Cytotoxic Effect of Freeze Dried Bovine Cartilage Powder and Platelet Rich Plasma (Prp) to Mesenchymal Stem Cell (Mscs)

Cartilage repair is a challenging clinical problem because the damage is an irreversible condition. Many studies had been performed using several kinds of natural or synthetic scaffold. Attempts to repair articular cartilage using scaffold usually found many problems, lacks the physical structure and mechanical properties necessary to ensure long-term efficacy to cartilage defect. Furthermore, scaffold frequently cause toxicity to the host. Therefore, this study was performed in vitro to test the toxicity effect of scaffold freeze dried bovine cartilage powder and platelets Rich Plasma (PRP). This research was conducted using pure experimental research design in 4 groups of animal stem cells which being added with scaffold freeze dried bovine cartilage scaffold provided with platelet rich plasma. This study using posttest only control group design. The result being processed with MTT assay and spectrophotometer for counting the viable stem cells. There was no significant difference in the amount of macrophage between control and the freeze dried bovine cartilage scaffold provided with PRP (p=0,128). With this result in the number of macrophages between the control with freeze dried bovine cartilage scaffold provided PRP, it can be concluded that these biomaterials have biocompatibility

Zinc oxide nanoparticles as selective killers of proliferating cells

Background: It has recently been demonstrated that zinc oxide nanoparticles (ZnO NPs) induce death of cancerous cells whilst having no cytotoxic effect on normal cells. However, there are several issues which need to be resolved before translation of zinc oxide nanoparticles into medical use, including lack of suitable biocompatible dispersion protocols and a better understanding being needed of the mechanism of their selective cytotoxic action. Methods: Nanoparticle dose affecting cell viability was evaluated in a model of proliferating cells both experimentally and mathematically. The key issue of selective toxicity of ZnO NPs toward proliferating cells was addressed by experiments using a biological model of noncancerous cells, ie, mesenchymal stem cells before and after cell differentiation to the osteogenic lineage. Results: In this paper, we report a biocompatible protocol for preparation of stable aqueous solutions of monodispersed zinc oxide nanoparticles. We found that the threshold of intracellular ZnO NP concentration required to induce cell death in proliferating cells is 0.4 ± 0.02 mM. Finally, flow cytometry analysis revealed that the threshold dose of zinc oxide nanoparticles was lethal to proliferating pluripotent mesenchymal stem cells but exhibited negligible cytotoxic effects to osteogenically differentiated mesenchymal stem cells. Conclusion: Results confirm the ZnO NP selective cytotoxic action on rapidly proliferating cells, whether benign or malignant

Interleukin-10 containing normal human serum inhibits granzyme B release but not perforin release from alloreactive and EBV-specific T cell clones

Interleukin-10 (IL-10), also known as cytokine synthesis inhibitory factor, has pleiotropic effects in immunoregulation and inflammation. It is capable of inhibiting synthesis of pro-inflammatory cytokines like interferon γ (IFNγ), IL-2, IL-3, tumor necrosis factor α(TNFα) and granulocyte macrophage colony stimulating factor (GM-CSF) made by cells such as macrophages and T helper Type 1 cells. We observed that normal human serum, derived from a healthy individual but containing large amounts of IL-10 (arbitrarily designated as "IL-10 serum"), inhibited cytotoxic activity and interfered with granzyme B release from alloreactive cytotoxic T cell (CTL) clones _in vitro_, but did not affect perforin release. The addition of normal human serum containing high levels of anti-IL-10 IgG (arbitrarily designated as "anti-IL-10 IgG serum") neutralized the inhibitory effects of IL-10 serum. Moreover, we have identified that cytotoxic activity and granzyme B release from an Epstein-Barr virus (EBV)-specific CTL clone was similarly inhibited in the presence of IL-10 serum, while perforin release was unaffected. Anti-IL-10 IgG serum also appeared to neutralize the inhibitory effect of IL-10 serum on an EBV-specific CTL clone

Targeting of Prosurvival Pathways as Therapeutic Approaches against Primary Effusion Lymphomas: Past, Present, and Future

Constitutively activated prosurvival pathways render cancer cells addicted to their effects. Consequently they turn out to be the Achilles’ heels whose inhibition can be exploited in anticancer therapy. Primary effusion lymphomas (PELs) are very aggressive non-Hodgkin’s B cell lymphomas, whose pathogenesis is strictly linked to Kaposi’s sarcoma herpesvirus (KSHV) infection. Here we summarized previous studies from our and other laboratories exploring the cytotoxic effect of drugs inhibiting the main prosurvival pathways activated in PEL cells. Moreover, the immunogenicity of cell death, in terms of dendritic cell (DC) activation and their potential side effect on DCs, is discussed

The Effect of Dimethyl Sulfone on Gastrointestinal Cancer Cell Lines (AGS, HepG2 and KYSE-30) and Flow Cytometric Analysis of the Cell Cycle

Background: Methylsulfonylmethane (MSM) is a dietary supplement to support healthy body. It is considered to be one of the least toxic substances in biology. Considering preventive effects of this substance on tumor onset and its low toxicity to healthy body we investigated in vitro cytotoxic effects of methylsulfonylmethane on cancer cell lines. Methods: AGS, HepG2 and KYSE-30 cancer cell lines were treated by MSM by initial concentration of 50 mg/ml with ¼ serial dilutions and incubated for 24, 48 and 72 hours. Cytotoxicity was examined through MTT, neutral red uptake and protein measurement assays. EB/AO staining was used for apoptotic cell detection. A DAPI staining method was used to analysis cell cycle by flow cytometry. Results: IC50 of the MSM on AGS, HepG2 and KYSE-30 cell lines were 28.04, 21.87 and 27.98 mg/ml after 72 hours respectively. The EB/AO staining showed an increase in apoptotic cells after MSM treatment. The flow cytometry cell cycle analysis showed a significant increase in cell density at G2/M phase. Conclusion: MSM had cytotoxic effect on cancer cell lines but HepG2 cell line was more susceptible. This study provides the evidence that MSM may induce cytotoxic effect on cancer cell lines by apoptosis and it could be of value in realm of chemotherapy

The cytotoxicity of fatty acid/α-lactalbumin complexes depends on the amount and type of fatty acid

peer-reviewedComplexes of the milk protein, α-lactalbumin, and the fatty acid, oleic acid, have previously been shown to be cytotoxic. Complexes of α-lactalbumin and five different fatty acids (vaccenic, linoleic, palmitoleic, stearic, and elaidic acid) were prepared and compared to those formed with oleic acid. All complexes were cytotoxic to human promyelocytic leukemia-derived (HL-60) cells but to different degrees depending on the fatty acid. The amount of fatty acid per α-lactalbumin molecule was found to correlate with the cytotoxicity; the higher the number of fatty acids per protein, the more cytotoxic the complex. Importantly, all the tested fatty acids were also found to be cytotoxic on their own in a concentration dependent manner. The cytotoxic effect of complexes between α-lactalbumin and linoleic acid, vaccenic acid, or oleic acid was further investigated using flow cytometry and found to induce cell death resembling apoptosis on Jurkat cells. Practical applications: Cytotoxic complexes of α-lactalbumin and several different fatty acids could be produced. The cytotoxicity of all the variants is similar to that previously determined for α-lactalbumin/oleic acid complexes.This work was supported by the Food Institutional Research Measure (FIRM, project no. 08RDTMFRC650) of the Department of Agriculture, Fisheries, and Food, Ireland

Bortezomib promotes KHSV and EBV lytic cycle by activating JNK and autophagy

KSHV and EBV are gammaherpesviruses strictly linked to human cancers. Even if the majority of cancer cells harbor a latent infection, the few cells that undergo viral replication may contribute to the pathogenesis and maintenance of the virus-associated malignancies. Cytotoxic drugs used for the therapies of cancers harboring virus-infection often have, as side effect, the activation of viral lytic cycle. Therefore it is important to investigate whether they affect viral reactivation and understand the underlying mechanisms involved. In this study, we found that proteasome inhibitor bortezomib, a cytotoxic drug that efficiently target gammaherpesvirus-associated B cell lymphomas, triggered KSHV or EBV viral lytic cycle by activating JNK, in the course of ER stress, and inducing autophagy. These results suggest that the manipulation of these pathways could limit viral spread and improve the outcome of bortezomib treatment in patients affected by gammaherpesvirus-associated lymphomas