PEG-PCL Nanocapsules Containing Curcumin: Validation of HPLC Method for Analyzing Drug-loading Efficiency, Stability Testing and Cytotoxicity on NIH-3T3 Cell Line

Abstract Curcumin (CUR) shows potential use for treating cancer. However, CUR has low solubility and reduced bioavailability, which limit its clinical effect. Therefore, the development of nanocarriers can overcome these problems and can ensure the desired pharmacological effect. In addition, it is mandatory to prove the quality, the efficacy, and the safety for a novel nanomedicine to be approved. In that sense, this paper aimed (a) to prepare CUR-loaded polyethylene glycol-poly(ε-caprolactone) nanocapsules; (b) to validate an analytical method by high performance liquid chromatography (HPLC) for quantifying CUR in these nanoformulations; (c) to evaluate the physicochemical stability of these formulations; and to investigate their cytotoxicity on NIH-3T3 mouse fibroblast cells. The HPLC method was specific to CUR in the loaded nanocapsules, linear (r = 0.9994) in a range of 10.0 to 90.0 µg.mL-1 with limits of detection and quantification of 0.160 and 0.480 µg.mL-1, respectively. Precision was demonstrated by a relative standard deviation lower than 5%. Suitable accuracy (102.37 ± 0.92%) was obtained. Values of pH, particle size, polydispersity index, and zeta potential presented no statistical difference (p > 0.05) for CUR-loaded nanoparticles. No cytotoxicity was observed against NIH-3T3 mouse embryo fibroblast cell line using both the tetrazolium salt and sulforhodamine B assays. In conclusion, a simple and inexpensive HPLC method was validated for the CUR quantification in the suspensions of nanocapsules. The obtained polymeric nanocapsules containing CUR showed suitable results for all the performed assays and can be further investigated as a feasible novel approach for cancer treatment.

In Vitro Study Of An Aqueous Extract Of Eclipta Alba Hassk. For Hepg2 Cell Line

It is the need of the day to identify the new anticancer herbal drug, which not only in possession of good anticancer effects but also cost effective. Here we are presenting such an anticancer Ayurvedic herb which is used since the centuries for the treatment of different diseases of diverse origin. Eclipta alba Hassk., also called as Bhringraj is very important medicinal herb in many medicinal formulations. Though it is commonly used for hair growth, many evidences found its hepatoprotective activity. Here we are presenting all aspects about Bhringraj in terms of qualitative and quantitative values and we have also tried to prove the anticancer activity of it for hepatic cancer. We have used the aqueous extract of Eclipta alba Hassk. for phytochemical analysis, TLC, HPLC analysis to test active chemical components in it. Extract showed presence of many active chemical components which were responsible for its anticancer activity. In vitro study we used the aqueous extract of Eclipta alba Hassk. for the evaluation of its effects on HepG2 (Human liver cancer cell line). The SRB assay results were used to evaluate the anti-cancer activity of the extract. The effects of whole plant extract on cancer cell line were studied. Percentage of cell growth and cell viability were calculated from tabulated result values of srb assay. The experiment revealed that the average percentage of growth inhibition was 68.74%. Cell viability SRB assay also showed significant growth inhibition, at the same time statistical analysis of SRB assay also proved significant results. The research performed here is very useful for set up of different extract studies of Bhringraj for its anticancer activity.

In vitro anticancer activity of extracts of Mentha Spp. against human cancer cells.

In vitro anticancer potential of methanolic and aqueous extracts of whole plants of Mentha arvensis, M. longifolia, M. spicata and M. viridis at concentration of 100 µg/ml was evaluated against eight human cancer cell lines — A-549, COLO-205, HCT-116, MCF-7, NCI-H322, PC-3, THP-1 and U-87MG from six different origins (breast, colon, glioblastoma, lung, leukemia and prostate) using sulphorhodamine blue (SRB) assay. Methanolic extracts of above-mentioned Mentha Spp. displayed anti-proliferative effect in the range of 70-97% against four human cancer cell lines, namely COLO-205, MCF-7, NCI-H322 and THP-1; however, aqueous extracts were found to be active against HCT-116 and PC-3. The results indicate that Mentha Spp. contain certain constituents with cytotoxic properties which may find use in developing anticancer agents.

In vitro cytotoxic activity of leaves extracts of Holarrhena antidysenterica against some human cancer cell lines.

In vitro cytotoxic potential of extracts (95% and 50% ethanolic extract and hot water extract at concentration of 100 µg/ml) from leaves of Holarrhena antidysenterica was evaluated against fourteen human cancer cell lines — A-549, COLO-205, DU-145, HeLa, HEP-2, IMR-32, KB, MCF-7, NCI-H23, OVCAR-5, SiHa, SK-N-MC, SW-620 and ZR-75-1 from nine different tissues (breast, colon, cervix, CNS, lung, liver, oral, ovary and prostate) using SRB assay. The 95% ethanolic extract displayed maximum anti-proliferative effect in the range of 73-92% against eight human cancer cell lines, while 50% ethanolic extract showed cytotoxic activity in the range of 70-94% against seven human cancer cell lines. However, the hot water extract did not show any activity. Among the fractions of 95% and 50% ethanolic extract, significant cytotoxic activity was found in the chloroform soluble fraction of 95% ethanolic extract at 100 µg/ml; it inhibited the growth in the range of 71-99% of seven human cancer cell lines from five different tissues viz., OVCAR-5 (ovary), HT-29 (colon), SK-N-MC (neuroblastoma), HEP-2 (liver), COLO-205 (colon), NIH-OVCAR-3 (ovary) and A-549 (lung). The cytotoxic activity of chloroform soluble fraction was found to be higher than 5-flurouracil, adriamycin, mitomycin-c and paclitaxel (anticancer drugs used as positive controls). Further in vivo studies and identification of active components from the chloroform fraction and their exact mechanism of action could be useful in designing new anticancer therapeutic agents.

Two colorimetric assays verify that calcium sulfate promotes proliferating activity of human gingival fibroblasts / 대한치과보철학회지

STATEMENT OF PROBLEM: The role of calcium sulfate in stimulating the growth of gingival soft tissue has been reported in few studies. Such a unique property of calcium sulfate could serve as a trouble-solving broker in compensating for the lack of soft tissues in various oral surgeries. PURPOSE: The purpose of this study was to compare the proliferating activities of human gingival fibroblasts seeded on various bone graft barrier materials of calcium sulfate, collagen, and polytetrafluorethylene (PTFE). MATERIAL AND METHODS: Two calcium sulfates (CAPSET(R) and CalForma(R), Lifecore Biomedical Inc., St. Paul, Minnesota, USA), a resorbable natural collagen (Bio-Gide(R), Geistlich Pharma Ag., Wolhusen, Switzerland), and a non-resorbable PTFE (TefGen-FD(R), Lifecore Biomedical Inc., St. Paul, Minnesota, USA) served as the human gingival fibroblasts'substrates and comprised the four experimental groups, whereas the untreated floors of culture plastics were used in the control group, in this study. Cells were trypsinized, seeded, and incubated for 48 h. The proliferating activities of fibroblasts were determined by XTT and SRB assay and absorbance (optical density, OD) was measured. One-way ANOVA was used to analyze the differences in the mean OD values between the groups of CAPSET, CalForma, Bio-Gide, TefGen, and the control (p<0.05). RESULTS: From the XTT assay, the mean OD value of the control group, the highest, was significantly greater than that of any of the four experimental groups followed by CalForma, CAPSET, TefGen, and Bio-Gide. Further, the mean OD value of CalForma, was significantly greater compared to that of Bio-Gide. From the SRB assay, Calforma showed the highest mean OD value, which was significantly greater than that of any other groups, followed by the control, CAPSET, Bio-Gide, and TefGen. The mean OD values of both the control and CAPSET were significantly greater compared to that of TefGen (p<0.05). CONCLUSION: Assessment of the viability and proliferation of cultured fibroblasts seeded and incubated for 48 h on various barrier-material substrates using XTT and SRB assay showed that calcium sulfate CalForma(R) promotes the proliferating activity of human gingival fibroblasts.

Induction of multidrug resistance in human breast cancer cells by exposure to chemotherapeutic drugs in vitro / 中国药理学通报

Aim To investigate the effect of exposure to high or low concentration chemotherapeutic drugs on multidrug resistance of human breast cancer cell MDA-MB-231.Method MDA-MB-231 was treated with high dose of adriamycin and paclitaxel or with low concentration of paclitaxe.SRB assay was used to determine the IC50 and RF.HE assay was used to evaluate the cellular morphology.The variations of P-gp and MDR1 were analyzed by immunocytochemistry and RT-PCR respectively.Results Cells survived only after treated with high dose of paclitaxel (MDA-MB-231/a).SRB assay showed that the growth rate of MDA-MB-231/a was the same as that of parent MDA-MB-231/w.The IC50 of the cells(MDA-MB-231/b)treated with low concentration of paclitaxel to several chemotherapeutic drugs was a little higher than that of MDA-MB-231/w.Immunocytochemistry showed that there was no difference between MDA-MB-231/a and MDA-MB-231/w in the expression of P-gp while the P-gp expression was a little higher in MDA-MB-231/b.RT-PCR assay showed that the MDR1 gene was over-expressed in MDA-MB-231/b.Conclusions The multidrug resistance cell lines can not be derived from MDA-MB-231/w by high dose of chemotherapeutic drugs.Induction of multidrug resistance response to chemotherapeutic drugs is related with transient exposure to low concentration of paclitaxel and this may be a way to establish the multidrug resistance model of MDA-MB-231 cells.