This study aimed to determine total phenolic content (TPC), total antioxidant activity (TAA), antioxidant vitamin composition (A, C, and E) of pomegranate fruit. In addition, two edible parts of pomegranate juice, pomegranate seed, and combination of them were compared based on antioxidant properties. TPC was determined by using Folin-Ciocalteu (FC) method based on colorimetric reduction. Ferric reduction ability power (FRAP assay) was used to test the antioxidant activity. Vitamin assessments were conducted by using high performance liquid chromatography (HPLC). Results for antioxidant vitamin composition in pomegranate juice (PJ) showed that the concentration of vitamin A was 22.8 ± 0.69 μg/100 g, vitamin C was 57.8 ± 0.59 mg/100 g, and vitamin E was 0.07 ± 0.01 mg/100 g. Besides, TPC in PJ, pomegranate seed (PS), and pomegranate seed-juice (PSJ) was 2502 ± 54, 165 ± 49, and 2696 ± 49 mg GAE/L, and TAA was 32 ± 5.1, 20 ± 2.8, and 47 ± 5.5 mmol/L respectively. This study revealed that PSJ contained high level of phenolic compounds, antioxidant activity, and vitamin C. In addition, TPC was as main contributor to antioxidant activities, and positively correlated with TAA (r2 = 0.91, p
Atherosclerosis is initiated by damage to the endothelium followed by accumulation of inflammation tissue, foam cell formation, migration and proliferation of vascular smooth muscle cell and rupture of the plaque causing formation of thrombus. The study aims to determine the anti-atherogenic or inflammatory effect of BVFCE and its underlying mechanism in in vivo model. Five groups of four rabbits were treated with normal diet (Group I), Berberis vulgaris fruit aqueous extract [25mg/kg(group II), 50mg/kg (group III) bodyweight], aspirin 30mg/kg bodyweight (group IV) and cholesterol 2% (group V) for 10 weeks of treatments. At the end of the study, the rabbits blood were collected and aorta harvested post euthanized. Serum of the blood tested for cytokines (interleukin- 6 and tumour necrosis factor – alpha while aortic plaque composition of intimal and medial cell population in aorta was assessed by macrophages specific antibody called monoclonal mouse antibody directed against RAM-11 using immunohistology-staining techniques. The fatty streak lesion were analysed macroscopically by Sudan IV and microscopically by using the Olympus Cell ^F Imaging software and served to compute intimal area. The total macrophage-rich (RAM-11-positive) area measured using a manual contrast-based, area analysis function of the Olympus Cell ^F Imaging software. Macrophage density calculated as the ratio between macrophage and intimal areas. The RAM-11– positive were cells identified by brown staining, were divided by the total number of intimal and medial cells. Macrophage infiltration into the aorta wall showed dose dependent inhibition RAM11- positive cells in the vessel wall from 1.1 ± 1.2% of total cells with Berberis vulgaris 25mg/kg to 0.58 ±1.8% with Berberis vulgaris extract (50mg/kg) while positive control showed 1.9 ± 1.1%. This study demonstrates that Berberis vulgaris fruit aqueous extract is associated with changes in rabbit aorta plaque composition that favor lesion stability, by a reduction in lipid content and inflammation, in this case, interleukin -6 and macrophages, which gave an effect of anti -inflammation.
K.alvarezii is a macro algae that claimed to have beneficial effect on reducing risk of cardiovascular disease. The present study investigated the effects of K. alvarezii on the histology of the heart in hypertensive and hypercholesterolemic induced rats. Thirty male Sprague-Dawley rats were divided into 5 groups: normal, negative control, positive control (Captopril 30mg/kg + Simvastatin 2 mg/kg), 5% and 10% K. alvarezii. All groups were fed with NaCl-high fat diet (NaCl-HFD) for 6 weeks except the normal group and treated for 4 weeks after the induction period. The body weight, blood pressure and total cholesterol were measured on week 0, 6 and 10. Body weight, blood pressure and total cholesterol was significantly increased after induction period compared to normal group (p
This study was conducted to compare the total antioxidant activity (TAA), total phenolic content (TPC) and total flavonoid content (TFC) from the different parts of papaya tree including their ripe and unripe fruit, seeds and the young leaves. Two methods namely DPPH radical scavenging activity and ß-carotene bleaching assay were used to determine the TAA, whereas TPC was determined by Folin-Ciocalteu’s method while TFC by aluminium trichloride (AlCl3). For these purposes, methanolic extracts (80%) were prepared. The results showed that the highest antioxidant activity through ß-carotene bleaching assay was observed in unripe fruit (90.67 ± 0.29%) followed by young leave, ripe fruit and the seed. In other hand, young leaves exhibited a significant higher scavenging effect compared to others and the dose required in reducing the absorbance of DPPH control solution by 50% (EC50) was calculated at 1.0 ± 0.08mg/ml. The EC50 values were 4.3 ± 0.01mg/ml, 6.5 ± 0.01mg/ml and 7.8 ± 0.06mg/ml for unripe fruit, ripe fruit and seeds respectively. Interestingly, both TPC and TFC also showed that young leaves had the highest antioxidant content (424.89 ± 0.22mg GAE/ 100 g dry weight and 333.14 ± 1.03mg rutin equivalent/ 100 g dry weight, respectively). Statistically, Pearson correlation showed there were positive correlations between TPC and TFC with antioxidant activity assayed by DPPH radical scavenging assay (r=0.846 and r=0.873, respectively). However there was no correlation between TPC and TFC with ß-carotene bleaching activity. In brief, taken into account all the parameters measured, antioxidants were highly remarkable in the sequence of young leaves > unripe fruit > ripe fruit > seed. Nevertheless, further investigation for isolation and identification of the phytoconstituents responsible for antioxidant activity is desirable.
Microinjection is a powerful tool to deliver various substances, such as nucleic acids, proteins, peptides, RNA, and synthetic molecules into mammalian cells mechanically. Through microinjection, a controlled amount of protein can be delivered into the target cells to elucidate the specific functional
effects in vitro. In this study, a series of protein microinjection optimization was performed in human breast cancer cells. The presence of Maltose Binding Protein (MBP) was microscopically monitored through indirect immunofluorescence assay. The optimization experimentation gave a high success rate when MBP protein was used at the minimum concentration of 1.5 mg/ml and at the injection pressures of 50 and 70 hPa. The average success rate of injections was 49.2±4.15% and 50.8±4.6%, while the average cell survivability was 50.98±4.67% and 49.72±5.48% at 50 and 70 hPa, respectively. The optimization of the MBP concentration and injection pressures successfully allowed an efficient delivery of precise protein dosage into breast cancer cells without any adverse effect. This microinjection optimization can be a practical guideline in any downstream applications of protein functional work.
Strobilanthes crispus (Acanthaceae) has been used traditionally as antidiabetic, diuretic, antilytic, and laxative and has been proven scientifically to possess high antioxidant activity, anti-AIDS, and anticancer properties. It is commonly consumed in the form of herbal tea. The ethnopharmacological value of this plant, such as the development of nutraceutical S. crispus herbal tea (fermented and unfermented) and assessment of their antihyperglycemic properties were investigated. The antidiabetic properties of S. crispus fermented and unfermented tea was carried out in normal and streptozotocin-induced hyperglycaemic rats for 21 days. Glucose and lipid profile (total cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol) were determined at day 0 (baseline), day 7, and day 21. The results showed that the hot water extract of both fermented and unfermented S. crispus tea reduced blood glucose in hyperglycaemic rats. S. crispus unfermented tea also reduced glucose level in normal rat. Both fermented and unfermented S. crispus tea also showed to improve lipid profile. Antioxidant and polyphenol content that present in the extracts might contribute to the antihyperglycemic and antilipidemic properties. Further study is needed to be carried out in pre-clinical and clinical environment to prove its efficacy in human.
Coracoid process (CP) bone quality and quantity are two important variables in many successful shoulder surgeries. The goal of this study was to determine the Bone Mineral Density (BMD) and morphometry of CP in Malaysian population using a non-invasive imaging data. Sixty-six (66) shoulders’ computed tomographic (CT) examinations of 132 potential sites were selected. BMD and morphometry of CP was evaluated using e-film. The present study found that there are significant (P
Two experiments were conducted to evaluate the effect of formaldehyde vaporization of a hatcher on the tracheal epithelium of chick embryos, and on the production performance and behaviour of commercial broiler chicks. In experiment 1, chick embryos were exposed to 23.5 ppm of formaldehyde vapour during the last 3 days of incubation. Tracheal samples were taken at 0, 6, 30 and 54 h after exposure to formaldehyde and examined by scanning electron microscopy for pathological changes. Observable lesions included excessive accumulation of mucus, matted cilia, loss of cilia and sloughing of the epithelium. The lesions were more severe in chicks exposed for 54 h as compared to those exposed for 6 or 30 h. In experiment 2, 60 chicks that had been exposed to formaldehyde vapour as above and 60 control chicks were used to investigate the effect of formaldehyde fumigation on production performance and behaviour. Formaldehyde vaporization resulted in higher weekly (days 0-6 and 21-27) and total (days 0-41) feed intake and poorer weekly (days 0-6, 7-13, 21-27 and 28-34) and overall (days 0-41) feed conversion ratios. Body weight, mortality and behaviour (eating, drinking, sitting and standing activities) were not affected by formaldehyde fumigation.
Our aim of this study was to develop a new methodology for constructing a bilayer human skin equivalent to create a more clinical compliance skin graft composite for the treatment of various skin defects. We utilized human plasma derived fibrin as the scaffold for the development of a living bilayer human skin equivalent: fibrin-fibroblast and fibrin-keratinocyte (B-FF/FK SE). Skin cells from six consented patients were culture-expanded to passage 1. For B-FF/FK SE formation, human fibroblasts were embedded in human fibrin matrix and subsequently another layer of human keratinocytes in human fibrin matrix was stacked on top. The B-FF/FK SE was then transplanted to athymic mice model for 4 weeks to evaluate its regeneration and clinical performance. The in vivo B-FF/FK SE has similar properties as native human skin by histological analysis and expression of basal Keratin 14 gene in the epidermal layer and Collagen type I gene in the dermal layer. Electron microscopy analysis of in vivo B-FF/FK SE showed well-formed and continuous epidermal-dermal junction. We have successfully developed a technique to engineer living bilayer human skin equivalent using human fibrin matrix. The utilization of culture-expanded human skin cells and fibrin matrix from human blood will allow a fully autologous human skin equivalent construction.
Chitosan has similar structure to glycosaminoglycans in the tissue, thus may be a good candidates as tissue engineering scaffold. However, to improve their cell attachment ability, we try to incorporate this natural polymer with collagen by combining it via cross-linking process. In this preliminary study we evaluate the cell attachment ability of chitosan-collagen scaffold versus chitosan scaffold alone. Chitosan and collagen were dissolved in 1% acetic acid and then were frozen for 24 hours before the lyophilizing process. Human skin fibroblasts were seeded into both scaffold and were cultured in F12: DMEM (1:1). Metabolic activity assay were used to evaluate cell attachment ability of scaffold for a period of 1, 3, 7 and 14 days. Scanning electron micrographs shows good cell morphology on chitosan-collagen hybrid scaffold. In conclusion, the incorporation of collagen to chitosan will enhance its cell attachment ability and will be a potential scaffold in tissue engineering.
Tricalcium phosphate/hydroxyapatite (TCP/HA), hydroxyapatite (HA), chitosan and calcium sulphate (CaSO4) were studied and evaluated for possible bone tissue engineered construct acting as good support for osteogenic cells to proliferate, differentiate, and eventually spread and integrate into the scaffold. Surface morphology visualized by SEM showed that scaffold materials with additional fibrin had more cell densities attached than those without, depicting that the presence of fibrin and collagen fibers were truly a favourite choice of cells to attach. In comparison of various biomaterials used incorporated with fibrin, TCP/HA had the most cluster of cells attached.
Neem, Azadirachta indica, is a plant from the family Meliaceae, known as "Pokok Semambu" in Malay community. It has been extensively used in India as traditional Ayurvedic and folklore minedicine for the treatment of various diseases. This study aimed to determine the distribution of selenium in the liver of rats during hepatocarcinogenesis when neem aqueous extract and dietary selenium was supplemented.
Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) evaluation were carried out in the in vivo skin construct using fibrin as biomaterial. To investigate its progressive remodeling, nude mice were grafted and the Extracellular Matrix (ECM) components were studied at four and eight weeks post-grafting. It was discovered that by 4 weeks of remodeling the skin construct acquired its native structure.
The strategy used to generate tissue-engineered bone construct, in view of future clinical application is presented here. Osteoprogenitor cells from periosteum of consenting scoliosis patients were isolated. Growth factors viz TGF-B2, bFGF and IGF-1 were used in concert to increase cell proliferation during in vitro cell expansion. Porous tricalcium phosphate (TCP)-hydroxyapatite (HA) scaffold was used as the scaffold to form 3D bone construct. We found that the addition of growth factors, greatly increased cell growth by 2 to 7 fold. TCP/HA proved to be the ideal scaffold for cell attachment and proliferation. Hence, this model will be further carried out on animal trial.
Our objective is to determine the quality of tissue engineered human skin via immunostaining, RT-PCR and electron microscopy (SEM and TEM). Culture-expanded human keratinocytes and fibroblasts were used to construct bilayer tissue-engineered skin. The in vitro skin construct was cultured for 5 days and implanted on the dorsum of athymic mice for 30 days. Immunostaining of the in vivo skin construct appeared positive for monoclonal mouse anti-human cytokeratin, anti-human involucrin and anti-human collagen type I. RT-PCR analysis revealed loss of the expression for keratin type 1, 10 and 5 and re-expression of keratin type 14, the marker for basal keratinocytes cells in normal skin. SEM showed fibroblasts proliferating in the 5 days in vitro skin. TEM of the in vivo skin construct showed an active fibrocyte cell secreting dense collagen fibrils. We have successfully constructed bilayer tissue engineered human skin that has similar features to normal human skin.
Biomaterial, an essential component of tissue engineering, serves as a scaffold for cell attachment, proliferation, and differentiation; provides the three dimensional (3D) structure and, in some applications, the mechanical strength required for the engineered tissue. Both synthetic and naturally occurring calcium phosphate based biomaterial have been used as bone fillers or bone extenders in orthopedic and reconstructive surgeries. This study aims to evaluate two popular calcium phosphate based biomaterial i.e., hydroxyapatite (HA) and tricalcium phosphate/hydroxyapatite (TCP/HA) granules as scaffold materials in bone tissue engineering. In our strategy for constructing tissue engineered bone, human osteoprogenitor cells derived from periosteum were incorporated with human plasma-derived fibrin and seeded onto HA or TCP/HA forming 3D tissue constructs and further maintained in osteogenic medium for 4 weeks to induce osteogenic differentiation. Constructs were subsequently implanted intramuscularly in nude mice for 8 weeks after which mice were euthanized and constructs harvested for evaluation. The differential cell response to the biomaterial (HA or TCP/HA) adopted as scaffold was illustrated by the histology of undecalcified constructs and evaluation using SEM and TEM. Both HA and TCP/HA constructs showed evidence of cell proliferation, calcium deposition, and collagen bundle formation albeit lesser in the former. Our findings demonstrated that TCP/HA is superior between the two in early bone formation and hence is the scaffold material of choice in bone tissue engineering.