Mouse Fibrosarcoma Research Articles

Synthesis and pharmacological study of novel indole based derivates as potential anti-cancer agents

Synthetic chemistry is an important tool for the development of the potent molecules against various ailments. Indole moiety plays a very crucial in the heterocyclic medicinal chemistry, especially in the treatment of the cancer. Vincristine and Vinblastine are two important naturally occurring molecules already used for the treatment of the cancer. In the current study, we first initially synthesised the 11 indole-based derivatives and then further put under the in-vitro cancer cell lines. From all the synthesised compounds, the molecule SS-1H emerged out as the most potent molecule especially against the mouse fibrosarcoma cell lines. This compound can further be modified for the development of a molecule.

Comparison of the accumulation manner of a macromolecular drug between two mouse tumour models: study with magnetic resonance imaging and the model macromolecular drug, gadolinium-conjugated dextran

A knowledge of the difference of spatio-temporal behaviour of nanomedicine in different type of tumour models is important to develop well-targeted nanomedicine for tumour. In this study, intratumoral accumulation of the model nanomedicine, gadolinium-conjugated dextran (Gd-Dex), was examined with magnetic resonance imaging in two tumour models; mouse sarcoma S180 and radiation-induced mouse fibrosarcoma RIF-1. From time-course of the distribution images, the plasma-to-tumour interstitial tissue transfer constant (Ktrans ) and fractional plasma volume (Vp ) were calculated and mapped. Gd-Dex preferentially distributed to the marginal region of S180 tumours immediately after its injection, and then started to accumulate in some parts of the central region. Ktrans and Vp values were large in the marginal region, while only Ktrans was large in some parts of the central region. In contrast, the distribution of Gd-Dex in RIF-1 tumours was fairly homogeneous, and may have resulted from the homogeneous distributions of Ktrans and Vp . The amounts of Gd-Dex that accumulated in entire tumours in both tumour models correlated with the volume of tumours; however, accumulation in large S180 tumours deviated from the correlation in the early phase. The differences in the manner and pharmacokinetics of nanomedicine among tumour models may affect the accumulation of the medicine.

Mitochondrial proteome analysis reveals that an augmented cytochrome c oxidase assembly and activity potentiates respiratory capacity in sarcoma

Mitochondrial oxidative phosphorylation (OXPHOS) is an obligatory process in sarcoma. Despite that, the metabolic programming of sarcoma mitochondria is still unknown. To obtain a comprehensive metabolic insight of mitochondria, we developed a mouse fibrosarcoma model by injecting 3-methylcholanthrene and compared mitochondrial proteomes between sarcoma and its contralateral normal muscle using mass spectrometry. Our study identified ∼449 proteins listed in the SwissProt databases, and all the data sets are available via ProteomeXchange with the identifier PXD044903. In sarcoma, 49 mitochondrial proteins were found differentially expressed, including 36 proteins up-regulated and 13 proteins down-regulated, with the significance of p-value <0.05 and the log2[fold change] > 1 and < −1 as compared to normal muscle. Our data revealed that various anaplerotic reactions actively replenish the TCA cycle in sarcoma. The comparative expression profile and Western blotting analysis of OXPHOS subunits showed that complex-IV subunits, MT-CO3 and COX6A1, were significantly up-regulated in sarcoma vs. normal muscle. Further, biochemical and physiological assays confirmed enhanced complex-IV specific enzymatic and supercomplex activities with a concomitant increase of oxygen consumption rate in sarcoma mitochondria compared to normal muscle. Validation with human post-operative sarcoma tissues also confirms an increased MT-CO3 expression compared to normal tissue counterparts. Thus, our data comprehensively analyses the mitochondrial proteome and identifies augmented complex-IV assembly and activity in sarcoma.

Bioactive Properties of Venoms Isolated from Whiptail Stingrays and the Search for Molecular Mechanisms and Targets.

The venom-containing barb attached to their 'whip-like' tail provides stingrays a defensive mechanism for evading predators such as sharks. From human encounters, dermal stingray envenomation is characterized by intense pain often followed by tissue necrosis occurring over several days to weeks. The bioactive components in stingray venoms (SRVs) and their molecular targets and mechanisms that mediate these complex responses are not well understood. Given the utility of venom-derived proteins from other venomous species for biomedical and pharmaceutical applications, we set out to characterize the bioactivity of SRV extracts from three local species that belong to the Dasyatoidea 'whiptail' superfamily. Multiple cell-based assays were used to quantify and compare the in vitro effects of these SRVs on different cell lines. All three SRVs demonstrated concentration-dependent growth-inhibitory effects on three different human cell lines tested. In contrast, a mouse fibrosarcoma cell line was markedly resistant to all three SRVs, indicating the molecular target(s) for mediating the SRV effects are not expressed on these cells. The multifunctional SRV responses were characterized by an acute disruption of cell adhesion leading to apoptosis. These findings aim to guide future investigations of individual SRV proteins and their molecular targets for potential use in biomedical applications.

Enhancement of antitumor response of staphylococcal enterotoxin C2 mutant 2M-118 by promoting cell-mediated antitumor immunity

BackgroundStaphylococcal enterotoxin C2 (SEC2) is used as an immunotherapeutic drug in China. However, SEC2 are limited due to its immunosuppressive and toxic effects. A SEC2 2M-118 (H118A/T20L/G22E) mutant generated by site-directed mutagenesis was studied to elucidate the underlying antitumor mechanism. MethodsThe effects of 2M-118 on mouse fibrosarcoma (Meth-A) cells and cytokine responses were tested in vitro using a transwell assay and ELISA, respectively. 2M-118 effect on immune function in tumor-bearing mice was tested. Cytokine levels and antitumor responses were measured using ELISA and flow cytometry, respectively. TUNEL staining and immunohistochemistry were employed to detect the tumor apoptosis and CD4+ and CD8+ tumor infiltrating lymphocytes (TILs) in tumor tissue. Results2M-118 demonstrated the growth inhibition on tumor cells, increase of cytokines production (IL-2, IFN-γ, and TNF-α) and splenocyte proliferation in vitro. 2M-118 effectively inhibited tumor development and increased lymphocytes and cytokines in a tumor-bearing mouse model. Additionally, 2M-118 regulated the tumormicroenvironment by reducing the number of myeloid-derived suppressor cells (MDSCs), increasing the number of TILs, and inducing tumorcell apoptosis. Conclusion2M-118 promotes immune function and enhances antitumor response. This indicates that 2M-118 could potentially be developed as a novel anti-tumor drug with-highefficiencyandlowtoxicity.

Mechanoregulation of Osteoclastogenesis-Inducing Potentials of Fibrosarcoma Cell Line by Substrate Stiffness.

A micro-physiological system is generally fabricated using soft materials, such as polydimethylsiloxane silicone (PDMS), and seeks an inflammatory osteolysis model for osteoimmunological research as one of the development needs. Microenvironmental stiffness regulates various cellular functions via mechanotransduction. Controlling culture substrate stiffness may help spatially coordinate the supply of osteoclastogenesis-inducing factors from immortalized cell lines, such as mouse fibrosarcoma L929 cells, within the system. Herein, we aimed to determine the effects of substrate stiffness on the osteoclastogenesis-inducing potential of L929 cells via cellular mechanotransduction. L929 cells showed increased expression of osteoclastogenesis-inducing factors when cultured on type I collagen-coated PDMS substrates with soft stiffness, approximating that of soft tissue sarcomas, regardless of the addition of lipopolysaccharide to augment proinflammatory reactions. Supernatants of L929 cells cultured on soft PDMS substrates promoted osteoclast differentiation of the mouse osteoclast precursor RAW 264.7 by stimulating the expression of osteoclastogenesis-related gene markers and tartrate-resistant acid phosphatase activity. The soft PDMS substrate inhibited the nuclear translocation of YES-associated proteins in L929 cells without reducing cell attachment. However, the hard PDMS substrate hardly affected the cellular response of the L929 cells. Our results showed that PDMS substrate stiffness tuned the osteoclastogenesis-inducing potential of L929 cells via cellular mechanotransduction.

Abstract 2835: Fibroblast activation protein (FAP)-targeted radiotherapy increases tumor CD8+ T cell infiltration and enhances response to PD-1 immune checkpoint blockade

Abstract Background: FAP is a membrane-bound protease under investigation as a pan-cancer target, given its high expression in tumors but limited expression in normal tissues. FAP-2286 is a FAP-targeted radiopharmaceutical in clinical development for multiple solid tumors that consists of two functional elements: a FAP-targeting peptide and a chelator used to attach radioisotopes for imaging and therapeutic use. Preclinically, we evaluated the potential modulation of the immune response and antitumor efficacy of FAP-2287, a murine surrogate for FAP-2286, conjugated to the β-emitting radionuclide lutetium-177 (177Lu) as a monotherapy and in combination with a programmed cell death protein 1-targeting antibody (anti-PD-1). Methods: C57BL/6 mice bearing MCA205 fibrosarcoma mouse FAP-expressing tumors (MCA205-mFAP) were treated with 177Lu-FAP-2287, anti-PD-1, or both. Tumor uptake of 177Lu-FAP-2287 was assessed by SPECT/CT scanning, while therapeutic efficacy was measured by tumor volume. Immune profiling of tumor infiltrates was evaluated through flow cytometry, RNA expression, and immunohistochemistry analyses. Results: 177Lu-FAP-2287 accumulated in MCA205-mFAP tumors with 8.2 %ID/g uptake at 3 hours post-injection (pi), which was maintained at 2.9 %ID/g by 24 hours pi. Treatment with 177Lu-FAP-2287 demonstrated significant tumor growth inhibition (TGI, [74%; 267 mm3; P &amp;lt; 0.01]) vs vehicle control (750 mm3). Significant TGI was also observed from anti-PD-1 (57%; 373 mm³; P &amp;lt; 0.01) and the combination (92%; 145 mm³; P &amp;lt; 0.001) vs vehicle control. No significant differences in TGI were observed between the three treatment groups. In flow cytometry analysis of tumors, 177Lu-FAP-2287 increased CD8+ T cell infiltration by 2-fold on day 8 pi, alone or in combination with anti-PD-1 (16.9% and 18.0% of leukocytes, respectively), compared with 7.5% for vehicle control (P &amp;lt; 0.01). In contrast, anti-PD-1 alone had a nonsignificant increase in CD8+ T cell infiltration of 10.8% vs vehicle control (P &amp;gt; 0.05). On day 13 pi, the combination had durable higher levels of CD8+ T cells, with 16.3% of leukocytes vs 5.0% for vehicle control (P &amp;lt; 0.01). Monotherapy 177Lu-FAP-2287 also displayed a trend towards higher levels of CD8+ T cells at Day 13 vs vehicle (11.9% of leukocytes; P &amp;gt; 0.05). The increase in CD8+ T cells was accompanied by changes in co-stimulatory molecules such as CD86, resulting in greater activation by antigen-presenting cells. Additional immune cell characterization will be presented. Conclusions: FAP-targeted radiopharmaceutical enhances PD-1-antibody-mediated TGI by increasing recruitment of tumor-infiltrating CD8+ T cells, which is enhanced and maintained in the presence of anti-PD-1. These findings provide a rationale for clinical studies of combined 177Lu-FAP-2286 radiotherapy and immune checkpoint blockade in FAP-positive tumors. Citation Format: Dirk Zboralski, Aileen Höhne, Esben Christensen, Matthias Paschke, Liliane Robillard, Andrew D. Simmons, Frank Osterkamp, Thomas C. Harding, Minh Nguyen. Fibroblast activation protein (FAP)-targeted radiotherapy increases tumor CD8+ T cell infiltration and enhances response to PD-1 immune checkpoint blockade [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2835.

In vivo anticancer activity of benzoxazine and aminomethyl compounds derived from eugenol.

Indonesia is the world's primary producer of clove. In order to find new utilization for clove and new biologically active compounds, eugenol, the main constituent of clove, has been converted to its derivatives. This study aims to examine in vivo anticancer activity of benzoxazine and aminomethyl compounds derived from eugenol. Fibrosarcoma was induced by injection of benzo(a)pyrene solution. The test compounds were given per oral at 20, 40, and 80 mg/Kg body weight, once a day for 30 days. As a result, all the tested compounds showed activity in reducing the cancer incidence rate. All the tested compounds were also found to reduce tumor weight. Benzoxazine derivatives gave slightly better activity compared to aminomethyl derivatives. The strongest activity was exhibited by 6-allyl-3-(furan-2- ylmethyl)-8-methoxy-3,4-dihydro-2H-benzo(e)(1,3)oxazine. All four benzoxazine and aminomethyl compounds derived from eugenol that were tested exhibited anticancer activity in mice fibrosarcoma.

Mannose-modified hyaluronic acid nanocapsules for the targeting of tumor-associated macrophages

Tumor-associated macrophages (TAMs), a class of immune cells that play a key role in tumor immunosuppression, are recognized as important targets to improve cancer prognosis and treatment. Consequently, the engineering of drug delivery nanocarriers that can reach TAMs has acquired special relevance. This work describes the development and biological evaluation of a panel of hyaluronic acid (HA) nanocapsules (NCs), with different compositions and prepared by different techniques, designed to target macrophages. The results showed that plain HA NCs did not significantly influence the polarization of M0 and M2-like macrophages towards an M1-like pro-inflammatory phenotype; however, the chemical functionalization of HA with mannose (HA-Man) led to a significant increase of NCs uptake by M2 macrophages in vitro and to an improved biodistribution in a MN/MNCA1 fibrosarcoma mouse model with high infiltration of TAMs. These functionalized HA-Man NCs showed a higher accumulation in the tumor compared to non-modified HA NCs. Finally, the pre-administration of the liposomal liver occupying agent Nanoprimer™ further increased the accumulation of the HA-Man NCs in the tumor. This work highlights the promise shown by the HA-Man NCs to target TAMs and thus provides new options for the development of nanomedicine and immunotherapy-based cancer treatments.Graphical abstract

357 (PB137) - A novel gut-restricted small molecule TLR2 agonist enhances immune checkpoint inhibitor efficacy in a preclinical mouse fibrosarcoma tumor model

Immune checkpoint inhibitor (ICI) therapy has revolutionized cancer treatment, but patient response remains variable. Substantial research has been focused on understanding the tumor-intrinsic and tumor-extrinsic factors underlying patient response. Recently, the gut microbiome emerged as a modulator of ICI therapy in both human patients and mouse tumor models. Antibiotic treatment has deleterious effects on ICI therapy efficacy and retrospective studies have identified microbiome signatures associated with ICI therapy responders and non-responders.

Abstract A016: Selective delivery of fibroblast activation protein conjugated dual phosphoinositide 3-kinase–AKT Kinase-mTOR inhibitor associated with decreased tumor proliferation and on-target toxicity in high-grade soft-tissue sarcomas

Abstract Soft-tissue sarcomas (STS) are a group of lethal mesenchymal tumors that are a major cause of cancer related morbidity for those under the age of 20.PI3K inhibitors have been found to be effective against subset of STS but are associated with significant toxicities limiting their use.We have thought to test a selective fibroblast activation protein (FAP) targeted delivery of PI3K/AKT/mTOR inhibitor omipalisib to STS in pre-clinical setting as FAP is overwhelmingly expressed agressive soft tissue malignancies. Experimental Design: Patient derived fibrosarcomas (aSMA-,FAP+,pAKT+) were isolated by flow cytometry.FAP(-)HT1080 fibrosarcoma cell lines were used as a negative control whereas methylcoanthrene induced mouse fibrosarcoma transfected with human FAP cDNA were used as positive control.Cell expression of FAP,AKT,mTOR was compared using SDS Page,IHC,and flow cytometry.FAP ligand coupled to omipalisib(FAPL-PI3Ki)was then evaluated in-vitro and in small animal models. Results: Cell lines with known FAP expression demonstrated significant uptake of FAPL conjugated NIR fluorochrome.Wild type cells(hFAP-) showed no ligand binding both in flow cytometry and fluorescent microscopy(p&amp;lt;0.05).Escalating doses of FAP-PI3Ki suppressed AKT/mTOR activity in HT-1080 hFAP cells with 10 nM being the smallest effective dose.In hFAP+cell lines 10 nM of FAP-PI3Ki inhibited the AKT/mTOR signaling but inhibition was not observed in hFAP- cell lines demonstrating the need for FAP presence for FAPL-PI3Ki activity(p&amp;lt;0.05).Pan non-targeted PI3Ki inhibited the AKT/mTOR pathway in both hFAP+ and hFAP-cell lines.Densitometric analysis demonstrated that in hFAP+ cells,FAP-PI3Ki at above 10 nM suppresses the AKT/mTOR activity with ratio of pAKT/AKT and pmTOR/mTOR decreasing 11 and 4-fold respectively(p&amp;lt;0.05).FAP-PI3Ki activity significantly reduced when hFAP+ cells were competitively inhibited with free FAPL(4.6x increase in AKT activity,p&amp;lt;0.05).hFAP+ cells treated with FAP-PI3Ki had significant decrease in cell migration.In small animal models,SCID Mice injected with 25 umol/kg of either FAP-PI3Ki and non-targeted PI3Ki showed marked elevation of serum insulin levels in the non-targeted PI3Ki group (0.91 ng/mL vs 0.58 ng/mL,p&amp;lt;0.05).Xenograft tumor digest demonstrated mice treated with FAP-PI3Ki had Akt/mTOR pathway downregulated only in hFAP+ cell lines and this was evident in mice which received 10 nM or higher dosing(pAkt/Akt ratio of 1.1 WT mice vs 0.55 in hFAP+ mice,p&amp;lt;0.05).There was near complete suppression of the mTOR expression in the hFAP+ treatment group (pmTOR/mTOR 1.05 vs 0.087,p&amp;lt;0.05).There was no treatment related mortality in FAPL-PI3Ki group as compared to non-targeted PI3Ki delivery (p&amp;lt;0.05). Conclusion: Our study demonstrated that FAP conjugated modified omipalisib binds FAP expressing soft-tissue sarcomas both in-vitro and in xenograft models in concordance with FAP receptor density.FAP-PI3Ki induces PI3K/AkT/mTOR pathway inhibition similar to non-targeted PI3K inhibitors without significant systemic toxicities observed in pan nontargeted PI3K inhibitors. Citation Format: Feredun Azari, Gregory T. Kennedy, Ashley Chang, Bilal Nadeem, Azra Din, Isvita Marfatia, Steven Albelda, Philip Low, Madduri Srinivasarao, Ramesh Mukkamala, Neil T. Sullivan, Evgeniy Eruslanov, Sunil Singhal. Selective delivery of fibroblast activation protein conjugated dual phosphoinositide 3-kinase–AKT Kinase-mTOR inhibitor associated with decreased tumor proliferation and on-target toxicity in high-grade soft-tissue sarcomas [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr A016.

The Efficacy of Radiation is Enhanced by Metformin and Hyperthermia Alone or Combined Against FSaII Fibrosarcoma in C3H Mice.

The effects of mild-temperature hyperthermia (MTH) and metformin, alone or in combination, on the efficacy of high-dose hypofractionated radiation against experimental tumors were investigated. FSaII fibrosarcoma grown subcutaneously in the hind legs of C3H mice was irradiated with a single 15 Gy dose using a 60Co irradiator. The radio frequency capacitive method was used to heat the tumors at 41.0°C for 30 min. Metformin was intraperitoneally (i.p.) administered daily to tumor-bearing mice at a dose of 150 mg/kg. The expression levels of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), and programmed cell death-ligand 1 (PD-L1) were determined by immunohistochemical staining of the excised tumor tissues. The apoptosis of tumor cells in vivo was quantified by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining and cleaved caspase-3 staining of the excised tumor tissues. Irradiation of tumors markedly increased the expression of HIF-1α, VEGF, and PD-L1, and MTH and metformin used either alone or in combination significantly abrogated the radiation-induced upregulation of these proteins. MTH and metformin alone or combined increased the radiation-induced apoptosis in tumor cells and enhanced the radiation-induced suppression of tumor growth. The findings indicated that the increased tumor response to 15 Gy irradiation by MTH and metformin alone or in combination was due, in part, to the abrogation of the radiation-induced upregulation of HIF-1α and its downstream targets VEGF and PD-L1.

Anti-tumour effect of combinations of three acids isolated from Taraxacum officinale

Taraxacum officinale (TO) is a well-known medicinal plant used in folk medicine for its variety of biological activities. In this study a methanolic extract from roots was used to examine its anti-tumour effect by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) viability assay on two mouse tumour cell lines, fibrosarcoma and hepatoma cell lines. Normal hepatocyte and fibroblast cell lines were used as a control. Furthermore, three active compounds were isolated from the extract, caffeic acid, chlorogenic acid, and ursolic acid, in order to investigate their cytotoxicity and possible interactions between them in their combinations on the same tumour and non-tumour cell lines. The anti-tumour effect of the TO extract was confirmed on the fibrosarcoma cell line in a dose dependent manner. The anti-proliferative acting of each acid was described on both cancer cell lines and for the first time the combinations of these acids were investigated and their common effect in the mixtures reported. Further experiments to determine the mechanism of action and examine their action with conventional chemotherapeutics as a potential adjuvant therapy to enhance the chemotherapeutic effect and improve patient health with its hepatoprotective activity could be encouraged.

Comparative effects of free doxorubicin, liposome encapsulated doxorubicin and liposome co-encapsulated alendronate and doxorubicin (PLAD) on the tumor immunologic milieu in a mouse fibrosarcoma model.

Background: We have previously shown that alendronate, an amino-bisphosphonate, when reformulated in liposomes, can significantly enhance the efficacy of cytotoxic chemotherapies and help remodel the immunosuppressive tumor microenvironment towards an immune-permissive milieu resulting in increased anticancer efficacy. In addition, we have previously shown that the strong metal-chelating properties of alendronate can be exploited for nuclear imaging of liposomal biodistribution. To further improve anticancer efficacy, a pegylated liposome formulation co-encapsulating alendronate and doxorubicin (PLAD) has been developed. In this study, we examined the effects of PLAD on the tumor immunologic milieu in a mouse fibrosarcoma model in which the tumor microenvironment is heavily infiltrated with tumor-associated macrophages (TAM) that are associated with poor prognosis and treatment resistance.Methods: Doxorubicin biodistribution, characterization of the tumor immunologic milieu, cellular doxorubicin uptake, and tumor growth studies were performed in Balb/c mice bearing subcutaneously implanted WEHI-164 fibrosarcoma cells treated intravenously with PLAD, pegylated liposomal doxorubicin (PLD), free doxorubicin, or vehicle.Results: PLAD delivery resulted in a high level of tumor doxorubicin that was 20 to 30-fold greater than in free doxorubicin treated mice, and non-significantly higher than in PLD treated mice. PLAD also resulted in increased uptake in spleen and slightly lower plasma levels as compared to PLD. Importantly, our results showed that PLAD, and to a lesser extent PLD, shifted cellular drug uptake to TAM and to monocytic myeloid-derived suppressor cells (MDSC), while there was no drug uptake in neutrophilic MDSC or lymphoid cells. Free doxorubicin cellular drug uptake was below detectable levels. PLAD, and to a lesser extent PLD, also induced significant changes in number and functionality of tumor-infiltrating TAM, MDSC, Treg, NKT, and NK cells that are consistent with enhanced antitumor immune responses in the tumor microenvironment. In contrast, free doxorubicin induced moderate changes in the tumor microenvironment that could promote (decreased Treg) or be detrimental to antitumor immune responses (decreased M1 TAM and NK cells). These immune modulatory effects are reflected in the therapeutic study which showed that PLAD and PLD inhibited tumor growth and significantly prolonged survival, while free doxorubicin showed little or no anticancer activity.Conclusion: We show that liposomal delivery of doxorubicin not only alters pharmacokinetics, but also dramatically changes the immune modulatory activity of the drug cargo. In addition, our data support that the PLAD nanotheranostic platform further enhances some immune changes that may act in synergy with its cytotoxic chemotherapy effects.

Anti-proliferative effect of Cistus laurifolius on human cervical adenocarcinoma (Hep2C), human muscle rhabdomyosarcoma (RD), mouse fibrosarcoma (Wehi 164) cell line

In the current study; it was tried to determine antiproliferative activity of extract obtained from Cistus laurifolius leaves on different cancer cell type such as human cervical adenocarcinoma cells (Hep2C), human muscle rhabdomyosarcoma cells (RD), mouse fibrosarcoma cells (Wehi 164) by MTT Assay. The results indicated that Cistus laurifolius inhibited, in a dose-dependent manner, the proliferation of Hep2C, RD, Wehi 164 cells. Maximum anti-proliferatif effects were observed in 1000 µg/ml, 500 µg/ml, 250µg/ml concentrations of extract. These findings suggest that Cistus laurifolius extract could reduce proliferatif effect of cancer cell lines. Their use as potential anticancer agent needs further studies.

One pot synthesis of luminescent Mn doped ZnSe nanoparticles and its silica based water dispersible formulation for targeted delivery of doxorubicin

The manganese doped zinc selenide nanoparticles (ZnSe:Mn NPs) were synthesized by thermolysis method using oleic acid and oleylamine as a capping agent, 1-octadecene as solvent. Coating of mesoporous silica was done on ZnSe:Mn (ZnSe:Mn@mSilica) which was further functionalized with amine functional groups by treating with (3-aminopropyl)trimethoxysilane. Further pegylation was done to achieve water dispersibility by conjugating carboxyl groups of poly(ethylene glycol) diacid with the amine groups. These pegylated NPs were subsequently treated with ethylenediamine followed by acrylic acid. Conjugation of tris-(hydroxymethyl-aminomethan) was performed by Michael-type addition reaction to afford ZnSe:Mn@mSilica-PEG-Tris-OH. These TRIS functionalized NPs exhibited broad emission ranging from 590-620 nm that is an indicative for their suitability in diagnosis and monitoring progress of cancer treatment. To explore the usefulness of increased surface area because of mesoporosity, doxorubicin was loaded on ZnSe:Mn@mSilica-PEG-Tris-OH NPs through silyl ether linkage and evaluated for cytotoxicity against WEHI-164 mouse fibrosarcoma and RAJI human hematopoietic origin cancer cell lines. A decrease in 12 % of cell viability of WEHI-164 cells while 30% decrease in RAJI cell lines (IC 50 ≈ 45 nM) were observed. This shows that our formulation has more cytotoxic in RAJI cancer cell lines than that of WEHI-164 cancer cells. These results revealed that the formulation has potential for the application in drug delivery and diagnosis in chemotherapeutics.

Drug Carriers Based on Graphene Oxide and Hydrogel: Opportunities and Challenges in Infection Control Tested by Amoxicillin Release.

Graphene oxide (GO) was proposed as an efficient carrier of antibiotics. The model drug, amoxicillin (AMOX), was attached to GO using a peptide linker (Leu-Leu-Gly). GO-AMOX was dispersed in a hydrogel to which the enzyme responsible for releasing AMOX from GO was also added. The drug molecules were released by enzymatic hydrolysis of the peptide bond in the linker. As the selected enzyme, bromelain, a plant enzyme, was used. The antibacterial nature of the carrier was determined by its ability to inhibit the growth of the Enterococcus faecalis strain, which is one of the bacterial species responsible for periodontal and root canal diseases. The prepared carrier contained only biocompatible substances, and the confirmation of its lack of cytotoxicity was verified based on the mouse fibrosarcoma cell line WEHI 164. The proposed type of preparation, as a universal carrier of many different antibiotic molecules, can be considered as a suitable solution in the treatment of inflammation in dentistry.

Differences in the Inhibitory Specificity Distinguish the Efficacy of Plant Protease Inhibitors on Mouse Fibrosarcoma.

Metastasis, the primary cause of death from malignant tumors, is facilitated by multiple protease-mediated processes. Thus, effort has been invested in the development of protease inhibitors to prevent metastasis. Here, we investigated the effects of protease inhibitors including the recombinant inhibitors rBbKI (serine protease inhibitor) and rBbCI (serine and cysteine inhibitor) derived from native inhibitors identified in Bauhinia bauhinioides seeds, and EcTI (serine and metalloprotease inhibitor) isolated from the seeds of Enterolobium contortisiliquum on the mouse fibrosarcoma model (lineage L929). rBbKI inhibited 80% of cell viability of L929 cells after 48 h, while EcTI showed similar efficacy after 72 h. Both inhibitors acted in a dose and time-dependent manner. Conversely, rBbCI did not significantly affect the viability of L929 cells. Confocal microscopy revealed the binding of rBbKI and EcTI to the L929 cell surface. rBbKI inhibited approximately 63% of L929 adhesion to fibronectin, in contrast with EcTI and rBbCI, which did not significantly interfere with adhesion. None of the inhibitors interfered with the L929 cell cycle phases. The synthetic peptide RPGLPVRFESPL-NH2, based on the BbKI reactive site, inhibited 45% of the cellular viability of L929, becoming a promising protease inhibitor due to its ease of synthesis.

Evaluation of anti-cancer activity of Tinospora cordifolia on experimentally induced spontaneous fibrosarcoma in Swiss albino mice

Tinospora cordifolia, an Indian medicinal plant, was used to evaluate anti-cancer activity against spontaneous fibrosarcoma in Swiss albino mice. The qualitative and quantitative phytochemical analysis was done in hydroethanolic extract using physical tests and GC-MS study, respectively. Hundred Swiss albino female mice of eight week age were divided into 5 groups containing 20 mice each. Group A acted as normal control without tumor. In four groups B-E, fibrosarcoma was induced by tissue grafting method. Group B was kept without any treatment and served as tumor control. Group C was treated with hydroethanolic extract of T. cordifolia stem (at the rate 400mg/kg) for 30 days. Group D was treated with standard chemotherapeutic drug Paclitaxel (at the rate 5 mg/kg) alternate days. Group E was treated with both hydroethanolic extract of T. cordifolia stem (at the rate 400mg/ kg) for 30 days and Paclitaxel drug (5 mg/kg)in alternate days. We assessed the mean tumor volume, tumor doubling time, sur-vivability and percent increase in lifespan. These parameters revealed the anti-tumor activity of T. cordifolia against fibrosarcoma. In treatment groups, the histological alterations in neoplastic tissues were alternate patches of multiplying and necrotic tissues, vacuolation in tissues, and apoptotic changes. In T. cordifolia and combined treatment changes were more marked better than paclitaxel treatment. Post observational period upto 90 days revealed a good survival rate in combined treatment followed by T. cordifolia and paclitaxel treatment. The present data strongly suggested that the Tinosporacordifolia extract has anti-tumor potential and the combination therapy can be a good strategy to combat fibrosarcoma.

Investigation of the radiopacity and cytotoxicity of albodent - novel strontium carbonate incorporated calcium silicate based dental cement

Introduction. Calcium silicate (CS) dental cements have numerous clinical indications in dentistry including pulp capping, root end surgery, perforation repair and apexification/apexogenesis treatment. Materials and methods. Novel CS based dental cement with incorporation of SrCO3 radiopacifier named ALBO-DENT was used as an experimental cement material while Portland cement (Aalborg, Denmark) and ProRoot MTA (Tulsa Dental, USA) were used as controls. The radiopacity evaluation was performed using digital Trophy Radiographic system with an intention to precisely determine the minimum of radiopaque agent needed to confer to ISO radiopacity requirement. Thereafter, biocompatibility of material was tested in in vitro conditions in mouse fibrosarcoma L929 cell culture treated with materials? extracts. Cell morphology was observed using phase-contrast microscopy, while cell viability was measured using crystal violet (CV) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assays. Results. Radiopacity evaluation revealed that 30%wt addition of SrCO3 was necessary to achieve satisfactory radiopacity (3.45 mm Al). Cytotoxicity analysis using CV and MTT assays revealed that pure extracts of ALBO-DENT presented superior biocompatibility when compared to PC and MTA controls while serial dilutions of experimental cements? extracts as well as that of PC and MTA did not influence L929 cell viability. Conclusions. Novel formulation of CS cement - ALBO-DENT presented satisfactory radiopacity and adequate biocompatibility.