Fluorescent Staining Method Research Articles

Carboplatin-loaded zeolitic imidazolate framework-8: Induction of antiproliferative activity and apoptosis in breast cancer cell.

The challenge with breast cancer is its ongoing high prevalence and difficulties in early detection and access to effective care. A solution lies in creating tailored metal-organic frameworks to encapsulate anticancer drugs, enabling precise and targeted treatment with less adverse effects and improved effectiveness. Zeolitic imidazolate framework-8 (ZIF-8) and carboplatin (CP)-loaded ZIF-8 were synthesized and characterized using various analytical techniques. High Resolution-transmission electron microscopy of ZIF-8 and CP@ZIF-8 indicates that the particles had a spherical shape and were nanosized. The drug release rate of CP is 98% under an acidic medium (pH 5.5) because of the dissolution of ZIF-8 into its coordinating ions, whereas 35% in a physiological medium (pH 7.4) with the addition of CP, the high porosity, and pore diameter of ZIF-8 decrease from 1243 to 1041m2/g. Breast cancer MCF-7 cells were shown greater IC50 in CP@ZIF-8 (15.01±3.03µg/mL) than free CP (34.98±4.25µg/mL) in an in vitro cytotoxicity assessment. The cytotoxicity of the CP@ZIF-8 against MCF-7 cells was studied using the methylthiazolyldiphenyl-tetrazolium bromide method. The morphological changes were examined using fluorescent staining (acridine orange-ethidium bromide and Hoechst 33258) methods. The comet assay assessed the DNA fragmentation (single-cell gel electrophoresis). The results from the study revealed that CP@ZIF-8 can be used in the treatment of breast cancer.

Novel Fluorescence-Based Methods to Determine Infarct and Scar Size in Murine Models of Reperfused Myocardial Infarction.

Determination of infarct and scar size following myocardial infarction (MI) is commonly used to evaluate the efficacy of potential cardioprotective treatments in animal models. However, histological methods to determine morphological features in the infarcted heart have barely improved since implementation while still consuming large parts of the tissue and offering little options for parallel analyses. We aim to develop a new fluorescence technology for determining infarct area and area at risk that is comparable to 2,3,5-triphenyltetrazolium chloride (TTC) staining but allows for multiple analyses on the same heart tissue. For early and late time points following MI, we compared classical histochemical approaches with fluorescence staining methods. Reperfused MI was induced in male mice, the hearts were extracted 24 h, 7-, 21-, or 28-days later and fluorescently stained by combining Hoechst and phalloidin. This approach allowed for clear visualization of the infarct area, the area at ischemic risk and the remote area not affected by MI. The combined fluorescence staining correlated with the classic TTC/Evans Blue staining 24 h after MI (r = 0.8334). In later phases (>7 d) post-MI, wheat germ agglutinin (WGA) is equally accurate as classical Sirius Red (r = 0.9752), Masson's (r = 0.9920) and Gomori's Trichrome (r = 0.8082) staining for determination of scar size. Additionally, feasibility to co-localize fluorescence-stained immune cells in specific regions of the infarcted myocardium was demonstrated with this protocol. In conclusion, this new procedure for determination of post-MI infarct size is not inferior to classical TTC staining, yet provides substantial benefits, including the option for unbiased software-assisted analysis while sparing ample residual tissue for additional analyses. Overall, this enhances the data quality and reduces the required animal numbers consistent with the 3R concept of animal experimentation.

FLUID: a fluorescence-friendly lipid-compatible ultrafast clearing method.

Many clearing methods achieve high transparency by removing lipid components from tissues, which damages microstructure and limits their application in lipid research. As for methods which preserve lipid, it is difficult to balance transparency, fluorescence preservation and clearing speed. In this study, we propose a rapid water-based clearing method that is fluorescence-friendly and preserves lipid components. FLUID allows for preservation of endogenous fluorescence over 60 days. It shows negligible tissue distortion and is compatible with various types of fluorescent labeling and tissue staining methods. High quality imaging of human brain tissue and compatibility with pathological staining demonstrated the potential of our method for three-dimensional (3D) biopsy and clinical pathological diagnosis.

A Comparison of Two Fluorescent Stains for the Evaluation of Bioceramic Sealer Penetration into the Tubular Dentine Using Confocal Microscopy

This study aimed to compare the percentage of bioceramic sealer penetration and maximum penetration sealer depending on the fluorescent staining method when evaluated with confocal laser scanning microscopy (CLSM). In total, 42 single-rooted premolars were instrumented. Two groups of 20 teeth each were filled using the single cone technique. TotalFill BC Sealer® mixed with Rhodamine B was used for the first group and TotalFill BC Sealer® mixed with Fluo-3 for the second group. Two teeth made up the control group. All teeth were sectioned at 3 and 6 mm from the apex and were analyzed with CLSM. The canal perimeter where the tubular penetration happened and the maximum penetration of each sample were calculated. There were no significant differences between the two groups in either the apical or medial sections. In terms of maximum penetration, Rhodamine B had greater tubular penetration in the apical sections, and it had similar results to Fluo-3 in the medial sections. TotalFill BC Sealer® has a higher tubular penetration in the medial slices than in the apical slices regardless of the fluorescent stain used. Rhodamine B has higher results than Fluo-3 in terms of the percentage of sealer penetration and maximum penetration.

Indocyanine Green Fluorescence-Guided Laparoscopic Anatomical Segmentectomy of Liver Segment 6: Surgical Strategy and Technical Details.

Because of the complex anatomy of the right posterior hepatic pedicle, there have been few reports on standardized laparoscopic portal territory staining-guided anatomical resection of liver segment 6 (LPTAR-S6). This study aimed to elucidate the indocyanine green (ICG) fluorescence staining methods for LPTAR-S6. LPTAR-S6 can be performed using positive and negative fluorescence staining approaches. We implemented these two approaches for patients with hepatocellular carcinoma. Descriptions of the surgical strategy and technical details are presented. Two patients safely underwent LPTAR-S6 using a preoperative three-dimensional reconstruction plan. The intraoperative ICG fluorescence staining effect was satisfactory, and the anatomical landmarks were fully exposed. A detailed preoperative three-dimensional reconstruction plan, complete intraoperative application of real-time laparoscopic ultrasound guidance, and ICG fluorescence staining can result in accurate transection of the liver parenchyma during LPTAR-S6.

The alpha2-adrenergic receptor agonist clonidine protects against cerebral ischemia/reperfusion induced neuronal apoptosis in rats.

Apoptosis is the crucial pathological mechanism following cerebral ischemic injury. Our previous studies demonstrated that clonidine, one agonist of alpha2-adrenergic receptor (α2-AR), could attenuate cerebral ischemic injury in a rat model of middle cerebral artery occlusion/reperfusion (MCAO/R). However, it's unclear whether clonidine exerts neuroprotective effects by regulating neuronal apoptosis. In this study, we elucidated whether clonidine can exert anti-apoptotic effects in cerebral ischemic injury, and further explored the possible mechanisms. Neurological deficit score was measured to evaluate the neurological function. TTC staining was used for the measurement of brain infarct size. Hematoxylin-Eosin (HE) staining was applied to examine the cell morphology. TUNEL and DAPI fluorescent staining methods were used to analyze the cell apoptosis in brain tissue. Fluorescence quantitative real-time PCR was performed to assess the gene expression of Caspase-3 and P53. Western blotting assay was applied to detect the protein expression of Caspase-3 and P53. The results showed that clonidine improved neurological function, reduced brain infarct size, alleviated neuronal damage, and reduced the ratio of cell apoptosis in the brain with MCAO/R injury. moreover, clonidine down-regulated the gene and protein expression of Caspase-3 and P53 which were over-expressed after MCAO/R injury. Whereas, yohimbine (one selective α2-AR antagonist) mitigated the anti-apoptosis effects of clonidine, accompanied by reversed gene and protein expression changes. The results indicated that clonidine attenuated cerebral MCAO/R injury via suppressing neuronal apoptosis, which may be mediated, at least in part, by activating α2-AR.

Insights into cellular interactions of characterized Mg–Al Layered Double Hydroxide on L929 cells

Layered double hydroxides are members of an anionic clay family, characterised by unique two-dimensional layered structures and lend versatility in various applications. These biocompatible compounds have the potential to get intercalated with biological compounds and physico-chemically adsorbed onto organic molecules. Thus, making them important candidates for pharmaceutical and biomedical purposes. This study aims to synthesise, characterise and investigate the cellular toxicity interactions of Mg–Al LDH towards the mouse fibroblast L929 cell line. The Mg–Al LDH was synthesized by a meticulous process of co-precipitation followed by the hydrothermal method to ensure a well-defined and stable structure for suitable biological application. Characterisation techniques like Dynamic Light Scattering, Zeta potential, Scanning Electron Microscopy, Fourier transform infrared, and X-ray diffraction analysis were employed to provide deeper insights into the physiochemical properties and structural integrity of the synthesized Mg–Al LDH. The investigation of cellular interactions with the L929 fibroblast cell line served to assess the biocompatibility and potential cytotoxic effects of Mg–Al LDH. This was observed by assessing the morphological changes and evaluating the cytotoxic effects of Mg–Al LDH by utilising various techniques like phase contrast microscopy, fluorescent staining, and Giemsa staining. The cellular metabolic activity was assessed by MTT assay, and the subcellular lysosomal alteration was examined using the fluorescent staining method by the acridine orange staining. The dose-dependent response observed in the cellular interaction underscores the importance of dosage considerations for potential biomedical applications. By elucidating the dose-response relationship, this study contributes valuable information for the safe and effective usage of LDH in biomedical contexts.

A two-photon fluorescent probe for highly selective detection of Cys over GSH and Hcy based on the Michael addition and transcyclization mechanism and its application in bioimaging and protein straining in SDS-PAGE

BackgroundCysteine (Cys), glutathione (GSH), and homocysteine (Hcy), as three major biothiols are involved in a variety of physiological processes and play a crucial role in plant growth. Abnormal levels of Cys can cause plants to fail to grow properly. To date, although a very large number of fluorescent probes have been reported for the detection of biothiols, very few of them can be used for the selective discrimination of Cys from GSH and Hcy due to their structural similarity, and only a few of them can be used for plant imaging. ResultsHere, three fluorescent probes (o-/m-/p-TMA) based on TMN fluorophore and the ortho-/meta-/para-substituted maleimide recognition groups were constructed to investigate the selective response effect of Cys. Compared to the o-/m-TMA, p-TMA can selectively detect Cys over GSH and Hcy with a rapid response time (10 min) and a low detection limit (0.26 μM). The theoretical calculation confirmed that the intermediate p-TMA-Cys-int has shorter interatomic reaction distances (3.827 Å) compared to o-/m-TMA-Cys (5.533/5.287 Å), making it more suitable for further transcyclization reactions. Additionally, p-TMA has been employed for selective tracking of exogenous and endogenous Cys in Arabidopsis thaliana using both single-/two-photon fluorescence imaging. Furthermore, single cell walls produced obvious two-photon fluorescence signals, indicating that p-TMA can be used for high-concentration Cys analysis in single cells. Surprisingly, p-TMA can be used as a fluorescent dye for protein staining in SDS-PAGE with higher sensitivity (7.49 μg/mL) than classical Coomassie brilliant blue (14.11 μg/mL). SignificanceThe outstanding properties of p-TMA make it a promising multifunctional molecular tool for the highly selective detection of Cys over GSH and Hcy in various complex environments, including water solutions, zebrafish, and plants. Additionally, it has the potential to be developed as a fluorescent dye for a simple and fast SDS-PAGE fluorescence staining method.

Influence of different pre-treatments on the resin infiltration depth into enamel of teeth affected by molar-incisor hypomineralization (MIH)

ObjectivesThis in vitro pilot study aimed to evaluate whether different pre-treatments (demineralization, deproteinization, (chemo-)mechanical reduction of the surface layer) influence the penetration depth of a resin infiltrant into MIH-affected enamel compared to initial carious lesions. MethodsThirty extracted human permanent molars with non-cavitated initial carious lesions (n = 5) or MIH (n = 25) were chosen and randomly assigned to six experimental groups: IC: initial caries; M: MIH; MN: MIH, 5.25% sodium hypochlorite; MM: MIH, microabrasion; MA: MIH, air abrasion; MAN: MIH, air abrasion and 5.25% sodium hypochlorite. A modified indirect dual fluorescence staining method was adopted to assess the penetration depth (PD) of the resin infiltrant and the lesion depth (LD) by confocal laser scanning microscopy (CLSM). Exemplarily, scanning electron microscopic (SEM) images were captured. The relationship between group assignment and penetration/lesion depth was estimated using a linear mixed model incorporating the tooth as random effect (two observations/tooth). The significance level was set at p < 0.05. ResultsFor MIH-affected molars, the mean PD (in µm; median, [minimum-maximum]) were M (178.2 [32.5–748.9]), MN (275.6 [105.3–1131.0]), MM (48.7 [0.0–334.4]), MA (287.7 [239.4–491.7]), and MAN (245.4 [76.1–313.5]). Despite the observed differences in PD between the groups, these could not be statistically verified (Bonferroni, p = 0.322). The percentage penetration was significantly higher for IC than for MIH groups (Bonferroni, p < 0.05). SignificanceCompared to IC, resin infiltration into MIH-affected enamel ist more variable. Different pre-treatments influence the resin penetration into developmentally hypomineralized enamel to a fluctuating level.

Molecular mechanism of Caulis Spatholobi in the treatment of Chronic Myeloid Leukemia based on Network Pharmacology and Experimental Verification.

This study aimed to predict the relevant targets of the treatment of chronic myeloid leukemia (CML) with Caulis Spatholobi by applying the network pharmacology method, and in vitro cell experiments were conducted to verify the mechanism of Caulis Spatholobi in the treatment of CML. TCMSP, ETCM, Genecards, and GisGeNET databases were used to obtain relevant targets of Caulis Spatholobi in the treatment of CML. Go and KEGG analyses were performed using the David database. Using Cytoscape 3.7.2, the "active compounds-targets-pathways" network was constructed. Further validation was carried out by pharmacological experiments in vitro. The proliferation and apoptosis of K562 cells were observed by the MTT method and Hoechst 33242 fluorescence staining method. The predicted targets and their related signal pathways were verified by western blotting. In this study, 18 active compounds and 43 potential targets were obtained. The results of the MTT method showed that compared with the normal control group, 62.5-500 μg/mL alcohol extract of Caulis Spatholobi had an obvious inhibitory effect on K562 and the IC50 value was less than 100 μg/mL. The Hoechst 33242 fluorescence staining method showed that the alcohol extract of Caulis Spatholobi could promote apoptosis. The results of western blotting showed that compared with the normal control group, the expressions of Bax and Caspase-3 proteins in the 62.5 and 125 μg/mL alcohol extract of Caulis Spatholobi groups were significantly up-regulated (P<0.05). The expression of Bcl-2 in the 125 μg/mL alcohol extract of the Caulis Spatholobi group was significantly down-regulated (P<0.01), and the expression of Bcl-2 in the 62.5 and 31.25 μg/mL alcohol extract of Caulis Spatholobi groups was also significantly down-regulated (P<0.05). It showed that the ethanol extract of Caulis Spatholobus could promote apoptosis by up-regulating the expression of Bax and caspase-3 and down-regulating the expression of the Bcl-2 protein. The treatment of Caulis Spatholobi for CML has the characteristics of multi-targets and multi-pathways. The results of in vitro pharmacological experiments demonstrated that its mechanism of action might be based on the expression of key target proteins, such as Caspase-3, Bcl-2, and Bax, thereby inhibiting cell proliferation and promoting cell apoptosis, which provides a scientific basis for the treatment of CML.

Designing nanostructured lipid carriers modified with folate-conjugated chitosan for targeted delivery of osthole to HT-29 colon cancer cells: investigation of anticancer, antioxidant, and antibacterial activities

AbstractThe present study proposed to design nanostructured lipid carriers (NLC) coated with chitosan (CS) conjugated folate (FA) for the targeted delivery of Osthole (OST) to the HT-29 colon cancer cell line and improve its anticancer capability. To assess the physicochemical characteristics of OST-loaded NLC decorated with CS-conjugated FA (OST-NCF-NPS), several techniques, including DLS, SEM, and FTIR, were applied. After determining the encapsulation efficiency of OST in CSFA-modified NLC-NPs, an MTT test was conducted to evaluate the cytotoxic effects of this nano platform on the HT-29 cancer cell line in comparison to normal HFF cells. Possible mechanisms of apoptosis in cancer cells treated with OST-NCF-NPs were examined using qPCR, flow cytometry, and AO/PI fluorescent staining methods. Moreover, the antioxidant capacity of these biosynthesized nanocarriers was determined using ABTS and DPPH methods, and their antibacterial potential was measured through disk diffusion, MIC, and MBC assays. According to the findings, OST-NCF-NPS had the ideal average size of 179.19 nm, low polydispersity (PI = 0.23), acceptable physical stability (ζ-potential = + 18.99 mV), and high entrapment efficiency (83.5%). The MTT data demonstrated the selective cytotoxicity of NPs toward cancerous cells compared to normal cells. Cell cycle and Annexin V/Propidium Iodide (AnV/PI) analysis indicated that OST-NCF-NPs increased the sub-G1 population and AnV/PI-positive cells. The occurrence of programmed cell death in the treated cells was also verified by altered expression of proapoptotic (BAX and caspase-3) and antiapoptotic (Bcl-2) genes. Furthermore, the NPs exhibited strong antibacterial activity, particularly against gram-negative bacteria, and high antioxidant effects in reducing ABTS and DPPH-free radicals. Graphical Abstract

Radix Astragali decoction improves liver regeneration by upregulating hepatic expression of aquaporin-9.

The therapeutic efficacy of liver injuries heavily relies on the liver's remarkable regenerative capacity, necessitating the maintenance of glycose/lipids homeostasis and oxidative eustasis during the recovery process. Astragali Radix, an herbal tonic widely used in China and many other countries, is believed to have many positive effects, including immune stimulation, nourishing, antioxidant, liver protection, diuresis, anti-diabetes, anti-cancer and expectorant. Astragali Radix is widely integrated into hepatoprotective formulas as it is believed to facilitate liver regeneration. Nevertheless, the precise molecular pharmacological mechanisms underlying this hepatoprotective effect remain elusive. To investigate the improving effects of Astragali Radix on liver regeneration and the underlying mechanisms. A mouse model of 70% partial hepatectomy (PHx) was employed to investigate the impact of Radix Astragali decoction (HQD) on liver regeneration. HQD was orally administered for 7 days before the PHx procedure and throughout the experiment. N-acetylcysteine (NAC) was used as a positive control for liver regeneration. Liver regeneration was assessed by evaluating the liver-to-body weight ratio (LW/BW) and the expression of representative cell proliferation marker proteins. Oxidative stress and glucose metabolism were analyzed using biochemical assays, Western blotting, dihydroethidium (DHE) fluorescence, and periodic acid-Schiff (PAS) staining methods. To understand the role of AQP9 as a potential molecular target of HQD in promoting liver regeneration, td-Tomato-tagged AQP9 transgenic mice (AQP9-RFP) were employed to determine the expression pattern of AQP9 protein. AQP9 knockout mice (AQP9-/-) were used to assess the specific targeting of AQP9 in the promotion of liver regeneration by HQD. HQD significantly upregulated hepatic AQP9 expression, alleviated liver injury and promoted liver regeneration in wild-type (AQP9+/+) mice after 70% PHx. However, the beneficial impact of HQD on liver regeneration was absent in AQP9 gene knockout (AQP9-/-) mice. Moreover, HQD facilitated the uptake of glycerol by hepatocytes, enhanced gluconeogenesis, and concurrently reduced H2O2 content and oxidative stress levels in AQP9+/+ but not AQP9-/- mouse livers. Additionally, main active substance of Radix Astragali, astragaloside IV (AS-IV) and cycloastragenol (CAG), demonstrated substantial upregulation of AQP9 expression and promoted liver regeneration in AQP9+/+ but not AQP9-/- mice. This study is the first to demonstrate that Radix Astragali and its main active constituents (AS-IV and CAG) improve liver regeneration by upregulating the expression of AQP9 in hepatocytes to increase gluconeogenesis and reduce oxidative stress. The study revealed novel molecular pharmacological mechanisms of Radix Astragali and provided a promising therapeutic target of liver diseases.

A novel technique for determining the distribution of impurities in crystals during cooling crystallization

This paper presents a novel fluorescence-staining method for effective solvent tagging within crystals, which can enhance impurity management.

Expanding the toolset of fluorescent covalent staining of biological samples by labeling carboxylate and phosphate groups.

Recently, fluorescent covalent staining methods have been developed for visualization of anatomical structures in cells and tissues. Coupled with expansion microscopy, these stains revealed various ultrastructural details. However, the covalently stainable chemical groups have been limited to amines, carbohydrates, and thiols. Here, we developed procedures for covalently labeling tissues for carboxylate and phosphate groups, utilizing carbodiimide crosslinker chemistry. In porcine kidney tissues, the carboxylate and phosphate stain provides 1.8-4.8-fold higher signal intensity than those from the three existing stains. In cancer cells, such stain allows 2-8-fold more accurate identification of nucleoli than the amine stain. In expansion microscopy samples, such stain reveals a variety of sub-cellular structures in tissues when combined with the amine stain. Such stain also allows imaging of lipid-based structures in cultured cells. With these advantages, this new covalent staining method further expands the toolset for fluorescent visualization of histology.

Furan promotes cytotoxic effects through DNA damage and cell apoptosis in Leydig cells

Furan is an organic chemical that can cause adverse effects on human health and is formed as a result of the thermal decomposition of many food components during cooking, storage, and processing techniques. Studies have shown that exposure to furan causes nephrotoxicity, hepatotoxicity, immunotoxicity, and reproductive toxicity. According to our current knowledge of the literature, the genotoxic mode of action of furan is highly controversial. The genotoxic effects of furan on the male reproductive system, however, have not been studied. In this study, the TM3 Leydig cell line was treated with 750, 1500, and 3000 μM concentrations of furan for 24 h. Following the completion of the exposure period, the cytotoxicity of furan in TM3 Leydig cells was assessed using a cell viability assay and a spectrophotometric measurement of lactate dehydrogenase (LDH) enzyme levels. The double fluorescence staining method was used to demonstrate furan-induced apoptosis, and DNA damage was shown using the micronucleus, comet, and chromosomal aberration assays. The result indicated that furan administration of Leydig cells resulted in an increase in structural chromosomal aberration, comet, and micronucleus formation, reduced cell viability, increased LDH activity, and a higher incidence of apoptotic cells. These findings revealed that furan induces DNA damage in TM3 Leydig cells, causing genotoxicity and DNA damage-induced cytotoxicity.

Preparation and Characterization of Allicin-Loaded Solid Lipid Nanoparticles Surface-Functionalized with Folic Acid-Bonded Chitosan: In Vitro Anticancer and Antioxidant Activities.

This investigation aimed to increase the bioavailability and anticancer effects of allicin (AC) by encapsulating it in solid lipid nanoparticles (SLN) decorated with chitosan (CS)-conjugated folic acid (FA). Nanoparticles (NPs) were synthesized by high-pressure homogenization, and then, Fourier-transform infrared (FTIR), Field-Emission Scanning Electron Microscopy (FESEM), Dynamic Light Scattering (DLS), and zeta potential methods were used to determine their physicochemical characteristics. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to assess the effect of toxicity and flow cytometry, while fluorescent staining methods were used to investigate the type of cell death. Real-time quantitative polymerase chain reaction (qPCR) was used to evaluate the expression levels of apoptotic genes: Bcl-2, and caspase-8. The presence of AC-SLN-CS-FA with a spherical morphology, an average size of 86.7 ± 9.4 nm, uniform distribution (0.31), a surface charge of +21.3 ± 13.3 mV, an encapsulation percentage of 86.3%, and a folate binding rate of 63% confirmed the success of the preparation method. Suppression of MCF-7 cancer cells and non-toxicity of AC-SLN-CS-FA on Human foreskin fibroblast (HFF) normal cells were confirmed by cytotoxic assay. The results of flow cytometry revealed that the cells were arrested in the sub-G1 phase, and the activation of the intrinsic apoptosis pathway was confirmed by the results of real-time qPCR. In general, AC-SLN-CS-FA has the potential to prevent free radicals and trigger apoptosis in cancer cells by activating the intrinsic apoptosis pathway; thus, making it a promising subject in preclinical research.

In-Vitro Cytotoxic, Antiproliferative and Apoptotic Activity of Siddha Formulation Nandhi Mezhugu (NM) Against Hela Cell Line.

Cervical cancer is the most common cancer among women of childbearing age. Nandhi Mezhugu is a Siddha herbo-mineral drug widely used to treat cancer. Due to a lack of scientific evidence, the present study was intended to evaluate the anti cancer activity of Nandhi Mezhugu in the HeLa cell line. The cells were cultured in Dulbecco's modified Eagle medium, then treated with different concentrations of the test drug (10 to 200µg/ml). The anti proliferative activity of the drug was evaluated using an MTT assay. Cell apoptosis and cell cycle were measured by flow cytometry and typical nuclear changes of apoptotic processes were observed under the microscope using the dual acridine orange/ethidium bromide fluorescent staining method. The study result showed that the percentage of cell viability decreased with an increase in the concentration of the test drug. The MTT assay data showed that the test drug Nandhi Mezhugu had the antiproliferative effect on cervical cancer cells with IC50 of 139.7 ± 13.87µg/ml. Further studies such as flow cytometry and dual staining method also revealed the apoptotic effect of the test drug. Nandhi Mezhugu can be effectively used as an anti cancer formulation to treat cervical cancer. Thus, the current study brings forth scientific evidence for the efficacy of Nandhi Mezhugu against the HeLa cell line. Further studies will be needed to prove the promising efficacy of Nandhi Mezhugu.

Pharmacological applications of metal nanoparticles derived from Caralluma sarkariae species

The present research is focused on synthesis of silver/gold nanoparticles using Caralluma sarkariae root extracts (CSR-AgNPs and CSR-AuNPs) used as capping/reducing agents and successfully carry out the pharmacological studies. The as-prepared CSR-AgNPs and CSR-AuNPs were systematically characterized using various analytical techniques. Moreover, the formation of CSR-AgNPs & CSR-AuNPs was confirmed by SPR values observed at 432 and 537 nm in UV–Visible spectroscopic. Morphological information of CSR-AgNPs & CSR-AuNPs were investigated using scanning electron microscope (SEM) and high-resolution transmission electron microscope (HR-TEM) analysis and as-synthesized nanomaterials were spherical in nature with mean size of 20 and 22 nm, respectively. The prepared nanoparticles clearly proven that the as-prepared nanoparticles exhibit high performance efficiencies in terms of in vitro antioxidant, anti-inflammatory, antibacterial and antifungal activities. The wound healing activity of as-fabricated CSR-AgNPs and CSR-AuNPs were carried out separately using three different methods such as in vitro scratch assay, MTT assay, and fluorescence staining methods. Furthermore, the fibroblast L929 cells were used for the wound healing activity of both nanoparticles and aqueous CSR extract in in vitro scratch assay. Moreover, the wide range of diseases could be treated by using the nanoparticles as-synthesized due to their outstanding pharmacological activities.

Güncel Tanı ve Tedavi Yaklaşımlarıyla MRONJ

ABSTRACT&#x0D; Osteonecrosis of the jaw bones is a persistent and rare pathology that develops with progressive bone destruction and bone necrosis as a result of impaired blood supply in the mandible and maxilla. The key point to the prevention and treatment of MRONJ is screening high-risk patients (eg, AAOMS stage 0) predisposed to the disease to detect it at an early stage to effectively prevent the risk of progression and occurrence. Although no exposed bone is seen in clinical examination, osteonecrosis can be diagnosed early by radiographic examination. Therefore, advanced imaging techniques such as MR, SPECT/PET and VELscope are important in the early diagnosis of MRONJ. There is no defined gold standard treatment in the literature and the treatment of MRONJ is often very difficult. Treatment strategies are mainly focused on minimizing the progression or formation of bone necrosis, eliminating pain, controlling infection and optimizing the patient's quality of life. The AAOMS recommends stage-based treatment planning in MRONJ patients. In addition to the treatment methods recommended by the AAOMS, there are research showing that some adjuvant treatments have a positive effect on recovery in MRONJ patients. For example, the use of platelet-rich plasma/fibrin in addition to surgical treatment increasing the success of surgery is one of them. Alternative treatment options are low-level laser therapy, surgical debridement with laser, surgical debridement under the guidance of fluorescent staining method, use of platelet concentrates, ozone and hyperbaric oxygen therapy, pentoxifylline, alpha-tocopherol, photo-bio modulation, use of parathormone or stem cell transplantation into the lesion. In this review, we aimed to update the knowledge, attitudes and behaviors of dentists about MRONJ and to shape the approach to MRONJ with the guidance of current literature.

Effective regeneration of rat sciatic nerve using nanofibrous scaffolds containing rat ADSCs with controlled release of rhNGF and melatonin molecules for the treatment of peripheral injury model

Different therapeutic strategies have been designed and developed for the repair and regeneration of peripheral nerve injury (PNI) tissue as a result of advancements in tissue engineering and regenerative medicine. Due to its versatility, controlled delivery and administration of multifunctional therapeutic agents can be regarded of as an effective strategy in treating nerve injury. In this study, melatonin (Mel) molecules and recombinant human nerve growth factor (rhNGF) were loaded on the surface and in the core of polycaprolactone/chitosan (PCL/CS) blended nanofibrous scaffold. To simulate the in vivo microenvironment, a dual-delivery three-dimensional (3-D) nanofibrous matrix was developed and the in vitro neural development of stem cell differentiation process was systematically examined. The microscopic technique with acridine orange and ethidium bromide (AO/EB) fluorescence staining method was used to establish the adipose-derived stem cells (ADSCs) differentiation and cell–cell communications, which demonstrated that the effective differentiation of the ADSCs with nanofibrous matrix. As investigated observations, ADSCs differentiation was further evident through cell migration assay and gene expression analysis. According to the biocompatibility analysis, the nanofibrous matrix did not trigger any adverse immunological reactions. Based on these characteristics, a 5-week in vivo investigation examined the potential of the developed nanofibrous matrix in the regeneration of sciatic nerve of rats. Additionally, compared to the negative control group, the electrophysiological and walking track analyses demonstrated improved sciatic nerve regeneration. This study demonstrates the nanofibrous matrix's ability to regenerate peripheral nerves.