has potential for use in future MR imaging studies of Alzheimer’s disease.

Ultrafast and Ultrasensitive Bacterial Detection in Biofluids: Leveraging Resazurin as a Visible and Fluorescent Spectroscopic Marker.

Here we report the application of chemometric analysis for modeling absorbance spectroscopy and fluorescence emission data from a resazurin-based assay targeting low-level bacterial detection in biofluids. Bacteria spiked samples were incubated with resazurin and absorbance and fluorescence data were collected at 30 min intervals. The absorbance data was subjected to Principal Component Analysis (PCA) and Partial Least Squares Regression (PLSR) and compared with the univariate fluorescence spectroscopy approach. The analysis demonstrated the multidimensional nature of the absorbance data, highlighting the appearance of the resorufin peak at the 2 h time point with a low bacterial inoculum of 0.01 CFU mL-1 across all the samples tested-water, urine and serum. The PLSR models supported the PCA data and exhibited strong predictive capabilities for water (RC2 = 0.937, RCV2 = 0.934), urine (RC2 = 0.899, RCV2 = 0.880) and serum (RC2 = 0.985, RCV2 = 0.967). Conversely, fluorescence is contingent upon resorufin existence, necessitating a prolonged waiting period postincubation with resazurin to verify the presence of bacteria, especially when contamination levels are low. Given the substantial global impact of bacteria-related infections, this method detects bacteria at low concentrations precisely and rapidly, improving efficiency and adaptability for point-of-care settings, promising swift diagnosis of bacterial infections, environmental monitoring, or food-quality control.

A low-cost device for measuring TEER at cultivation of epithelial/endothelial cells on inserts.

Background: The barrier properties of epithelial and endothelial cells are usually studied in vitro when cells are cultured on mesh inserts in culture plates, and it is necessary to assess the state of the cell monolayer on the membrane of the inserts before the experiment. Typically, monolayer density is analyzed by passing a fluorescent label through the insert with cells or by measuring the transendothelial resistance (TEER) of the cell layer. Many studies use both methods of assessing monolayer integrity in parallel, depending on the purpose of the study. The TEER method also allows to record early changes in the monolayer state under the action of various substances. Aim of this work is to demonstrate the possibility of TEER measurements using the proposed device (conductometer) made from freely available components using endothelial/epithelial cells as an example. Materials and Methods. A device (conductometer) for measuring TEER, created on the basis of easily accessible components, is presented. The device was tested by culturing two cell lines – hybridoma of endothelial origin Ea.hy926 and human colon adenocarcinoma Caco-2. Caco-2 cells were cultured for 22 days and Ea.hy926 cells were cultured for 7 days. The integrity of the cell monolayer and the density of intercellular contacts were evaluated by the TEER value determined by the proposed conductometer, as well as by the fluorescein permeability of the cell monolayer. Results: The results of TEER measurements using the proposed device and at the same time, the fluorescein permeability of the cell monolayer during the cultivation of Caco-2 and EA.hy926 cells are presented. For Caco-2 cells from the moment of 100% confluence the TEER value gradually increased reaching maximum values by 20-21 days, after which it decreased slightly. The permeability values decreased as the cells were cultured, indicating the formation of dense contacts. For EA.hy926 cells the rise in TEER values are observed on day 3 and decrease was observed on day 7. The results of TEER and permeability obtained by the proposed conductometer have a strong inverse correlation for both cell lines and are in good agreement with each other. Conclusions. The presented device, made on the basis of simple and affordable components, is similar to commercially available devices and can be used to study the integrity and density of a monolayer in the cultivation of epithelial/endothelial cells, to study the processes of trans/paracellular transport under the action of various substances, as well as in experiments with the co-cultivation of various cell lines.

Overcoming fixation and permeabilization challenges in flow cytometry by optical barcoding and multi-pass acquisition.

The fixation and permeabilization of cells are essential for labeling intracellular biomarkers in flow cytometry. However, these chemical treatments often alter fragile targets, such as cell surface and fluorescent proteins (FPs), and can destroy chemically-sensitive fluorescent labels. This reduces measurement accuracy and introduces compromises into sample workflows, leading to losses in data quality. Here, we demonstrate a novel multi-pass flow cytometry approach to address this long-standing problem. Our technique utilizes individual cell barcoding with laser particles, enabling sequential analysis of the same cells with single-cell resolution maintained. Chemically-fragile protein markers and their fluorochrome conjugates are measured prior to destructive sample processing and adjoined to subsequent measurements of intracellular markers after fixation and permeabilization. We demonstrate the effectiveness of our technique in accurately measuring intracellular FPs and methanol-sensitive antigens and fluorophores, along with various surface and intracellular markers. This approach significantly enhances assay flexibility, enabling accurate and comprehensive cellular analysis without the constraints of conventional one-time measurement flow cytometry. This innovation paves new avenues in flow cytometry for a wide range of applications in immuno-oncology, stem cell research, and cell biology.

Triarylmethanol Derivatives with Ultralong Organic Room-Temperature Phosphorescence.

Triphenylmethyl-based compounds such as rhodamines and fluoresceine representing an old and well-known class of triphenylmethane dyes, are widely used in fluorescent labeling of bioimaging. Inspired by ultralong room temperature phosphorescence of triphenylphosphine derivatives, herein we report a methoxy substituted triarylmethanol ((4-methoxyphenyl)diphenylmethanol, LJW-1) exhibits ultralong room temperature phosphorescence (RTP) under ambient condition with afterglows of about 7 seconds. Its multiple C-H···π intermolecular interactions, C-H···O intermolecular interactions, hydrogen bond and π-π interactions are beneficial for forming rigid environment in the aggregated state which is evidently an important factor in the appearance of excellent RTP. Different substituents on triarylmethanol result in compounds with different lifetimes varying from 7 µs to 818 ms. The substituent effects on the phosphorescence of triarylmethanol derivatives provide an efficient method for the design of organic RTP materials and may be enlightening the phosphorescence research of triarylmethanol derivatives.

Impact of neonatal hyperoxia on the development of the coronary vascular bed: an experimental model for the study of cardiomyopathy associated with premature birth

Abstract Introduction Very preterm birth (PT; <32 weeks of gestation), nearly 2% of all births, leads to short-term complications such as bronchopulmonary dysplasia and retinopathy of prematurity that are driven by compromised organ vascular development. Adults born preterm display an increased risk of ischaemic cardiomyopathy and heart failure. Our group has shown that neonatal rats exposed to hyperoxia, simulating the PT neonatal conditions, develop cardiomyocytes hypertrophy, fibrosis and cardiac dysfunction, or Oxygen-Induced Cardiomyopathy (OIC). Whether cardiac changes also associate with alterations in vascular development in the left ventricle (LV) remains unknown. Purpose We hypothesized that neonatal exposure to hyperoxia impact the development of the LV vascular network. Methods Male pups were exposed to 80% O2 (OIC) or room air (CTRL) from day 3 (P3) to P10 of life. Hearts were collected at P10, 4- and 16 weeks. Arteriolar length and capillary density per cardiomyocyte were measured using stereology (elastin) and immunofluorescence (WGA, α-SMA, CD31). High-resolution 3D reconstruction of the vascular network was achieved by combining fluorescent labelling of coronaries (anti-α-SMA) and capillaries (anti-CD31) with a tissue-cleaning protocol (iDisco+), allowing detailed morphometric characterization of the vascular network. The expression of vascular endothelial growth factors and receptors (VEGFA, VEGFB, VEGFR 1 and 2, NRP1, and PIGF) were quantified by RT-qPCR and Western blots. Differences between groups were analyzed with Student's t-test (P < 0.05) (N = 6 per group). Results At P10, we observed a notable rise in the number of capillaries per cardiomyocyte (0.88 ± 0.05 vs. 0.62 ± 0.02 mm2) and a significant increase in VEGFA (+76%) and NRP1 (+38%) expression in the LV from OIC vs. CTRL rats. By 4 weeks, the OIC group showed a significant reduction in arteriolar length vs. CTRL (6.51 ± 0.76 vs. 9.77 ± 1.12 m/cm3), coupled with an increase in the number of capillaries per cardiomyocyte (1.11 ± 0.05 vs. 0.85 ± 0.03 mm2). VEGFA was decreased (-14%) and PLGF increased (+24%) in the LV of OIC rats. At 16 weeks, the OIC condition displayed a significant increase in the number of capillaries per cardiomyocyte (1.37 ± 0.03 vs. 0.98 ± 0.07 mm2) and a decrease in VEGFA (-23%), along with reduction in its receptors VEGFR1 (-27%), VEGFR2 (-20%) and NRP1 (-17%). Conclusion OIC resulting from neonatal hyperoxia exposure associates with vascular changes in the LV that persist to adulthood. These changes are characterized by a reduction in arteriolar length and an increase in the number of capillaries, accompanied by modulation of the expression of pro-angiogenic VEGFA and receptors. The impact of these vascular alterations on the susceptibility of the LV to ischemic or hypertension-related damage is to be explored. These findings suggest a crucial mechanistic pathway for understanding the risk of cardiac pathologies associated with preterm birth.

Adaptive-modulated fast fluctuation super-resolution microscopy

Fluorescence microscopy has significantly advanced biological imaging at the nanoscale, particularly with the advent of super-resolution microscopy (SRM), which transcends the Abbe diffraction limit. Most cutting-edge SR methods require high-precision optical setups, which constrain the widespread adoption of SRM. Fluorescence fluctuation-based SRM (FF-SRM) can break the diffraction limit without complex optical components, making it particularly well-suited for biological imaging. However, conventional FF-SRM methods, such as super-resolution optical fluctuation imaging (SOFI), still require specific fluorescent molecular blinking properties. Instead of enhancing the intrinsic blinking characteristics by finding specific fluorescent markers, employing optical methods such as spatial light modulation to adjust the excitation light field allows for easier and more flexible matching of the on-time ratio with the analysis of temporal stochastic intensity fluctuations. Nevertheless, the specific parameters of the modulation patterns have not been thoroughly explored, despite their crucial influence on the reconstruction quality. Herein, we propose adaptive-modulated fast fluctuation super-resolution microscopy. Our method demonstrates theoretically and experimentally that restricting the size of modulation units in a certain range ensures better image quality with fewer artifacts and signal losses. We find it still significantly effective when applied to other FF-SRM. Overall, the further development of the adaptive modulation technique has made it more stable in behavior and maintained high-quality imaging, presenting broader prospects for super resolution imaging based on statistical analysis.

Methods for Introdusing Fluorescent Labels into Polysaccharides

Polysaccharides are widely represented in plants and bacteria, where they are distinguished by their extraordinary structural diversity and the performance of various functions. To study the functions of polysaccharides, their fluorescent labeling is needed. This review discusses methods for introducing fluorescent labels into polysaccharides by chemical modification of certain functional groups of these complex biopolymers, as well as using the so-called bioorthogonal reactions, which allow labeling in a cell without affecting its viability. In addition to modification with organic dyes, the possibility of using quantum dots and coordination compounds of lanthanides is also discussed.

Mitochondrial Bioenergetics of Functional Wound Closure is Dependent on Macrophage-Keratinocyte Exosomal Crosstalk.

Tissue nanotransfection (TNT)-based fluorescent labeling of cell-specific exosomes has shown that exosomes play a central role in physiological keratinocyte-macrophage (mϕ) crosstalk at the wound-site. Here, we report that during the early phase of wound reepithelialization, macrophage-derived exosomes (Exomϕ), enriched with the outer mitochondrial membrane protein TOMM70, are localized in leading-edge keratinocytes. TOMM70 is a 70 kDa adaptor protein anchored in the mitochondrial outer membrane and plays a critical role in maintaining mitochondrial function and quality. TOMM70 selectively recognizes cytosolic chaperones by its tetratricopeptide repeat (TPR) domain and facilitates the import of preproteins lacking a positively charged mitochondrial targeted sequence. Exosomal packaging of TOMM70 in mϕ was independent of mitochondrial fission. TOMM70-enriched Exomϕ compensated for the hypoxia-induced depletion of epidermal TOMM70, thereby rescuing mitochondrial metabolism in leading-edge keratinocytes. Thus, macrophage-derived TOMM70 is responsible for the glycolytic ATP supply to power keratinocyte migration. Blockade of exosomal uptake from keratinocytes impaired wound closure with the persistence of proinflammatory mϕ in the wound microenvironment, pointing toward a bidirectional crosstalk between these two cell types. The significance of such bidirectional crosstalk was established by the observation that in patients with nonhealing diabetic foot ulcers, TOMM70 is deficient in keratinocytes of wound-edge tissues.

On-site detection of OTA and AFB1 based on branched hybridization chain reaction coupled with lateral flow assay

Mycotoxins are widely prevalent in various agricultural commodities, whose excessive consumption can pose significant risks to human health. In this study, we developed a facile mycotoxin detection platform based on branched hybridization chain reaction coupled with lateral flow assay. Ochratoxin A/Aflatoxin B1 bind to aptamers triggering the release of initiators, which leads to bHCR amplification and forms three-dimensional dendritic DNA nanostructures. Using the functionalized quantum dots as a fluorescent label, by leveraging smartphones and handheld ultraviolet lamps, the qualitative and quantitative detection of OTA and AFB1 can be achieved with a significantly enhanced sensitivity level, surpassing that of commercial test strips by 2–3 orders of magnitude. The visual detection limits for OTA and AFB1 were 30 pg/mL and 4 pg/mL, respectively. This approach eliminates the necessity for enzyme catalysis or the preparation and purification of antibodies and/or hapten, thereby reducing testing expenses and streamlining operational procedures. Moreover, substituting aptamer and nucleic acid sequences can effectively expand the scope of detection targets. Consequently, the as-proposed strategy exhibits great potential as a versatile technique, suitable for various analytical scenarios due to its sensitivity, accuracy, simplicity, and portability.

Chemo-Click: Receptor-Controlled and Bioorthogonal Chemokine Ligation for Real-Time Imaging of Drug-Resistant Leukemic B Cells.

Drug resistance in B cell leukemia is characterized by the coexpression of CXCR5 and CXCR3 chemokine receptors, making it a valuable biomarker for patient stratification. Herein, we report a novel platform of activatable chemokines to selectively image drug-resistant leukemic B cells for the first time. The C-terminal derivatization of the human chemokines CXCL13 and CXCL10 with bioorthogonal tetrazine-BODIPY and BCN groups retained binding and internalization via their cognate CXCR5 and CXCR3 receptors and enabled rapid fluorescence labeling of CXCR5+ CXCR3+ resistant B cells─but not drug-susceptible leukemic cells─via intracellular chemokine ligation. This modular chemical approach offers a versatile strategy for real-time immunophenotyping of cell populations with distinct chemokine profiles and will accelerate the design of new precision medicine tools to advance personalized therapies in blood tumors.

Specificity determinants of pathogens in forest

Abstract Host‐specific pathogens have long been suggested to act as a major driver of species diversity in tropical forests. However, determining the degree of host specificity of potential pathogens coupled to key cellular characteristics of infection is difficult and time‐consuming. These challenges have delayed progress in relating the functional specificity of pathogenic fungi to ecological consequences. We tested the pathogenicity of 27 (of 215) fungi that were isolated from surface sterilized roots of seedlings from four common tree species in a diverse subtropical forest. Inoculation experiments showed that six fungi exhibited strong pathogenicity on the host seedlings. Five of these only infected their specific hosts (i.e. host‐specific pathogen species). Green fluorescent protein labelling revealed that in three host‐specific pathogens fungal hyphae were able to grow into the vascular tissues only in their specific host plant, in contrast to other fungal‐host combinations that exhibit non‐pathogenic interactions. This fluorescent labelling technique allows direct tracking of the progress of fungal infection in different host tissues. Synthesis. By coupling the green fluorescent protein technique with standard host inoculation experiments, we determined the developmental differences between infections by pathogenic and non‐pathogenic fungi in a relatively simple and straightforward way. Our work provides a useful tool for rapidly screening and categorizing host–fungal interactions, reflecting the functional basis of host specificity of pathogenic fungi. More broadly, this technique can contribute to understanding the roles of pathogens in species coexistence and biodiversity maintenance in forest communities.

Measurements of Phagocytosis in Trabecular Meshwork Cells.

The restriction in the movement of aqueous humor through the trabecular meshwork is known to play an important role in the pathogenesis of glaucomas. Among the functions of the trabecular meshwork that help to prevent this restriction and maintain the movement of aqueous humor is phagocytosis. Cells in the trabecular meshwork actively phagocytose pigment, cellular debris, and other materials in order to help keep the drainage pathway clear. A decrease in phagocytosis has thus been implicated in the development of glaucomas. In this chapter, we outline two methods using fluorescent markers that can be used to measure the phagocytic properties of trabecular meshwork cells in culture.

Determination of triterpenic acids in Ribes stenocarpum maxim by HPLC-APCI-MS with 2-(12,13-dihydro-7H-dibenzo[a,g]carbazol-7-yl)ethyl4-methylben-zenesulfonate labelling agent

The study utilized a fluorescent labeling reagent called 2-(12,13-dihydro-7H-dibenzo[a,g]carbaz-ol-7-yl)ethyl4-methylben-zenesulfonate (DDCETS) to label of triterpenic acids and the approach was used to measure the amount of triterpenic acids in different organs of the Ribes stenocarpum Maxim. (RSM) from several areas in Qinghai Province. The analytical potential of the reagent was evaluated using six triterpene acids, namely maslinic acid, corosolic acid, betulinic acid, betulonic acid, oleanolic acid and ursolic acid. The results showed that the method successfully determined that the main components of triterpenoids in RSM from different regions of Qinghai Province were oleanolic acid and ursolic acid. The highest content of oleanolic acid and ursolic acid was 2037.59 mg/Kg in branches and 2088.97 mg/Kg in fruits. The least distributed organs of oleanolic acid and ursolic acid in the RSM in each region were seeds, with the lowest content of 460.88 mg/Kg in Dongxia Township, Datong County. And when analyzed in relation to the geographical conditions of the diverse origins, the fruits and leaves of the RSM were more abundant in both types of triterpene acids in areas with lower altitude, temperature and precipitation.

Beyond conventional therapy: Synthesis of multifunctional nanoparticles for rheumatoid arthritis therapy

Abstract Rheumatoid arthritis (RA) is a persistent inflammatory illness that causes joint destruction and dysfunction due to the activation of macrophages and the generation of reactive oxygen species. Current therapy choices frequently limit the effectiveness of targeting the inflammatory areas. To reduce inflammation and oxidative stress in RA, this research will create and assess multifunctional nanoparticles that selectively target inflammatory cells and deliver therapeutic medicines. Tannic acid, ferric chloride hexahydrate, methotrexate (MTX), and bovine serum albumin were conjugated using sonication and centrifugation to create the nanoparticles. Folic acid was added to improve the ability to target. Transmission electron microscopy, dynamic light scattering (DLS), UV-vis spectroscopy, and in vitro release experiments were used to characterize the nanoparticles. RAW 264.7 macrophage cells were used to test the cellular uptake of the nanoparticles using confocal microscopy and fluorescence-activated cell sorting (FACS). TFMBP-FA achieved 65.56%, and TFMBP reached 68.96%, indicating a high drug delivery rate for the synthesized nanoparticles. Confocal microscopy showed that the TFMBP-FA group had a greater density of fluorescent markers, indicating that the cells effectively targeted and absorbed the inflammatory environment. These results imply that the created nanoparticles may improve how medications are delivered during RA therapy.