Stability In Biofluids Research Articles

Exploring dysregulated miRNAs in ALS: implications for disease pathogenesis and early diagnosis.

Amyotrophic lateral sclerosis(ALS) is a progressive neurodegenerative disease marked by motor neuron degeneration, leading to muscle weakness and paralysis, with no effective treatments available. Early diagnosis could slow disease progression and optimize treatment. MicroRNAs(miRNAs) are being investigated as potential biomarkers due to their regulatory roles in cellular processes and stability in biofluids. However, variability across studies complicates their diagnostic utility in ALS. This study aims to identify significantly dysregulated miRNAs in ALS through meta-analysis to elucidate disease mechanisms and improve diagnostic strategies. We systematically searched PubMed, Google Scholar, and the Cochrane Library, following predefined inclusion and exclusion criteria. The primary effect measure was the standardized mean difference (SMD) with a 95% confidence interval, analyzed using a random-effects model. Additionally, we used network pharmacology to examine the targets of dysregulated miRNAs and their roles in ALS pathology. Analysing 34 studies, we found significant upregulation of hsa-miR-206, hsa-miR-133b, hsa-miR-23a, and hsa-miR-338-3p, and significant downregulation of hsa-miR-218, hsa-miR-21-5p, and hsa-let-7b-5p in ALS patients. These miRNAs are involved in ALS pathophysiology, including stress granule formation, nuclear pore complex, SMCR8 and Sig1R dysfunction, histone methyltransferase complex alterations, and MAPK signaling perturbation, highlighting their critical role in ALS progression. This study identifies several dysregulated miRNAs in ALS patients, offering insights into their role in the disease and potential as diagnostic biomarkers. These findings enhance our understanding of ALS mechanisms and may inform future diagnostic strategies. Validating these results and exploring miRNA-based interventions are crucial for improving ALS diagnosis and treatment outcomes.

Polyphenol-stabilized coacervates for enzyme-triggered drug delivery

Stability issues in membrane-free coacervates have been addressed with coating strategies, but these approaches often compromise the permeability of the coacervate. Here we report a facile approach to maintain both stability and permeability using tannic acid and then demonstrate the value of this approach in enzyme-triggered drug release. First, we develop size-tunable coacervates via self-assembly of heparin glycosaminoglycan with tyrosine and arginine-based peptides. A thrombin-recognition site within the peptide building block results in heparin release upon thrombin proteolysis. Notably, polyphenols are integrated within the nano-coacervates to improve stability in biofluids. Phenolic crosslinking at the liquid-liquid interface enables nano-coacervates to maintain exceptional structural integrity across various environments. We discover a pivotal polyphenol threshold for preserving enzymatic activity alongside enhanced stability. The disassembly rate of the nano-coacervates increases as a function of thrombin activity, thus preventing a coagulation cascade. This polyphenol-based approach not only improves stability but also opens the way for applications in biomedicine, protease sensing, and bio-responsive drug delivery.

Scalable biomimetic sensing system with membrane receptor dual-monolayer probe and graphene transistor arrays.

Affinity-based biosensing can enable point-of-care diagnostics and continuous health monitoring, which commonly follows bottom-up approaches and is inherently constrained by bioprobes' intrinsic properties, batch-to-batch consistency, and stability in biofluids. We present a biomimetic top-down platform to circumvent such difficulties by combining a "dual-monolayer" biorecognition construct with graphene-based field-effect-transistor arrays. The construct adopts redesigned water-soluble membrane receptors as specific sensing units, positioned by two-dimensional crystalline S-layer proteins as dense antifouling linkers guiding their orientations. Hundreds of transistors provide statistical significance from transduced signals. System feasibility was demonstrated with rSbpA-ZZ/CXCR4QTY-Fc combination. Nature-like specific interactions were achieved toward CXCL12 ligand and HIV coat glycoprotein in physiologically relevant concentrations, without notable sensitivity loss in 100% human serum. The construct is regeneratable by acidic buffer, allowing device reuse and functional tuning. The modular and generalizable architecture behaves similarly to natural systems but gives electrical outputs, which enables fabrication of multiplex sensors with tailored receptor panels for designated diagnostic purposes.

Blood RNA-Seq profiling reveals a set of circular RNAs differentially expressed in frail individuals

BackgroundFrailty is an intermediate and reversible geriatric syndrome that often precedes dependence. Therefore, its identification is essential to prevent dependence. Several molecules have been proposed as biomarkers of frailty, but none of them have reached clinical practice. Recently, circular RNAs have emerged as new non-coding RNAs. Their regulatory role together with their high stability in biofluids makes them good candidates as biomarkers for various processes, but, to date, no study has characterized the expression of circRNA in frailty.ResultsWe studied RNA from leukocytes of 35 frails and 35 robust individuals. After RNA-Sequencing, circRNA detection was performed by CIRI2 and Circexplorer2 and differential expression analysis by DESeq2. Validation was performed by Quantitative-PCR. Linear Discriminant Analysis was performed to determine the best circRNA combination to discriminate frail from robust. In addition, CircRNA candidates were studied in 13 additional elder donors before and after a 3-month physical intervention. We found 89 differentially expressed circRNAs (p-value<0.05, FC>|1.5|) with frailty. Upregulation of hsa_circ_0007817, hsa_circ_0101802 and hsa_circ_0060527 in frail individuals was validated. The combination of hsa_circ_0079284, hsa_circ_0007817 and hsa_circ_0075737 levels showed a great biomarker value with a 95.9% probability of correctly classifying frail and robust individuals. Moreover, hsa_circ_0079284 levels decreased after physical intervention in concordance with an improvement in frailty scores.ConclusionsThis work describes for the first time a different expression pattern of circular RNA (circRNAs) between frail and robust individuals. Moreover, the level of some circRNAs is modulated after a physical intervention. These results suggest that they could be used as minimally invasive biomarkers of frailty.

Wearable, Sensing-Controlled, Ultrasound-Based Microneedle Smart System for Diabetes Management.

An intelligent closed-loop system that senses glucose and automatically delivers insulin can provide enhanced glycemic control for diabetes management. Here, we report for the first time on the development of a wearable, sensing-controlled, ultrasound-based closed-loop microneedle smart system for diabetes management. Polylactic acid (PLA) hollow microneedles were fabricated by soft lithography that can maintain excellent mechanical strength and stability in biofluids, followed by the deposition of sensing electrodes on the outer layer. An ultrasonic insulin pump was constructed by integrating a lead zirconate titanate piezoelectric (PZT) ring and a biocompatible stainless-steel sheet with hundreds of tapered holes on top of the inner layer of the microneedles. Both the sensor and the pump were powered and controlled by a printed circuit board (PCB). When the sensing device detects interstitial glucose levels above normal, the closed-loop control algorithm triggers the ultrasonic pump to deliver insulin into the interstitial fluid through the microneedle hollow channels of the PLA microneedles. The ultrasound from the pump can cause the vaporization of insulin in the subcutaneous tissue fluid, accelerate the diffusion rate, and improve the efficiency of insulin treatment and utilization. The system has successfully demonstrated effective control of glucose levels in diabetic rats. The glucose sensor showed a high sensitivity of 0.212 μA/mM at 0-28 mM with a detection limit of 14 μM, and the mean absolute relative difference (MARD) was 9.96% with an average error of 1.6 mM. The flow rate of the ultrasonic pump was 120 μL/min, and the glucose-lowering rate was 4 mM/h. This work may open a new paradigm for the development of intelligent systems for diabetes management, as well as a wide range of practical applications in diabetes patients.

Comparison of microstructure and tensile behavior of hydroxyapatite-coated PEEK meshes and cellulose-based fabrics and mats

Polymeric biomaterials, such as polyether ether ketone (PEEK) and cellulose, have been explored as scaffolds for bone tissue engineering in the past decade. In this study, microstructure and mechanical behavior of uncoated and hydroxyapatite (HA)-coated polymeric meshes, fabrics, and mats were investigated. Commercially available monofilament PEEK meshes, and cellulose fabrics and mats were selected, then coated with a customized low temperature sol–gel method (≤ 150°C). Adhesive HA coating consisting of HA, β-TCP, and CaO with nanorod structure was derived. After HA coating, porosity of substrates (except filter-paper cellulose mats) decreased by up to 43%, indicating effective coating. Both uncoated and HA-coated substrates’ degradation rate in phosphate-buffered saline decreased after day 3. This is a result of ion precipitation or calcium compounds formation, indicating potential stability in biofluids for an extended period of time. Regarding tensile test results, highest tensile strength and elongation at break were obtained for PEEK meshes (approximately 80 MPa and 35%, respectively), as a result of the bulk material properties. HA coating did not significantly affect the tensile properties of the specimens, except for cellulose mats with an initial porosity of 77% (tensile modulus increased by 270% and strength by 210%). The increase in tensile properties could be attributed to increased rigidity, resulting from the adhesion between HA coating and cellulose fibers. Overall, HA-coated cellulose mats and PEEK meshes show promise as customizable, flexible scaffolds for implant applications and bone regeneration for future work.

Diagnostic and Prognostic Circulating MicroRNA in Acute Stroke: A Systematic and Bioinformatic Analysis of Current Evidence.

Background and PurposeStroke is the second leading cause of death and disability worldwide and its diagnosis, and assessment of prognosis, remains challenging. There is a need for improved diagnostic and prognostic biomarkers. MicroRNAs (miRNAs) play important roles in the post-transcriptional regulation of gene expression and their secretion and remarkable stability in biofluids highlights their potential as sensitive biomarkers in the diagnosis and prognosis of acute stroke. MethodsWe carried out a systematic review to assess current evidence supporting the potential of miRNAs to act as unique diagnostic and prognostic biomarkers in blood samples collected from patients suffering acute stroke within 24 hours of symptoms onset. ResultsWe identified 22 studies eligible for inclusion with 33 dysregulated miRNAs having diagnostic potential in the acute phase of the disease. We identified miR-16, miR-126, and miR-335 as having the highest sensitivity as diagnostic and prognostic biomarkers in acute ischaemic stroke and present original bioinformatic and pathway enrichment analysis of putative miRNA–target interactions. ConclusionsmiRNAs represent unique biomarkers which have a promising future in stroke diagnosis and prognosis. However, there is a need for more standardized and consistent methodology for the accurate interpretation and translation of miRNAs as novel specific and sensitive biomarkers into clinical practice.

Rational Design of DNA Framework-Based Hybrid Nanomaterials for Anticancer Drug Delivery.

Engineered DNA frameworks have been extensively exploited as affinity scaffolds for drug delivery. However, few studies focus on the rational design to comprehensively improve their stability, internalization kinetics, and drug loading efficiency. Herein, DNA framework-based hybrid nanomaterials are rationally engineered by using a molecular docking tool, where the framework acts as a template to support conjugated polymers. The hybrid materials exhibit high stability in biofluids owning to the multiple interactions between DNA and cationic conjugated polymer. Through molecular docking, it is found that a specific structure of the conjugated polymer at major grooves of DNA gives rise to a unique pocket for small-molecular drug doxorubicin (DOX) yielding lower binding energy than conventional DOX binding sites. This increases the binding affinity of DOX, allowing for high drug loading content and efficiency, and preventing drug leakage under physiological condition. As a proof of concept, the hybrid nanomaterials equipped with aptamer are used to carry DOX and antisense oligonucleotide G3139, which effectively inhibits solid tumor growth and shows negligible side effects on mice. It is anticipated that this approach would find broad applications in hybrid materials design and precise medicine.

Generic Method for Designing Self-Standing and Dual Porous 3D Bioscaffolds from Cellulosic Nanomaterials for Tissue Engineering Applications.

Three-dimensional scaffolds (3D) with controlled shape, dual porosity and long-term mechanical and dimensional stability in biofluids are of interest as biotemplates in tissue engineering. Herein, self-standing and lightweight cellulose-based biogenic scaffolds with a spatially structured morphology, macropores and interconnected micropores were fabricated using a combination of direct ink writing 3D printing and freeze-drying techniques. This was achieved by developing a water-based and low-cost bicomponent ink based on commercially available nanofibrillated cellulose (NFC) and carboxymethyl cellulose (CMC). Physical cross-linking through dehydrothermal treatment significantly increased the surface hardness, indentation modulus, compression strength, as well as the dimensional stability of the scaffolds in biofluids, in comparison to untreated materials. However, no differences in the spectra of solid state nuclear magnetic resonance or infrared were observed for dehydrothermal treated samples, suggesting that the increase of mechanical properties and dimensional stability is based on the physical cross-linking of functional groups both at the interface between NFC and CMC. The supramolecular structure of the polymers was well-preserved as disclosed by X-ray diffraction measurements. The cross-linked scaffolds showed high proliferation, viability, and attachment of human bone tissue derived osteoblast cells (hFOB). The simple and straightforward avenue proposed here for the design of cellulose-based fibrous inks and dual porous scaffolds from the commercially available materials and without the need of any additional cross-linkers should pave the way for the development of implantable, degradable scaffolds and cell-laden biomaterials for bone tissue regeneration and 3D bioprinting applications.

Circular RNA profiling reveals that circular RNAs from ANXA2 can be used as new biomarkers for multiple sclerosis

Multiple sclerosis is an autoimmune disease, with higher prevalence in women, in whom the immune system is dysregulated. This dysregulation has been shown to correlate with changes in transcriptome expression as well as in gene-expression regulators, such as non-coding RNAs (e.g. microRNAs). Indeed, some of these have been suggested as biomarkers for multiple sclerosis even though few biomarkers have reached the clinical practice. Recently, a novel family of non-coding RNAs, circular RNAs, has emerged as a new player in the complex network of gene-expression regulation. MicroRNA regulation function through a 'sponge system' and a RNA splicing regulation function have been proposed for the circular RNAs. This regulating role together with their high stability in biofluids makes them seemingly good candidates as biomarkers. Given the dysregulation of both protein-coding and non-coding transcriptome that have been reported in multiple sclerosis patients, we hypothesised that circular RNA expression may also be altered. Therefore, we carried out expression profiling of 13.617 circular RNAs in peripheral blood leucocytes from multiple sclerosis patients and healthy controls finding 406 differentially expressed (P-value < 0.05, Fold change > 1.5) and demonstrate after validation that, circ_0005402 and circ_0035560 are underexpressed in multiple sclerosis patients and could be used as biomarkers of the disease.

MicroRNAs in Ischemic Heart Disease: From Pathophysiology to Potential Clinical Applications.

Despite rapid advances in cardiovascular research and therapeutic strategies, ischemic heart disease (IHD) remains the leading cause of mortality worldwide. MicroRNAs (miRNAs) are small, noncoding RNAs which post transcriptionally regulate gene expression. In the past few years, miRNAs have emerged as key tools for the understanding of the pathophysiology of IHD, with potential uses as new biomarkers and therapeutic targets. Several studies report a regulatory role of miRNAs, with regard to fundamental components of IHD pathogenesis and progression, such as lipoprotein metabolism, atherogenesis, vascular calcification, platelet function, and angiogenesis. Due to their high stability in biofluids, circulating miRNAs have attracted attention as promising biomarkers of IHD, especially in cardiovascular risk prediction and the diagnosis of myocardial infarction. Furthermore, experimental studies have demonstrated the potential of miRNA-targeted therapy in improving hyperlipidemia, atherosclerosis, and angiogenesis. In this review, the current knowledge on the role of miRNAs in IHD and translational perspectives of their use is discussed.

BIOSYNTHESIS OF GOLD NANOPARTICLES BY BIOSORPTION USING NEOSARTORYA UDAGAWAE: CHARACTERIZATION AND INVITRO EVALUATION

Objective: The present study was aimed to investigate gold nanoparticles synthesized by fungal isolate Neosartorya udagawae and determination of their stability in biofluids to probe their aptness in drug delivery applications.Methods: In this procedure, gold nanoparticles were prepared by biosynthesis using seven days old culture of Neosartorya udagawae and aqueous chloroauric acid. After the complete reaction, the fungal biomass was subjected to UV-Vis, XRD, FT-IR Spectrum analysis, TEM, Zeta potential, SEM and EDX analysis.Results: Intra/extracellular synthesis of gold nanoparticles was confirmed by a sharp peak at 526 nm in UV spectroscopy. SEM, TEM analysis demonstrates the spherical shape of AuNPs with an average diameter of 50 nm and XRD confirm the crystalline gold nanoparticles. FTIR analysis reveals the presence of the protein shell around the gold nanoparticles. The zeta potential value of AuNPs was-36mV which confirmed the stability of nanoparticles dispersion. Gold nanoparticles have shown high stability in biofluids of Bovine Serum Albumin and Phosphate Buffer Saline at pH-5, pH-7and pH-9 which mimic the human colonic biological environment.Conclusion: The fungal synthesis of AuNPs has been experimentally demonstrated and their stability in BSA, 10% NaCl and PBS at pH-7. This might be a promising option for drug delivery applications in carcinogenic colon disorders in human beings.

Au@pNIPAM SERRS Tags for Multiplex Immunophenotyping Cellular Receptors and Imaging Tumor Cells.

Detection technologies employing optically encoded particles have gained much interest toward clinical diagnostics and drug discovery, but the portfolio of available systems is still limited. The fabrication and characterization of highly stable surface-enhanced resonance Raman scattering (SERRS)-encoded colloids for the identification and imaging of proteins expressed in cells are reported. These plasmonic nanostructures are made of gold octahedra coated with poly(N-isopropylacrylamide) microgels and can be readily encoded with Raman active dyes while retaining high colloidal stability in biofluids. A layer-by-layer polyelectrolyte coating is used to seal the outer surface of the encoded particles and to provide a reactive surface for covalent conjugation with antibodies. The targeted multiplexing capabilities of the SERRS tags are demonstrated by the simultaneous detection and imaging of three tumor-associated surface biomarkers: epidermal growth factor receptor (EGFR), epithelial cell adhesion molecule (EpCAM), and homing cell adhesion molecule (CD44) by SERRS spectroscopy. The plasmonic microgels are able to discriminate tumor A431 (EGFR+/EpCAM+/CD44+) and nontumor 3T3 2.2 (EGFR-/EpCAM-/CD44+) cells while cocultured in vitro.

Studying the Effects of Cysteine Residues on Protein Interactions with Silver Nanoparticles.

Studies of protein and organothiol interactions with silver nanoparticles (AgNPs) are important for understanding AgNP nanotoxicity, antimicrobial activity, and material fabrications. Reported herein is a systematic investigation of the effects of both reduced and oxidized protein cysteine residues on protein interactions with AgNPs. The model proteins included wild-type and mutated protein GB3 variants that contain 0, 1, or 2 reduced cysteine residues, respectively. Bovine serum albumin (BSA) that contains a total of 34 oxidized (disulfide-linked) cysteine residues and one reduced cysteine residue was also included. Protein cysteine content has no detectable effect on the kinetics of protein/AgNP binding. However, only proteins that contain reduced cysteine residues induce significant AgNP dissolution. Proteins can slow down, but do not prevent the AgNP dissolution induced by subsequently added organothiols. The insights provided in this work are important to the mechanistic understanding of AgNP stability in biofluids that are rich in proteins and amino acid thiols.

Quantitative assessment of nanoparticle surface hydrophobicity and its influence on pulmonary biocompatibility

To date, the role of nanoparticle surface hydrophobicity has not been investigated quantitatively in relation to pulmonary biocompatibility. A panel of nanoparticles spanning three different biomaterial types, pegylated lipid nanocapsules, polyvinyl acetate (PVAc) and polystyrene nanoparticles, were characterized for size, surface charge, and stability in biofluids. Surface hydrophobicity of five nanoparticles (50–150nm) was quantified using hydrophobic interaction chromatography (HIC) and classified using a purpose-developed hydrophobicity scale: the HIC index, range from 0.00 (hydrophilic) to 1.00 (hydrophobic). This enabled the relationship between the nanomaterial HIC index value and acute lung inflammation after pulmonary administration to mice to be investigated. The nanomaterials with low HIC index values (between 0.50 and 0.64) elicited little or no inflammation at low (22cm2) or high (220cm2) nanoparticle surface area doses per animal, whereas equivalent surface area doses of the two nanoparticles with high HIC index values (0.88–0.96) induced neutrophil infiltration, elevation of pro-inflammatory cytokines and adverse histopathology findings. In summary, a HIC index is reported that provides a versatile, discriminatory, and widely available measure of nanoparticle surface hydrophobicity. The avoidance of high (HIC index>~0.8) surface hydrophobicity appears to be important for the design of safe nanomedicines for inhalation therapy.

In vitro toxicity of nanoceria: effect of coating and stability in biofluids

Due to the increasing use of nanometric cerium oxide in applications, concerns about the toxicity of these particles have been raised and have resulted in a large number of studies. We report here on the interactions between 7 nm anionically charged cerium oxide particles and living mammalian cells. By a modification of the particle coating including low-molecular weight ligands and polymers, two generic behaviours are compared: particles coated with citrate ions that precipitate in biofluids and particles coated with poly(acrylic acid) that are stable and remain nanometric. We find that nanoceria covered with both coating agents are taken up by mouse fibroblasts and localized into membrane-bound compartments. However, flow cytometry and electron microscopy reveal that as a result of their precipitation, citrate-coated particles interact more strongly with cells. At cerium concentration above 1 mM, only citrate-coated nanoceria (and not particles coated with poly(acrylic acid)) display toxicity and moderate genotoxicity. The results demonstrate that the control of the surface chemistry of the particles and its ability to prevent aggregation can affect the toxicity of nanomaterials.

Diisocyanate mediated polyether modified gelatin drug carrier for controlled release

Gelatin is an extensively studied biopolymer hydrogel drug carrier due to its biocompatibility, biodegradability and non-toxicity of its biodegraded products formed in vivo. But with the pristine gelatin it is difficult to achieve a controlled and desirable drug release characteristics due to its structural and thermal lability and high solubility in aqueous biofluids. Hence it is necessary to modify its solubility and structural stability in biofluids to achieve controlled release features with improved drug efficacy and broader carrier applications. In the present explorations an effort is made in this direction by cross linking gelatin to different extents using hitherto not studied isocyanate terminated poly(ether) as a macrocrosslinker prepared from poly(ethylene glycol) and isophorone diisocyanate in dimethyl sulfoxide. The crosslinked samples were analyzed for structure by Fourier transform-infrared spectroscopy, thermal behavior through thermogravimetric analysis and differential scanning calorimetry. The cross linked gelatins were biodegradable, insoluble and swellable in biofluids. They were evaluated as a carrier for in vitro drug delivery taking theophylline as a model drug used in asthma therapy. The crosslinking of gelatin decreased the drug release rate by 10–20% depending upon the extent of crosslinking. The modeled drug release characteristics revealed an anomalous transport mechanism. The release rates for ampicillin sodium, 5-fluorouracil and theophylline drugs in a typical crosslinked gelatin carrier were found to depend on the solubility and hydrophobicity of the drugs, and the pH of the fluid. The observed results indicated that this material can prove its mettle as a viable carrier matrix in drug delivery applications.

Comparative Investigations on In Vitro Serum Stability of Polymeric Micelle Formulations

Stability of polymeric micelles upon injection is essential for a drug delivery system but is not fully understood. We optimized an analytical test allowing quantification of micellar stability in biofluids and applied it to a variety of block copolymer micelles with different hydrophobic block architechtures. Polymeric micelles were prepared from four different polymers and investigated via encapsulation of two fluorescent dyes. Samples were incubated in human serum; changes in Foerster Resonance Energy Transfer (FRET) were recorded as a function of time. This fluorescence-based approach was supported semi-quantitatively by results from Asymmetrical Flow Field-Flow-Fractionation (AF4). After incubation experiments, micellar stability was determined by calculation of two stability-indicating parameters: residual micellar fractions (RMFs) and in vitro serum half-lives. Both parameters showed that PEG-PVPy micelles rapidly destabilized after 3h (RMF < 45%), whereas PEG-PLA, PEG-PLGA and PEG-PCL micelles were far more stable (RMFs 65 to 98%). This FRET-based assay is a valuable tool in evaluating and screening serum stability of polymeric micelles and revealed low serum stability of PEG-PVPy micelles compared to polyester-based micelles.

Activity of water-soluble turmeric extract using hydrophilic excipients

Turmeric ( Curcuma longa) is extensively used in a number of ailments by the traditional medical practitioners. Traditionally turmeric is extracted with different non-edible organic solvents or supercritical fluid extraction using carbon dioxide and suffers from many drawbacks as it fails to achieve aqueous solubility, acceptable content of active principle in edible form and stability in biofluids. In the present work we report application of food-grade polyglycolized glycerides to obtain water-soluble extract of turmeric, which contains lipophilic curcuminoids in the aqueous phase. Extraction of turmeric using amphiphilic vehicles such as Gelucire 44/14, Gelucire 50/13, and PEG 6000 was able to achieve aqueous solubility. Gelucire 44/14 gives the maximum concentration of curcuminoids than extraction with Gelucire 50/13 and PEG 6000, as quantified with RP-HPLC. Turmeric extract was further screened for in vitro free radical scavenging assays, ROS scavenging ability and cell proliferation activity in cell line.