In Vivo Studies and Flow Cytometric Investigation on Anticancer Potential of Selenium Nanoparticles Synthesized via Aqueous Extract of Clerodendron phlomidis

Nowadays,doctors and nutritionists recommend individuals incorporate selenium-rich foods such as nuts,cereals,and mushrooms into their regular diet to maintain fitness and overall health.Selenium nanoparticles(SeNPs)exhibit strong chemopreventive capabilities.The anticipations for SeNPs with enhanced and tunable bioactive activities have led to a keen interest in phytofabrication.In this study,the aqueous extract of Clerodendron phlomidis plant leaves was utilized for the synthesis of SeNPs.In traditional Indian medicine,this plant extract is recognized as a significant anti-diabetic agent.The flavonoids tetrahydroxylflavone,7-hydroxyflavanone,and 6,4’-dimethyl-7-acetoxy-scutellarein present in this plant leaf extract demonstrate excellent anticancer activity.These secondary metabolites exhibit the ability to reduce sodium selenite into SeNPs.At a concentration of 13μg/mL,the synthesized SeNPs effectively inhibited the proliferation of the HepG2 cell line.The results suggest that the SeNPs possess promising anti-cancer potential against liver cancer and can be considered as a therapeutic agent for liver cancer treatment.Additionally,the cell cycle arrest induced by SeNPs was further confirmed by the fluorescence-activated cell sorting(FACS)method,indicating that SeNPs could efficiently differentiate cancer cells from normal cells.Notably,it showed a significant improvement in diethylnitrosamine(DEN)-induced Swiss Wistar rat groups.This scientific investigation highlights the high anti-cancer potential of SeNPs,positioning them as a promising therapeutic agent for liver cancer treatment.

MACROPHAGES IN DEGRADATION OF COLLAGEN / HYDROXYLAPATITE(CHA), BETA-TRICALCIUM PHOSPHATE CERAMICS (TCP) ARTIFICIAL BONE GRAFT AN IN VIVO STUDY

The macrophages mediated biodegradation of two biomaterials, collagen / hydroxylapatite (CHA) and beta-tricalcium phosphate ceramics (TCP), was studied in 24 male Kunming mice and 20 male C57BL / 6 mice with histopathologic, histochemical and ultrastructural observation. It was demonstrated that macrophages infiltrated after CHA, TCP were implanted. The macrophages could be differentiated from fibroblasts and the other infiltrated cells for special cellular profile and strong acid phosphatase activity. Morphologically, monocyte macrophages and infused multinuclear giant cell degraded CHA and TCP by phagocytosis and extracellular resorption. The carbonic anhydrase activity of macrophages was demonstrated by histochemical technique. It suggested that macrophages secreted H+ and accomplished the decalcification of calcium phosphate compound of CHA and TCP. We conclude that macrophages are the main mediating cells which degraded CHA and TCP intracellularly and extracellularly.

MET inhibitor tepotinib antagonizes multidrug resistance mediated by ABCG2 transporter:In vitro and in vivo study

Overexpression of ABCG2 transporter in cancer cells has been linked to the development of multidrug resistance(MDR), an obstacle to cancer therapy. Our recent study uncovered that the MET inhibitor,tepotinib, is a potent reversal agent for ABCB1-mediated MDR. In the present study, we reported for the first time that the MET inhibitor tepotinib can also reverse ABCG2-mediated MDR in vitro and in vivo by directly binding to the drug-binding site of ABCG2 and reversibly inhibiting ABCG2 drug efflux activity, therefore enhancing the cytotoxicity of substrate drugs in drug-resistant cancer cells. Furthermore, the ABCB1/ABCG2 double-transfected cell model and ABCG2 gene knockout cell model demonstrated that tepotinib specifically inhibits the two MDR transporters. In mice bearing drug-resistant tumors, tepotinib increased the intratumoral accumulation of ABCG2 substrate drug topotecan and enhanced its antitumor effect. Therefore, our study provides a new potential of repositioning tepotinib as an ABCG2 inhibitor and combining tepotinib with substrate drugs to antagonize ABCG2-mediated MDR.

A rened analytical model for reconstruction problems in diuse reectance spectroscopy

A rened analytical model of spatially resolved diffuse reectance with small source-detector separations(SDSs)for the in vivo skin studies is proposed.Compared to the conventional model developed by Farrell et al.,it accounts for the limited acceptance angle of the detectorber.The rened model is validated in the wide range of optical parameters by Monte Carlo simulations of skin diffuse reectance at SDSs of units of mm.Cases of uniform dermis and two-layered epidermis-dermis structures are studied.Higher accuracy of the rened model compared to the conventional one is demonstrated in the separate,constraint-free reconstruction of absorption and reduced scattering spectra of uniform dermis from the Monte Carlo simulated data.In the case of epidermis-dermis geometry,the recovered values of reduced scattering in dermis are overestimated and the recovered values of absorption are underestimated for both analytical models.Presumably,in the presence of a thin mismatched topical layer,only the effective attenuation coe±cient of the bottom layer can be accurately recovered using a diffusion theorybased analytical model while separate reconstruction of absorption and reduced scattering fails due to the inapplicability of the method of images.These-ndings require implementation of more sophisticated models of light transfer in inhomogeneous media in the recovery algorithms.

An overview on recent in vivo biological application of cerium oxide nanoparticles

Cerium oxide nanoparticles(CNPs)possess a great potential as therapeutic agents due to their ability to self-regenerate by reversibly switching between two valences+3 and+4.This article reviews recent articles dealing with in vivo studies of CNPs towards Alzheimer’s disease,obesity,liver inflammation,cancer,sepsis,amyotrophic lateral sclerosis,acute kidney injury,radiation-induced tissue damage,hepatic ischemia reperfusion injury,retinal diseases and constipation.In vivo anti-cancer studies revealed the effectiveness of CNPs to reduce tumor growth and angiogenesis in melanoma,ovarian,breast and retinoblastoma cancer cell-induced mice,with their conjugation with folic acid,doxorubicin,CPM,or CXC receptor-4 antagonist ligand eliciting higher efficiency.After conjugation with triphenylphosphonium or magnetite nanoparticles,CNPs were shown to combat Alzheimer’s disease by reducing amyloid-β,glial fibrillary acidic protein,inflammatory and oxidative stress markers in mice.By improving muscle function and longevity,the citrate/EDTA-stabilized CNPs could ameliorate amyotrophic lateral sclerosis.Also,they could effectively reduce obesity in mice by scavenging ROS and reducing adipogenesis,triglyceride synthesis,GAPDH enzyme activity,leptin and insulin levels.In CCl4-induced rats,stress signaling pathways due to inflammatory cytokines,liver enzymes,oxidative and endoplasmic reticulum messengers could be attenuated by CNPs.Commercial CNPs showed protective effects on rats with hepatic ischemia reperfusion and peritonitis-induced hepatic/cardiac injuries by decreasing oxidative stress and hepatic/cardiac inflammation.The same CNPs could improve kidney function by diminishing renal superoxide,hyperglycemia and tubular damage in peritonitis-induced acute kidney injury in rats.Radiation-induced lung and testicular tissue damage could be alleviated in mice,with the former showing improvement in pulmonary distress and bronchoconstriction and the latter exhibiting restoration in spermatogenesis rate and spermatid/spermatocyte number.Through enhancement of gastrointestinal motility,the CNPs could alleviate constipation in both young and old rats.They could also protect rat from light-induced retinal damage by slowing down neurodegenerative process and microglial activation.

Understanding the Recovery of the Intervertebral Disc:A Comprehensive Review of In Vivo and In Vitro Studies

Within the consistent daily rhythm of human life,intervertebral discs endure a variety of complex loads beyond the influences of gravity and muscle forces,leading to significant morphological changes(in terms of volume,area,and height)as well as biomechanical alterations,including an increase in disc stiffness and a decrease in intradiscal pressure.Remarkably,the discs demonstrate an ability to regain their original morphological and biomechanical characteristics after a period of nocturnal rest.The preservation of normal disc function is critically dependent on this recovery phase,which serves to forestall premature disc degeneration.This phenomenon of disc recovery has been extensively documented through numerous in vivo studies employing advanced clinical techniques such as Magnetic Resonance Imaging(MRI),stadiometry,and intradiscal pressure measurement.However,the findings from in vitro studies present a more complex picture,with reports varying between full recovery and only partial recuperation of the disc properties.Moreover,research focusing on degenerated discs in vitro has shed light on the quantifiable impact of degeneration on the disc ability to recover.Fluid dynamics within the disc are considered a primary factor in recovery,yet the disc intricate multiscale structure and its viscoelastic properties also play key roles.These elements interact in complex ways to influence the recovery mechanism,particularly in relation to the overall health of the disc.The objective of this review is to collate,analyze,and critically evaluate the existing body of in vivo and in vitro research on this topic,providing a comprehensive understanding of disc recovery processes.Such understanding offers a blueprint for future advancements in medical treatments and bionic engineering solutions designed to mimic,support,and enhance the natural recovery processes of intervertebral discs.

Dynamic changes in proprotein convertase 2 activity in cortical neurons after ischemia/reperfusion and oxygen-glucose deprivation

In this study, a rat model of transient focal cerebral ischemia was established by performing 100 minutes of middle cerebral artery occlusion, and an in vitro model of experimental oxygen-glucose deprivation using cultured rat cortical neurons was established. Proprotein convertase 2 activity gradually decreased in the ischemic cortex with increasing duration of reperfusion. In cultured rat cortical neurons, the number of terminal deoxynucleotidyl transferase-mediated 2＇-deoxyuridine 5＇-triphosphate-biotin nick end labeling-positive neurons significantly increased and proprotein convertase 2 activity also decreased gradually with increasing duration of oxygen-glucose deprivation. These experimental findings indicate that proprotein convertase 2 activity decreases in ischemic rat cortex after reperfusion, as well as in cultured rat cortical neurons after oxygen-glucose deprivation. These changes in enzyme activity may play an important pathological role in brain injury.

Kaempferol attenuates knee osteoarthritis via inhibiting cartilage apoptosis in mice

Objective:To study the mechanism of kaempferol in the intervention of knee osteoarthritis(KOA)in mice by inhibiting cartilage apoptosis.Methods:Firstly,the target genes of kaempferol were retrieved via TCMSP,and the genes related to KOA were obtained by GeneCards,OMIM,PharmGKB,TTD,and Drugbank databases.Then GO enrichment analysis and KEGG signaling pathway analysis were also performed.Subsequently,18 male C57 mice were randomly divided into the sham operation group,the model group,and the kaempferol group(50 mg/kg).Except for the sham operation group,the KOA mouse model was induced by destabilization of medial meniscus surgery.The sham operation group and model group were given the same amount of normal saline daily for 8 weeks while the kaempferol group was given 50 mg/kg kaempferol intragastrically.Results:A total of 63 targets of kaempferol were found that included 35 common target genes with KOA.GO and KEGG analyses showed that biological processes such as extrinsic apoptotic signaling pathway and response to oxidative stress,as well as signaling pathways such as cell apoptosis and regulation of TNF were closely related to common target genes.Molecular docking results also showed kaempferol has good binding properties with predicted targets Bcl-2,BAX,and CASP3.Compared with the model group,the pathological changes of cartilage in the kaempferol group were significantly reduced,OARSI scores were significantly decreased(P<0.001),and cartilage area was increased(P<0.01).In addition,Western blot analysis showed that kaempferol significantly decreased the protein expression of BAX and CASP3(P<0.01,P<0.05),and increased the protein expression of BCL-2(P<0.05).Conclusion:The treatment of KOA with kaempferol has the characteristics of multi-target and multi-pathway,and the mechanism may be related to the regulation of key genes such as Bcl-2,BAX,and CASP3 to inhibit cartilage apoptosis.

Preparation and evaluation of sustained-release diltiazem hydrochloride pellets

In this study,diltiazem hydrochloride(DTZ)pellets were prepared successfully by extrusionespheronization method.Then methacrylic acid and ethylcellulose coating formulations were employed to make the DTZ pellets sustained release.The pellets with different coatings were investigated by in vitro dissolution tests.At last,the pellets with the best coating copolymer were subjected to pharmacokinetic studies in beagle dogs.The dissolution profiles of pellets coated with Eudragit
NE30D were similar to Herbesser
,one of the marketed sustained release capsules.In the bioavailability study,the principal pharmacokinetic parameters of self-made pellets and the marketed ones were comparable;the relative bioavailability of DTZ sustained release capsules compared with Herbesser
was 98.5
36.4%.All the data indicated self-made sustained pellets could prolong the release of DTZ,decrease the fluctuation of drug level in vivo,and increase the compliance of patients.

NiTi laser textured implants with improved in vivo osseointegration:An experimental study in rats

Laser surface texturing is a versatile approach for manufacturing implants with suitable surfaces for os-seointegration.This work explores the use of laser to fabricate NiTi textured implants,testing two dif-ferent groove-based designs.Their performance was evaluated in vivo through implantation in Sprague Dawley rats’femur,being then analyzed after 4 and 12 weeks of implantation.Push-out experiments and histological characterization allowed to assess bone-implant bond and osseointegration and to compare the laser textured solutions with non-textured NiTi.Histology showed that,at 4 weeks of implantation,mainly immature woven bone was present whilst at 12 weeks a more mature bone had developed.Con-sidering the largest implantation time(12 weeks),results showed extraction forces considerably higher for textured implants(G2 and G3).Moreover,when comparing G2 and G3,it was found that G2(having the highest textured surface area)displayed the maximum extraction force among all groups,with an increase of 212%when compared to non-textured implants(G1).These results prove that the design and manufacturing technology are effective to promote an im-proved bone-implant bond,aiming the development of orthopedic implants.

Simultaneous size characterization and mass quantification of the in vivo core-biocorona structure and dissolved species of silver nanoparticles

Size characterization of silver nanoparticles with biomolecule corona（AgNP@BCs） and mass quantification of various silver species in organisms are essential for understanding the in vivo transformation of Ag NPs. Herein, we report a versatile method that allows simultaneous determination of the size of AgNP@BCs and mass concentration of various silver species in rat liver. Both particulate and ionic silver were extracted in their original forms from the organs by alkaline digestion, and analyzed by size exclusion chromatography combined with inductively coupled plasma mass spectrometry（SEC-ICP-MS）. While the silver mass concentrations were quantified by ICP-MS with a detection limit of 0.1 μg/g, the effective diameter of AgNP@BCs was determined based on the retention time in SEC separation with size discrimination of 0.6-3.3 nm. More importantly, we found that the BC thickness of AgNP@BCs is core size independent, and a linear correlation was found between the effective diameter and core diameter of AgNP@BCs in extracted tissues, which was used to calibrate the core diameter with standard deviations in the range of 0.2-1.1 nm. The utility of this strategy was demonstrated through application to rat livers in vivo. Our method is powerful for investigating the transformation mechanism of Ag NPs in vivo.

Intranasal delivery of nanostructured lipid carriers,solid lipid nanoparticles and nanoemulsions:A current overview of in vivo studies

The management of the central nervous system(CNS)disorders is challenging,due to the need of drugs to cross the blood-brain barrier(BBB)and reach the brain.Among the various strategies that have been studied to circumvent this challenge,the use of the intranasal route to transport drugs from the nose directly to the brain has been showing promising results.In addition,the encapsulation of the drugs in lipid-based nanocarriers,such as solid lipid nanoparticles(SLNs),nanostructured lipid carriers(NLCs)or nanoemulsions(NEs),can improve nose-to-brain transport by increasing the bioavailability and site-specifc delivery.This review provides the state-of-the-art of in vivo studies with lipid-based nanocarriers(SLNs,NLCs and NEs)for nose-to-brain delivery.Based on the literature available from the past two years,we present an insight into the different mechanisms that drugs can follow to reach the brain after intranasal administration.The results of pharmacokinetic and pharmacodynamics studies are reported and a critical analysis of the differences between the anatomy of the nasal cavity of the different animal species used in in vivo studies is carried out.Although the exact mechanism of drug transport from the nose to the brain is not fully understood and its effectiveness in humans is unclear,it appears that the intranasal route together with the use of NLCs,SLNs or NEs is advantageous for targeting drugs to the brain.These systems have been shown to be more effective for nose-to-brain delivery than other routes or formulations with non-encapsulated drugs,so they are expected to be approved by regulatory authorities in the coming years.

In Vivo Toxicity Assessment of Gold Nanoparticles in Drosophila melanogaster

The growing use of nanomaterials in commercial goods and novel technologies is generating increasing questions about possible risks for human health and environment, due to the lack of an in-depth assessment of their potential toxicity. In this context, we investigated the effects of citrate-capped gold nanoparticles （AuNPs） on the model system Drosophila melanogaster upon ingestion. We observed a significant in vivo toxicity of AuNPs, which elicited clear adverse effects in treated organisms, such as a strong reduction of their life span and fertility, presence of DNA fragmentation, as well as a significant overexpression of the stress proteins. Transmission electron microscopy demonstrated the localization of the nanoparticles in tissues of Drosophila. The experimental evidence of high in vivo toxicity of a nanoscale material, which is widely considered to be safe and biocompatible in its bulk form, opens up important questions in many fields, including nanomedicine, material science, health, drug delivery and risk assessment.

Chinese Medicine for Treating Diabetic Nephropathy

Diabetic nephropathy is one of the main causes of renal end-stage disease. The pathogenesis of diabetic nephropathy is complex. The current treatment is only for a particular cause without multi-target therapeutic drugs. Chinese medicine is a great treasure with multi-component complex drugs interacting with multiple targets and functions. This paper reviewed the protective effect of Chinese medicine for treating diabetic nephropathy in clinical studies, in vivo studies, and in vitro studies. The possible mechanisms, the major compounds and active crude drugs were also summarized. It was shown that Chinese medicine could not only relieve several symptoms and improve the quality of life, but also reduce the levels of proteinuria and kidney damage, and further improve renal function via multiple pathways based on the whole human system. Moreover, there were no reports of severe adverse reactions during the treatment.