Abstract:The Chinese Hamster ovary (CHO) cell line is widely used for measuring drug cytotoxicity and resistance. Therefore, the effects of two major Ayurvedic drugs (W. somnifera root and E. officinalis fruits) on the short and long-term growth of these cells were investigated. A standard 96-well plate assay was used to measure short-term growth. For assessment of long-term growth, the colony formation assay (CFA) was used, which measures clonogenic potential. This assay is the best measure of the cytotoxicity of anti… Show more
“…This assay is based on cell metabolic activity where the reduction of yellow tetrazolium salt to orange formazan dye only occurs in viable cells by mitochondrial dehydrogenases; this can be measured by absorbance. It is important to bear in mind that the XTT assay aims to demonstrate immediate cytotoxic effect on cultured cells, whereas the clonogenic survival assay shows whether other damage has occurred to cells that interfere with or stop their proliferative capacity at a later time (Sumantran et al, 2007). The results of this study showed the absence of cytotoxic effects from HA/ PLGA and Bonefill ® eluates, which are in accordance with literature demonstrating HA, bovine bone, and PLGA as biocompatible materials (Galia et al, 2008;Cieślik et al, 2009;Trif et al, 2015).…”
Bone substitutes based on hydroxyapatite (HA) and Bonefill ® (BO-inorganic bovine bone) associated with poly(lactic-co-glycolic acid) (PLGA) (HA/PLGA and BO/PLGA) were evaluated concerning cytotoxicity, genotoxicity and mutagenicity as potential candidates for bone repair. The materials were developed and provided by Bionnovation Biomedical Products Ltda. Eluates from these bone substitutes were prepared for toxicity evaluations using eukaryotic cell cultures. HA/PLGA was used as a comparison for Bonefill ®. Cell viability was evaluated by XTT assay and surviving fraction was calculated for clonogenic survival. Additionally, tail moment was used to assess genotoxicity (comet assay). The frequencies of binucleated cells with micronucleus (FBMN), micronucleus (FMN), nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) were analysed by cytokinesis-block micronucleus assay (CBMN assay). Results showed no statistical difference in cell viability compared with negative control (NC) The eluates did not promote delayed cytotoxicity whereas the surviving fraction rate for cultured cells was similar to NC. Furthermore, no genotoxicity or mutagenicity effects were observed for cultured cells with the Bonefill/PLGA and HA/PLGA eluates. In conclusion, the negative cytotoxicity, genotoxicity and mutagenicity results indicate that these bone substitutes presented interesting preliminary results as potential biomaterials for bone repair.
“…This assay is based on cell metabolic activity where the reduction of yellow tetrazolium salt to orange formazan dye only occurs in viable cells by mitochondrial dehydrogenases; this can be measured by absorbance. It is important to bear in mind that the XTT assay aims to demonstrate immediate cytotoxic effect on cultured cells, whereas the clonogenic survival assay shows whether other damage has occurred to cells that interfere with or stop their proliferative capacity at a later time (Sumantran et al, 2007). The results of this study showed the absence of cytotoxic effects from HA/ PLGA and Bonefill ® eluates, which are in accordance with literature demonstrating HA, bovine bone, and PLGA as biocompatible materials (Galia et al, 2008;Cieślik et al, 2009;Trif et al, 2015).…”
Bone substitutes based on hydroxyapatite (HA) and Bonefill ® (BO-inorganic bovine bone) associated with poly(lactic-co-glycolic acid) (PLGA) (HA/PLGA and BO/PLGA) were evaluated concerning cytotoxicity, genotoxicity and mutagenicity as potential candidates for bone repair. The materials were developed and provided by Bionnovation Biomedical Products Ltda. Eluates from these bone substitutes were prepared for toxicity evaluations using eukaryotic cell cultures. HA/PLGA was used as a comparison for Bonefill ®. Cell viability was evaluated by XTT assay and surviving fraction was calculated for clonogenic survival. Additionally, tail moment was used to assess genotoxicity (comet assay). The frequencies of binucleated cells with micronucleus (FBMN), micronucleus (FMN), nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) were analysed by cytokinesis-block micronucleus assay (CBMN assay). Results showed no statistical difference in cell viability compared with negative control (NC) The eluates did not promote delayed cytotoxicity whereas the surviving fraction rate for cultured cells was similar to NC. Furthermore, no genotoxicity or mutagenicity effects were observed for cultured cells with the Bonefill/PLGA and HA/PLGA eluates. In conclusion, the negative cytotoxicity, genotoxicity and mutagenicity results indicate that these bone substitutes presented interesting preliminary results as potential biomaterials for bone repair.
“…Moreover, cell numbers were significantly reduced on exposure to positive control camptothecin at 0.1 μM as depicted in Figure b, whereas the cell number observed was higher in the case of NDRM as compared to nHA (Figure c), indicating the cell survival and viability of normal cells on treatments. Colony formation was reported as an in vitro quantitative technique to look at the potential of a single cell to grow into a large colony through clonal expansion . This assay also measures cell survival, and it is normally employed as a sensitive model for measuring long-term cytotoxicity.…”
The aim of this study was to prepare a novel dental restorative material (NDRM) and to understand its cell viability behavior. The hydroxyapatite (HA) nanopowder was synthesized using a wet chemical precipitation method using calcium hydroxide and orthophosphoric acid as precursors. The as-prepared HA nanopowder was annealed at different temperatures to get a pure compound with a Ca/P ratio close to 1.67. The optimal temperature was found to be 600 °C, whereas at a higher temperature, HA starts decomposing into CaO. The preparation of NDRM was conducted in two steps. The first step comprises the preparation of HA nanopowder-and gelatin (G)-based film using microwave heating. In the second step, the homogenized mixture of the HA-G film was mixed with different amounts of acrylic acid to form a self-flowable NDRM paste. Further, both these materials (HA nanopowder and NDRM) were characterized using FTIR, XRD, and SEM−EDX analyses. The FTIR and XRD results show the peaks corresponding to natural bone apatite and therefore confirm the formation of HA. EDX results showed the presence of Ca and P in HA nanopowder and NDRM with Ca/P ratios of 1.79 and 1.63, respectively. Synthesized NDRM was also analyzed for its in vitro cytotoxic and reproductive viability potential against normal cells using MTT and clonogenic assay. The analysis showed significantly higher cellular viability on the treatment with NDRM when compared to HA nanopowder as well as no colony suppression by both materials was observed on the normal cell line (fR2) even after exposure for 24 h, indicating its nontoxicity. The synthesized NDRM therefore can be considered as a promising candidate for dental caries restoration applications.
“…Figure 11(b) shows the surviving fraction obtained by the clonogenic cell survival assay of the β-TCP/ MCPM and β-TCP/MCPM/Si cements. This assay measures the proliferative ability of single cells to form a colony and is routinely used as a sensitive model for assessing long-term cytotoxicity [38]. Considering the XTT results, CHO-K1 cells treated with C1 and C2 exhibited a proliferation rate lower than the NC.…”
Calcium phosphate cement has been widely investigated as a bone graft substitute due to its excellent self-setting ability, biocompatibility, osteoconductivity and moldability. In addition, mesoporous materials have been studied as potential materials for application in medical devices due to their large surface area, which is capable of loading numerous biological molecules, besides being bioactive. In this study, bone β-TCP-MCPM-based injectable cement with mesoporous silica particles was synthesized and characterized in terms of its mechanical properties, microstructure, porosity, injectability, in vitro bioactivity and degradability; together with toxicity effects in CHO-K1 cell culture. The results showed that the β-TCP-MCPM cement is bioactive after soaking in simulated body fluid solution, and mesoporous silica particles provided better physicochemical properties compared with silica-free cement. Toxicity assays showed low CHO-K1 cell viability after treatment with more concentrated extracts (200 mg ml). However, this behavior did not compromise the reproductive capacity and did not promote significant DNA damage in those cells. In conclusion, the β-TCP-MCPM cement associated with mesoporous silica might be considered as a potential bone substitute for the repair and regeneration of bone defects.
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