Formulation Of Green Tea Extract Research Articles

Anticancer effect of green tea extract (GTE)-Loaded pH-responsive niosome Coated with PEG against different cell lines

Despite its excellent anti-cancer capacity in green tea extract (GTE), its applications are constrained by its low bioavailability solubility and poor bioavailability. This study aimed to design and introduce a new, stable plasma, pH-sensitive niosomal (Nio) formulation of GTE using a thin-film hydration process. Instead of cholesterol, cholesterol hemisuccinate (CHEMS) was added to generate pH-sensitivity properties and, using polyethylene glycol (PEG) (Nio / GTE / PEG / CHEMS), pH-sensitivity and plasma stability properties were integrated simultaneously into one delivery device. GC/MS, UV–vis, DLS, optical microscopy, SEM, and TEM techniques were used for the characterization of formulations. The release of formulations with pH-sensitivity was assessed in vitro in two different buffers (pH 5 and 7.4). In vitro pH-sensitivity cytotoxicity experiment was done on three different MCF-7, HepG2, and HL-60 cancer cell lines and one normal cell line (hGF) to guarantee specificity against cancer cells in the formulation. The GTE’s mass assay with GC shows many anticancer polyphenols are in the sample. The pH-responsive formulation provided positive advantages, including pH-sensitive sustained-release (about 77 percent after 24 h at pH 5) followed Higuchi release kinetics, smooth morphology of the globular surface, high trapping efficiency (81 percent), nanometer diameter (240 nm), and high stability after 3 weeks. Exposure of cancer cells to Nio / GTE / PEG / CHEMS has demonstrated excellent performance against MCF-7, HepG2, and HL-60 cell lines by strong cell internalization and synergistic toxic effect (> 50 percent; compared to free GTE). Interestingly, when treated with the formulations, the viability percentage of the normal cell line (hGF) was higher than the cancer cells, which suggests the higher formulation selectivity against cancer cells. Stable plasma, pH-sensitive niosomes will improve bioavailability and strengthen GTE's cytotoxic effect.

EVALUATION OF ETHOSOMAL EPIGALLOCATECHIN-3-GALLATE GEL FORMULATIONS AS AN ANTIOXIDANT AND THERMOGENIC AGENT

Objective: The aim of this study was to investigate in vitro and in vivo behavior of the prepared ethosomal formulations of green tea extract, which in turn would result in enhancing their therapeutic efficacy as antioxidants and thermogenic agents in obese mice.
 Methods: Fat Wistar rats were acclimated for at least 7 d in environmentally controlled cages, then they were divided into five groups: 1st received only distilled water, 2nd received an oral dose of green tea extract, in 3rd group green tea extract loaded gel base was applied on the previously shaven dorsal side of rats and in the other 2 groups selected green tea extract loaded Ethosomal gel base were applied on the previously shaven dorsal side of rats. Total antioxidant capacity by ferric reducing ability of plasma (FRAP) method, catalase enzyme activity, malondialdehyde (MDA) and thiobarbituric acid reactive substance (TBARs) levels, and real-time qPCR of UCP2 and 3 were then determined.
 Results: Formulation of green tea extract as ethosomal preparations exhibited a controlled release rate due to the reservoir action of ethosomes. The levels of TBARs and MDA were lower in groups supplemented with green tea extracts compared to control group and in groups (3-5) received it transdermal (P<0.0001 in all 3 groups) was lower than group received it orally with P = 0.0081. The messenger RNA levels of UCPs 2 and 3 in BAT were increased in those 3 groups.
 Conclusion: The results support the efficiency of ethosomal gel in penetrating the lipid rich biological membrane. The in vivo study confirms the antioxidant and thermogenic behavior of transdermal applied green tea extract.

Topical green tea formulation with anti-hemorrhagic and antibacterial effects.

Objective(s):Potentially preventable death from uncontrolled hemorrhage clearly indicates the importance of simple, fast and efficient ways to achieving hemostasis. The aim of this study was to develop a topical formulation of green tea extract for reducing bleeding that can be helpful in hemorrhage control.Materials and Methods:Hydroalcoholic extract of green tea was isolated from Camellia sinensis and formulated in polyvinyl alcohol (PVA) to achieve two concentrations of 2% and 4% v/v. Folin-Ciocalteau assay was used to determine the total amount of tannins in extract. Rheological behavior of solutions was investigated by measuring viscosity at shear rates of 0–200 sec−1. Quantitative and qualitative microbial limit tests and minimum inhibitory concentration (MIC) assay were done. The effect of formulations on bleeding time was evaluated in an animal model. Results:The total amount of tannin in green tea extract was 3.8% w/w and addition of green tea significantly increased the viscosity of PVA. The results of MIC assay showed that PVA could not inhibit the growth of bacteria, while, 716 µg/ml of green tea and 2860 µg/ml of green tea/PVA 4% inhibited the growth of Staphylococcus aureus and Pseudomonas aeruginosa. In an animal study both 2% and 4% formulations were able to stop hemorrhage approximately at an equal time compared with tranexamic acid (TXA) 50 mg/ml as a control and the lowest bleeding time was 6.4±0.51 sec for green tea/PVA 4%.Conclusion:Based on our results, the topical formulation of green tea extract in PVA has a great potential for anti-hemorrhage applications.

Theaphenon E prevents fatty liver disease and increases CD4+ T cell survival in mice fed a high-fat diet.

Obesity is a major cause of non-alcoholic fatty liver disease (NAFLD). NAFLD is an epidemic affecting nearly 34% of the adult population in the US. As a chronic inflammatory disease, NAFLD influences the immune system by dysregulating T-cell activity. Remedies for the adverse effects on the immune system are urgently needed. We studied Theaphenon E (TE), a standardized formulation of green tea extract, on the adverse effects of NAFLD in C57BL/6J mice fed a high fat diet (HFD). Mice received HFD, low fat diet (LFD) or HFD+2% TE for 35 weeks. Hepatic lipid accumulation, cell proliferation, apoptosis and CD4+T lymphocytes were measured throughout the bioassay. The hepatic composition of fatty acids was determined. The effects of epigallocatechin gallate (EGCG) metabolites on lipid accumulation in mouse and primary human liver cells were studied. Unlike mice receiving HFD, mice on HFD+2% TE maintained normal liver to body weight ratios with low levels of alanine and aspartate aminotransferase (ALT and AST). Hepatic lipid accumulation was observed in HFD mice, accompanied by increased proliferation, reduced apoptosis and loss of CD4+ T lymphocytes. TE significantly inhibited lipid accumulation, decreased proliferation, induced apoptosis and increased CD4+ T cell survival in HFD mice. It was found that the EGCG metabolite EGC-M3 reduced lipid accumulation in mouse and human hepatocytes. Linoleic acid showed the largest increase (2.5-fold) in livers of mice on a HFD and this increase was significantly suppressed by TE. Livers of HFD-fed mice showed lipid accumulation, increased proliferation, reduced apoptosis, elevated linoleic acid and loss of CD4+ T cells. TE effectively ameliorated all of these adverse effects.

Prevention of Lipid Peroxidation-derived Cyclic DNA Adduct and Mutation in High-Fat Diet-induced Hepatocarcinogenesis by Theaphenon E.

Obesity is associated with cancer risk and its link with liver cancer is particularly strong. Obesity causes non-alcoholic fatty liver disease (NAFLD) that could progress to hepatocellular carcinoma (HCC). Chronic inflammation likely plays a key role. We carried out a bioassay in the high-fat diet (HFD)-fed C57BL/6J mice to provide insight into the mechanisms of obesity-related HCC by studying γ-OHPdG, a mutagenic DNA adduct derived from lipid peroxidation. In an 80-week bioassay, mice received a low-fat diet (LFD), high-fat diet (HFD), and HFD with 2% Theaphenon E (TE) (HFD+TE). HFD mice developed a 42% incidence of HCC and LFD mice a 16%. Remarkably, TE, a standardized green tea extract formulation, completely blocked HCC in HFD mice with a 0% incidence. γ-OHPdG measured in the hepatic DNA of mice fed HFD and HFD+TE showed its levels increased during the early stages of NAFLD in HFD mice and the increases were significantly suppressed by TE, correlating with the tumor data. Whole-exome sequencing showed an increased mutation load in the liver tumors of HFD mice with G>A and G>T as the predominant mutations, consistent with the report that γ-OHPdG induces G>A and G>T. Furthermore, the mutation loads were significantly reduced in HFD+TE mice, particularly G>T, the most common mutation in human HCC. These results demonstrate in a relevant model of obesity-induced HCC that γ-OHPdG formation during fatty liver disease may be an initiating event for accumulated mutations that leads to HCC and this process can be effectively inhibited by TE. Cancer Prev Res; 11(10); 665-76. ©2018 AACR.

Gambaran Histopatologi Organ Hati dan Ginjal Mencit Balb/c setelah Pemberian Krim Ekstrak Teh Hijau (Camellia sinensis L.)

Background:Development of green tea extract formulation with the addition of enhancers to increase the ability of epigallocatechin gallate to penetrate the skin layers has been done. Objective: The purpose of this study was to determine the effect of green tea extract cream that containing enhancers on the profile of kidney and liver histopathology. Methods: The study used 4 groups which each group was treated with different concentrations of extract (2,0%; 2,5%; 3,0%; 3,5%) and 1 control group. After 24 hours and 14 days of application of the cream, the mice were sacrificed. The liver and kidneys were weighed and made preparations histopathology. Results: The results of the study showed that there similar (p>0,05) of the weight ratio of the liver and kidneys of Balb/c mice after 24 hours and 14 days. As well the histopathological test indicated that there similar (p>0,05) of the liver and kidneys after 24 hours and 14 days of administration of green tea extract cream. Conclusions: The levels concentration of green tea extract in cream (2,0%; 2,5%; 3,0%; 3,5%) that contain enhancers do not affect the liver and kidney histopathological of Balb/c mice.

Gambaran Histopatologi Organ Hati dan Ginjal Mencit Balb/c setelah Pemberian Krim Ekstrak Teh Hijau (Camellia sinensis L.)

Background:Development of green tea extract formulation with the addition of enhancers to increase the ability of epigallocatechin gallate to penetrate the skin layers has been done. Objective: The purpose of this study was to determine the effect of green tea extract cream that containing enhancers on the profile of kidney and liver histopathology. Methods: The study used 4 groups which each group was treated with different concentrations of extract (2,0%; 2,5%; 3,0%; 3,5%) and 1 control group. After 24 hours and 14 days of application of the cream, the mice were sacrificed. The liver and kidneys were weighed and made preparations histopathology. Results: The results of the study showed that there similar (p>0,05) of the weight ratio of the liver and kidneys of Balb/c mice after 24 hours and 14 days. As well the histopathological test indicated that there similar (p>0,05) of the liver and kidneys after 24 hours and 14 days of administration of green tea extract cream. Conclusions: The levels concentration of green tea extract in cream (2,0%; 2,5%; 3,0%; 3,5%) that contain enhancers do not affect the liver and kidney histopathological of Balb/c mice.

Oxidative stress-induced Akt downregulation mediates green tea toxicity towards prostate cancer cells

Green tea consumption has been shown to possess cancer chemopreventive activity. Polyphenol E (PE) is a widely used standardized green tea extract formulation. This study was designed to investigate the impact of PE on prostate cancer cells (PC3), analyze the potential signals involved and elucidate whether anti- or pro-oxidant effects may be implicated. Treatment of PC3 cells with 30 and 100μg/ml PE significantly decreased cell viability and proliferation. At the tested concentrations, PE did not exert any antioxidant activity, eliciting instead a pro-oxidant effect at concentrations 30 and 100μg/ml, which was consistent with the observed PE cytotoxicity. PE-induced cell death was associated with mitochondrial dysfunction and downregulation of Akt activation, thus suggesting their implication in the PE-elicited cell dysfunction. Cell exposure to the ROS scavenger N-Acetyl Cysteine prevented PE-induced ROS increase, pAkt impairment, and cell death, clearly indicating the causative role of ROS in the observed phenomena. Failure of PE to induce PC3 damage in cells overexpressing Akt further confirms its implication in the PE-elicited cell death. Our findings showed an association between the antiproliferative and the pro-oxidant effect elicited by PE on PC3 cells and delineates a molecular signaling pattern potentially implicated in the toxicity of PE towards prostate cancer cells.

Abstract CT122: A presurgical study of lecithin formulation of green tea extract in women with early breast cancer

Abstract Background: Epidemiologic data support an inverse association between green tea intake and breast cancer (BC) risk. Greenselect® Phytosome® (GSP) is a lecithin formulation of a caffeine-free green tea catechin extract. The purpose of the study was to determine the tissue distribution of Epigallocatechin 3-O-gallate (EGCG) and its effect on cell proliferation and other biomarkers in BC patients undergoing surgery. Methods: Twelve early BC patients received GSP, 300 mg daily for 4 weeks prior to surgery. The EGCG levels were measured before (free-) and after (total-) enzymatic hydrolysis by HPLC-MS/MS in plasma, urine, BC tissue and surrounding normal tissue. Fasting blood samples were taken at baseline, before the last administration and 2 hours later. Women were asked to avoid any tea for at least 14 days prior the beginning of the study and for the entire duration of the study. Results: All patients were fully compliant and completed the treatment program. No toxicity was observed. Free-EGCG and Total-EGCG were undetectable in baseline samples except for 1 patient with total-EGCG of 1 ng/mL in plasma. Despite a high between-subject variability, total EGCG was detectable in all tumor tissue samples collected (n = 8). Repeated administration of GSP achieved levels of total-EGCG ranging from 17 to 121 ng/mL in plasma, and up to 8 ng/g in BC tissue. Median total-EGCG concentration was higher in the tumor as compared to the adjacent normal tissue (P = 0.01).Urine EGCG levels correlated inversely with concentration in normal tissue (P = 0.01). Free-EGCG concentrations ranged from 8 to 65 ng/mL in plasma, but were under the limit of quantitation in tumor tissue and present in traces in adjacent normal breast tissue. Conclusions: Compared to other green tea extracts, GSP taken orally, appears to increase bioavailability of EGCG which selectively accumulates in breast tumor tissue. Biomarkers of activity will be presented at the conference. Median (Q1-Q3) concentrations after supplementation:Urine-final ng/mLPlasma-end ng/mLPlasma-end-2h ng/mLBreast-K ng/gBreast-N ng/gTotal-EGCG4.7 (3.3, 5.4)20.3 (14.7, 30.7)89.0 (50.9, 111.4)3.2 (2.7, 4.6)0.50 (0.5,2.3)end: before the last administration; end-2h: 2 hours after the last administration; value below the detectable level were imputed as 0.5. K: cancer tissue; N: normal tissue; P-value from Wilcoxon-rank test. Citation Format: Matteo Lazzeroni, Aliana Guerrieri-Gonzaga, Sara Gandini, Harriet Johansson, Davide Serrano, Pietro Caldarella, Gianmatteo Pagani, Paolo Arnone, Giancarlo Pruneri, Valentina Aristarco, Antonella Puccio, Serena Mora, Antonella Riva, Giovanna Petrangolini, Paolo Morazzoni, Bernardo Bonanni. A presurgical study of lecithin formulation of green tea extract in women with early breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr CT122.

Green tea extract formulation may promote wound healing

Wound repair is a natural reaction to injury, which results in restoration of tissue integrity. In this study the healing effect of green tea extract formulation was investigated on open skin wound of rabbit. Active ingredients were extracted from green tea using water and glycerin (4:1) vehicle. Three cream formulations containing 2, 5 and 10% of green tea extract were prepared in the base. A full tickness wound was made on the left flank rabbits. Green tea creams were applied on wound area twice daily. Control groups were treated with the base only or phenytoin cream 1%. Healing was determined quantitatively as the reduction in wound area. The wound healing profile of green tea treated groups were significantly better than non-treated groups. The best healing effect was observed in the green tea cream, which exert the lowest period for healing. Such effect was significantly different from phenytoin or the base. The quantity of tissue repairing was confirmed by histological examination. Green tea formulation exhibited a considerable potency for wound healing (P

Outcomes of 3% Green Tea Emulsion on Skin Sebum Production in Male Volunteers

This study was aimed to depict potential effects of stable formulation (water in oil emulsion), containing 3% green tea (Camellia sinensis L) extract on skin sebum production in healthy human volunteers. For this purpose formulation was designed using 3% ethanolic green tea extract and Abil®EM90 was used as an emulsifier. Formulation was applied to the cheeks of healthy human volunteers (n=10) for a period of 8 weeks. Measurements for skin sebum production were considered using Sebumeter MPA 5. Results were compiled and any effect produced by the formulation was justified statistically. It was observable that statistically significant (p < 0.5%) results were found for skin sebum production after long term application of the formulation. 3% formulation of green tea extract was ideal in all aspects and can be experienced in skin disorders like acne to further investigate its effects in unhealthy volunteers.

Pharmacokinetics of Green Tea Catechins in Extract and Sustained-Release Preparations

Catechins are a major constituent of green tea. For green tea to have cancer therapeutic benefit, catechin concentrations in the range of 100 nM are required continuously until apoptosis (programmed cell death) is induced. To prolong elevated plasma and interstitial concentrations of catechins, a sustained-release formulation of green tea extract was tested and compared to a commercial green tea extract (Tegreen97®). Sustained-release formulations are usually developed in the pharmaceutical industry to slowly deliver the compound over a period of time and increase the dosing interval. Plasma and interstitial fluid (ISF) pharmacokinetics of catechins were determined following an oral dose in the rat. The sustained-release formulation profile included multiple smaller peaks of total catechins in both plasma and ISF. Interstitial fluid profiles of green tea extract indicate that higher catechins concentration and longer duration in tissue than in blood may make a sustained-release form unnecessary.

Antitumor Effect of a Combination of Lysine, Proline, Arginine, Ascorbic Acid, and Green Tea Extract on Pancreatic Cancer Cell Line MIA PaCa-2

Current treatment of pancreatic cancer is generally associated with poor prognosis, even if diagnosed early, owing to its aggressive rate of metastasis and non-responsiveness to chemotherapy and radiotherapy. Matrix metalloproteinases (MMPs) have received much attention in recent years for their role in various malignancies, and have been implicated in tumor invasion, metastasis, and angiogenesis. Reported antitumor properties of ascorbic acid, lysine, proline, and green tea extract prompted us to investigate the effect of a combination of lysine, proline, arginine, ascorbic acid, and green tea extract on pancreatic cancer cell line MIA PaCa-2 for viability, MMP expression, invasion, and morphology. Viability was evaluated based on cell proliferation by MTT assay and MMP expression in condition media by gelatinase zymography. Invasion through Matrigel was assayed and morphology was observed by hematoxylin and eosin (H+E)staining. Data was analyzed by independent sample "t" test. The nutrient mixture (NM) did not inhibit cell proliferation at 10 microg/mL and exhibited a dose-dependent antiproliferative effect with maximum inhibition of 38% over the control at 1000 microg/mL. Zymography demonstrated production of only MMP-9, which showed a dose-dependent decreased expression that was abolished at 100 microg/mL of NM. Invasion through Matrigel was inhibited at 10, 50, 100, and 500 microg/mL by 66%, 66%, 87% and 100%, respectively. H&E staining did not indicate changes even at the highest concentration of NM. Our results suggest that the formulation of green tea extract, lysine, proline, and ascorbic acid, tested as a promising adjunct to standard treatment of pancreatic cancer, by inhibiting MMP expression and invasion without toxic effects important parameters in cancer metastasis.