Search results for: in vivo study

Authors: N. O. Muniz, F. A. Vechietti, L. Treccani, K. Rezwan, Luis Alberto dos Santos

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Developing new technologies in the manufacture of biomaterials is a major challenge for researchers in the tissue engineering area. Many in vitro and in vivo studies have revealed the significance of the porous structure of the biomaterials on the promotion of bone ingrowth. The use of Rapid Prototyping in the manufacture of ceramics in the biomedical area has increased in recent years and few studies are conducted on obtaining alumina pieces. The aim of this work was the study of alumina pieces obtained by 3D printing and uniaxial dry pressing (DP) in order to evaluate porosity achieved by this two different techniques. Also, the influence of the powder drying process was determined. The row alumina powders were drying by freeze drying and oven. Apparent porosity, apparent density, retraction after thermal treatment were evaluated. The porosity values obtained by DP, regardless of method of drying powders, were much lower than those obtained by RP as expected. And for the prototyped samples, the method of powder drying significantly influenced porosities, reached 48% for drying oven versus 65% for freeze-drying. Therefore, the method of 3D printing, using different powder drying, allows a better control over the porosity.

Keywords: rapid prototyping, freeze-drying, porosity, alumina

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Authors: Mardiati Zain, W. S. N. Rusmana, Erpomen, Malik Makmur, Ezi Masdia Putri

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Background and Aim: The leaves of the Leucaenaleucocephala tree have potential as a nitrogen source for ruminants. Leucaena leaf meal as protein supplement has been shown to improve the feed quality of ruminants. The effects of different levels of Leucaena leucocephala supplementation as substitute of concentrate on fiber digestibility and in vitro rumen fermentation characteristics were investigated. This research was conducted in vitro. The study used a randomized block design consisting of 3 treatments and 5 replications. The treatments were A. 40% rice straw ammoniated + 60% concentrate, B. 40% rice straw ammoniated + 50% concentrate + 10% Leucaena leuchephala, C. 40% rice straw ammoniated + 40% concentrate + 20% Leucaena leuchephala, Result: The results showed that the addition of Leucaena leucocephala increased the digestibility of Neutral detergent Fiber NDF and Acid Detergent Fiber (ADF) (p < 0.05). In this study, rumen NH3, propionate, amount of escape protein and total Volatyl Fatty Acid (VFA) were found increased significantly at treatment B. No significant difference was observed in acetate and butyrate production. The populations of total protozoa and methane production had significantly decreased (P < .05) in supplemented group. Conclusion: Supplementation of leuchaena leucochepala on completed feed based on ammoniated rice straw in vitro can increase fiber digestibility, VFA production and decreased protozoa pupulataion and methane production. Supplementation of 10% and 20% L. leucochepala were suitable to be used for further studies, therefore in vivo experiment is required to study the effects on animal production.

Keywords: digestibility, Leucaena leucocephala, complete feed, rice straw ammoniated

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Authors: Stephanie Newman

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Niemann-Pick Type C disease is a rare, usually fatal lysosomal storage disorder. Although clinically characterized by progressive neurodegeneration, there is also evidence of altered innate immune responses such as neuroinflammation that promote disease progression. We have initiated an investigation into whether phagocytosis, an important innate immune activity and the process by which particles are ingested is defective in NPC. Using an in vitro assay, we have shown that NPC macrophages have a deficiency in the phagocytosis of different particles. We plan to investigate the mechanistic basis for impaired phagocytosis, the contribution that this deficiency makes to disease pathology, and whether therapies that have shown in vivo benefit are able to restore phagocytic activity.

Keywords: Niemann Pick Disease C, phagocytosis, innate immunity, lysosomal storage disorder

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Authors: Elena Petersen, Inna Kornienko, Svetlana Guryeva, Sergey Simakov

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In vitro organoid cultivation requires the simultaneous provision of necessary vascularization and nutrients perfusion of cells during organoid development. However, many aspects of this problem are still unsolved. The functionality of vascular network intergrowth is limited during early stages of organoid development since a function of the vascular network initiated on final stages of in vitro organoid cultivation. Therefore, a microchannel network should be created in early stages of organoid cultivation in hydrogel matrix aimed to conduct and maintain minimally required the level of nutrients perfusion for all cells in the expanding organoid. The network configuration should be designed properly in order to exclude hypoxic and necrotic zones in expanding organoid at all stages of its cultivation. In vitro vascularization is currently the main issue within the field of tissue engineering. As perfusion and oxygen transport have direct effects on cell viability and differentiation, researchers are currently limited only to tissues of few millimeters in thickness. These limitations are imposed by mass transfer and are defined by the balance between the metabolic demand of the cellular components in the system and the size of the scaffold. Current approaches include growth factor delivery, channeled scaffolds, perfusion bioreactors, microfluidics, cell co-cultures, cell functionalization, modular assembly, and in vivo systems. These approaches may improve cell viability or generate capillary-like structures within a tissue construct. Thus, there is a fundamental disconnect between defining the metabolic needs of tissue through quantitative measurements of oxygen and nutrient diffusion and the potential ease of integration into host vasculature for future in vivo implantation. A model is proposed for growth prognosis of the organoid perfusion based on joint simulations of general nutrient diffusion, nutrient diffusion to the hydrogel matrix through the contact surfaces and microchannels walls, nutrient consumption by the cells of expanding organoid, including biomatrix contraction during tissue development, which is associated with changed consumption rate of growing organoid cells. The model allows computing effective microchannel network design giving minimally required the level of nutrients concentration in all parts of growing organoid. It can be used for preliminary planning of microchannel network design and simulations of nutrients supply rate depending on the stage of organoid development.

Keywords: 3D model, consumption model, diffusion, spheroid, tissue organoid

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Authors: Murtada Ahmed Oshi, Jin-Wook Yoo

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Layer-by-layer (LBL) coating has gained popularity for drug delivery of therapeutic drugs. Herein we described a novel approach for enhancing the therapeutic efficiency of the locally administered dexamethasone (Dex) for inflammatory bowel disease (IBD). We utilized a LBL-coating technique on Dex microcrystals (DexMCs) with multiple layers of polyelectrolytes composed of poly (allylamine hydrochloride) (PAH), poly (sodium 4-styrene sulfonate) (PSS) and Eudragit® S100 (ES). The successful deposition of the layers onto DexMCs surfaces were confirmed through zeta potential measurement and confocal laser scanning microscopy. The surface morphology was investigated through scanning electron microscopy. The drug encapsulation efficiency was 95% with a mean particle size of 2 µm and negative surface charge (-40 mV). Moreover, in vitro drug release study showed a minimum release of the drug ( 15%) at an acidic condition during initial first 5 h, followed by sustained-release at an alkaline condition. For in vivo study, LBL-DxMCs were administered orally to ICR mice suffering from dextran sulfate sodium-induced colitis. LBL-DxMCs substantially enhanced anti-IBD activities as compared to DxMCs. Macroscopic, histological and biochemical (tumor necrosis factor-α, interleukin-6 and myeloperoxidase) examinations revealed marked improvements of colitis signs in the mice treated with LBL-DxMCs compared with those treated with DxMCs. Overall, LBL-DxMCs could be a suitable candidate for the treatment of IBD.

Keywords: dexamethasone, inflammatory bowel disease, LBL-coating, polyelectrolytes

Procedia PDF Downloads 177

Authors: Richard Nyanzi, Maupi E. Letsoalo, Jacobus N. Eloff, Piet J. Jooste

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Candida albicans naturally occurs in the gastrointestinal tract (GIT) of more than 50% of humans. Overgrowth of the fungus causes several forms of candidiasis including oral thrush. Overgrowth tends to occur in immunocompromised humans such as diabetic, cancer and HIV patients. Antifungal treatment is available, but not without shortcomings. In this study, inhibitory activity of five probiotic Lactobacillus strains was demonstrated against the growth of seven clinical strains of Candida albicans by co-culturing of the organisms in a maize gruel (MG) medium. Phenotypic tests, molecular techniques and phylogenetic analysis have enabled precise identification of the organisms used in the study. The quantitative pour plate technique was used to enumerate colonies of the yeasts and the lactobacilli and the Kruskal-Wallis test and ANOVA tests were employed to compare the distributions of the colonies of the organisms. The cereal medium, containing added carbon sources, was inoculated with a Candida and a Lactobacillus strain in combination and incubated at 37 °C for 168 h. Aliquots were regularly taken and subjected to pH determination and colony enumeration. Certain Lactobacillus strains proved to be inhibitory and also lethal to some Candida albicans strains. A low pH due to Lactobacillus acid production resulted in significant low Candida colony counts. Higher Lactobacillus colony counts did not necessarily result in lower Candida counts suggesting that inhibitory factors besides low pH and competitive growth by lactobacilli contributed to the reduction in Candida counts. Such anti-Candida efficacy however needs to be confirmed by in vivo studies.

Keywords: candida albicans, oral thrush, candidiasis, lactobacillus, probiotics

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Authors: H.Karnezi, K. Tierney

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Childhood fears that persist over time and interfere with the children’s normal functioning may have detrimental effects on their social and emotional development. Cognitive behavior therapy is considered highly effective in treating fears and anxieties. However, given that many childhood fears are based on fantasy, the applicability of CBT may be hindered by cognitive immaturity. Furthermore, a lack of motivation to engage in therapy is another commonly encountered obstacle. The purpose of this study was to introduce and evaluate a more developmentally appropriate intervention model, specifically designed to provide phobic children with the motivation to overcome their fears. To this end, principles and techniques from cognitive and behavior therapies are incorporated into the ‘Drama in Education’ model. The Cognitive Behaviour Drama (CBD) method involves using the phobic children’s creativity to involve them in the therapeutic process. The children are invited to engage in exciting fictional scenarios tailored around their strengths and special interests. Once their commitment to the drama is established, a problem that they will feel motivated to solve is introduced. To resolve it, the children will have to overcome a number of obstacles culminating in an in vivo confrontation with the fear stimulus. The study examined the application of the CBD model in three single cases. Results in all three cases shown complete elimination of all fear-related symptoms. Preliminary results justify further evaluation of the Cognitive Behaviour Drama model. It is time and cost-effective, ensuring the clients' immediate engagement in the therapeutic process.

Keywords: phobias, autism, intervention, drama

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Authors: Saba Atefyekta

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Aim: Control of bacterial bioburden in wounds is an important step for minimizing the risk of wound infection. An antimicrobial hydrogel wound dressing is developed out of soft polymeric hydrogels that contain antimicrobial peptides (AMPs). Such wound dressings can bind and kill all types of bacteria, even the resistance types at the wound site. Methods: AMPs are permanently bonded onto a soft nanostructured polymer via covalent attachment and physical entanglement. This improves stability, rapid antibacterial activity, and, most importantly, prevents the leaching of AMPs. Major Findings: Antimicrobial analysis of antimicrobial hydrogels using in-vitro wound models confirmed >99% killing efficiency against multiple bacterial trains, including MRSA, MDR, E. Coli. Furthermore, the hydrogel retained its antibacterial activity for up to 4 days when exposed to human serum. Tests confirmed no release of AMPs, and it was proven non-toxic to mammalian cells. An in-vivo study on human intact skin showed a significant reduction of bacteria for part of the subject’s skin treated with antibacterial hydrogels. A similar result was detected through a qualitative study in veterinary trials on different types of surgery wounds in cats, dogs, and horses. Conclusions: Antimicrobial hydrogels wound dressings developed by permanent attachment of AMPs can effectively and rapidly kill bacteria in contact. Such antibacterial hydrogel wound dressings are non-toxic and do not release any substances into the wound.

Keywords: antibacterial wound dressing, antimicrobial peptides, post-surgical wounds, infection

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Authors: Cheng-Chih Tsai, Yu-Hsuan Liu, Cheng-Ying Ho, Chun-Chin Huang

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The aim of this study evaluated the in vitro and in vivo antimicrobial activity of selected lactic acid bacteria (LAB) against Uropathogenic Escherichia coli (UPEC) for prevention and amelioration of UTIs. We screened LAB strains with antimicrobial effects on UPEC using a well-diffusion assay, bacterial adherence to the uroepithelium cell line SV-HUC-1 (BCRC 60358), and a coculture inhibition assay. The results showed that the 7 LAB strains (Lactobacillus paracasei, L. salivarius, two Pediococcus pentosaceus strains, two L. plantarum strains, and L. crispatus) and the fermented probiotic products produced by these multi-LAB strains exhibited potent zones of inhibition against UPEC. Moreover, the LAB strains and probiotic products adhered strongly to the uroepithelium SV-HUC-1 cell line. The growth of UPEC strains was also markedly inhibited after co-culture with the LAB strains and probiotic products in human urine. In addition, the enhanced levels of IL-6, IL-8 and lactic acid dehydrogenase were significantly decreased by treatments with the LAB strains and probiotic products in UPEC-induced SV-HUC-1 cells. Furthermore, oral administration of probiotic products reduced the number of viable UPEC in the urine of UPEC-challenged BALB/c mice. Taken together, this study demonstrates that probiotic supplementation may be useful as an adjuvant therapy for the treatment of bacterial-induced urinary tract infections.

Keywords: lactic acid bacterium, SV-HUC-1 uroepithelium, urinary tract infection, uropathogenic Escherichia coli, BALB/c mice

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Authors: Venkatalakshmi Ranganathan, Ong Hsin Ju, Tan Yinn Ming, Lim Kien Sin, Wong Man Ting, Venkata Srikanth Meka

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The mucoadhesive buccal tablet is an oral drug delivery system which attached to the buccal surface for direct drug absorption into the systemic circulation and the unidirectional drug release is ensured by formulating a hydrophobic backing layer. The objective of present study was to formulate mucoadhesive atenolol bilayer buccal tablets by using sodium alginate, hydroxyethyl cellulose, and xanthan gum as mucoadhesive polymer and the technique applied was direct compression method. Ethyl cellulose was used as backing layer of the tablet. FTIR and DSC analysis were carried out to identify the drug polymer interactions. The prepared tablets were evaluated for physicochemical parameters, ex vivo mucoadhesion time and in-vitro drug release. The formulated tablets showed the average surface pH 6-7 which is favourable for oral mucosa. The formulation containing sodium alginate showed more than 90 % of drug release at the end of the 7 hours in vitro dissolution studies. The formulation containing xanthan gum showed more than 8 hours of mucoadhesion time and all formulation exhibited non fickian release kinetics. The present study indicates enormous potential of erodible mucoadhesive buccal tablet containing atenolol for systemic delivery with an added advantage of circumventing the hepatic first pass metabolism.

Keywords: atenolol, mucoadhesion, in vitro drug release, direct compression, ethyl cellulose

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Authors: Agustine Melviana, Rizkita Esyanti

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Stevia plant contains steviol glycosides which is estimated to be 300 times sweeter than sucrose. However in Indonesia, conventional (in vivo) propagation of Stevia rebaudiana was not effective due to a poor result. Therefore, alternative methods to propagate S. rebaudiana plants is needed, one of it is using in vitro method. Multiplication with a large quantity of stevia biomass in relatively short period can be conducted by using TIS RITA® (Recipient for Automated Temporary Immersion System). The objective of this study was to evaluate the effect of immersion period of the medium on growth and the medium bioconversion into the production of shoot biomass. The study was conducted to determine the effect of different intensity period of medium to enhance biomass of stevia shoots. Shoot culture of S. rebaudiana was grown in full strength MS medium supplemented with 1 ppm Kinetin. RITA® bioreactors were set up with two different immersion periods, 15 min (RITA® 15) and 30 min (RITA® 30), scheduled every 6 hours and incubated for 21 days. The result indicated that immersion period affected the biomass and growth rate (µ). Thirty-minutes immersion showed greater percentage of shoot multiplication (93.44 ± 0.83%), percentage of leaf growth (85.24 ± 5.99%), growth rate (0.042 ± 0.001 g/day), and productivity (0.066 g/L medium/day) compared to that immersed in RITA® 15 min (76.90 ± 4.85%; 79.73 ± 7.76; 0.045 ± 0.004 g/day, and 0.045 g/L medium/day respectively). Enhancement of biomass in RITA® 30 reached 1,702 ± 0,114 gr, whereas in RITA® 15 only 0,953 ± 0,093 gr. Additionally, the pattern of sucrose, mineral, and inorganic compounds consumption followed the growth of plant biomass for both systems. In conclusion, the bioconversion efficiency from medium to biomass in RITA® 30 is better than RITA® 15.

Keywords: intensity period, shoot culture, Stevia rebaudiana, TIS RITA®

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Authors: Yahia Massinissa, Henhouda Affaf, Yahia Mouloud

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The aim of this study was to investigate the toxicity; analgesic and anti-pyretic properties of standardized HA methanolic extract (HAMeOH) in vivo. The acute toxicity study was performed on rats while adopting the OECD-420 Guidelines (fixed dose procedure). Assessment of analgesic activity was performed in rats with two analgesic models. One was acetic acid induced writhing response and the other formalin-induced paw licking. The anti-pyretic effect was tested by brewer’s yeast induced fever in rats. For the acute toxicity test, the higher dose administration of 2000 mg/kg bw. of Hyoscyamus albus did not produce any toxic signs or deaths in rats. There were no significant differences (p>0.05) in the body and organ weights between control and treated groups. The (LD50) of Hyoscyamus albus was higher than 2000 g/kg bw. In subacute toxicity study, no mortality and toxic signs were observed with the doses of 100 and 200 mg/kg bw. of extracts of for 28 consecutive days. These analgesic experimental results indicated that HAMeOH (100 mg/kg and 200 mg/kg) decreased the acetic acid-induced writhing responses and HAMeOH (100 mg/kg and 200 mg/kg) decreased the licking time in the second phase of the formalin test. Moreover, in the model of yeast induced elevation of the body temperature HAMeOH showed dose-dependent lowering of the body temperature up to 3h at both the doses these results obtained, were comparable to that of paracetamol. The present findings indicate that the leaves of Hyoscyamus albus L. possess potent analgesic and antipyretic activity.

Keywords: Hyoscyamus albus, methanolic extract, toxicity, analgesic activity, antipyretic activity, formalin test

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Authors: A. Adetutu, T. M. Awodugba, O. A. Owoade

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The development of resistance to currently known conventional anti-typhoid drugs has necessitated search into cheap, more potent and less toxic anti-typhoid drugs of plant origin. Therefore, this study investigated the anti-typhoid activity of fractions of A. indica in Salmonella typhi infected rats. Leaves of A. indica were extracted in methanol and fractionated into n-hexane, chloroform, ethyl-acetate, and aqueous fractions. The anti-salmonella potentials of fractions of A. indica were assessed via in-vitro inhibition of S. typhi using agar well diffusion, Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and biofilm assays. The biochemical and haematological parameters were determined by spectrophotometric methods. The histological analysis was performed using Haematoxylin and Eosin staining methods. Data analysis was performed by one-way ANOVA. Results of this study showed that S. typhi was sensitive to aqueous and chloroform fractions of A. indica, and the fractions showed biofilm inhibition at concentrations of 12.50, 1.562, and 0.39 mg/mL. In the in-vivo study, the extract and chloroform fraction had significant (p < 0.05) effects on the number of viable S. typhi recovered from the blood and stopped salmonellosis after 6 days of treatment of rats at 500 mg/kg b.w. Treatments of infected rats with chloroform and aqueous fractions of A. indica normalized the haematological parameters in the animals. Similarly, treatment with fractions of the plants sustained a normal antioxidant status when compared with the normal control group. Chloroform and ethyl-acetate fractions of A. indica reversed the liver and intestinal degeneration induced by S. typhi infection in rats. The present investigation indicated that the aqueous and chloroform fractions of A. indica showed the potential to provide an effective treatment for salmonellosis, including typhoid fever. The results of the study may justify the ethno-medicinal use of the extract in traditional medicine for the treatment of typhoid and salmonella infections.

Keywords: Azadirachta indica L, salmonella, typhoid, leave fractions

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Authors: Jing Zhang, Yvonne Reinwald, Nick Poulson, Alicia El Haj, Chung See, Mike Somekh, Melissa Mather

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Biomedical imaging of native tissue using light offers the potential to obtain excellent structural and functional information in a non-invasive manner with good temporal resolution. Image contrast can be derived from intrinsic absorption, fluorescence, or scatter, or through the use of extrinsic contrast. A major challenge in applying optical microscopy to in vivo tissue imaging is the effects of light attenuation which limits light penetration depth and achievable imaging resolution. Recently Selective Plane Illumination Microscopy (SPIM) has been used to map the 3D distribution of fluorophores dispersed in biological structures. In this approach, a focused sheet of light is used to illuminate the sample from the side to excite fluorophores within the sample of interest. Images are formed based on detection of fluorescence emission orthogonal to the illumination axis. By scanning the sample along the detection axis and acquiring a stack of images, 3D volumes can be obtained. The combination of rapid image acquisition speeds with the low photon dose to samples optical sectioning provides SPIM is an attractive approach for imaging biological samples in 3D. To date all implementations of SPIM rely on the use of fluorescence reporters be that endogenous or exogenous. This approach has the disadvantage that in the case of exogenous probes the specimens are altered from their native stage rendering them unsuitable for in vivo studies and in general fluorescence emission is weak and transient. Here we present for the first time to our knowledge a label-free implementation of SPIM that has downstream applications in the clinical setting. The experimental set up used in this work incorporates both label-free and fluorescent illumination arms in addition to a high specification camera that can be partitioned for simultaneous imaging of both fluorescent emission and scattered light from intrinsic sources of optical contrast in the sample being studied. This work first involved calibration of the imaging system and validation of the label-free method with well characterised fluorescent microbeads embedded in agarose gel. 3D constructs of mammalian cells cultured in agarose gel with varying cell concentrations were then imaged. A time course study to track cell proliferation in the 3D construct was also carried out and finally a native tissue sample was imaged. For each sample multiple images were obtained by scanning the sample along the axis of detection and 3D maps reconstructed. The results obtained validated label-free SPIM as a viable approach for imaging cells in a 3D gel construct and native tissue. This technique has the potential use in a near-patient environment that can provide results quickly and be implemented in an easy to use manner to provide more information with improved spatial resolution and depth penetration than current approaches.

Keywords: bioimaging, optics, selective plane illumination microscopy, tissue imaging

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Authors: Małgorzata Drzewiecka, Tomasz Śliwiński, Maciej Radek

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The inhibition of histone deacetyles (HDACs) holds promise as a potential anti-cancer therapy because histone and non-histone protein acetylation is frequently disrupted in cancer, leading to cancer initiation and progression. Additionally, histone deacetylase inhibitors (HDACi) such as class I HDAC inhibitor - valproic acid (VPA) have been shown to enhance the effectiveness of DNA-damaging factors, such as cisplatin or radiation. In this study, we found that, using of VPA in combination with talazoparib (BMN-637 – PARP1 inhibitor – PARPi) and/or Dacarabazine (DTIC - alkylating agent) resulted in increased DNA double strand break (DSB) and reduced survival (while not affecting primary melanocytes )and proliferation of melanoma cells. Furthermore, pharmacologic inhibition of class I HDACs sensitizes melanoma cells to apoptosis following exposure to DTIC and BMN-637. In addition, inhibition of HDAC caused sensitization of melanoma cells to dacarbazine and BMN-637 in melanoma xenografts in vivo. At the mRNA and protein level histone deacetylase inhibitor downregulated RAD51 and FANCD2. This study provides that combining HDACi, alkylating agent and PARPi could potentially enhance the treatment of melanoma, which is known for being one of the most aggressive malignant tumors. The findings presented here point to a scenario in which HDAC via enhancing the HR-dependent repair of DSBs created during the processing of DNA lesions, are essential nodes in the resistance of malignant melanoma cells to methylating agent-based therapies.

Keywords: melanoma, hdac, parp inhibitor, valproic acid

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Authors: Alok Pal Jain, Santram Lodhi

Abstract:

Martynia annua Linn. (Martyniaccae) is traditionally used in inflammation and applied locally to tuberculosis glands of camel’s neck. The leaves used topically to bites of venomous insects and wounds of domestic animals. Chemical examination of Martynia annua leaves revealed the presence of glycosides, tannins, proteins, phenols and flavonoids. The present study was aimed to evaluate the anti-arthritic activity of methanolic extract of Martynia annua leaves. Methanolic extract of Martynia annua leaves was tested by using in vivo collagen-induced arthritis mouse model to investigate the anti-rheumatoid arthritis activity. In addition, antioxidant effect of methanolic extract was determined by the estimation of antioxidants level in joint tissues. The severity of arthritis was assessed by arthritis score and edema. Levels of cytokines TNF-α and IL-6, in the joint tissue homogenate were measured using ELISA. A high dose (250 mg/kg) of methanolic extract was significantly reduced the degree of inflammation in mice as compared with reference drug. Antioxidants level and malondialdehyde (MDA) in joint tissue homogenate found significantly (p < 0.05) higher. Methanolic extract at dose of 250 mg/kg modulated the cytokines production and suppressed the oxidative stress in the mice with collagen-induced arthritis. This study suggested that Martynia annua might be alternative herbal medicine for the management of rheumatoid arthritis.

Keywords: Martynia annua, collagen, rheumatoid arthritis, antioxidants

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Authors: A. Almohammedi, A. J. Hudson, N. M. Storey

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Following ischaemia/reperfusion injury, as in a myocardial infraction, cardiac myocytes undergo oxidative stress which leads to several potential outcomes including; necrotic or apoptotic cell death or dysregulated calcium homeostasis or disruption of the electron transport chain. Several studies have shown that nitric oxide donors protect cardiomyocytes against ischemia and reperfusion. However until present, the mechanism of cardioprotective effect of nitric oxide donor in isolated ventricular cardiomyocytes is not fully understood and has not been investigated before using Raman spectroscopy. For these reasons, the aim of this study was to develop a novel technique, pre-resonance Raman spectroscopy, to investigate the mechanism of cardioprotective effect of nitric oxide donor in isolated ventricular cardiomyocytes exposed to metabolic inhibition and re-energisation. The results demonstrated the first time that Raman microspectroscopy technique has the capability to monitor the metabolic inhibition of cardiomyocytes and to monitor the effectiveness of cardioprotection by nitric oxide donor prior to metabolic inhibition of cardiomyocytes. Metabolic inhibition and reenergisation were used in this study to mimic the low and high oxygen levels experienced by cells during ischaemic and reperfusion treatments. A laser wavelength of 488 nm used in this study has been found to provide the most sensitive means of observe the cellular mechanisms of myoglobin during nitric oxide donor preconditioning, metabolic inhibition and re-energisation and did not cause any damage to the cells. The data also highlight the considerably different cellular responses to metabolic inhibition to ischaemia. Moreover, the data has been shown the relationship between the release of myoglobin and chemical ischemia where that the release of myoglobin from the cell only occurred if a cell did not recover contractility.

Keywords: ex vivo biospectroscopy, Raman spectroscopy, biophotonics, cardiomyocytes, ischaemia / reperfusion injury, cardioprotection, nitric oxide donor

Procedia PDF Downloads 329

Authors: Yahia Massinissa, Afaf Benhouda, Mouloud Yahia

Abstract:

Objective: The aim of this study was to investigate the toxicity; analgesic and anti-pyretic properties of standardized HA methanolic extract (HAMeOH) in vivo. Methods: The acute toxicity study was performed on rats while adopting the OECD-420 Guidelines (fixed dose procedure). Assessment of analgesic activity was performed in rats with two analgesic models. One was acetic acid induced writhing response and the other formalin-induced paw licking. The anti-pyretic effect was tested by Brewer’s yeast induced fever in rats. Results: For the acute toxicity test, the higher dose administration of 2000 mg/kg bw. of H.albus did not produce any toxic signs or deaths in rats. There were no significant differences (p>0.05) in the body and organ weights between control and treated groups. The (LD50) of 'H. albus' was higher than 2000 g/kg bw. In subacute toxicity study, no mortality and toxic signs were observed with the doses of 100 and 200 mg/kg bw. of extracts of for 28 consecutive days. These analgesic experimental results indicated that HAMeOH (100 mg/kg and 200 mg/kg) decreased the acetic acid-induced writhing responses and HAMeOH (100 mg/kg and 200 mg/kg) decreased the licking time in the second phase of the formalin test. Moreover, in the model of yeast-induced elevation of the body temperature HAMeOH showed dose-dependent lowering of the body temperature up to 3h at both the doses these results obtained, were comparable to that of paracetamol. Conclusion: The present findings indicate that the leaves of Hyoscyamus albus L. possess potent analgesic and antipyretic activity.

Keywords: Hyoscyamus albus, Umbilicus rupestris, secondary metabolites, NMR with protons, pharmacobiologic activities, methanolic extract

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Authors: Eric Yi-Hsiu Huang, Meng-Chun Kao, Wen-Chuan Kuo

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For every laparoscopic surgery, a safe pneumoperitoneumcreation (gaining access to the peritoneal cavity) is the first and essential step. However, closed pneumoperitoneum is usually obtained by blind insertion of a Veress needle into the peritoneal cavity, which may carry potential risks suchas bowel and vascular injury.Until now, there remains no definite measure to visually confirm the position of the needle tip inside the peritoneal cavity. Therefore, this study established an image-guided Veress needle method by combining a fiber probe with optical coherence tomography (OCT). An algorithm was also proposed for determining the exact location of the needle tip through the acquisition of OCT images. Our method not only generates a series of “live” two-dimensional (2D) images during the needle puncture toward the peritoneal cavity but also can eliminate operator variation in image judgment, thus improving peritoneal access safety. This study was approved by the Ethics Committee of Taipei Veterans General Hospital (Taipei VGH IACUC 2020-144). A total of 2400 in vivo OCT images, independent of each other, were acquired from experiments of forty peritoneal punctures on two piglets. Characteristic OCT image patterns could be observed during the puncturing process. The ROC curve demonstrates the discrimination capability of these quantitative image features of the classifier, showing the accuracy of the classifier for determining the inside vs. outside of the peritoneal was 98% (AUC=0.98). In summary, the present study demonstrates the ability of the combination of our proposed automatic identification method and OCT imaging for automatically and objectively identifying the location of the needle tip. OCT images translate the blind closed technique of peritoneal access into a visualized procedure, thus improving peritoneal access safety.

Keywords: pneumoperitoneum, optical coherence tomography, automatic identification, veress needle

Procedia PDF Downloads 104

Authors: Rania Hanafi Mahmoud Said, Rasha Mohamed Taha

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Background: A recent study has explored the potential of an approach to treating infected root canals using a combination of Mineral Trioxide Aggregate (MTA) and Tetracycline-loaded Nanochitosan. Material and methods: Forty dogs were included in the study, with infected periapical areas induced by leaving access openings in their teeth for four months. Bacteriological samples from the infected root canals were collected and managed anaerobically to identify and count the different microorganisms present. The most common microorganisms detected were Prevotella oris, Fusobacterium nucleatum, Streptococcus viridans, Enterococcus faecalis, Clostridium subterminale, Porphyromonas gingivalis, and Peptostreptococcus anaerobius. The dogs were divided into four groups based on the sealant used to treat the infected periapical areas: Group I: Negative control (no treatment) Group II: Positive control (MTA only) Group III: MTA + tetracycline Group IV: MTA + tetracycline loaded on nanochitosan Results: Periapical areas in Group IV showed significantly more bone healing than those in Groups I, II, and III. The newly formed bone was evaluated radiographically, histologically, and immunohistochemically using Osteopontin (OSP) antibodies. Data collected was statistically analysed using SPSS software at a 0.05 significance level. Conclusion: The study concluded that the combined use of Tetracycline-loaded Nanochitosan and MTA presents a promising approach for the treatment of infected root canals. The potent antimicrobial activity of Tetracycline-loaded Nanochitosan, along with the biocompatibility and desirable properties of MTA, may synergistically contribute to improved clinical outcomes in endodontic therapy. This study has important implications for the clinical management of infected root canals. The combination of Tetracycline-loaded Nanochitosan and MTA could provide a more effective and efficient means of treating these challenging cases. Further research is needed to confirm these findings in humans and to optimize the treatment protocol.

Keywords: mineral trioxide aggregate, tetracycline-loaded nanochitosan, periapical infection, osteopontine

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Authors: Anil P. Dewani, Ravindra L. Bakal, Anil V. Chandewar

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Present work demonstrates the applicability of high-performance liquid chromatography (HPLC) with UV detection for the determination of malondialdehyde as malondialdehyde-thiobarbituric acid complex (MDA-TBA) in-vivo in rats. The HPLC-UV method for MDA-TBA was achieved by isocratic mode on a reverse-phase C18 column (250mm×4.6mm) at a flow rate of 1.0mLmin−1 followed by UV detection at 278 nm. The chromatographic conditions were optimized by varying the concentration and pH followed by changes in percentage of organic phase optimal mobile phase consisted of mixture of water (0.2% Triethylamine pH adjusted to 2.3 by ortho-phosphoric acid) and acetonitrile in ratio (80:20 % v/v). The retention time of MDA-TBA complex was 3.7 min. The developed method was sensitive as limit of detection and quantification (LOD and LOQ) for MDA-TBA complex were (standard deviation and slope of calibration curve) 110 ng/ml and 363 ng/ml respectively. The method was linear for MDA spiked in plasma and subjected to derivatization at concentrations ranging from 100 to 1000 ng/ml. The precision of developed method measured in terms of relative standard deviations for intra-day and inter-day studies was 1.6–5.0% and 1.9–3.6% respectively. The HPLC method was applied for monitoring MDA levels in rats subjected to chronic treatment of ciprofloxacin (CFL) (5mg/kg/day) for 21 days. Results were compared by findings in control group rats. Mean peak areas of both study groups was subjected for statistical treatment to unpaired student t-test to find p-values. The p value was < 0.001 indicating significant results and suggesting increased MDA levels in rats subjected to chronic treatment of CFL of 21 days.

Keywords: MDA, TBA, ciprofloxacin, HPLC-UV

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Authors: Muhammad nor Farhan Sa'At, Xin Y. Lim, Terence Y. C. Tan, Siti Hajar M. Rosli, Syazwani S. Ali, Ami F. Syed Mohamed

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INTRODUCTION: Hemp (Cannabis sativa subsp. sativa), commonly used for industrial purposes, differs from marijuana by containing lower levels of delta-9-tetrahydronannabidiol- the principal psychoactive constituent in cannabis. Due to its non-psychoactive nature, there has been growing interest in hemp’s therapeutic potential, which has been investigated through pre-clinical and clinical study modalities. OBJECTIVE: To provide an overview of the current landscape of hemp research, through recent scientific findings specific to the pharmacological effects of the medicinal hemp plant and its derived compounds. METHODS: This review was conducted through a systematic search strategy according to the preferred reporting items for systematic review and meta-analysis-ScR (PRISMA-ScR) checklist on electronic databases including MEDLINE, OVID (OVFT, APC Journal Club, EBM Reviews), Cochrane Library Central and Clinicaltrials.gov. RESULTS: From 65 primary articles reviewed, there were 47 pre-clinical studies related to medicinal hemp. Interestingly, the hemp derivatives showed several potential activities such as anti-oxidative, anti-hypertensive, anti-inflammatory, anti-diabetic, anti-neuroinflammatory, anti-arthritic, anti-acne, and anti-microbial activities. Renal protective effects and estrogenic properties were also exhibited in vitro. CONCLUSION: Medicinal hemp possesses various pharmacological effects tested in vitro and in vivo. Information provided in this review could be used as tool to strengthen the study design of future clinical trial research.

Keywords: Preclinical, Herbal Medicine, Hemp, Cannabis

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Authors: Haile F. Darge, Hsieh C. Tsai

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The tumor microenvironment affects the therapeutic outcomes of cancer disease. In a malignant tumor, overexpression of vascular endothelial growth factor (VEGF) provokes the production of pathologic vascular networks. This results in a hostile tumor environment that hinders anti-cancer drug activities and profoundly fuels tumor progression. In this study, we develop a strategy of sequential sustain release of the anti-angiogenic drug: Bevacizumab(BVZ), and anti-cancer drug: Doxorubicin(DOX) which had a synergistic effect on cancer treatment. Poly (D, L-Lactide)- Poly (ethylene glycol) –Poly (D, L-Lactide) (PDLLA-PEG-PDLLA) thermo-sensitive hydrogel was used as a vehicle for local delivery of drugs in a single platform. The in vitro release profiles of the drugs were investigated and confirmed a relatively rapid release of BVZ (73.56 ± 1.39%) followed by Dox (61.21 ± 0.62%) for a prolonged period. The cytotoxicity test revealed that the copolymer exhibited negligible cytotoxicity up to 2.5 mg ml-1 concentration on HaCaT and HeLa cells. The in vivo study on Hela xenograft nude mice verified that hydrogel co-loaded with BVZ and DOX displayed the highest tumor suppression efficacy for up to 36 days with pronounce anti-angiogenic effect of BVZ and with no noticeable damage on vital organs. Therefore, localized co-delivery of anti-angiogenic drug and anti-cancer drugs by the hydrogel system may be a promising approach for enhanced chemotherapeutic efficacy in cancer treatment.

Keywords: anti-angiogenesis, chemotherapy, controlled release, thermo-sensitive hydrogel

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Authors: Daniela Istrati, Alina Morosan, Maria Stanca, Bogdan Purcareanu, Adrian Fudulu, Laura Olariu, Alice Buteica, Ion Mindrila, Rodica Cristescu, Dan Eduard Mihaiescu

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Our present work is related to the synthesis, characterization and applications of new nanocomposite materials based on silica mesoporous nanocompozites systems. The nanocomposite support was obtained by using a specific step–by–step multilayer structure buildup synthetic route, characterized by XRD (X-Ray Difraction), TEM (Transmission Electron Microscopy), FT-IR (Fourier Transform-Infra Red Spectrometry), BET (Brunauer–Emmett–Teller method) and loaded with Salvia officinalis plant extract obtained by a hydro-alcoholic extraction route. The sustained release of the target compounds was studied by a modified LC method, proving low release profiles, as expected for the high specific surface area support. The obtained results were further correlated with the in vitro / in vivo behavior of the nanocomposite material and recommending the silica mesoporous nanocomposites as good candidates for biomedical applications. Acknowledgements: This study has been funded by the Research Project PN-III-P2-2.1-PTE-2016-0160, 49-PTE / 2016 (PROZECHIMED) and Project Number PN-III-P4-ID-PCE-2016-0884 / 2017.

Keywords: biomedical, mesoporous, nanocomposites, natural products, sustained release

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Authors: Sun Il Yu, Jung Hyun Joo, Hwa Sung Shin

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Astrocytes are known as dominant cells in CNS and play a role as a supporter of CNS activity and regeneration. Recently, three-dimensional culture of astrocytes were actively applied to understand in vivo astrocyte works. Electrospun nanofibers are attractive for 3D cell culture system because they have a high surface to volume ratio and porous structure, and have already been used for 3D astrocyte cultures. In this research, the stiffness of cellulose acetate (CA) nanofiber was controlled by heat treatment. As stiffness increased, astrocyte cell viability and adhesion increased. Reactivity of astrocyte was also upregulated in stiffer CA nanofiber in terms of GFAP, an intermediate filament protein. Finally, we demonstrated that stiffness-controllable CA is attractive for astrocyte tissue engineering.

Keywords: astrocyte, cellulose acetate, nanofiber, tissue scaffold

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Authors: Fayzan Ahmed, Saad Bin Saeed, Abdul Qadir Jangda

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Gafchromic™ sheet are extensively used for the QA of intensity modulated radiation therapy and other in-vivo dosimetry. Intra-sheet Non-uniformity of scanner as well as film causes undesirable fluctuations which are reflected in dosimetry The aim of this study is to define a systematic and robust method to investigate the intra-sheet uniformity of the unexposed Gafchromic Sheets and the region of interest (ROI) of the scanner. Sheets of lot No#: A05151201 were scanned before and after the expiry period with the EPSON™ XL10000 scanner in the transmission mode, landscape orientation and 72 dpi resolution. ROI of (8’x 10’ inches) equal to the sheet dimension in the center of the scanner is used to acquire images with full transmission, block transmission and with sheets in place. 500 virtual grids, created in MATALB® are imported as a macros in ImageJ (1.49m Wayne Rasband) to analyze the images. In order to remove the edge effects, the outer 86 grids are excluded from the analysis. The standard deviation of the block transmission and full transmission are 0.38% and 0.66% confirming a higher uniformity of the scanner. Expired and non-expired sheets have standard deviations of 2.18% and 1.29%, show that uniformity decreases after expiry. The results are promising and indicates a good potential of this method to be used as a uniformity check for scanner and unexposed Gafchromic sheets.

Keywords: IMRT, film dosimetry, virtual grids, uniformity

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Authors: Chaoran Liu

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Bioactive polysaccharides and polysaccharide-protein complex derived from mushrooms and fungi have a wide range of immunomodulatory activity with low side-effects and have therefore the potential to be developed as an adjuvant in cancer therapies. Mushrooms sclerotium is rich in polysaccharides and the polysaccharides isolated from the sclerotium of Polyporus rhinocerus have shown potent in vivo and in vitro immunomodulatory effects. Macrophages are considered to be an important component of the innate immune response against bacterial infection and cancer. To better understanding the immunomodulatory effects and its underlying mechanisms of sclerotial water-soluble polysaccharides extracted from P. rhinocerus on macrophages, the objectives of this study are to purify the water-soluble novel sclerotial polysaccharides and to characterize the structure and properties as well as to study the detailed molecular mechanisms of the in vitro immunomodulating effects in murine macrophages. The hot water-soluble fraction PRW from the sclerotium of P. rhinocerus was obtained using solvent extraction. PRW was further fractionated by membrane ultrafiltration to a give a fraction (PRW1) with molecular mass less than 50 kDa. PRW1 was characterized to be a polysaccharide-protein complex composed of 45.7% polysaccharide and 44.2% protein. The chemical structure of the carbohydrate moiety of PRW1 was elucidated by GC and FTIR to be mainly beta-D-glucan with trace amount of galactose and mannose. The immunomodulatory effects of PRW1 on murine RAW 264.7 macrophages were demonstrated in terms of the increase in nitric oxide production and cytokine production. Mechanistically, PRW1 initiates ERK phosphorylation to activate macrophages within 15 min and significantly improves the expression level of inducible NOS (iNOS) from 6 h after treatment. In summary, this study indicates that PRW1 is a potent immunomodulatory agent for macrophages and suggests that mushroom sclerotia from Polyporus rhinocerus requires for further investigation in cancer research.

Keywords: Polyporus rhinocerus, mushroom sclerotia, Polysaccharide-Protein Complex, macrophage activation

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Authors: Sharif Abdelghany

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Polymeric nanoparticles have been widely investigated as a controlled release drug delivery platform for the treatment of tuberculosis (TB). These nanoparticles were also readily internalised into macrophages, leading to high intracellular drug concentration. In this study two anti-TB drugs, amikacin and moxifloxacin were encapsulated into PLGA nanoparticles. The novelty of this work appears in: (1) the efficient encapsulation of two hydrophilic second-line anti-TB drugs, and (2) intramacrophage delivery of this synergistic combination potentially for rapid treatment of multi-drug resistant TB (MDR-TB). Two water-oil-water (w/o/w) emulsion strategies were employed in this study: (1) alginate coated PLGA nanoparticles, and (2) alginate entrapped PLGA nanoparticles. The average particle size and polydispersity index (PDI) of the alginate coated PLGA nanoparticles were found to be unfavourably high with values of 640 ± 32 nm and 0.63 ± 0.09, respectively. In contrast, the alginate entrapped PLGA nanoparticles were within the desirable particle size range of 282 - 315 nm and the PDI was 0.08 - 0.16, and therefore were chosen for subsequent studies. Alginate entrapped PLGA nanoparticles yielded a drug loading of over 10 µg/mg powder for amikacin, and more than 5 µg/mg for moxifloxacin and entrapment efficiencies range of approximately 25-31% for moxifloxacin and 51-59% for amikacin. To study macrophage uptake efficiency, the nanoparticles of alginate entrapped nanoparticle formulation were loaded with acridine orange as a marker, seeded to THP-1 derived macrophages and viewed under confocal microscopy. The particles were readily internalised into the macrophages and highly concentrated in the nucleus region. Furthermore, the anti-mycobacterial activity of the drug-loaded particles was evaluated using M. tuberculosis-infected macrophages, which revealed a significant reduction (4 log reduction) of viable bacterial count compared to the untreated group. In conclusion, the amikacin-moxifloxacin alginate entrapped PLGA nanoparticles are promising for further in vivo studies.

Keywords: moxifloxacin and amikacin, nanoparticles, multidrug resistant TB, PLGA

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Authors: Jayarama Reddy, Anand S., H., Sundaram, Jeldi Hemachandran

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Forty two strains of Trichoderma sp. were isolated from cultivated lands around Bangalore and analyzed for their antagonistic potential against Macrophomina phaseolina. The potential of biocontrol agents ultimately lies in their capacity to control pathogens in vivo. Bioefficacy studies were hence conducted using chickpea (Cicer arientum c.v. Annigeri) as an experimental plant by the roll paper towel method. Overall the isolates T6, T35, T30, and T25 showed better antagonistic potential in addition to enhancing plant growth. The production of chitinases to break down the mycelial cell walls of fungal plant pathogens has been implicated as a major cause of biocontrol activity. In order to study the mechanism of biocontrol against Macrophomina phaseolina, ten better performing strains were plated on media, amended with colloidal chitin and Sclerotium rolfsii cell wall extract. All the isolates showed chitinolytic activity on day three as well as day five. Production of endochitinase and exochitinase were assayed in liquid media using colloidal chitin amended broth. Strains T35 and T6 displayed maximum endochitinase and exochitinase activity. Although all strains exhibited cellulase activity, the quantum of enzyme produced was higher in T35 and T6. The results also indicate a positive correlation between enzyme production and bioefficacy.

Keywords: biocontrol, bioefficacy, cellulase, chitinase

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Authors: Vijayakameswara Rao Neralla, Jueun Jeon, Jae Hyung Park

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To develop a potential nanocarrier for diagnosis and treatment of rheumatoid arthritis (RA), we prepared a hyaluronic acid (HA)-5β-cholanic acid (CA) conjugate with an acid-labile ketal linker. This conjugate could self-assemble in aqueous conditions to produce pH-responsive HA-CA nanoparticles as potential carriers of the anti-inflammatory drug methotrexate (MTX). MTX was rapidly released from nanoparticles under inflamed synovial tissue in RA. In vitro cytotoxicity data showed that pH-responsive HA-CA nanoparticles were non-toxic to RAW 264.7 cells. In vivo biodistribution results confirmed that, after their systemic administration, pH-responsive HA-CA nanoparticles selectively accumulated in the inflamed joints of collagen-induced arthritis mice. These results indicate that pH-responsive HA-CA nanoparticles represent a promising candidate as a drug carrier for RA therapy.

Keywords: rheumatoid arthritis, hyaluronic acid, nanocarrier, self-assembly, MTX

Procedia PDF Downloads 270