Metabolomics of Respiratory Diseases.

Metabolomics is an expanding field of systems biology that is gaining significant attention in respiratory research. As a unique approach to understanding and diagnosing diseases, metabolomics provides a snapshot of all metabolites present in biological samples such as exhaled breath condensate, bronchoalveolar lavage, plasma, serum, urine, and other specimens that may be obtained from patients with respiratory diseases. In this article, we review the rapidly expanding field of metabolomics in its application to respiratory diseases, including asthma, chronic obstructive pulmonary disease (COPD), pneumonia, and acute lung injury, along with its more severe form, adult respiratory disease syndrome. We also discuss the potential applications of metabolomics for monitoring exposure to aerosolized occupational and environmental materials. With the latest advances in our understanding of the microbiome, we discuss microbiome-derived metabolites that arise from the gut and lung in asthma and COPD that have mechanistic implications for these diseases. Recent literature has suggested that metabolomics analysis using nuclear magnetic resonance (NMR) and mass spectrometry (MS) approaches may provide clinicians with the opportunity to identify new biomarkers that may predict progression to more severe diseases which may be fatal for many patients each year.

Exposure to heavy metals alters the surface topology of alveolar macrophages and induces respiratory dysfunction among Indian metal arc-welders.

BACKGROUND: Despite the available clinico-epidemiological evidence of heavy metal-associated respiratory health hazards among metal arc-welders, experimental confirmation of such an association is lacking. METHODS: In this study, we recruited 15 metal arc-welders and 10 referent workers without direct exposure. We assessed respiratory health through a questionnaire and spirometry; estimated manganese, nickel and cadmium levels in blood, urine and induced sputum; performed differential counts of sputum leucocytes and measured plasma malondialdehyde (MDA). We used atomic force and scanning electron microscopy to assess the physical property of the alveolar macrophages (AMs) obtained from induced sputum and analysed cell surface deposition of heavy metals using energy dispersion X-ray analysis (EDX). Sputum cellular DNA damage was assessed by DNA-laddering assay. RESULTS: There was a higher body burden of manganese and nickel in the metal arc-welders than the referents. Among major spirometric indices, only the forced mid-expiratory flow rates (FEF25-75) were reduced in the welders compared with the referents (63.4 ± 14.7 vs. 89.2 ± 26.7, p < 0.01); this reduction was associated with both heavy metal levels (ß: -41.8, 95% CI: -78.5% to -5.1%) and plasma MDA (-0.37; -0.68 to -0.06). In metal arc-welders, significant physical and morphological changes were observed in AMs through microscopic evaluation while EDX analyses demonstrated higher deposition of heavy metals on the AM cell surface than the referents. We also observed a higher degree of DNA damage in the sputum cells of the exposed workers than the referents. CONCLUSION: Heavy metal exposure-induced adverse respiratory effects among metal arc-welders are mediated through haematological and cytological interactions.

A study of the expression and localization of toll-like receptors 2 and 9 in different grades of cervical intraepithelial neoplasia and squamous cell carcinoma.

TLRs are important molecules of innate immune response, those play central role in host pathogen interaction and recognition through pathogen associated molecular patterns (PAMPs). Previous studies have indicated the role of TLRs in many human malignancy and cervical cancer in terms of viral recognition and inflammatory changes in-vivo. The objective of this study was to evaluate the expression and localization of toll-like receptor (TLR) 2 and TLR9 in preinvasive and invasive cervical cancer patients and to investigate its use as a probable diagnostic tool for better management cervical cancer. This single institution study includes individuals with normal, precancerous lesions, cervical intraepithelial neoplastic (CIN) and invasive squamous cell carcinoma (SCC) of the cervix. Upon confirmation by histopathology, fluorescence based immunohistochemistry was performed in all patients for TLR2 and TLR9, followed by semi-quantitative estimation of the staining intensity and grade of expression. The expression pattern of TLR2 and TLR9 does not vary greatly from normal to precancerous lesions, but a significant variation was observed in advance stages, i.e. squamous cell carcinoma of the uterine cervix. Additionally the expression increased marginally in higher grades. In spite of their low difference in expression along different stages of cervical cancer, both TLR2 and TLR9 could detect the disease at an advance stages as depicted by the receiver operator characteristics curve analysis.

A simple one pot purification of bacterial amylase from fermented broth based on affinity toward starch-functionalized magnetic nanoparticle.

Surface-functionalized adsorbant particles in combination with magnetic separation techniques have received considerable attention in recent years. Selective manipulation on such magnetic nanoparticles permits separation with high affinity in the presence of other suspended solids. Amylase is used extensively in food and allied industries. Purification of amylase from bacterial sources is a matter of concern because most of the industrial need for amylase is met by microbial sources. Here we report a simple, cost-effective, one-pot purification technique for bacterial amylase directly from fermented broth of Bacillus megaterium utilizing starch-coated superparamagnetic iron oxide nanoparticles (SPION). SPION was prepared by co-precipitation method and then functionalized by starch coating. The synthesized nanoparticles were characterized by transmission electron microscopy (TEM), a superconducting quantum interference device (SQUID, zeta potential, and ultraviolet-visible (UV-vis) and Fourier-transform infrared (FTIR) spectroscopy. The starch-coated nanoparticles efficiently purified amylase from bacterial fermented broth with 93.22% recovery and 12.57-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the molecular mass of the purified amylase was 67 kD, and native gel showed the retention of amylase activity even after purification. Optimum pH and temperature of the purified amylase were 7 and 50°C, respectively, and it was stable over a range of 20°C to 50°C. Hence, an improved one-pot bacterial amylase purification method was developed using starch-coated SPION.

Localized surface plasmon resonance-based DNA detection in solution using gold-decorated superparamagnetic Fe3O4 nanocomposite.

In the field of nucleic acid-based biosensor technology, DNA-conjugated nanocomposites have attracted much attention due to their unique properties and multimodal applicability. However, quantitative estimation of sequence-specific oligonucleotide in a simpler way is still a challenge. Precise positioning of DNA probes over the surface of the nanocomposite can overcome problems such as steric hindrance of the surface-bound molecules to enable further sensing as well as nonspecific folding of the DNA molecule over the surface of the gold (Au) nanolayer. Considering such objectives, we have developed glutathionated Fe3O4@Au core/shell nanocomposite, fabricated with DNA molecules and applied for sensing complementary oligo spectrophotometrically, using the localized surface plasmon resonance (LSPR) properties of the synthesized nanocomposite. When hybridization experiments were performed with 10 to 100 fM complementary DNA and DNA-conjugated nanocomposite, a strong linear relationship was observed between DNA concentration and surface plasmon resonance (SPR). Discrimination even at the single-base level was also observed when further experiments were performed with complementary DNA, but with a sequential decrease of bases from the single level to the fifth level.

Potential application of superparamagnetic nanoparticles for extraction of bacterial genomic DNA from contaminated food and environmental samples.

BACKGROUND: Isolation of high-molecular-weight DNA is essential for many molecular biology applications. Owing to the presence of polymerase chain reaction (PCR) inhibitors, there is a scarcity of suitable protocols for PCR-ready DNA extraction from food and natural environments. The conventional chemical methods of DNA extraction are time consuming and laborious and the yield is very low. Thus the aim of this research was to develop a simple, rapid, cost-effective method of genomic DNA extraction from food (milk and fruit juice) and environmental (pond water) samples and to detect bacterial contaminants present in those samples. RESULTS: This approach is efficient for both Gram-positive and Gram-negative bacteria from all the studied samples. Herein super paramagnetic bare iron oxide nanoparticles were implemented for bacterial genomic DNA isolation. The method was also compared to the conventional phenol-chloroform method in the context of quality, quantity and timing process. This method took only half an hour or less to obtain high-molecular-weight purified DNA from minimum bacterial contamination. Additionally, the method was directly compatible to PCR amplification. CONCLUSION: The problem of availability of suitable generalized methods for DNA isolation from various samples including food and environmental has been solved by a nanobiotechnological approach that may prove to be extremely useful in biotechnological applications.

Reproducibility of the "International Academy of Cytology Yokohama System for Reporting Breast Cytology" - A Retrospective Analysis of 70 Cases.

Background: Fine-needle aspiration cytology (FNAC) is the most practiced initial method for evaluation of breast lesions. The International Academy of Cytology Yokohama System for Reporting Breast (IAC YSRB) Fine-Needle Aspiration Biopsy Cytopathology has been developed to standardize the reporting system. However, literature available on the inter-observer reproducibility of 5 IAC YSRB categories is limited. Aim: We investigated the inter-observer reproducibility of the IAC YSRB system. Method and Materials: A total of 70 consecutive specimens obtained from FNAC of breast lesions were reviewed retrospectively by 3 experienced cytopathologists who allotted 1 to 5 IAC YSRB categories. Results: The percent overall agreement between observers was 70.48% and the free marginal kappa was 0.63, which signifies substantial agreement. After combining "Suspicious" and "Malignant" categories, overall agreement was 80.95% and free marginal kappa became 0.75. Conclusions: Inter-observer agreement of three (70.48%) cytopathologists was substantial. Agreement can be improved by combining certain categories, especially "Suspicious" and "Malignant". Technical quality limitation plays a significant role in a proportion of cases, mainly the "Atypical" and "Suspicious of malignancy" categories. Application of the IAC YSRB system in day-to-day practice will increase the inter-observer agreement.

Effect of iron oxide and gold nanoparticles on bacterial growth leading towards biological application.

BACKGROUND: Nanoparticle-metal oxide and gold represents a new class of important materials that are increasingly being developed for use in research and health related activities. The biological system being extremely critical requires the fundamental understanding on the influence of inorganic nanoparticles on cellular growth and functions. Our study was aimed to find out the effect of iron oxide (Fe3O4), gold (Au) nanoparticles on cellular growth of Escherichia coli (E. coli) and also try to channelize the obtained result by functionalizing the Au nanoparticle for further biological applications. RESULT: Fe3O4 and Au nanoparticles were prepared and characterized using Transmission electron microscopy (TEM) and Dynamic Light Scattering (DLS). Preliminary growth analysis data suggest that the nanoparticles of iron oxide have an inhibitory effect on E. coli in a concentration dependant manner, whereas the gold nanoparticle directly showed no such activity. However the phase contrast microscopic study clearly demonstrated that the effect of both Fe3O4 and Au nanoparticle extended up to the level of cell division which was evident as the abrupt increase in bacterial cell length. The incorporation of gold nanoparticle by bacterial cell was also observed during microscopic analysis based on which glutathione functionalized gold nanoparticle was prepared and used as a vector for plasmid DNA transport within bacterial cell. CONCLUSION: Altogether the study suggests that there is metal nanoparticle-bacteria interaction at the cellular level that can be utilized for beneficial biological application but significantly it also posses potential to produce ecotoxicity, challenging the ecofriendly nature of nanoparticles.

Toll-Like Receptor 9 Expression Levels in Breast Carcinoma Correlate with Improved Overall Survival in Patients Treated with Neoadjuvant Chemotherapy and Could Serve as a Prognostic Marker.

Aim: Toll-like receptor 9 (TLR9) can recognize the DNA fragments released from chemotherapy-treated cancer cells in tumor tissues and induce an inflammatory response. The aim of the present study was to evaluate the survivability benefit of TLR9 expression levels as a potential prognostic marker in breast cancer patients treated with neoadjuvant chemotherapy (NACT). Methods: To study the expression of TLR9 in breast tumor, immunohistochemical (IHC) analyses were performed on two patient cohorts, with NACT (n = 19) and without NACT (n = 23). To corroborate the findings from the in-house cohort, we also used publicly available datasets including SurvExpress (GSE 20685) and the Kaplan-Meier plotter tool (GSE 16446) to analyze the relationship between the expression of TLR9 and overall survivability for NACT. Results: The IHC analyses of our inhouse cohort demonstrated that TLR9 was expressed in both malignant breast epithelial cancer cells as well as in the adjacent stromal cells. The IHC results also indicated that, the percentage of malignant epithelial cells (54.76%) expressing TLR9 was higher than in the adjacent stromal compartment (11.9%). We also observed an increase in the expression levels of TLR9 in the patients who were given NACT (p = 0.0379). Further, the analysis of publicly available datasets demonstrated that elevated TLR9 expression was related to increased overall survival in patients treated with NACT. Conclusions: In this study, we show for the first time that elevated TLR9 tissue expression levels in breast cancer may serve as a prognostic marker for patients treated with NACT and could potentially be used to select the neoadjuvant regime.

Implication of therapeutic outcomes associated with molecular characterization of paediatric aplastic anaemia.

OBJECTIVES: Severe aplastic anemia is characterized by a hypocellular bone marrow and peripheral cytopenia. Mesenchymal stem cells (MSCs) play a crucial role in haematopoietic stem cells (HSCs) development and the development of microenvironment suitable for hematopoiesis. Molecular characterization of telomere maintenance pathway and gene expression profiling of MSCs can be important for the therapeutic interventions among paediatric aplastic anaemia patients. METHODS: The study involved paediatric aplastic anaemia patients (n = 10) and age matched paediatric healthy donors (n = 8). Peripheral blood samples were collected from the individuals. Average leucocyte telomere length and gene expression of the telomere maintenance genes were determined by quantitative real time PCR. Microarray based gene expression profiles (GSE33812) of MSCs for five paediatric aplastic anaemia patients were analyzed compared to five healthy controls and the data was downloaded from the GEO database. RESULTS: The telomere length was significantly shorter among paediatric AA patients compared to age matched healthy donors. Interestingly, one subgroup (n = 2) of paediatric AA patients has moderate telomere length comparable to age matched healthy donors. Based on the gene expression analysis of telomere maintenance pathway, TERF2 was significantly downregulated among paediatric patients with shorter telomere length but not among paediatric patients with moderate telomere length. Gene expression profiling of MSCs revealed three differentially expressed genes (GAS2L3, MK167 and TMSB15A) among the patients and was associated with therapeutic outcome. CONCLUSION: Telomere length estimation and gene expression patterns of the MSCs and telomere length maintenance pathway may serve as a potential biomarker and could be associated with therapeutic choice of paediatric aplastic anaemia patients.

A study of CD10 positivity of stromal cells in core needle biopsy specimen of breast cancer patients and its relation with histological grade and lymphovascular invasion.

BACKGROUND: In core needle biopsy (CNB) often the histological grade of invasive breast carcinoma is under-estimated due to heterogeneity of epithelial component. Stroma is relatively homogenous throughout the tumor and strong CD10 stromal positivity is proposed to be associated with high tumor grade. AIMS AND OBJECTIVES: The aim of this work was to study the expression of CD10 in stromal cells of invasive carcinoma of breast, no specific type (NST) in CNB specimens, and analyze its association with final histological grade and lymphovascular invasion (LVI). MATERIALS AND METHODS: A total of 50 cases of invasive carcinoma of breast, NST were studied for 18 months. CNB specimens were graded according to modified Scarff-Bloom-Richardson (SBR) system and CD10 positivity was assessed in stromal cells. Mastectomy specimens were also similarly graded. Relation of stromal CD10 positivity with histological grading and LVI was studied. STATISTICS: Associations between the variables were studied by Chi-square test. A value of P < 0.05 was considered to be statistically significant. RESULTS: On CNB 46% patients had a grade 2 tumor, followed by 30% grade 3 and 24% grade 1 tumor. Strong CD10 positivity was seen in 40% cases, 32% showed weak positivity and 28% were negative for CD10 in stromal cells in CNB specimen. On evaluation of mastectomy specimen 48% of the patients had a grade 2 tumor, followed by 40% grade 3 tumor and 12% grade 1 tumor. Strong CD10 positivity was found to be significantly associated with final grade 3 tumor (P < 0.001) and LVI (P = 0.005). CONCLUSIONS: There was underestimation of histological grade on CNB, while strong stromal CD10 positivity in CNB was significantly associated with final grade 3 tumor and LVI.

A report of two cases of dedifferentiated endometrioid carcinoma: A newly described underrecognized tumor of poor prognosis.

Dedifferentiated endometrioid carcinoma or dedifferentiated endometrioid adenocarcinoma (DEAC) is defined by the presence of undifferentiated carcinoma with endometrioid carcinoma. Undifferentiated component can be misinterpreted as solid component of high-grade endometrioid carcinoma or sarcomatous component of malignant mixed mullerian tumor. We present two cases of DEAC. Two postmenopausal women underwent hysterectomy for vaginal bleeding. Microscopically, sections from the endometrial tumors showed a biphasic growth consisting of an undifferentiated component and a glandular component with sharp transition between the two components. The undifferentiated component showed focal positivity for cytokeratin and vimentin, while glandular component was diffusely positive for cytokeratin and negative for vimentin expression.

Tumor Cell Representation by an Improvised Technique of Fine-Needle Aspiration Specimen Acquisition and Cell Block Preparation: Our Experience in Lung Cancer Cases in a Peripheral Center of Eastern India.

BACKGROUND: Being a minimally invasive diagnostic technique, Fine-Needle Aspiration Cytology (FNAC) has become the first-line test and corresponding aspirated material has become the target specimen for diagnosis and ancillary tests in lung carcinoma. Although the role of Cell Blocks (CBs) in diagnosis and in ancillary testing is well recognized in literature, limited attention has been paid to specimen procurement and triage in the preparation of CBs. In the present scenario, CBs are not consistently optimal because of its low cellularity. AIMS: This study is aimed to describe an improvised technique of specimen acquisition and cell block preparation in CT-guided FNACs of lung carcinoma cases in a resource-constrained center and to assess its efficacy for optimal representation of cellularity, morphology, and architecture. MATERIALS AND METHODS: Total 85 lung carcinoma cases undergoing CT-guided FNAC in our center from February 2017 to January 2018 were included in this study. 4 to 5 direct smears and subsequent CBs were made from material obtained by single pass. Cellularity of smears and corresponding cell blocks were assessed and categorized according to a scoring system (score 1 to 3 for number of cells <50, 50-100, >100, respectively). Preserved architecture and morphology were also assessed in smears and CBs. RESULTS: The evaluated samples showed a cellularity score 3 in 65.4%CBs and score 2 in 24.7% CBs. Overall, 90.1% cell blocks had acceptable cellularity. Cell morphology was preserved in all CBs of acceptable cellularity, except for two adenocarcinoma, one squamous cell carcinoma, and one small cell carcinoma blocks. Cellular architecture was also preserved in all CBs of acceptable cellularity. CONCLUSIONS: This simple improvised technique of CB preparation optimized its cellularity, morphology, and architectural preservation, even after adequate cellular FNA smears.

Appreciation of Pattern in Diagnosis of Lung Adenocarcinoma from Cytology Specimen: Our Experience with Fine Needle Aspiration Cytology and Cell Block in a Resource Constraint Setup.

BACKGROUND AND AIMS: Advent of personalised treatment needs correct diagnosis of lung adenocarcinoma with its molecular subtyping. Minimal use of special stain or immunohistochemistry (IHC) in small specimens save material for molecular testing. Various histologic patterns in adenocarcinoma (ADC) subtypes have different prognostic implications and current recommendation is to describe these patterns in small specimens. Aim of this study was to diagnose adenocarcinoma from cytology specimens depending on adenocarcinoma pattern on fine needle aspiration smears and cell blocks. We also studied the additional role of cell blocks as a platform for special stain and IHC. MATERIALS AND METHODS: Conventional smears and cell block (CB) preparation were examined from transthoracic CT guided FNA samples of suspicious lung malignancy cases. Clear defining architectural pattern and cytomorphological features in favour of adenocarcinoma were evaluated and mucin stain and IHC were used as and when required. RESULTS: A total of 86 cases were included in this study, of which 83 cases were diagnosed as adenocarcinoma, 52 (62.5%) showed clear cut evidence of adenocarcinoma from smears and CBs. CB morphology alone aided the diagnosis in 12. Various ADC patterns in combination or alone were appreciated in these 64 cases. Sixteen needed mucin stain and 3 needed IHC for diagnosis. Forty one were ADC with solid pattern of which 39 showed high nuclear grade. CONCLUSION: Adequately cellular FNA smears and corresponding cell blocks of optimal quality can aid effectively in diagnosing adenocarcinoma and appreciating its pattern. Therefore, it would minimize the need for special stain and/or IHC with preservation of more material for molecular testing.

Reproducibility of "the Papanicolaou Society of Cytopathology system for reporting respiratory cytology" - A retrospective analysis of 101 cases of CT-guided FNAC.

INTRODUCTION: Guided fine-needle aspiration cytology is a popular investigative procedure in diagnosing pulmonary lesions. The Papanicolaou Society of Cytopathology (PSC) has already outlined a categorical system for reporting respiratory cytology. Though each category has a known malignancy risk, their inter observer reproducibility have not been well documented. This study was directed towards establishing the reproducibility of this categorical system in diagnosing pulmonary lesions. METHOD: One hundred and one consecutive cytology specimens obtained by CT-guided FNA from lung lesions were independently reviewed by 3 experienced cytopathologists, who allotted each case to 1 of 6 PSC categories. Statistical analysis for percent overall agreement was done using Fleiss' Kappa. RESULT: Percent overall agreement was 71.29% and free marginal kappa was 0.66. On combining categories "suspicious" and "malignant" percent overall agreement was 79.54% and free marginal kappa was 0.74. CONCLUSION: There was substantial agreement among the observers as regards reproducibility of categories which can improve if we combine certain categories, especially "suspicious" and "malignant."

Galactocele of adult male breast: A cytopathologist's perspective.

Galactocele, although a common cytological diagnosis in females, is not previously reported as a cause of breast enlargement in adult males. Hyperprolactinemia is the principal cause of galactocele in male breast. Besides drug induced hyperprolactinemia, other anatomical lesions of hypothalamo-pituitary region and different medical conditions like cirrhosis and chronic kidney disease are to be considered along with a full evaluation of features revealing hypogonadism in case of galactocele. Aspirated milk from the male breast is the primary clue for this detailed investigation process. Here we are presenting the first case of galactocele of the male breast due to hypogonadotropic hypogonadism with idiopathic hyperprolactinemia.

The destructive spontaneous ingression of tunable silica nanosheets through cancer cell membranes.

Robust inorganic graphene analogues with atomic level sharp edges have seldom been investigated to decipher the interaction of two-dimensional materials with the cell membrane. Silica nanosheets (NSs) with four different thicknesses between 2.9 nm and 11.1 nm were synthesized by microwave irradiation and these colloidal NSs were able to spontaneously penetrate the cell membrane leaving membrane perforations at their sites of entry. The NS-ingression was most effective with the thinnest NSs, when studied in vitro. The atomistic details of the NS-membrane interaction were revealed by molecular dynamics (MD) simulations, which showed that the extraction of phospholipids was most favored when NSs were oriented vertically with respect to the membrane surface. While the folic acid modified NSs demonstrated a riveting tendency to penetrate the cancer cell membrane, co-modification with doxorubicin (DOX) unexpectedly reduced their capability. Migrating away from a conventional drug delivery approach, here we show that silica NSs can kill cancer cells primarily by mechanical scalpelling. Targeted ingress could be achieved through antibody conjugation on the NSs and thus only cancerous HeLa cells are affected by this treatment, leaving the normal HEK-293 cells intact. This destructive ingress through limited oxidative stress offers a previously unexplored route to treat fatal diseases without the necessity of transporting expensive drugs or radiation therapy, thereby bypassing deleterious side effects on healthy cells.

Cytology Microarray on Cell Block Preparation: A Novel Diagnostic Approach in Fluid Cytology.

BACKGROUND: The cytological examination of serous body effusions to diagnose and stage malignancy is well accepted in clinical medicine. Conventional smear (CS) and cell block (CB) study has to be complemented with immunohistochemistry (IHC) for a definitive diagnosis of malignancy and also to differentiate it from reactive mesothelial cells. Cytology microarray (CMA) is a modification of tissue microarray which involves core needle biopsy of multiple cell blocks and embedding it in a single block. AIM: The aim of this study was to assess the effectiveness of IHC technique in CMA for rapid diagnosis of malignancy and to reduce the cost of testing. MATERIALS AND METHODS: In this study, 82 pleural fluids were collected and subjected to CS and CB study followed by IHC in CMA blocks. Six commonly used antibodies were applied to confirm malignancy and diagnose the primary. RESULTS: Nineteen cases were diagnosed as malignancy by CB method. MOC-31 confirmed adenocarcinoma deposit in 67% cases of which 44% were proved to be of lung primary by TTF1. CONCLUSIONS: IHC on CMA blocks of effusion fluids is a very effective technique that can significantly reduce the cost of testing by >70%.

Enhancement of anticancer activity and drug delivery of chitosan-curcumin nanoparticle via molecular docking and simulation analysis.

Computational analyses followed by traditional wet-bench experiments have become a method of choice due to successful results. To enhance the solubility and bioavailability of curcumin within chitosan nanoparticle, we have exploited computational methodologies i.e. docking, BBD-RSM and MD simulation for the polymer selection, NPs' formulation, optimization and their stability confirmation in an aqueous medium, respectively. Formulated CSCur NPs were assessed for in-vitro release, which exhibited a sustained release pattern and four-fold higher cytotoxic activity in a nanoparticulated system. Enhanced uptake, apoptotic effect of CSCur NPs were established by morphological changes in cells as observed by fluorescence microscopy and FE-SEM. DNA damage, cell-cycle blockage and elevated ROS levels further confirm the anticancer activity of the CSCur NPs following apoptotic pathways. In-vivo study on Danio rerio, for uptake and toxicity reveal the particle's biocompatibility and nontoxicity. Therefore, CSCur NPs could be the potential formulation for a safe chemotherapeutic drug for cancer.