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A field experiment was conducted to study the effect of organic manures and growth regulators on the growth and yield of two varieties of strawberry namely ‘Sweet Charlie’ and ‘Winter Dawn’. Six treatments were taken combining three organic manures viz. Vermicompost @ 3.0 t/ha, Mustard oil cake @ 1.0 t/ha and Neem cake @ 1.0 t/ha and two concentrations of gibberellic acid (GA3) viz. 75 ppm and 100 ppm along with a control. Foliar application of GA3 was carried out at 40 and 60 days after planting whereas organic manures were applied as basal dose. Results of the study suggested that higher doses i.e. 100 ppm of GA3 along with vermicompost exhibited more vegetative growth whereas 75 ppm GA3 resulted in higher fruit set and yield in both the varieties. It was found that vermicompost @ 3.0 t/ha combined with 100 ppm GA3 recorded the highest plant height (24.7 cm and 21.4 cm) and numbers of leaves per plant (46.0 and 68.7) in both Sweet Charlie and Winter Dawn varieties, respectively. Whereas, highest fruit diameter (3.3cm and 3.4cm), fruit length (4.6cm and 4.8cm), fruit weight (18.2 g and 17.9 g), number of fruits per plant (24.6 and 32.0), yield per plant (447.8 g and 572.1 g) and yield per hectare (18.80 t and 24.03 t) were recorded under vermicompost @ 3.0 t/ha in combination with 75 ppm GA3 in both Sweet Charlie and Winter Dawn varieties, respectively. It was observed that Winter Dawn variety produced a 28.0% higher yield as compared to Sweet Charlie under the best treatment i.e. vermicompost @ 3.0 t/ha in combination with 75 ppm GA3.

The effect of gibberellic acid concentrations on fruit characters and yield of strawberry was studied on two varieties, namely ‘Sweet Charlie’ and ‘Winter Dawn’ during November, 2018 to March, 2019. Foliar application of three different concentration of GA3 viz. 50 ppm, 75 ppm and 100 ppm were done at 40 days and 60 days after planting. Experimental results showed that GA3 @ 75 ppm had the best effect to increase yield components and yield of both the varieties. Under GA3 @ 75 ppm, Winter Dawn exhibited higher fruit diameter (3.38cm) and fruit length (4.74cm) as compared to Sweet Charlie (3.28cm fruit diameter and 4.60cm fruit length). Though the fruit weight was slightly higher in Sweet Charlie (18.36g) as against Winter Dawn (17.90g) under the best treatment i.e. 75 ppm GA3 the yield recorded was higher in Winter Dawn (573.62g/plant and 24.04t/ha) as against Sweet Charlie (445.43g/plant and18.73t/ ha) due to higher number of fruits (32.44/plant) produced by Winter Dawn as compared to Sweet Charlie (24.94/plant). So, Winter Dawn variety produced 28.35% higher yield as compared to Sweet Charlie under GA3 @ 75 ppm treatment.

We report a significant improvement of adipose-derived mesenchymal stem cells' (ADMSCs) biocompatibility and proliferation on hierarchically patterned porous honey-incorporated silk fibroin scaffolds fabricated using a combination of soft lithography and freeze-drying techniques. Parametric variations show enhanced surface roughness, swelling, and degradation rate with good pore interconnectivity, porosity, and mechanical strength for soft-lithographically fabricated biomimetic microdome arrays on the 2% honey silk fibroin scaffold (PHSF2) as compared to its other variants, which eventually made PHSF2 more comparable to the native environment required for stem cell adhesion and proliferation. PHSF2 also exhibits sustained honey release with remarkable antibacterial efficacy against methicillin-resistant

Photosensitizer-functionalized reduced graphene oxide (rGO) nanoparticles are promising materials for photodynamic therapy in cancer management. In this study, rGO is synthesized by a green route employing glucose as the reducing agent and functionalized with photosensitizer, protoporphyrin IX (PPIX) in a convenient, single-step procedure. PPIX-functionalized rGO exhibits photodynamic effect against cancer cells (HeLa) at 0.001 mg mL-1 under visible light illumination (635 nm). A 50% elimination of HeLa cells after 5 min irradiation is observed while very low phototoxicity (80% cell viability) is noted against normal dermal fibroblast cells. A positive correlation with ROS accumulation and increased expression of caspase-3 in PPIX-functionalized rGO-treated cancer cells is also established. The results evidence a simple and cost-effective route for developing photosensitizer-functionalized rGO for effective and selective killing of cancer cells.

In recent days, vanadium complexes and nanoparticles have received sustainable attention owing to their vast applications in different fields. In the present study, we report a facile approach for the synthesis of irregular dumbbell shaped vanadium pentoxide nanoparticles (V2O5 NPs: 30-60 nm) via the polyol-induced microwave irradiation process along with calcination. The as-synthesized nanoparticles were characterized using various physico-chemical techniques (e.g. XRD, TEM, FT-IR, DLS and XPS). The cell viability assay showed that V2O5 NPs could efficiently inhibit the proliferation of different cancer cells (B16F10, A549, and PANC1), depicting their anti-proliferative activity. However, V2O5 NPs did not exert significant cytotoxicity to the normal cells (CHO, HEK-293 and NRK-49F), suggesting their biocompatible nature. Interestingly, these nanoparticles inhibited the proliferation and migration of the endothelial cells (HUVECs and EA.hy926) and disrupted the blood vasculature in a chick embryo model, indicating their anti-angiogenic properties. The mechanistic study revealed that the effective internalization of V2O5 NPs generated intracellular reactive oxygen species (ROS) which in turn up-regulated p53 protein and down-regulated survivin protein in cancer cells, leading to the apoptosis process. Furthermore, the administration of V2O5 NPs to melanoma bearing C57BL6/J mice significantly increased their survivability as compared to the control untreated tumor bearing mice, exhibiting the therapeutic potential of the nanoparticles against melanoma. Additionally, the in vivo toxicity study demonstrated no toxic effect in mice upon sub-chronic exposure to V2O5 NPs. Altogether, we strongly believe that V2O5 NPs could intrinsically provide a new direction for alternative therapeutic treatment strategies for melanoma and other cancers by employing their anti-angiogenic properties in the future.

Angiogenesis is a crucial biological process of development of blood vessels from pre-existing vasculature, which helps in several physiological functions including embryonic development, hair growth, ovulation, menstruation, tissue repair, and regeneration. Contrastingly, it is also imperative in various pathological conditions like cardiovascular/ischemic diseases, rheumatoid arthritis, cancers, ocular/retinal diseases, and others. These disease conditions are often treated by manipulating angiogenesis using different pro-angiogenic or antiangiogenic factors/molecules through either promoting or inhibiting this complex process, respectively. However, these conventional angiogenic treatment strategies fall short in attaining the desired therapeutic effect due to several limitations including low bioavailability, rapid clearance, high cost, nonspecificity, drug resistance and side effects. Therefore, it is high time for the advancement of different pro- and antiangiogenic materials that could overcome aforesaid limitations, followed by their effective use for the therapy of angiogenesis related diseases. Recently, nanotechnology has drastically advanced in various areas of biology and medicine including therapeutic angiogenesis. Globally, many research groups including ours explored various inorganic metal nanomaterials that could efficiently manipulate the angiogenesis process either by augmenting or inhibiting it. The extensive investigation of the mechanisms underlying nanomaterials-mediated manipulation of angiogenesis is also well-documented. In the present review article, we intend to introduce the recent developments of inorganic nanomedicine manipulating angiogenesis with major focus on pro-angiogenic nanomaterials and their therapeutic applications along with associated challenges and future directions.

Tobacco smoking is considered as one of the major risk factors for development of oral cancer. In vitro studies indicate that cigarette smoke initiates transformation of epithelial cells toward development of oral cancer through altering mitochondrial metabolic pathways. However the present in vitro models need to be improved to correlate these molecular changes with epithelial transformations. In present study, we investigated the association of mitochondrial metabolic events with oral cancer progression under cigarette smoke extract (CSE). In this regard, an in vitro model of oral keratinocyte cell line (MOE1A) was developed by exposing them with different concentrations of CSE. Alterations in cellular phenomena were confirmed by Fourier-transform infrared spectroscopy (FTIR) study, which indicated changes in important functional groups of CSE-induced oral cells. Enhanced reactive oxygen species (ROS) of exposed cells altered the mitochondrial metabolic activities in terms of increased mitochondrial mass and DNA content. Further, mitochondrial heme-metabolism was investigated and real-time PCR study showed altered expression of important genes like ALAS1, ABCB6, CPOX, FECH, HO-1. Both transcriptomic and proteomic studies showed up- and down-regulation of important biomarkers related to cellular cancer progression. Overall data suggest that CSE alters mitochondrial heme metabolic pathway and initiates cancer progression through modifying cellar biomarkers in oral epithelial cells.

Redox signaling plays a vital role in regulating various cellular signaling pathways and disease biology. Recently, nanomedicine (application of nanotechnology in biology and medicine) has been demonstrated to regulate angiogenesis through redox signaling. A complete understanding of redox signaling pathways influenced angiogenesis/antiangiogenesis triggered by therapeutic nanoparticles is extensively reviewed in this article. Recent Advances: In recent times, nanomedicines are regarded as the Trojan horses that could be employed for successful drug delivery, gene delivery, peptide delivery, disease diagnosis, and others, conquering barriers associated with conventional theranostic approaches.Physiological angiogenesis is a tightly regulated process maintaining a balance between proangiogenic and antiangiogenic factors. The redox signaling is one of the main factors that contribute to this physiological balance. An aberrant redox signaling cascade can be caused by several exogenous and endogenous factors and leads to reduced or augmented angiogenesis that ultimately results in several disease conditions.Redox signaling-based nanomedicine approach has emerged as a new platform for angiogenesis-related disease therapy, where nanoparticles promote angiogenesis via controlled reactive oxygen species (ROS) production and antiangiogenesis by triggering excessive ROS formation. Recently, investigators have identified different efficient nano-candidates, which modulate angiogenesis by controlling intracellular redox molecules. Considering the importance of angiogenesis in health care a thorough understanding of nanomedicine-regulated redox signaling would inspire researchers to design and develop more novel nanomaterials that could be used as an alternative strategy for the treatment of various diseases, where angiogenesis plays a vital role.

We have investigated the effect of piezoelectric actuating voltage on cell behaviour after drop on demand inkjet printing using mouse 3T3 cells as a model cell line. Cell viability after printing was assessed using a live/dead assay, Alamar Blue as an assay for cell proliferation, and propidium iodide (PI) and Texas Red labeled dextran molecular probes to assess cell membrane integrity. No significant difference was found for the cell death rate compared between an unprinted control population and after printing at 80, 90 and 100 V respectively. However, cell proliferation was lower than that of the control population at all time points post-printing. Cell membrane integrity was quantified using PI and dextran probes of mean molecular weight of 3, 10, 40 and 70 kDa. Total membrane damage (assessed by PI) increased with increasing piezoelectric actuator driving voltage and this was always greater than the unprinted control cells. The uptake of the labeled dextran only occurs after inkjet printing and was never observed with the control cells. The largest dextran molecular probe of 70 kDa was only taken up by cells after printing using the lower printing voltages of 80 and 90 V and was absent after printing at 100 V. At the two lower printing voltages, the membrane damage is recovered and no dextran molecule penetrate the cells 2 hours after printing. However, printing at 100 V leads to an increased uptake of 3 and 10 kDa dextran molecules, the retention of membrane porosity and continued uptake of these 3 and 10 kDa dextran for greater than 2 hours post-printing. We hypothesize that the change in membrane porosity with increasing actuation voltage can be explained by distinct nucleation and growth stages for pore formation in response to printing stress.

Zinc oxide nanoparticles (ZnO NPs) are currently among the most promising nanomaterials for theranostics. However, they suffer from some drawbacks that could prevent their application in nanomedicine as theranostic agents. The doping of ZnO NPs can be effectively exploited to enhance the already-existing ZnO properties and introduce completely new functionalities in the doped material. Herein, we propose a novel synthetic approach for iron-doped ZnO (Fe:ZnO) NPs as a multifunctional theranostic nanoplatform aimed at cancer cell treatment. Pure ZnO and Fe:ZnO NPs, with two different levels of iron doping, were synthesized by a rapid wet-chemical method and analyzed in terms of morphology, crystal structure and chemical composition. Interestingly, Fe:ZnO NPs featured bioimaging potentialities thanks to superior optical properties and novel magnetic responsiveness. Moreover, iron doping provides a way to enhance the electromechanical behavior of the NPs, which are then expected to show enhanced therapeutic functionalities. Finally, the intrinsic therapeutic potentialities of the NPs were tested in terms of cytotoxicity and cellular uptake with both healthy B lymphocytes and cancerous Burkitt’s lymphoma cells. Furthermore, their biocompatibility was tested with a pancreatic ductal adenocarcinoma cell line (BxPC-3), where the novel properties of the proposed iron-doped ZnO NPs can be potentially exploited for theranostics.

Umbilical cord-derived mesenchymal stem cells (UCDMSC) are attractive candidates for cell-based regenerative medicine. However, they are susceptible to replicative senescence during repetitive passaging for in-vitro expansion and induced senescence in an oxidative, inflammatory microenvironment in vivo. Aim of this study is to investigate if honey-incorporated matrices can be employed to reduce senescence of UCDMSC. Matrices were prepared by electrospinning solutions of honey with poly-vinyl alcohol (PVA). PVA:honey matrices exhibited free radical scavenging activity. Culture of UCDMSC on PVA:honey matrices showed improvement in cell proliferation compared to pure PVA nanofibres. Expression of vimentin indicated that mesenchymal phenotype is preserved after culturing on these matrices. Further, UCDMSC were serially subcultured and cells of two passages (P2 and P6) were evaluated for reactive oxygen species (ROS) load and senescence parameters. P6 cells showed a higher ROS load and β-galactosidase (β-gal) positive senescent cells compared to P2. However, culturing on PVA:honey substrates significantly reduced both ROS and β-gal markers compared to cells on PVA substrates. Honey contains several antioxidant and anti-inflammatory components, which can reduce the ROS-related senescence. Thus, it is concluded that honey containing nanofibres can be effective substrates for stem cell-based wound healing and regenerative medicine.

This work ensemble the structural features of Porous Silicon (PS) layers which are being fabricated employing modified stain etch method. Additionally, this study also categorizes the optimal etching parameters which are extensively used to grow Porous Silicon (PS) layers enabling Metal Induced Vapor Phase Stain Etching methodology. Various growth conditions are considered to fabricate uniform porous structures. The structural morphology of PS layers has been studied in scanning electron microscopy (SEM) that reveals macroporous nature of pore morphology supported by investigation of optical properties. This has been supported by characterizing ultraviolet-visible (UV-VIS) spectrophotometer absorption spectra. Surface and optical investigations discover uniform porous structure which augments the size of silicon substrate. This augmented surface of planar silicon substrate can be used as sensing platform for different kinds of sensors.

The design of peptide-based therapeutics is generally based on the replacement of l-amino acids with d-isomers to obtain improved therapeutic efficiency. However, d-isomers are expensive and frequently induce undesirable immune responses. In the present work, we demonstrate that an intra-mitochondrially self-assembling amphiphilic peptide exhibits analogous activity in both d- and l-isomeric forms. This outcome is in contrast to the general observation considering higher therapeutic efficiencies of d-isomers compared with l-analogues. This suggests that l-peptides overcome proteolytic degradation during intra-mitochondrial self-assembly both in vitro and in vivo.

Angiogenesis is a vital process that deals with the generation of new blood vessels from pre-existing vasculature and is well known to regulate various physiological as well as pathophysiological processes. We demonstrated that zinc oxide nanoflowers (ZONF) exhibited pro-angiogenic properties in endothelial cells through the production of intracellular reactive oxygen species (ROS), especially H2O2 (hydrogen peroxide). The immense importance of angiogenesis in ischemic and cardiovascular diseases highlights an urgent need to comprehend the detailed molecular mechanisms underlying the ZONF induced angiogenesis process. However, the exact mechanism and signaling pathways behind nanoflowers mediated angiogenesis still remain unclear. In the present study, we report that ZONF induce angiogenesis through MAPK/Akt/eNOS mediated nitric oxide formation, which further acts in a cGMP dependent manner. We strongly believe that exploration of the molecular mechanism and signaling pathways of ZONF driven angiogenesis would be helpful for the advancement of alternative and efficient treatment strategies for ischemic and cardiovascular diseases using a nanomedicine approach.

Background: The importance of inhibiting the kinases of the DDR pathway for radiosensitizing cancer cells is well established. Cancer cells exploit these kinases for their survival, which leads to the development of resistance towards DNA damaging therapeutics. Objective: In this article, the focus is on targeting the key mediator of the DDR pathway, the ATM kinase. A new set of quinoline-3-carboxamides, as potential inhibitors of ATM, is reported. Methods: Quinoline-3-carboxamide derivatives were synthesized and cytotoxicity assay was performed to analyze the effect of molecules on different cancer cell lines like HCT116, MDA-MB-468, and MDA-MB-231. Results: Three of the synthesized compounds showed promising cytotoxicity towards a selected set of cancer cell lines. Western Blot analysis was also performed by pre-treating the cells with quercetin, a known ATM upregulator, by causing DNA double-strand breaks. SAR studies suggested the importance of the electron-donating nature of the R group for the molecule to be toxic. Finally, Western-Blot analysis confirmed the down-regulation of ATM in the cells. Additionally, the PTEN negative cell line, MDA-MB-468, was more sensitive towards the compounds in comparison with the PTEN positive cell line, MDA-MB-231. Cytotoxicity studies against 293T cells showed that the compounds were at least three times less toxic when compared with HCT116. Conclusion: In conclusion, these experiments will lay the groundwork for the evolution of potent and selective ATM inhibitors for the radio- and chemo-sensitization of cancer cells.

Neuritogenesis, a complex process of the sprouting of neurites, plays a vital role in the structural and functional restoration of cerebral ischemia-injured neuronal tissue. Practically, there is no effective long-term treatment strategy for cerebral ischemia in clinical practice to date due to several limitations of conventional therapies, facilitating the urgency to develop new alternative therapeutic approaches. Herein, for the first time we report that pro-angiogenic nanomaterials, zinc oxide nanoflowers (ZONF), exhibit neuritogenic activity by elevating mRNA expression of different neurotrophins, following PI3K/Akt-MAPK/ERK signaling pathways. Further, ZONF administration to global cerebral ischemia-induced Fischer rats shows improved neurobehavior and enhanced synaptic plasticity of neurons via upregulation of Neurabin-2 and NT-3, revealing their neuroprotective activity. Altogether, this study offers the basis for exploitation of angio-neural cross talk of other pro-angiogenic nano/biomaterials for future advancement of alternative treatment strategies for cerebral ischemia, where neuritogenesis and neural repair are highly critical.

A colorimetric and fluorescence turn-off probe 10-(4-azido phenyl)-5,5-difluoro-5h-dipyrrolo[1,2-c:1’,2’-f][1,3,2] diazaborinin-4-ium-5-uide, 1, for selective detection of H2S is reported. The probe displayed a robust decrease in fluorescence intensity with high sensitivity, specificity and least toxicity to detect exogenous H2S and also in live normal human oral fibroblast cells loaded with probe 1. The detection limit of probe 1 being 0.17 µm for H2S. A colorimetric and fluorescence turn-off BODIPY probe 1, for selective detection of exogenous H2S in live normal human oral fibroblast cells loaded with probe 1 and poisonous H2S gas formed under biodegradation in sewage waters and water tanks and corroded pipe lines. The detection limit of probe 1 being 0.17 μM

We report potentiation of healing efficacy of alginate by value addition at its structural level. Dual crosslinked (ionically and covalently) sodium alginate hydrogel coupled with honey (HSAG) brings about an intermediate stiffness in the fabric, confers consistent swelling property and limits erratic degradation of the polymer which ultimately provides conducive milieu to cellular growth and proliferation. In this work honey concentrations in HSAGs are varied from 2% to 10%. FTIR, XRD and nanoindentation studies on the HSAGs exhibited physicochemical integrity. In vitro degradation study provided the crucial finding on 4% HSAG having controlled degradation rate up to 12 days with a weight loss of 87.36 ± 1.14%. This particular substrate also has an ordered crystalline surface morphology with decent cellular viability (HaCaT and 3T3) and antimicrobial potential against Methicillin Resistant Staphylococcus aureus (MRSA) and Escherichia coli. The in vivo wound contraction kinetics on murine models (4% HSAG treated wound contraction: 94.56 ± 0.1%) has been monitored by both invasive (histopathology) and noninvasive (Swept Source Optical Coherence Tomography) imaging and upon corroborating them it evidenced that 4% HSAG treated wound closure achieved epithelial thickness resembling to that of unwounded skin. Thus, the work highlights structurally modified alginate hydrogel embedded with honey as a potential antimicrobial healing agent.

The process of angiogenesis, involving generation of new blood vessels from the existing ones, is vital for the supply of oxygen and nutrients to various tissues of body system. Angiogenesis is directly associated with several physiological and pathological processes. It is well-established that impairment in angiogenesis process results in various fatal conditions. Recently, few research groups including ours demonstrated therapeutic angiogenesis through nanomedicine approach using metal oxide/hydroxide nanoparticles. However, there is still a thorough necessity for the development of novel, eco-friendly, pro-angiogenic nanomaterials. Hence, in the present study we demonstrate the in vitro and in vivo pro-angiogenic properties of terbium hydroxide nanorods (THNRs) synthesized using an advanced microwave irradiation method, along with the detailed molecular signaling cascade underlying THNRs induced angiogenesis. The in vivo wound healing and nonimmunogenicity of the THNRs have been validated in the mouse models. We thus strongly believe that the present study establishing the pro-angiogenic properties of THNRs will aid in the development of alternative treatment strategies for wound healing along with cardiovascular and ischemic diseases, where angiogenesis is the chief target.

Edible mushrooms are natural sources for numerous bioactive properties which can protect human body from oxidative damages, hypertension, abnormal cell growth etc., The present study disclosed that, bioactive properties of mushroom proteins are significantly enhanced by adopting enzymatic hydrolysis. This study established the mushroom protein hydrolysates (MPHs), produced from freshly cultivated edible mushroom, Pleurotus ostreatus, could be the supreme source of both bioactive and nutraceutical compounds. Here, both unhydrolysed mushroom extract (UME) and proteinase K treated MPHs of different degrees of hydrolysis DH% (55%, 60%, 65%, 70% and 80%) under optimized condition were evaluated for antioxidative, angiotensin rennin converting enzyme (ACE) inhibitory and antiproliferative activities at different concentrations (2.5, 5 and 10 mg/ml). Among these, 65% DH at 10 mg/ml exhibited profound antioxidative (DPPH, ABTS, hydroxyl radical scavenging, metal chelating, linoleic acid auto-oxidation, FRAP, and reducing power) and ACE inhibitory activities. Maximum amount of total phenolic and flavonoid content was also obtained in this case. MTT assay confirmed the remarkable antiproliferative properties of MPHs against human cervical carcinoma cell line (HeLa). Presence of a wide range of phenolic (i.e., gallic, trans-cinnamic, caffeic, ferulic, aspartic and vanillic acid) and flavonoid (quercetin) compounds in both UME and MPHs were identified through HPLC.

Nanosized reduced graphene oxide (rGO) is found in active microcarbon used in popular face cream from the manufacturers like Ponds, Nevia, and Garnier which, under visible light exposure, gets activated by aerial oxygen to generate reactive oxygen species (ROS) harmful to skin.

In the last decade, gold nanoparticles have emerged as promising agents for in vitro bio-sensing and in vivo cancer theranostics. However, different investigations have reported widely varying cytotoxicity and uptake efficiency of gold nanoparticles depending upon their size. Therefore, more extensive studies are needed to standardize these biological effects as a function of size on a particular cell line. In addition, to obtain robust confirmation on the correlation of a size to biological effect, thorough mechanistic study must also be performed. In this study, the size dependent biological activities of gold nanoparticles on osteosarcoma cells is investigated towards exploring their potential theranostic application in bone cancer, for which very scarce literature reports are available. Tris-assisted citrate based method was optimized to synthesize stable gold naoparticles of 40–60 nm sizes. Nanoparticles were characterized through UV–Vis spectroscopy, field emission scanning electron microscope (FESEM) and dynamic light scattering (DLS). Increasing concentrations of gold nanoparticles (AuNPs) of 46 nm size, enhanced the rate of reactive oxygen species (ROS)-induced apoptosis in MG63 cells by disrupting their mitochondrial membrane potential. Considerably higher cell death was observed for 46 and 60 nm AuNPs compared to 38 nm at all concentrations of 200, 400 and 800 ng/mL. Further, molecular signatures of cellular apoptosis under nanoparticle treatment were optically assessed through surface enhanced Raman scattering (SERS). A significant Raman enhancement in cancer cells under treatment of larger gold nanoparticles (46 and 60 nm) at fixed wavelength of 785 nm and laser power of 8.0 mW was evident. In corroboration with molecular biology techniques, SERS observation confirmed the size-dependent apoptotic phenomena in osteosarcoma cells under treatment of gold nanoparticles. Study demonstrates a facile, non-active targeting approach for detection of size-dependent AuNP-induced apoptosis in osteosarcoma cells through label-free SERS method.

In the perspective of dental restorative applications, co-doped bioceramics have not been explored much. From the clinical perspective, a successful dental implant is expected to interact with peri-prosthetic bones, gingival tissue, and surrounding connective tissues. The interaction of implant and implant coating materials with bone tissue is well studied. However, their interaction with surrounding epithelial components needs scientific validation. In this context, the present study aims at quantitative evaluation of the electrical properties of Fe/Sr co-doped biphasic calcium phosphate (BCP) samples and assessment of their cytocompatibility with epithelial (vero) cells. Sr/Fe co-doped BCPs were prepared by sol-gel synthesis technique, with different dopant concentration. Impact of co-doping on conductivity was assessed and interestingly an increase in conductivity with dopant amount was recorded in different co-doped BCPs. Cellular study showed the significant ( p = 0.01) increase in both cellular viability and functionality with increasing conductivity of samples. Higher epithelial cell adhesion indicates that (Sr/Fe) co-doped BCP would be favorable for faster epithelial sealing and also would reduce the chances of infection. Real-time PCR and immunofluorescence studies indicated that the expression of the epithelial marker (E-cadherin) significantly ( p = 0.01) increased in 10, 30 and 40 mol% co-doped samples in comparison to undoped BCP. In contrast to E-cadherin, fold change of β-catenin remains unchanged amongst the co-doped ceramics, implying the absence of tumorigenic potential of (Sr/Fe) co-doped BCP. In addition, immune-fluorescence signatures for cellular polarity are established from enhanced expression PARD3 protein, which has major relevance for cellular morphogenesis and cell division. Summarizing, the present study establishes the efficacy of Sr/Fe co-doped BCPs as a dental implant coating material and its ability to modulate vero cell functionality.

Impedimetric biosensor is commonly applied for analysis of a wide variety of physical and chemical properties. Impedimetric techniques have been employed in monitoring different catalytic enzymatic reactions for identification of binding of specific proteins, nucliec acids, antibodies, cell receptors, or other biomolecules. In this process, impedance of any system under examination is measured over a wide range of alternating current frequencies. Other than different biomecules, this method is also important for assessment of fuel cells coating as well as in corrosion phenomena. The biological reactions occurring on the surface of electrodes is monitored through impedimetric measurements. Generally, biological molecules like enzymes, antibodies, nucleic acids, cells, etc., are immobilized on the surface of impedimetric biosensor electrodes. Various research groups have explored the interaction between the biomolecules with the conductive or semiconductive surface of biosensors. Impedimetric sensors are available in different forms depending on the target molecules under investigation. Some types of biosensors are enzyme- and protein-based sensor, immuno-biosensors, aptamer-based biosensors, cell-based sensors, etc. In this chapter, we will discuss these biosensor types, their applications in biomedical fields, and recent developments in the area of impedimetric biosensors.

Nanotechnology brings an extensive alteration in modern diagnostic and therapeutic treatment strategies for different diseases. In view of a significant implication of metals/metal ions in different physiological processes, scientists had earlier developed various kinds of metal nanoparticles for diverse biomedical applications. However, there is a growing concern of toxicity and biosafety of medicinally important metal nanoparticles, which necessitates the fabrication of metal nanoparticles implementing green chemistry principle. Recently, several research groups have developed green-synthesized metal nanoparticles using polysaccharides (e.g., starch, cellulose, pectin, glucan, xanthan, dextrose, etc.) of different bio-resources (e.g., plant, bacteria, fungus, etc.) due to various advantages of green approach such as simple, fast, easy, safer, cost effective, and less toxic route as compared to other conventional chemical methods. This chapter describes the overview of recent advancement of green-synthesized metal nanoparticles employing polysaccharides for different biomedical applications including drug delivery, anticancer activity, antimicrobial activity, wound healing, bio-imaging, bio-sensing, blood coagulation, etc. Considering the promising clinical applications, in vivo toxicity studies of these nanomaterials are also discussed in a brief manner. Finally, we conclude with different challenges and future prospects of green-synthesized metal nanoparticles for their versatile biomedical applications.

FTIR spectroscopy and Raman spectroscopy of biological analytes are increasingly explored as screening tools for early detection of cancer. In the present study, an integrated analysis of the FTIR and Raman spectra obtained from exfoliated cells is adopted to improve discrimination of normal, pre-cancerous and cancerous conditions. Multiple spectra were obtained from 13 normal, 13 pre-cancer and 10 cancer patients in both modes. Compared to normal patients, significant differences were observed at 1550, 1580, 1640, 2370, 2330, 2950-3000 and 3650-3750 cm-1 (FTIR) and 520, 640, 785, 827, 850, 935, 1003, 1175, 1311 cm-1 and 1606 cm-1 (Raman) vibrations of the other two. The increase in DNA, protein and lipid content with malignancy was more clearly elucidated by examining both spectra. Principal component analysis (PCA)-linear discriminant analysis (LDA) with 10-fold cross validation of the FTIR and Raman spectral data sets showed efficient discrimination between normal and pathological conditions while overlapping was seen between the two pathologies. The PCA-LDA model of the dual spectra yielded a classification accuracy of 98% in comparison with either FTIR (85%) or Raman (82%) in a spectrum-wise comparison. In the patient-wise approach (mean of all spectra from a patient), the overall classification efficiency was 73%, 80% and 87% for FTIR, Raman and integrated spectral approaches respectively. Moreover, the efficiency of the integrated FTIR-Raman PCA-LDA model as a prediction tool was tested to screen susceptible individuals (11 cigarette smokers) using the dual spectra acquired from these individuals. The study presents proof-of-concept for adopting a large-scale, follow-up trial of the approach for mass screening purposes.

Stem-cell (SC) chirality or left-right (LR) asymmetry is an essential attribute, observed during tissue regeneration. The ability to control the LR orientation of cells by biophysical manipulation is a promising approach for recapitulating their inherent function. Despite remarkable progress in tissue engineering, the development of LR chirality in SCs has been largely unexplored. Here, we demonstrate the role of substrate stiffness on the LR asymmetry of cultured mesenchymal stem cells (MSCs). We found that MSCs acquired higher asymmetricity when cultured on stiffer PCL/collagen matrices. To confirm cellular asymmetry, different parameters such as the aspect ratio, orientation angle and intensity of polarized proteins (Par) were investigated. The results showed a significant (p < 0.01) difference in the average orientation angle, the cellular aspect ratio, and the expression of actin and Par proteins in MSCs cultured on matrices with different stiffnesses. Furthermore, a Gaussian support-vector machine was applied to classify cells cultured on both (2% and 10% PCL/Collagen) matrices, with a resulting accuracy of 96.2%. To the best of our knowledge, this study is the first that interrelates and quantifies MSC asymmetricity with matrix properties using a simple 2D model.

The present study investigates the efficiency of electrospun montmorillonite-impregnated cellulose acetate nanofiber membranes (MMT-CA-NFM) for adsorptive removal of ciprofloxacin (CIP). Surface morphology, nanofiber characteristics and pHzpc of NFM were analysed by FE-SEM, EDS and pH drift method, respectively. The surface morphology of NFM showed random arrangements of nanofiber. Experiments were performed in batch mode to examine the effect of contact time, adsorbent dose, initial CIP concentration, pH, electrolyte, temperature and agitation speed. It was observed that 60 min contact was sufficient to reach reaction equilibrium and corresponding percentage removal was 76 % when CIP concentration, MMT-CA-NFM dose and solution pH were 10 mg/L, 4 g/L and 6.0 respectively. Equilibrium of the adsorption was described well by both Langmuir and Freundlich isotherm models. The sorption capacity was determined to be 13.8 mg/g. The sorption most closely followed Elovich kinetics model among the models tested. The pH range 6–7.5, where the zwitter ionic nature of CIP was dominant, was found to be most favourable for adsorption. Decrease of ΔG with increase in temperature demonstrated that CIP removal was favourable at higher temperature and positive values of ΔH confirmed endothermic nature of reaction. In continuous mode of operation using fixed bed column, the height of mass transfer zone was found to be 14.96 cm and adsorption rate and capacity constant in continuous mode were also calculated. The regeneration experiments indicated that 10 mM NaOH solution can desorb CIP from exhausted NFM and adsorbent demonstrated good reusability for removal of the antibiotic studied.

Two different silica based (MSU-2 and MCM-41) curcumin loaded mesoporous materials V3 and V6 were synthesized and characterized by several physico-chemical techniques. Release kinetic study revealed the slow and sustained release of curcumin from those materials in blood simulated fluid (pH: 7.4). The materials V3 and V6 were found to be biocompatible in non-cancerous CHO cell line while exhibiting significant cytotoxicity in different cancer cells (human lung carcinoma cells: A549, human breast cancer cells: MCF-7, mouse melanoma cells: B16F10) compared to pristine curcumin indicating the efficacy of the mesoporous silica materials based drug delivery systems (DDSs). The generation of intracellular reactive oxygen species (ROS) and down regulation of anti-apoptotic protein leading to the induction of apoptosis were found to be the plausible mechanisms behind the anti-cancer activity of these DDSs. These results suggest that curcumin-loaded drug delivery system may be successfully employed as an alternative treatment strategy for cancer therapeutics through a nanomedicine approach in near future.

A rapid method for qualitative and quantitative assaying of l-lactate in milk is described. The method is based on oxidation of lactate by lactate oxidase with the simultaneous formation of H2O2 which subsequently oxidises 2, 4, 6-tribromo 3-hydroxy benzoic acid (TBHBA) in the presence of peroxidase. Oxidised TBHBA then reacts with 4-amino antipyrine to obtain magenta colour (λmax = 523 nm). In the qualitative assay, colour change becomes visible in raw milk samples spiked with 300 mg lactate/L. The absorbance increases linearly up to 1000 mg/L. The method can be used for rapid assessment of the hygienic quality of milk.

Prior studies reported significant anticancer activities of ceramides. However, anticancer activities of homoserine based ceramides have not been tested. With a view to compare the anticancer activity of ceramides and homoceramides, in the present study, we have synthesized four serine based and four homoserine based C8-ceramide analogues. Since many cancer cells have shown resistance to ceramides, curcumin is now being used in combination with ceramides because of its ability to reverse multidrug resistance. Aimed at targeting curcumin-ceramide combination to tumor endothelial cells, herein we have used a tumor vasculature targeting liposomes of a newly synthesized pegylated RGDGWK-lipopeptide. Importantly, the liposomal formulations of the homoserine based C8-ceramide analogue containing oleyl chain showed more promising antineoplastic activities under both in vitro and systemic settings than the liposomal formulations of commercially available C8-ceramide. Findings in the mouse tumor growth inhibition study revealed synergistic therapeutic benefit from simultaneous delivery of curcumin and a homoserine based ceramide containing oleyl chain to tumor vasculature. Results in RT-PCR and Western blot experiments suggest that inhibition of solid tumor growth is mediated via inhibition of PI3K-Akt signaling pathway. The present structure-activity study is the first report to demonstrate therapeutic promise of curcumin-homoserine based ceramide combination in antiangiogenic cancer therapy.

Accelerating the clearance of intracellular protein aggregates through elevation of autophagy represents a viable approach for the treatment of neurodegenerative diseases. In our earlier report, we have demonstrated the enhanced degradation of mutant huntingtin protein aggregates through autophagy process induced by europium hydroxide nanorods [EHNs: Eu(III)(OH)3], but the underlying molecular mechanism of EHNs mediated autophagy was unclear. The present report reveals that EHNs induced autophagy does not follow the classical AKT-mTOR and AMPK signaling pathways. The inhibition of ERK1/2 phosphorylation using the specific MEK inhibitor U0126 partially abrogates the autophagy as well as the clearance of mutant huntingtin protein aggregates mediated by EHNs suggesting that nanorods stimulate the activation of MEK/ERK1/2 signaling pathway during autophagy process. In contrast, another mTOR-independent autophagy inducer trehalose has been found to induce autophagy without activating ERK1/2 signaling pathway. Interestingly, the combined treatment of EHNs and trehalose leads to more degradation of mutant huntingtin protein aggregates than that obtained with single treatment of either nanorods or trehalose. Our results demonstrate the rational that further enhanced clearance of intracellular protein aggregates, needed for diverse neurodegenerative diseases, may be achieved through the combined treatment of two or more autophagy inducers, which stimulate autophagy through different signaling pathways.

The presence of leaky vasculature and the lack of lymphatic drainage of small structures by the solid tumors formulate nanoparticles as promising delivery vehicles in cancer therapy. In particular, among various nanoparticles, the mesoporous silica nanoparticles (MSN) exhibit numerous outstanding features, including mechanical thermal and chemical stability, huge surface area and ordered porous interior to store different anti-cancer therapeutics with high loading capacity and tunable release mechanisms. Furthermore, one can easily decorate the surface of MSN by attaching ligands for active targeting specifically to the cancer region exploiting overexpressed receptors. The controlled release of drugs to the disease site without any leakage to healthy tissues can be achieved by employing environment responsive gatekeepers for the end-capping of MSN. To achieve precise cancer chemotherapy, the most desired delivery system should possess high loading efficiency, site-specificity and capacity of controlled release. In this review we will focus on multimodal decorations of MSN, which is the most demanding ongoing approach related to MSN application in cancer therapy. Herein, we will report about the recently tried efforts for multimodal modifications of MSN, exploiting both the active targeting and stimuli responsive behavior simultaneously, along with individual targeted delivery and stimuli responsive cancer therapy using MSN.

In the present paper, we synthesized and characterized four N-donor polypyridyl copper(II) complexes (C1-C4); [Cu(mono-CN-PIP)2]2+ (C1), [Cu(tri-OMe-PIP)2]2+ (C2), [Cu(di-CF3-PIP)2]2+ (C3) and [Cu(DPPZ)2]2+ (C4). The (Calf-Thymus) CT-DNA binding studies depicted that the complexes could interact with DNA via intercalative mode. All the complexes, particularly C3 and C4 attenuated the proliferation as well as migration of various cancer cells, indicating their anti-cancer and anti-metastatic activity. Additionally, chick embryo angiogenesis (CEA) assay exhibited the inhibition of vascular sprouting in presence of C3 and C4, suggesting their potential in inhibiting the blood vessel growth. Mechanistic studies revealed that the complexes induced the excessive production of cellular reactive oxygen species (ROS) leading to apoptosis through up regulation of p53 and downregulation of Bcl-xL, which might be the plausible mechanisms underlying their anti-cancer properties. To understand the feasibility of practical application of anti-cancer copper complexes C3 and C4, in vivo sub-chronic toxicity study (4 weeks) was performed in C57BL6 mice and the results exhibited almost non-toxic effects induced by these complexes in terms of haematology and serum biochemical analyses, suggesting their biocompatible nature. The current study provides the basis for future advancement of other novel biocompatible metal complexes that could be employed for the therapy of different cancers.

Combining hydrated polymeric networks containing physically or covalently cross-linked three-dimensional, swollen structures with micro- and nanoparticles (carbon, glass, silica, ceramic, polymeric, or metallic) to form a composite structure is emerging as an attractive means to impart superior functionality for various biomedical applications such as biosensing and drug delivery. The blending results in synergism in various properties such as mechanical strength of the polymer gel while mutually benefitting dispersed particle system by reducing their aggregation. In this chapter, advances in such particle-loaded gel systems are provided with discussion on mechanical, optical, and stimuli-responsive properties. The applications of microparticle-loaded gels and nanoparticle-loaded gels are then expounded separately to give the reader a broad view of the current state of the art and applicability.

Synthetic polymer gels possess unique combination of acceptable biocompatibility with tailorable mechanical and degradation properties. They can be synthesized by several methods with a wide compositional range. Their applications are thus witnessing rapid growth in drug delivery, regenerative medicine, microfluidics, bioprinting, stem cell delivery, etc. Scaffolds fabricated from such gels can mimic the natural microenvironment and provide variety of biophysical cues for cellular signaling. In this chapter, we provide a brief on properties of common synthetic gels such as poly(ethylene glycol), poly(vinyl alcohol), poly(methyl methacrylate), poly(hydroxyethylmethyl acrylate), polyurethanes, poly(amino acids), and poly(vinyl pyrrolidone). The important properties of several other synthetic polymer gels are also tabulated. The major application areas of the synthetic gels are also presented to the reader.

The aim of this study was to microencapsulate the reuterin produced by Lactobacillus reuteri BPL-36 strain for its long-term efficacy against food-borne pathogen Listeria monocytogenes. Lactobacillus reuteri BPL-36 strain previously isolated from a human infant fecal sample in lab was selected for the present study based on its ability to produce reuterin. The organism displayed a broad-spectrum antimicrobial activity. Reuterin concentration of 89.63 mM was obtained in the MRS–glycerol medium after 16 h incubation at 37 °C. The reuterin concentration required to inhibit the growth of Pseudomonas aeruginosa, Escherichia coli O157: H7, Salmonella typhi, Staphylococcus aureus, and Listeria monocytogenes was found to be 1.0, 2.0, 2.0, 4.0, and 10.0 AU/mL, respectively. Microencapsulation of reuterin to enhance long-term efficacy against food-borne pathogens was done. Results in this study indicated that the release characteristics of reuterin from the encapsulated particles were pH dependent. The release characteristics were unaffected by the storage of encapsulated reuterin at 4 °C for 2 weeks. The anti-listerial efficacy of the encapsulated reuterin was tested against L. monocytogenes in the BHI medium adjusted to pH 5.0 with a reuterin content equivalent to 16 mM, similar to un-encapsulated (free) reuterin. Encapsulated reuterin demonstrated enhanced efficacy against L. monocytogenes for longer duration of time when compared with un-encapsulated (free) reuterin. The present work demonstrated a novel antimicrobial delivery system that ensured much better capability of inhibiting the growth of L. monocytogenes throughout 24 h incubation at 37 °C.

Docetaxel (DTX) is an anticancer drug that is used alone and in combination with other drugs to treat tumours. However, it suffers from the drawback of non-specific cytotoxicity. To improve the therapeutic potential of DTX, we report the synthesis of cRGDfK peptide-conjugated succinoyl-TPGS (tocopheryl polyethylene glycol succinate) nanomicelles for targeted delivery of DTX. Among RGD (Arg-Gly-Asp) peptides, cRGDfK peptide shows specificity towards α v β 3 integrin receptors that are most commonly over-expressed in tumour cells. To cRGDfK peptide, succinoylated TPGS was synthesised and conjugated to cRGDfK peptide using a carbodiimide reaction. Peptide-conjugated DTX loaded nanomicelles (PDNM) displayed small particle size with a narrow distribution, controlled drug release and high physicochemical stability. Cytotoxicity, cellular uptake, apoptosis and anti-angiogenic comparisons of unconjugated nanomicelles to PDNM in DU145 human prostate cancer cells and HUVECs (Human Umblical Vein Endothelial Cells) clearly revealed the importance of the cRGDfK peptide in enhancing the drug delivery performance of nanomicelles. From the Clinical Editor Common to many chemotherapeutic agents for cancer, systemic toxicity remains a big concern. In this article, the authors attempted to address this issue by conjugating RGD based peptides to Docetaxel, which would target integrins expressed on tumor cell surface. The experimental data revealed enhanced drug delivery.

The identification of a new class of luminogens based on pyrrolo-quinoxaline embedded coumarin (PQC) structures is reported. The reaction relies on the gold-catalyzed hydroarylation reaction of appropriately substituted aryl alkynoates that produces molecules that exhibit solvatochromism and aggregation-induced emission enhancement (AIEE) phenomena. The application of PQCs for live cell imaging has also been demonstrated.

A series of four new mononuclear copper(II) polypyridyl complexes (1-4) have been designed, developed, and thoroughly characterized by several physicochemical techniques. The CT-DNA binding properties of 1-4 have been investigated by absorption, emission spectroscopy, and viscosity measurements. All the complexes especially 1 and 4 exhibit cytotoxicity toward several cancer cell lines, suggesting their anticancer properties as observed by several in vitro assays. Additionally, the complexes show inhibition of endothelial cell (HUVECs) proliferation, indicating their antiangiogenic nature. In vivo chick embryo angiogenesis assay again confirms the antiangiogenic properties of 1 and 4. The formation of excessive intracellular ROS (H2O2 and O2(•-)) and upregulation of BAX induced by copper(II) complexes may be the plausible mechanisms behind their anticancer activities. The present study may offer a basis for the development of new transition metal complexes through suitable choice of ligands for cancer therapeutics by controlling tumor angiogenesis.

Angiogenesis, a process resulting in the formation of new capillaries from the pre-existing vasculature plays vital role for the development of therapeutic approaches for cancer, atherosclerosis, wound healing, and cardiovascular diseases. In this report, the synthesis, characterization, and angiogenic properties of graphene oxide (GO) and reduced graphene oxide (rGO) have been demonstrated, observed through several in vitro and in vivo angiogenesis assays. The results here demonstrate that the intracellular formation of reactive oxygen species and reactive nitrogen species as well as activation of phospho-eNOS and phospho-Akt might be the plausible mechanisms for GO and rGO induced angiogenesis. The results altogether suggest the possibilities for the development of alternative angiogenic therapeutic approach for the treatment of cardiovascular related diseases where angiogenesis plays a significant role.

Angiogenesis, a process involving the growth of new blood vessels from the pre-existing vasculature, plays a crucial role in various pathophysiological conditions. We have previously demonstrated that europium hydroxide [Eu(III)(OH)3] nanorods (EHNs) exhibit pro-angiogenic properties through the generation of reactive oxygen species (ROS) and mitogen activated protein kinase (MAPK) activation. Considering the enormous implication of angiogenesis in cardiovascular diseases (CVDs) and cancer, it is essential to understand in-depth molecular mechanisms and signaling pathways in order to develop the most efficient and effective alternative treatment strategy for CVDs. However, the exact underlying mechanism and cascade signaling pathways behind the pro-angiogenic properties exhibited by EHNs still remain unclear. Herein, we report for the first time that the hydrogen peroxide (H2O2), a redox signaling molecule, generated by these EHNs activates the endothelial nitric oxide synthase (eNOS) that promotes the nitric oxide (NO) production in a PI3K (phosphoinositide 3-kinase)/Akt dependent manner, eventually triggering angiogenesis. We intensely believe that the investigation and understanding of the in-depth molecular mechanism and signaling pathways of EHNs induced angiogenesis will help us in developing an effective alternative treatment strategy for cardiovascular related and ischemic diseases where angiogenesis plays an important role.

Reduced graphene oxide (rGO) generates reactive oxygen species (ROS) under visible light in air via a singlet oxygen–superoxide anion radical pathway which readily kills Enterobacter sp. The rGO+ intermediate reacts with a hydroxyl ion to produce graphene oxide (GO) as a coating on the surface of rGO resulting in enhanced fluorescence and a slow down in photo-induced ROS formation. GO is not toxic but on ageing it gets a surface coating of rGO and shows toxicity.

Metabolic alterations of oral epithelial cells under oxidative stress are important signatures for early diagnosis of oral cancer. Amongst different metabolic alterations, non-invasive photo-diagnostic methods have been extensively used for determining cellular heme metabolism and accumulation of protoporphyrin IX (PpIX) under administration of suitable photosensitizer. In this study, we report these metabolic alterations by direct analysis of oral exfoliated cells obtained from individuals with prolonged smoking habit without the exogenous administration of any photosensitizer. The relative expression level of relevant biomolecules of study groups were compared with clinically diagnosed and histopathologically confirmed leukoplakia (OLPK) and oral squamous cell carcinoma (OSCC) patients. The energy imbalance and variation in 'redox ratio' were examined through spectroscopic studies which showed an increasing trend (p < 0.001) in smokers to OSCC groups in comparison to nonsmoker control. Gene expression of important intermediates of the heme metabolic pathway (viz. 5-aminolevulinate synthase 1 (ALAS1), Ferrochelatase (FECH), hemeoxygenase 1 (HO-1) and ATP binding cassette subfamily G member 2 (ABCG2)) which affect production of PpIX was assessed. Relative mRNA level of ALAS1 and HO1 was upregulated whereas mRNA level of other genes (viz. FECH and ABCG2) were found to be downregulated in smokers as well as in cancer groups. Outcome of different spectroscopic studies on exfoliated cells (viz. fluorescence, atomic absorption and Fourier transform infrared spectroscopy) corroborated with the expression of biomarkers related to cellular endogenous metabolism related to heme cycle. This study indicates significant alterations in endogenous metabolic products, and cellular functional groups in oral epithelial cells among the study groups. Our study reports a strong possibility of diagnosis of early cancer signatures amongst habitual smokers by direct and non-invasive assessment of metabolic status of oral epithelial cells without exogenous administration of photosensitizers. The knowledge accrued from the study may guide clinicians in precise detection of precancer trend in the susceptible population through a noninvasive rapid screening method.