3T3-L1 Cell Line Research Articles

Effects of Cigarette Smoke on Adipose and Skeletal Muscle Tissue: In Vivo and In Vitro Studies.

Chronic obstructive pulmonary disease (COPD), often caused by smoking, is a chronic lung disease with systemic manifestations including metabolic comorbidities. This study investigates adaptive and pathological alterations in adipose and skeletal muscle tissue following cigarette smoke exposure using in vivo and in vitro models. Mice were exposed to cigarette smoke or air for 72 days and the pre-adipose cell line 3T3-L1 was utilized as an in vitro model. Cigarette smoke exposure decreased body weight, and the proportional loss in fat mass was more pronounced than the lean mass loss. Cigarette smoke exposure reduced adipocyte size and increased adipocyte numbers. Adipose macrophage numbers and associated cytokine levels, including interleukin-1β, interleukine-6 and tumor necrosis factor-α were elevated in smoke-exposed mice. Muscle strength and protein synthesis signaling were decreased after smoke exposure; however, muscle mass was not changed. In vitro studies demonstrated that lipolysis and fatty acid oxidation were upregulated in cigarette smoke-exposed pre-adipocytes. In conclusion, cigarette smoke exposure induces a loss of whole-body fat mass and adipose atrophy, which is likely due to enhanced lipolysis.

Rosmarinus Officinalis L, Eriobotrya Japonica and Olea Europaea L Attenuate Adipogenesis in 3T3-L1-Derived Adipocytes and Inflammatory Response in LPS-Induced THP-1-Derived Macrophages

This study aims to in vitro evaluate the impact of three traditionally used anti-obesity medicinal plant extracts on inflammatory mediators in the THP-1 cell line and adipocyte differentiation and fat accumulation in the 3T3-L1 cell line. After extract preparation, an in vitro evaluation of cell viability using MTT on THP-1 and 3T3-L1, NO production levels using Griess reagent and cytokines production in THP-1 cells, and adipogenicity evaluation using Oil Red O on 3T3-L1 cells were performed. O. europaea showed the highest cytostatic on THP-1 cells with IC50 of 290.6 µg/mL, while R. officinalis was the highest on 3T3-L1 cells with IC50 of 486.6 µg/mL. All extracts significantly reduced dose-dependently nitric oxide (NO) production by THP-1-derived macrophages from 80 µM to control levels after treatment with 125 µg/mL. Similarly, all extracts decreased the levels of TNF-α and IL-6 production dose-dependently, with the highest effects reached at 250 µg/mL. all extracts increased the production levels of IL-10 from 32 pg/mL to 106.8 pg/mL, 83.5 pg/mL and 87 pg/mL at 125µg/mL, respectively. After treatment with plant extracts, the adipocyte differentiation and fat accumulation in 3T3-L1 were reduced to 20% of the control values. All three medicinal plants exhibit their traditionally known anti-obesity effects through cytostatic effects, modulation of proinflammatory and anti-inflammatory cytokine production levels, and reduction of adipocyte differentiation and fat accumulation.

Peanut Shell Extract and Luteolin Regulate Lipid Metabolism and Induce Browning in 3T3-L1 Adipocytes.

Peanut shells are agricultural waste products that require utilization. The freeze-dried ethanolic peanut shell extract (PSE) contained 10.01 ± 0.55 mg/g of luteolin (LUT) with a total polyphenol content of 18.11 ± 0.88 mg GAE/g. Thus, LUT is one of the major polyphenolic components in PSE. Although PSE displays antibacterial and neurotrophic activities, minimal research is available addressing its potential role in lipid metabolism. This study investigated the role of PSE in terms of inhibiting adipogenesis, accelerating lipolysis, and promoting lipid browning using the 3T3-L1 cell line. Without affecting cell viability, high concentrations of PSE and LUT prevented adipogenesis by reducing the mRNA levels of C/EBPα, PPARγ, and SREBP1-c, and increasing the protein levels of pACC and pAMPK. Moreover, PSE and LUT induced lipolysis by activating lipolytic proteins, and enhanced the protein expressions of the brown adipocyte-specific markers, UCP1, PGC-1α, and SIRT1 in fully differentiated 3T3-L1 adipocytes. Increased mitochondrial biosynthesis provided additional evidence in favor of these findings. Due to their anti-obesity properties, it is proposed that PSE and LUT could be used as potential dietary supplements.

Metronidazole nanosuspension loaded dissolving microarray patches: An engineered composite pharmaceutical system for the treatment of skin and soft tissue infection.

Bacteroides fragilis is one of the most common causative group of microorganisms that is associated with skin and soft tissue infections (SSTI). Metronidazole (MTZ) is the drug of choice used in the treatment of SSTI caused by the bacterium. However, owing to its physiochemical properties, MTZ have limited skin permeation, which render the drug unsuitable for the treatment of deep-rooted SSTIs. One strategy to overcome this limitation is to reformulate MTZ into nanosuspension which will then be loaded into dissolving microarray patches (MAPs) for the treatment of SSTIs caused by B. fragilis. Herein, we report for the first time on the preparation and optimisation of MAP loaded with MTZ nanosuspension (MTZ-NS). After screening a range of polymeric surfactants, we identified that Soluplus® resulted in the formation of MTZ-NS with the smallest particle size (115nm) and a narrow PDI of 0.27. Next, the MTZ-NS was further optimised using a design of experiments (DoE) approach. The optimised MTZ-NS was then loaded into dissolving MAPs with varying MTZ-NS content. Furthermore, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and cell proliferation assays along with LIVE/DEAD™ staining on the 3T3L1 cell line showed that the MTZ-NS loaded dissolving MAPs displayed minimal toxicity and acceptable biocompatibility. In vitro dermatokinetic studies showed that the MTZ-NS loaded MAPs were able to deliver the nitroimidazole antibiotic across all strata of the skin resulting in a delivery efficiency of 95% after a 24-hour permeation study. Lastly, agar plating assay using bacterial cultures of B. fragilis demonstrated that MTZ-NS loaded MAP resulted in complete bacterial inhibition in the entire plate relative to the control group. Should this formulation be translated into clinical practice, this pharmaceutical approach may provide a minimally invasive strategy to treat SSTIs caused by B. fragilis.

In-vitro Study of Trachyspermum ammi extract against Diabetes Mellitus

Diabetes resulting from the low secretion of insulin and insulin resistance, is a chronic metabolic disorder. Medication from plant materials for treatment of Diabetes Mellitus is advised as good source for novel drug or leading to preparation of novel drug. We investigated the antidiabetic property of fractions and subfractions obtained from the ethyl acetate extract of Trachyspermum ammi seeds. TLC analysis of crude extract was done. Then column chromatography was used to isolate the fractions of T. ammi. Primary screening of fractions was done by Starch-Agar Gel Diffusion assay for identification of Alpha-amylase inhibitors. Then after chromatographic separation of subfractions, their secondary screening was carried out by Glucose-Uptake assay in Yeast cells. Finally the in vitro Anti-diabetic activity of crude and screened subfraction of ethyl acetate extract of T. ammi was evaluated in 3T3-L1 cell lines. The results presented the antidiabetic activity of T. ammi and their fractions along with subfractions. Further in vitro cell line assay also proved the antidiabetic activity of T. ammi. It is suggested that the T. ammi had antidiabetic activity by inhibiting the α-amylase enzyme, also by facilitating the glucose utilization in adipose tissues and skeletal muscles.
 HIGHLIGHTS
 
 TLC analysis of crude extract of Trachyspermum ammi was performed
 Starch-Agar Gel Diffusion assay for identification of Alpha-amylase inhibitors was done
 Glucose-Uptake assay in Yeast cells was carried out
 In-vitro Anti-diabetic activity of ethyl acetate extract of ammi was evaluated in 3T3-L1 cell lines
 
 GRAPHICAL ABSTRACT

Genetic association-based functional analysis detects HOGA1 as a potential gene involved in fat accumulation.

Although there are a number of discoveries from genome-wide association studies (GWAS) for obesity, it has not been successful in linking GWAS results to biology. We sought to discover causal genes for obesity by conducting functional studies on genes detected from genetic association analysis. Gene-based association analysis of 917 individual exome sequences showed that HOGA1 attains exome-wide significance (p-value < 2.7 × 10–6) for body mass index (BMI). The mRNA expression of HOGA1 is significantly increased in human adipose tissues from obese individuals in the Genotype-Tissue Expression (GTEx) dataset, which supports the genetic association of HOGA1 with BMI. Functional analyses employing cell- and animal model-based approaches were performed to gain insights into the functional relevance of Hoga1 in obesity. Adipogenesis was retarded when Hoga1 was knocked down by siRNA treatment in a mouse 3T3-L1 cell line and a similar inhibitory effect was confirmed in mice with down-regulated Hoga1. Hoga1 antisense oligonucleotide (ASO) treatment reduced body weight, blood lipid level, blood glucose, and adipocyte size in high-fat diet-induced mice. In addition, several lipogenic genes including Srebf1, Scd1, Lp1, and Acaca were down-regulated, while lipolytic genes Cpt1l, Ppara, and Ucp1 were up-regulated. Taken together, HOGA1 is a potential causal gene for obesity as it plays a role in excess body fat development.

Chemical composition of pumpkin (Cucurbita maxima) seeds and its supplemental effect on Indian women with metabolic syndrome

This study aimed to investigate the effect of pumpkin ( Cucurbita maxima ) seed supplementation on the anthropometric measurements, biochemical parameters, and blood pressure (BP) of Indian women with metabolic syndrome (MetS). Initially, in vitro antioxidant activities of pumpkin seeds extract were assessed using standard methods. In vitro alpha-amylase, alpha-glucosidase, and dipeptidyl peptidase IV (DPP-IV) inhibition effects, along with glucose uptake assay using 3T3-L1 cell lines were performed to determine the antidiabetic effects of the seeds extract. Fatty acids and phytoconstituents were identified using gas chromatography-mass spectrometry (GC–MS). Indian women aged 30–50 years, having MetS were assigned either to intervention (n = 21) or control (n = 21) group on a random basis. Participants in the intervention group received 5 g of pumpkin seeds for 60 days. Participants in both intervention and control were advised to follow certain dietary guidelines throughout the study. Pumpkin seeds extract exhibited not only strong reducing power but also scavenged DPPH and ABTS ●+ free radicals with low IC 50 values. Pumpkin seeds inhibited alpha-amylase, alpha-glucosidase, and DPP-IV enzymes at varying concentrations with IC 50 values of 138, 22, and 246 µg/mL, respectively. Furthermore, glucose uptake was enhanced by 213% at 300 ng/mL on the 3T3-L1 cell line. GC–MS analysis showed the presence of propyl piperidine, flavone, oleic acid, and methyl esters of fatty acids in the seed extract. On comparing the changes in mean reduction/ increment in the anthropometric measurements as well as biochemical parameters and BP between the groups, significant difference ( P = 0.012) was observed only for fasting plasma glucose. Findings of the present study highlight the role of pumpkin seeds as a cost-effective adjunct in treating MetS.

Halophila beccarii extract ameliorate glucose uptake in 3T3-L1 adipocyte cells and improves glucose homeostasis in streptozotocin-induced diabetic rats

The regulation of carbohydrate metabolizing enzymes is an effective way of reducing blood glucose levels and improving glycogen synthesis during the management of type 2 diabetes. The present investigation was conducted to explain the detailed mechanism with which a Seagrass, Halophila beccarii extract (HBE) enhances the glucose uptake in the 3T3-L1 adipocyte cell culture system in invitro. HBE stimulates the glucose uptake by the translocation of glucose transporter 4 (GLUT4) on to plasma cell membrane through induction of insulin receptor substrate 1 (IRS-1)/protein kinase B (Akt) signaling pathways. To assess the effect of HBE on T2DM, we used invivo experimental diabetes rat models induced with streptozotocin (STZ) to perform oral GTT and ITT. Furthermore, we assessed the enzymatic profile of Glycolysis, Pentose phosphate pathway, and gluconeogenesis from liver tissue homogenate. After long-term exposure with HBE, our results confirmed, that HBE improves the glucose uptake in 3T3-L1 cell lines by up-regulation of glucose transporter type 4 (GLUT4) through uptake of glucose by the adipocytes. The resulting data indicated that HBE had a great potentiality in preventing diabetes and maintaining glucose homeostasis through improving glucose uptake. The present data also showed that HBE with its insulin mimetic activity activates glycogen synthesis and enhances glucose utilization by regulating the carbohydrate metabolic enzymes. The similarity between HBE and insulin indicates that the HBE follows the mechanisms same as the insulin signaling pathway to show the antidiabetic activity.

Dual Beneficial Effects of Methylnissolin-3-O-β-d-Glucopyranoside on Obesity-Induced Inflammatory Responses in Adipocyte-Macrophage Co-Culture.

Methylnissolin-3-O-β-d-glucopyranoside (MNG) is a pterocarpan analog, which protects EA.hy926 cells against oxidative damage through the Nrf2/HO-1 pathway. However, the effects of MNG on obesity-induced inflammatory responses in adipocyte-macrophage co-culture remain unclear. A differentiated murine preadipocyte cell line (3T3-L1) was co-cultured with a murine macrophage cell line (RAW264.7). Intracellular lipid accumulation was determined using Oil Red O staining. Western blotting was performed to investigate the expression of adipogenesis- and inflammation-associated proteins. Cell culture supernatants were assayed using ELISA kits to measure the levels of proinflammatory cytokines such as interleukin 6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1). MNG inhibited lipid accumulation and the production of IL-6 and MCP-1 in the 3T3-L1 and RAW264.7 cell co-culture. Moreover, MNG inhibited the protein expression of CCAAT/enhancer-binding protein alpha (C/EBPα), C/EBPβ, peroxisome proliferator-activated receptor γ (PPARγ), cyclooxygenase 2 (COX-2), and inducible nitric oxide synthase (iNOS) under the same co-culture conditions. MNG also inhibited IL-6 and MCP-1 production compared with the co-culture control. These findings demonstrate that MNG inhibited lipid accumulation and inflammatory response by downregulating IL-6 and MCP-1 production and protein expression of C/EBPβ, C/EBPα, PPARγ, COX-2, and iNOS in co-culture conditions with 3T3-L1 and RAW264.7 cells. These results suggest that MNG may be beneficial in preventing obesity-related inflammatory status.

L-Glutamic acid loaded collagen chitosan composite scaffold as regenerative medicine for the accelerated healing of diabetic wounds

Diabetic wounds (DWs) are characterized by prolonged inflammation, which poses a significant challenge for clinicians and researchers to promote healing. In this study, we fabricate L-Glutamic acid (LGA) loaded collagen/chitosan (COL-CS) composite scaffold for the accelerated healing of DW. The characterization outcomes of the composite scaffold revealed that a crosslinked scaffold holds optimum porosity, low matrix degradation, and sustained drug release in contrast to a non-crosslinked scaffold. In vitro , LGA composite scaffolds have not exhibited any toxicity on 3T3L1 cell lines. In vivo , the LGA composite scaffold has shown significantly (p < 0.001), higher rates of wound contraction than those in control and COL-CS scaffold treated groups. In addition, MMP-9 levels were also significantly reduced in LGA composite scaffold-treated group compared with those in the control and COL-CS scaffold treated group. Thus, the LGA composite scaffold may serve as a promising therapy in DW due to its unique modulatory effect on inflammatory biomarker MMP-9.

Effects of physiological and supraphysiological concentrations of cannabidiol on pre‐adipocyte proliferation

Cannabidiol (CBD) is one of the non‐psychotropic endocannabinoids from the Cannabis sativa plant. Studies support that CBD can modulate inflammation and lipid production in various cell lines, but its anti‐obesity effects are not well defined. Specifically, the effects of CBD on differentiation and proliferation of adipocytes are still unknown. Excessive fat accumulation can result in obesity, a worldwide public health problem with an increasing trend. Obesity increases the risk of developing chronic diseases. The purpose of this study was to determine the effects of different concentrations of CBD (0.2 [physiological] and 20 μM [supraphysiological]) on proliferation of pre‐adipocytes (3T3‐L1 cell line). Cells were cultured at 37°C in a humidified atmosphere (5% CO2) until they became pre‐confluent. Three groups of cells were set up: control (cells in complete growth media [CGM]), CGM supplemented with 0.2 μM CBD, and CGM supplemented with 20 μM CBD. Cells were plated in 96‐well plates in octuplicates in media containing the corresponding treatment (C, 0.2 μM CBD, or 20 μM CBD). Cells were incubated for 6, 12, 24 and 48 h, at which time points cell proliferation was measured using an MTS assay. At the 24 and 48 hour time points, cells treated with 20 μM CBD had higher cell proliferation (24 hours: 0.299 ± 0.006; 48 hours: 0.449±0.016) compared to the 0.2 μM group (24 hours: 0.262± 0.006; 48 hours: 0.217± 0.016) and the control group (24 hours: 0.249± 0.006; 48 hours: 0.312± 0.016) (p&lt;0.0001, full‐factorial ANOVA). Interestingly, cells treated with 0.2 μM CBD had decreased proliferation at 48 hours compared to control cells, while there was no difference in proliferation between these two groups at 24 hours. Our results suggest that physiological and supraphysiological concentrations of CBD affect pre‐adipocyte proliferation differently and that further research is needed to clarify these effects.

Increased Endogenous Production of Pro‐inflammatory Adipokines by 3T3‐L1 Adipocytes Differentiated with Arachidonic Acid

Inflammatory factors derived from adipose tissue (AT) can set the stage for chronic, low‐grade inflammation. This meta‐inflammation can predispose the individual to further inflammatory‐based disorders and diseases. Polyunsaturated fatty acids (PUFAs) are known to have immune modulating effects, with omega‐6 (n‐6) PUFAs typically being associated with increased production of inflammatory molecules, and n‐3 PUFAs having been shown to be anti‐inflammatory, in both in vivo and in vitro studies. The 3T3‐L1 cell line is an embryonic mouse fibroblast line that can be differentiated into an adipocyte phenotype making it a suitable line for studies on the effects of PUFAs on inflammatory molecule production by AT. The aim of this research was to determine the effects of n‐6 and n‐3 PUFAs on the release of inflammatory adipokines in lipopolysaccharide (LPS)‐challenged and LPS‐unchallenged adipocytes. Fibroblasts were differentiated into adipocytes with the addition of 100 μM of linoleic acid (LA; n‐6), arachidonic acid (ARA; n‐6) or eicosapentaenoic acid (EPA; n‐3) throughout the differentiation period, followed by a 6 h, 1 μg/ml LPS challenge. Cell‐conditioned media was collected before and after the LPS challenge for interleukin (IL)‐6 and prostaglandin E2 (PGE2) quantification via enzyme‐linked immunosorbent assays. Differentiation of adipocytes with n‐6 ARA caused a significant increase in PGE2 levels compared to control cells, or cells differentiated with LA or EPA, both after LPS challenge (two‐way ANOVA; p&lt;0.0001) and before LPS challenge (two‐way ANOVA; p&lt;0.0001). We did not observe differences in PGE2 levels between LPS‐challenged and non‐challenged cells differentiated with ARA (matched pairs t‐test; p=0.42) and EPA (p=0.57), while control (p=0.0005) and LA‐treated cells (p=0.0003) produced higher amounts of PGE2 after LPS stimulation. Interestingly, quantified levels of IL‐6 across treatments did not differ before and after LPS stimulation (matched pairs t‐test; p=0.75), although, as observed with PGE2, there was a significant increase in IL‐6 production by ARA‐differentiated cells compared to control, LPS, LA, and EPA‐treated cells (two‐way ANOVA; p=0.004). In conclusion, use of ARA in the differentiation process of 3T3‐L1 fibroblasts into a final adipocyte phenotype increases their endogenous (non‐LPS stimulated) production of IL‐6 and PGE2.

Ameliorative Effect of Ocimum forskolei Benth on Diabetic, Apoptotic, and Adipogenic Biomarkers of Diabetic Rats and 3T3-L1 Fibroblasts Assisted by In Silico Approach.

Diabetes mellitus (DM) is a complicated condition that is accompanied by a plethora of metabolic symptoms, including disturbed serum glucose and lipid profiles. Several herbs are reputed as traditional medicine to improve DM. The current study was designed to explore the chemical composition and possible ameliorative effects of Ocimum forskolei on blood glucose and lipid profile in high-fat diet/streptozotocin-induced diabetic rats and in 3T3-L1 cell lines as a first report of its bioactivity. Histopathological study of pancreatic and adipose tissues was performed in control and treatment groups, along with quantification of glucose and lipid profiles and the assessment of NF-κB, cleaved caspase-3, BAX, and BCL2 markers in rat pancreatic tissue. Glucose uptake, adipogenic markers, DGAT1, CEBP/α, and PPARγ levels were evaluated in the 3T3-L1 cell line. Hesperidin was isolated from total methanol extract (TME). TME and hesperidin significantly controlled the glucose and lipid profile in DM rats. Glibenclamide was used as a positive control. Histopathological assessment showed that TME and hesperidin averted necrosis and infiltration in pancreatic tissues, and led to a substantial improvement in the cellular structure of adipose tissue. TME and hesperidin distinctly diminished the mRNA and protein expression of NF-κB, cleaved caspase-3, and BAX, and increased BCL2 expression (reflecting its protective and antiapoptotic actions). Interestingly, TME and hesperidin reduced glucose uptake and oxidative lipid accumulation in the 3T3-L1 cell line. TME and hesperidin reduced DGAT1, CEBP/α, and PPARγ mRNA and protein expression in 3T3-L1 cells. Moreover, docking studies supported the results via deep interaction of hesperidin with the tested biomarkers. Taken together, the current study demonstrates Ocimum forskolei and hesperidin as possible candidates for treating diabetes mellitus.

Strychnos Potatorum L. Seed Polysaccharide-Based Stimuli-Responsive Hydrogels and Their Silver Nanocomposites for the Controlled Release of Chemotherapeutics and Antimicrobial Applications.

Natural Strychnos potatorumL. (SPL) polysaccharide-based dual-responsive semi-IPN-type (SPL-DMA) hydrogels have been fabricated using dimethylaminoethyl methacrylate by simple free radical polymerization. Furthermore, a facial and eco-friendly method has been developed for the green synthesis of silver nanoparticles on SPL-DMA hydrogel templates (SPL-DMA-Ag) using an aqueous leaf extract of Carissa spinarum (as a bioreducing agent). SPL-DMA and SPL-DMA-Ag were characterized using Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), thermogravimetry analysis (TGA), differential scanning calorimetry (DSC), scanning electron microscopy (SEM), transmission electron microscopy (TEM), dynamic light scattering (DLS), and evaluated network parameters. 5-Fluorouracil and doxorubicin were successfully encapsulated, and in vitro drug release studies were performed at pH values of 1.2 and 7.4 and at 25 and 37 °C. To understand the drug release mechanism of SPL-DMA hydrogels, various kinetic parameters were calculated. Biocompatibility and anticancer activities of SPL-DMA hydrogels were proved by an antioxidant activity study and in vitro cell viability studies against HeLa and 3T3-L1 cell lines. SPL-DMA-Ag hydrogels were used for antibacterial application.

Investigation of wound healing in vitro of specie Brachymenium exile (Dozy &amp; Molk.) Bosch &amp; Sande Lac.

The Brachymenium exile is the species of Bryaceae family, the family of bryophytes best represented in cerrado biome. The potential for biotechnological and biopharmaceutical applications of bryophytes have been investigated sharply. The aimed to investigate the cicatrizing potential of ethanolic extract of B. exile by scratch assay. The in vitro study was using the spreading and migration capabilities of fibroblast cell line using the extract of B. exile. the extract of B. exile allowed the cellular migration in 76% more that the control and not citotoxic assay in fibroblast cell line (3T3-L1). The extract of B. exile showed activity potential wound healing in vitro assay.

Development of Chitosan Silver Nanocomposites: Its Characteristic Study and Toxicity Effect against 3T3-L1 Cell Line

Nanocomposite from the natural source is opened a wide area for the researchers to find a natural remedy to replace the chemicals or harmful products in all the fields of agricultural, food and medical fields. Here the biopolymer (chitosan) was extracted from the two white rot fungi of Pleurotus floridanus and Pleurotus djamor, and biologically synthesized with 1mM AgNO3 solution. Synthesized chitosan nanocomposite was characterized with UV-Visible study, FTIR, FESEM, XRD, EDAS for the confirmation based upon the peaks, functional group, crystalline nature, size, morphology and the percentage of elements respectively. Toxicity study was carried out using 3T3 L1 (Mouse embryo fibroblast Cell Line) normal Cell Line to find out the cytotoxicity effect of the chitosan nanocomposite and found that the nanocomposites were non toxic to the Cell Line.

Anticancer and Cytotoxicity Activity of Native and Modified Black Rice Flour on Colon Cancer Cell Lines.

This study is intended to evaluate the cytotoxicity of native and dual-modified black rice flour against the colon cancer cell line (HCT116) and mouse embryo cell line (3T3-L1) by using the MTT assay. The modification techniques applied to prepare rice flour samples were enzymatic modification and heat moisture treatment. In this study, the IC50 of native black rice flour and modified black rice flour was 255.78 µg/mL and 340.85 µg/mL, respectively. The result confirms that the native black rice flour has significant cytotoxic and anticancer potential against human colon cancer cells. In addition, the IC50 of native black rice flour and modified black rice flour on the 3T3-L1 cell line was found to be 345.96 µg/mL and 1106.94 µg/mL, respectively. The results showed that the native black rice flour had weak cytotoxicity, and modified black rice flour was nontoxic in both the cell lines. The active component of phytochemicals present in black rice flour has a potential role in preventing colon cancer.

Buchu (Agathosma betulina and A. crenulata): Rightfully Forgotten or Underutilized?

Today, the term buchu refers to the two species in commerce, Agathosma betulina (P.J.Bergius) Pillans and Agathosma crenulata (L.) Pillans (Rutaceae). Its traditional use in urinary tract infections and related ailments made it a popular remedy, specifically in the US, in 19th century, but with the advent of antibiotics it became largely obsolete. Recent focus is on technological use and on the essential oil for use in the perfume and food-flavouring industry. A review of the scarce pharmacological research revealed moderate antimicrobial activity for a leaf extract but not the essential oil of both species in the MIC assay. In the 5-lipoxygenase (5-LO) assay the essential oil of both species revealed IC50 values of 50.37 ± 1.87 μg/ml and 59.15 ± 7.44 μg/ml, respectively. In another study 98% inhibitory activity was determined for 250 μg/ml of an ethanolic extract of A. betulina on cyclooxygenase (COX)-1 and a 25% inhibitory activity on COX-2. Analgesic activity of an ethanolic extract of A. betulina was shown in mice. Moderate antioxidant activity was determined for methanol:dichlormethane extracts of A. betulina and A. crenulata and an aqueous extract of A. betulina showed a Trolox equivalent antioxidant capacity (TEAC) of 11.8 µM Trolox. Recent in vitro studies with a commercial aqueous extract of buchu revealed increased uptake of glucose added to 3T3-L1 cell line, significant inhibition of the respiratory burst of neutrophils and monocytes, reduction in the expression of adhesion molecules and inhibition of the release of IL-6 and TNF-α. In diabetic rats the ingestion of aqueous buchu extract completely normalized the glucose level and in rats receiving a high fat diet the consumption of aqueous buchu extract resulted in less weight gain and less intraperitoneal fat gain as well as reduction of elevated blood pressure to normal associated with cardioprotective effects. Limitations in the hitherto conducted research lie in the undisclosed composition of the buchu extracts used and the difficulty in extrapolating data from animal studies to humans. Health claims for buchu products need to be substantiated by randomized, double-blind and placebo-controlled studies. Only then can they be promoted for their true therapeutic potential.

Fabrication of 3D scaffolds based on fully biobased unsaturated polyester resins by microstereo-lithography

Additive Manufacturing (AM) technologies are an effective route to fabricate tailor made scaffolds for tissue engineering (TE) and regenerative medicine, with microstereo-lithography (µSLA) being one of the most promising techniques to produce high quality 3D structures. Here, we report the crosslinking studies of fully biobased unsaturated polyesters (UPs) with 2-hydroxyethyl methacrylate (HEMA) as the unsaturated monomer (UM), using thermal and µSLA crosslinking processes. The resulting resins were fully characterized in terms of their thermal and mechanical properties. Determination of gel content, water contact angle, topography and morphology analysis by atomic force microscopy and scanning electron microscopy were also performed. The results show that the developed UP resins (UPRs) have promising properties for µSLA. In vitro cytotoxicity assays performed with 3T3-L1 cell lines showed that the untreated scaffolds exhibited a maximum cellular viability around 60%, which was attributed to the acidic nature of the UPRs. The treatment of the UPRs and scaffolds with ethanol (EtOH) improved the cellular viability to 100%. The data presented in this manuscript contribute to improve the performance of biobased UPs in AM.

Evaluation of the anti-dyslipidemic effect of spice fixed oils in the in vitro assays and the high fat diet-induced dyslipidemic mice

Dyslipidemia is a condition in which the lipid levels are altered in the body. The fixed oils (FOs) from spices, Alpinia galanga, Cinnamomum zeylanicum, Trigonella foenum-graecum, Foeniculum vulgare, and Myristica fragrans were evaluated as potential sources of anti-dyslipidemic activity. The nutraceutical compound fractions such as polyphenols, tocopherols, γ-oryzanol, and phytosterols were extracted from spice FOs, tested individually and in combination for cytotoxicity and their effect on lipid accumulation in 3T3-L1 and Hep-G2 cell lines. Further, the best performing FOs in the in vitro studies were fed to high fat diet-induced dyslipidemic C57BL/6 mice and studied its activity in the liver and adipose tissue. The in vitro assays showed no cytotoxicity in the studied FOs up to 200 μg/mL concentration. Out of all the studied nutraceutical compound fractions, the γ-oryzanol and phytosterols fractions showed a reduction in the accumulation of lipid droplets in 3T3-L1 cell lines. Moreover, the reconstituted nutraceutical fractions from C. zeylanicum, T. foenum-graecum, and F. vulgare FOs exhibited a dose dependent inhibition of lipid accumulation compared to the fractions of A. galanga and M. fragrans FOs. The in vivo studies confirmed the anti-dyslipidemic activity of C. zeylanicum FO that showed an improved lipid profile, antioxidant enzymes, and reduced lipid droplets in the liver and adipose tissue. The histopathological images of the liver and adipose tissue showed a comparable visualisation in Atorvastatin, and C. zeylanicum fed group mice. Hence, the spice FOs are potentially efficacious anti-dyslipidemic functional food ingredients and could be safer than statins.