Search results for: mice bone

Authors: Keivan Jamshidi, Afshin Zahedi

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Reactive amyloidosis is a condition that complicates a long list of chronic inflammation, chronic infectious, malignant, and hereditary disorders. In the present study the propensity for amyloidogenesis in male and female rats on spatio-temporal pattern was evaluated. For this purpose a total of 40 male and female Swiss mice, obtained from Pasteur Institute Tehran, after being weighted were randomly divided into 4 groups including 2 treatment groups [ 10 male (Group A1) and 10 female (Group B1) each], and 2 control groups [10 male (Group A2) and 10 female (Group B2) each]. At the end of 3rd, 5th and 7th weeks of experiment 3 mice were randomly selected and euthnised. Spleen samples of each animal were obtained and preserved in 10% neutral buffer formalin. Sample were then processed through different stages of dehydration, clearing and impregnation and finally embedded in paraffin blocks. Sections of 5µm thickness then cut and stained by alkaline Congo red techniques. The data obtained from polarized microscopic quantitative analysis did show significant differences between groups A1 and B1. A preferential expression of reactive amyloidosis is concluded in male, indicating sex differences in amyloidosis.

Keywords: amyloidosis, amyloidogenesis, mice, gender

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Authors: Yu-Shan Wu, Po-Liang Lai, I-Ming Chu

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Poly(methylmethacrylate)(PMMA) is the most frequently used bone void filler for vertebral augmentation in osteoporotic fracture. PMMA bone cement not only exhibits strong mechanical properties but also can fabricate according to the shape of bone defect. However, the adhesion between the PMMA-based cement and the adjacent bone is usually weak and as PMMA bone cement is inherently bioinert. The combination of bioceramics and polymers as composites may increase cell adhesion and improve biocompatibility. The nano-hydroxyapatite(HAP) not only plays a significant role in maintaining the properties of the natural bone but also offers a favorable environment for osteoconduction, protein adhesion, and osteoblast proliferation. However, defects and cracks can form at the polymer/ceramics interface, resulting in uneven distribution of stress and subsequent inferior mechanical strength. Surface-modified HAP nano-crystals were prepared by chemically grafting poly(ε-caprolactone)(PCL) on surface-modified nano-HAP surface to increase the affinity of polymer/ceramic phases .Thus, incorporation of surface-modified nano-hydroxyapatite (EC-HAP) may not only improve the interfacial adhesion between cement and bone and between nanoparticles and cement, but also increase biocompatibility. In this research, PMMA mixing with 0, 5, 10, 15, 20, 25 and 30 wt% EC-HAP were examined. MC3T3-E1 cells were used for the biological evaluation of the response to the cements in vitro. Morphology was observed using scanning electron microscopy (SEM). Mechanical properties of HAP/PMMA and EC-HAP/PMMA cement were investigated by compression test. Surface wettability of the cements was measured by contact angles.

Keywords: bone cement, biocompatibility, nano-hydroxyapatite, polycaprolactone, PMMA, surface grafting

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Authors: Mahesh Pal, Tripti Mishra, Chandana Rao, Dalip Upreti

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Cancer has been considered to be a very dreadful disease. Holmskioldia sanguinea is a large climbing shrub found in the Himalayas at an altitude of 5,000 ft and preliminary investigation showed the excellent yield of andrographolide and subjected for the anticancer activity. Protective effect of Holmskioldia sanguinea leaf ethanolic extract has been investigated against Ehrlich ascites carcinoma (EAC) and Daltons ascites lymphoma (DAL) in Swiss albino mice to evaluate the possible mechanism of action. The enzymatic antioxidant status was studied on tumor bearing mice, which shows the potential of the compound to possess significant free radical scavenging property and revealed significant tumor regression and prolonged survival time. The isolated bioactive molecule andrographolide from Holmskioldia sanguinea yields (2.5%) in subject to HPTLC/HPLC analysis. The cellular defense system constituting the superoxide dismutase, catalyses was enhanced whereby the lipid peroxidation content was restricted to a larger extent. The Holmskioldia sanguinea is a new source of andrographolide and demonstrated the potency in treatment of cancer.

Keywords: Holmskioldia sanguinea, tumor, mice, andrographolide

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Authors: Pureun-Haneul Lee, Byeong-Gon Kim, Sun-Hye Lee, An-Soo Jang

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Background: Nanoparticles may pose adverse health effects due to particulate matter inhalation. Nanoparticle exposure induces cell and tissue damage, causing local and systemic inflammatory responses. The inflammasome is a major regulator of inflammation through its activation of pro-caspase-1, which cleaves pro-interleukin-1β (IL-1β) into its mature form and may signal acute and chronic immune responses to nanoparticles. Objective: The aim of the study was to identify whether nanoparticles exaggerates inflammasome pathway leading to airway inflammation and hyperresponsiveness in an allergic mice model of asthma. Methods: Mice were treated with saline (sham), OVA-sensitized and challenged (OVA), or titanium dioxide nanoparticles. Lung interleukin 1 beta (IL-1β), interleukin 18 (IL-18), NACHT, LRR and PYD domains-containing protein 3 (NLRP3) and caspase-1 levels were assessed with Western Blot. Caspase-1 was checked by immunohistochemical staining. Reactive oxygen species were measured for the marker 8-isoprostane and carbonyl by ELISA. Results: Airway inflammation and hyperresponsiveness increased in OVA-sensitized/challenged mice and these responses were exaggerated by TiO2 nanoparticles exposure. TiO2 nanoparticles treatment increased IL-1β and IL-18 protein expression in OVA-sensitized/challenged mice. TiO2 nanoparticles augmented the expression of NLRP3 and caspase-1 leading to the formation of an active caspase-1 in the lung. Lung caspase-1 expression was increased in OVA-sensitized/challenged mice and these responses were exaggerated by TiO2 nanoparticles exposure. Reactive oxygen species was increased in OVA-sensitized/challenged mice and in OVA-sensitized/challenged plus TiO2 exposed mice. Conclusion: Our data demonstrate that inflammasome pathway activates in asthmatic lungs following nanoparticles exposure, suggesting that targeting the inflammasome may help control nanoparticles-induced airway inflammation and responsiveness.

Keywords: bronchial asthma, inflammation, inflammasome, nanoparticles

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Authors: Ritu Shitak

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Humans are social mammals, of the primate order. Our biology, behaviour, and pathologies are unique to us. In our desire to understand, reduce solitary confinement one source of information is the many reports of social isolation of other social mammals, especially primates. A behavioural study was conducted in the department of pharmacology at Indira Gandhi Medical College, Shimla in Himachal Pradesh province in India using white albino mice. Different behavioural parameters were observed by using open field, tail suspension, tests for aggressive behaviour and social interactions and the effect of isolation was studied. The results were evaluated and the standard statistics were applied. The said study was done to establish facts that isolation itself impairs social behaviour and can lead to alcohol dependence as well as related drug dependence.

Keywords: social isolation, albino mice, drug dependence, isolation on social behaviour

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Authors: John O. Akindoyo, M. D. H. Beg, Suriati Binti Ghazali, Nitthiyah Jeyaratnam

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Regeneration of bone due to the many health challenges arising from traumatic effects of bone loss, bone tumours and other bone infections is fast becoming indispensable. Over the period of time, some approaches have been undertaken to mitigate this challenge. This includes but not limited to xenografts, allografts, autografts as well as artificial substitutions like bioceramics, synthetic cements and metals. However, most of these techniques often come along with peculiar limitation and problems such as morbidity, availability, disease transmission, collateral site damage or absolute rejection by the body as the case may be. Hydroxyapatite (HA) is very compatible and suitable for this application. However, most of the common methods for HA synthesis are expensive and environmentally unfriendly. Extraction of HA from bio-wastes have been perceived not only to be cost effective, but also environment-friendly. In this research, HA was produced from bio-waste: namely bovine bones through a combination of hydrothermal chemical processes and ordinary calcination techniques. Structure and property of the HA was carried out through different characterization techniques (such as TGA, FTIR, DSC, XRD and BET). The synthesized HA was found to possess similar properties to stoichiometric HA with highly desirable thermal, degradation, structural and porous properties. This material is unique for its potential minimal cost, environmental friendliness and property controllability. It is also perceived to be suitable for tissue and bone engineering applications.

Keywords: biomaterial, biopolymer, bone, hydroxyapatite

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Authors: Hernandez Pardo Diego F., Guiza Arguello Viviana R., Coy Echeverria Ana, Viejo Abrante Fernando

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The biological hydroxyapatite obtained from bovine bone arouses great interest in its application as a material for bone regeneration due to its better bioactive behavior in comparison with synthetic hydroxyapatite. For this reason, the objective of the present investigation was to determine the effect of chemical and thermal treatments in obtaining biological bovine hydroxyapatite of high purity and crystallinity. Two different chemical reagents were evaluated (NaOH and HCl) with the aim to remove the organic matrix of the bovine cortical bone. On the other hand, for analyzing the effect of thermal treatment temperature was ranged between 500 and 1000°C for a holding time of 4 hours. To accomplish the above, the materials before and after the chemical and thermal treatments were characterized by elemental compositional analysis (CHN), infrared spectroscopy by Fourier transform (FTIR), RAMAN spectroscopy, scanning electron microscopy (SEM), thermogravimetric analysis (TGA) and X-ray diffraction (XRD) and energy dispersion X-ray spectroscopy (EDS). The results allowed to establish that NaOH is more effective in the removal of the organic matrix of the bone when compared to HCl, whereas a thermal treatment at 700ºC for 4 hours was enough to obtain biological hydroxyapatite of high purity and crystallinity.

Keywords: bovine bone, hydroxyapatite, biomaterials, thermal treatment

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Authors: Austin Belfiori, Kevin Rinek, Zach Barcroft, Jennifer Berglind

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Diet influences the composition of the gut microbiota and host's health. Disruption of the balance among the microbiota, epithelial cells, and resident immune cells in the intestine is involved in the pathogenesis of inflammatory bowel disease (IBD). To study the role of gut microbiota in intestinal inflammation, the microbiome of control mice (C57BL6) given a gluten-containing standard diet versus C57BL6 mice given the gluten-free (GF) feed (n=10 in each group) was examined. All mice received the 3% DSS for 5 days. Throughout the study, feces were collected and processed for DNA extraction and MiSeq Illumina sequencing of V4 region of bacterial 16S rRNA gene. Alpha and beta diversities and compositional differences at phylum and genus levels were determined in intestinal microbiota. The mice receiving the GF diet showed a significantly increased abundance of Firmicutes and a decrease of Bacteroides and Lactobacillus at phylum level. Therefore, the gluten free diet led to reductions in beneficial gut bacteria populations. These findings indicate a role of wheat gluten in dysbiosis of the intestinal microbiota.

Keywords: gluten, colitis, microbiota, DSS, dextran sulphate sodium

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Authors: Vimal Kumar Dewangan, T. S. Sampath Kumar, Mukesh Doble, Viju Daniel Varghese

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The apatite-based, i.e., calcium-deficient hydroxyapatite (CDHAp) bone cement is well-known potential bone graft/substitute in orthopaedics due to its similar chemical composition with natural bone minerals. Therefore, an easy approach was attempted to fabricate the apatite-based (CDHAp) bone cement with improved injectability, bioresorbability, and macroporosity. In this study, the desired bone cement was developed by mixing the solid phase (consisting of wet chemically synthesized nanocrystalline hydroxyapatite and commercially available (synthetic) tricalcium phosphate) and the liquid phase (consisting of cement binding accelerator with few biopolymers in a dilute acidic solution) along with a liquid porogen as polysorbate or a solid porogen as mannitol (for comparison) in an optimized liquid-to-powder ratio. The fabricated cement sets within clinically preferred setting time (≤20 minutes) are better injectable (>70%) and also stable at ~7.3-7.4 (physiological pH). The CDHAp phased bone cement was resulted by immersing the fabricated after-set cement in phosphate buffer solution and other similar artificial body fluids and incubated at physiological conditions for seven days, confirmed through the X-ray diffraction and Fourier transform-infrared spectroscopy analyses. The so-formed synthetic apatite-based bone cement holds the acceptable compressive strength (within the range of trabecular bone) with average interconnected pores size falls in a macropores range (~50-200μm) inside the cement, verified by scanning electron microscopy (SEM), mercury intrusion porosimetry and micro-CT analysis techniques. Also, it is biodegradable (degrades ~19-22% within 10-12 weeks) when incubated in artificial body fluids under physiological conditions. The biocompatibility study of the bone cement, when incubated with MG63 cells, shows a significant increase in the cell viability after 3rd day of incubation compared with the control, and the cells were well-attached and spread completely on the surface of the bone cement, confirmed through SEM and fluorescence microscopy analyses. With this all, we can conclude that the developed synthetic macroporous apatite-based bone cement may have the potential to become promising material used as a trabecular bone substitute.

Keywords: calcium deficient hydroxyapatite, synthetic apatite-based bone cement, injectability, macroporosity, trabecular bone substitute

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Authors: Aftab Ahmed, Ghulam Mehboob

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The object of presentation is to highlight the importance of condition which is a very rare genetic disorder although Paget’s disease is common but its juvenile type is very rare and a late presentation due to very slow onset and lack of earlier standard management. We present a case of 25 years old male with a chronic history of bone pain and a slow onset of mild swelling, later on diagnosed as juvenile Paget disease of bone. Rarity of this condition with inaccessibility for standard health treatment can lead to a significant delay in presentation and its management. There have been 50 reported cases worldwide according to Genetic Home Reference. There is increased osteoclastic activity along with osteoblastic activity related to gene alteration and osteoprotegrin deficiency. Morbidity of disease is very significant which lead children to become immobilize.

Keywords: juvenile, Paget’s disease, bone, Northern Area of Pakistan

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Authors: Sang-Sun Yoon, Yea-Hyun Leem, Sangmee Ahn Jo

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The N-acetylated-alpha-linked-acidic (NAAG) peptidase inhibitor 2-phosphonomethyl pentanedioic acid (2-PMPA) has demonstrated to be neuroprotective against glutamate-mediated neuron degeneration and neurological disorders such as ischemia. Several studies have demonstrated impaired psychiatric function by altered glutamate carboxypeptidase II expression, although 2-PMPA has not yet been studied. Thus, we investigated effect of 2-PMPA on depressive-like phenotype using C57BL/6 mice. Treatment of 2-PMPA (10 mg/kg for 6 days/daily, ip injection) on C57BL/6 naïve mice showed depressive-like symptoms such as decreased social preference, but did not affect the immobility measured by tail suspension test. Reduction of phosphorylated cAMP-responsive element binding (p-CREB) known as a representative marker of depressive-like behavior was observed in layer 1 and piriform cortex subregions of the prefrontal cortex of 2-PMPA-treated mice. The immunoreactivity of metabotropic glutamate receptors 5 (mGluR5) that mediate phosphorylation of CREB was also decreased in layer 1 and piriform cortex subregions of the prefrontal cortex of 2-PMPA injected mice. Thus, our results suggest that dysregulation of the GCPII or NAAG by 2-PMPA treatment is likely to be associated with pathogenesis of depression and further studies are needed to understand whether the reduced NAAG level or enhanced glutamate level in the brain is involved in this response.

Keywords: depression, GCPII, 2-PMPA, p-CREB, mGluR5

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Authors: P. Shrivastava, S. Vijayalakshmi, A. K. Singh, S. Dalai, R. Teotia, P. Sharma, J. Bellare

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This paper focuses on the synthesis and application of nanobioactive glass for bone regeneration studies. Nanobioactive glass has been synthesized by sol gel method having a combination of silicon, calcium and phosphorous in the molar ratio of 75:21:4. The prepared particles were analyzed for surface morphology by FEG SEM and FEG TEM. Physiochemical properties were investigated using ICP AES, FTIR spectroscopy and X-ray diffraction (XRD) techniques. To ascertain their use for therapeutic use, biocompatibility evaluation of the particles was done by performing soaking studies in SBF and in vitro cell culture studies on MG63 cell lines. Cell morphology was observed by FE SEM and phase contrast microscopy. Nanobioactive glasses (NBG) thus prepared were of 30-200 nm in size, which makes them suitable for nano-biomedical applications. The spherical shape of the particles imparts high surface to volume ratio, promoting fast growth of hydroxyapatite (HA), which is the mineral component of bone. As evaluated by in vitro cell culture studies the NBG was found to enhance the surface activation which enhances osteoblast adhesion. This is an essential parameter to improve bone tissue integration, thereby making nanobioactive glass therapeutically suitable for correcting bone defects.

Keywords: biocompatibility, bone tissue engineering, hydroxyapatite, nanobioactive glass

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Authors: Ling-Ling E., Hong-Chen Liu, Dong-Sheng Wang, Fang Su, Xia Wu, Zhan-Ping Shi, Yan Lv, Jia-Zhu Wang

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Objective: The objective of the present study is to evaluate the capacity of a tissue-engineered bone complex of recombinant human bone morphogenetic protein 2 (rhBMP-2) mediated dental pulp stem cells (DPSCs) and nano-hydroxyapatite/collagen/poly(L-lactide)(nHAC/PLA) to reconstruct critical-size alveolar bone defects in New Zealand rabbit. Methods: Autologous DPSCs were isolated from rabbit dental pulp tissue and expanded ex vivo to enrich DPSCs numbers, and then their attachment and differentiation capability were evaluated when cultured on the culture plate or nHAC/PLA. The alveolar bone defects were treated with nHAC/PLA, nHAC/PLA+rhBMP-2, nHAC/PLA+DPSCs, nHAC/PLA+DPSCs+rhBMP-2, and autogenous bone (AB) obtained from iliac bone or were left untreated as a control. X-ray and a polychrome sequential fluorescent labeling were performed post-operatively and the animals were sacrificed 12 weeks after operation for histological observation and histomorphometric analysis. Results: Our results showed that DPSCs expressed STRO-1 and vementin, and favoured osteogenesis and adipogenesis in conditioned media. DPSCs attached and spread well, and retained their osteogenic phenotypes on nHAC/PLA. The rhBMP-2 could significantly increase protein content, alkaline phosphatase (ALP) activity/protein, osteocalcin (OCN) content, and mineral formation of DPSCs cultured on nHAC/PLA. The X-ray graph, the fluorescent, histological observation and histomorphometric analysis showed that the nHAC/PLA+DPSCs+rhBMP-2 tissue-engineered bone complex had an earlier mineralization and more bone formation inside the scaffold than nHAC/PLA, nHAC/PLA+rhBMP-2 and nHAC/PLA+DPSCs, or even autologous bone. Implanted DPSCs contribution to new bone were detected through transfected eGFP genes. Conclutions: Our findings indicated that stem cells existed in adult rabbit dental pulp tissue. The rhBMP-2 promoted osteogenic capability of DPSCs as a potential cell source for periodontal bone regeneration. The nHAC/PLA could serve as a good scaffold for autologous DPSCs seeding, proliferation and differentiation. The tissue-engineered bone complex with nHAC/PLA, rhBMP-2, and autologous DPSCs might be a better alternative to autologous bone for the clinical reconstruction of periodontal bone defects.

Keywords: nano-hydroxyapatite/collagen/poly (L-lactide), dental pulp stem cell, recombinant human bone morphogenetic protein, bone tissue engineering, alveolar bone

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Authors: Kathy Stein, Mariola Lysson, Anja Schmidt, Beatrix Schumak, Sabine Specht, Hicham Bouabe, Jürgen Heesemann, Axel Roers, Joerg C. Kalff, Sven Wehner

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Objective: Postoperative ileus (POI) is a common complication of abdominal surgery. Monocyte infiltration is a hallmark of POI. The polarization of macrophages/monocytes in this process is not well understood. We aimed to investigate if and how M2 macrophage/monocyte differentiation is involved in POI pathogenesis. Design: POI was induced by intestinal manipulation (IM). C57Bl/6, CCR2-/-, IL-10 reporter (ITIB), IL-10-/- and LysMcre/IL-10fl/fl mice underwent IM. At various points in time leukocyte influx, gene and protein expression of cytokines, chemokines and M2 differentiation markers and intestinal motility were analyzed. Results: IM induced the postoperative expression of the M2 markers Arginase-1 and YM-1, predominantly in F4/80+Ly6C+ monocytes. Gene expression analyses indicated an IL-10-dependent, IL-4-independent M2 polarization of these monocytes. IL-10 dependency of M2 differentiation was confirmed in IL-10 deficient mice. Leukocytes, in the order of infiltrating monocytes, neutrophils, and resident macrophages were the main IL-10 producers during POI. IL-10 producing monocytes as well as M2 marker expression were almost absent in CCR2-deficient mice. However, postoperative IL-10 expression was not altered in CCR2-/- mice. The loss of M2 polarized monocytes neither protected CCR2-/- mice from nor affected resolution of POI. In contrast, IL-10 deficiency reduced postoperative neutrophil numbers and ameliorated POI. IL-10Ra expression was strongly induced in neutrophils but not in monocytes. Conclusion: We conclude that IL-10 counteracts POI resolution by activating IL-10Ra-expressing neutrophils in the late phase of disease while IL-10-dependent M2 differentiation is not pivotal to POI manifestation and resolution.

Keywords: interleukin-10, macrophages, neutrophils, postoperative ileus

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Authors: S. Alireza Mirghasemi, Elly Trepman, Mohammad Saleh Sadeghi, Narges Rahimi Gabaran, Shervin Rashidinia

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Bone marrow edema syndrome (BMES) is an uncommon and self-limited syndrome characterized by atraumatic extremity pain with unknown of etiology. Symptom onset may include sudden or gradual swelling and pain at rest or during activity, usually at night. This syndrome mostly affects middle-aged men and younger women who have pain in the lower extremities. The most common sites involved with BMES, in decreasing order of frequency, are the bones about the hip, knee, ankle, and foot. The diagnosis of BMES is made with magnetic resonance imaging to exclude other causes of bone marrow edema. The correct diagnosis often is delayed because of the low prevalence and nonspecific signs in the foot and ankle. This delay may intensify bone pain and impair patient function and quality of life. The goal of BMES treatment is to relieve pain and shorten disease duration. Treatment options are limited and may include symptomatic treatment, pharmacologic treatment, and surgery.

Keywords: transient osteoporosis, bone marrow edema syndrome, iloprost, bisphosphonates

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Authors: Adriana Garcia-Gurrola, Carlos Adrian Peña Natividad, Ana Laura Martinez Martinez, Alberto Abraham Escobar Puentes, Estefania Ochoa Ruiz, Aracely Serrano Medina, Abraham Wall Medrano, Simon Yobanny Reyes Lopez

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Type 2 diabetes mellitus (T2DM) is a metabolic disease characterized by elevated blood glucose levels. Quercetin is a natural flavonoid with a hypoglycemic effect, but reported data are inconsistent due mainly to the structural instability and low solubility of quercetin. Nanoencapsulation is a distinct strategy to overcome the intrinsic limitations of quercetin. Therefore, this work aims to develop a quercetin nano-formulation based on biopolymeric starch nanoparticles to enhance the release and hypoglycemic effect of quercetin in T2DM induced mice model. Starch-quercetin nanoparticles were synthesized using high-intensity ultrasonication, and structural and colloidal properties were determined by FTIR and DLS. For in vivo studies, CD1 male mice (n=25) were divided into five groups (n=5). T2DM was induced using a high-fat and high-sugar diet for 32 weeks and streptozotocin injection. Group 1 consisted of healthy mice fed with a normal diet and water ad libitum; Group 2 were diabetic mice treated with saline solution; Group 3 were diabetic mice treated with glibenclamide; Group 4 were diabetic mice treated with empty nanoparticles; and Group 5 was diabetic mice treated with quercetin nanoparticles. Quercetin nanoparticles had a hydrodynamic size of 232 ± 88.45 nm, a PDI of 0.310 ± 0.04 and a zeta potential of -4 ± 0.85 mV. The encapsulation efficiency of nanoparticles was 58 ± 3.33 %. No significant differences (p = > 0.05) were observed in biochemical parameters (lipids, insulin, and peptide C). Groups 3 and 5 showed a similar hypoglycemic effect, but quercetin nanoparticles showed a longer-lasting effect. Histopathological studies reveal that T2DM mice groups showed degenerated and fatty liver tissue; however, a treated group with quercetin nanoparticles showed liver tissue like that of the healthy mice group. These results demonstrate that quercetin nano-formulations based on starch nanoparticles are effective alternatives with hypoglycemic effects.

Keywords: quercetin, diabetes mellitus tipo 2, in vivo study, nanoparticles

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Authors: Azin Zargham, Gholamreza Rouhi, Allahyar Geramy

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One of the most prevalent types of alveolar bone loss is horizontal bone loss (HBL) in which the bone height around teeth is reduced homogenously. In the presence of HBL the magnitudes of forces during orthodontic treatment should be altered according to the degree of HBL, in a way that without further bone loss, desired tooth movement can be obtained. In order to investigate the appropriate orthodontic force system in the presence of HBL, a three-dimensional numerical model capable of the simulation of orthodontic tooth movement was developed. The main goal of this research was to evaluate the effect of different degrees of HBL on a long-term orthodontic tooth movement. Moreover, the effect of different force magnitudes on orthodontic tooth movement in the presence of HBL was studied. Five three-dimensional finite element models of a maxillary lateral incisor with 0 mm, 1.5 mm, 3 mm, 4.5 mm and 6 mm of HBL were constructed. The long-term orthodontic tooth tipping movements were attained during a 4-weeks period in an iterative process through the external remodeling of the alveolar bone based on strains in periodontal ligament as the bone remodeling mechanical stimulus. To obtain long-term orthodontic tooth movement in each iteration, first the strains in periodontal ligament under a 1-N tipping force were calculated using finite element analysis. Then, bone remodeling and the subsequent tooth movement were computed in a post-processing software using a custom written program. Incisal edge, cervical, and apical area displacement in the models with different alveolar bone heights (0, 1.5, 3, 4.5, 6 mm bone loss) in response to a 1-N tipping force were calculated. Maximum tooth displacement was found to be 2.65 mm at the top of the crown of the model with a 6 mm bone loss. Minimum tooth displacement was 0.45 mm at the cervical level of the model with a normal bone support. Tooth tipping degrees of models in response to different tipping force magnitudes were also calculated for models with different degrees of HBL. Degrees of tipping tooth movement increased as force level was increased. This increase was more prominent in the models with smaller degrees of HBL. By using finite element method and bone remodeling theories, this study indicated that in the presence of HBL, under the same load, long-term orthodontic tooth movement will increase. The simulation also revealed that even though tooth movement increases with increasing the force, this increase was only prominent in the models with smaller degrees of HBL, and tooth models with greater degrees of HBL will be less affected by the magnitude of an orthodontic force. Based on our results, the applied force magnitude must be reduced in proportion of degree of HBL.

Keywords: bone remodeling, finite element method, horizontal bone loss, orthodontic tooth movement.

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Authors: Levylee Bautista, Dawn Grace Santos, Aishwarya Veluchamy, Jesusa Santos, Ghafoor Haque, Jr. I, Rodolfo Rafael

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Sesbania grandiflora is widely used in traditional medicine to treat a wide range of ailments. Assessment of its toxic properties is hence crucial when considering public health protection because exposure to plant extracts may pose adverse effects on consumers. This study aimed to investigate the acute intraperitoneal toxicity of S. grandiflora flower methanolic extract (SGFME) in Swiss albino mice. Four different concentrations (11.25, 22.5, 40, and 90 mg/kg) of SGFME were administered intraperitoneally and immediate behavioral and clinical signs were observed. All concentrations of SGFME-treated mice exhibited gasping and faster respiratory rate, writhing, reddening and fanning of the ears, paralysis of the hind leg, and mortality. Such reactions may be attributed to the histamine and saponin content of S. grandiflora. Results of this study suggests that intraperitoneal administration of SGFME produced significant adverse effect in mice, therefore, caution should be exercised in using it as herbal remedy since there is little control over its quality.

Keywords: acute toxicity test, histamine, medicinal plants, Sesbania grandiflora

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Authors: Mohamed A. Dkhil, Denis Delic, Saleh Al-Quraishy

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Coccidiosis causes considerable economic loss in the poultry industry. The current study aimed to investigate the response of goblet cells as well as the induced tissue damage during Eimeria papilata infection. Mice were infected with sporulated E. papillata oocyts. On day 5 post-infection, the fecal output was determined. Also, the jejunum was prepared for the histological, histochemical and molecular studies. Our results revealed that the intestinal coccidian infection with E. papillata induced a marked goblet cell hypoplasia and depleted mucus secretion. Also, the infection was able to alter the jejuna architecture and increased the apoptotic cells inside the villi. In addition, the real time PCR results indicated that, the inflammatory cytokines TNF-α, iNOS, IFN-y and IL-1β were significantly up-regulated. In contrast, the mRNA expression patterns of IL-6 in response to E. papillata infection did not differ significantly between control and infected mice. Moreover, the mRNA expression of TLR4 was significantly up-regulated, whereas the expression of MUC2 is significantly down-regulated upon infection. Further studies are required to understand the regulatory mechanisms of goblet cells related genes.

Keywords: goblet cells, Eimeria papillata, mice, jejunum

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Authors: Joanna Maj, Awais Hussain, Lyndsey Vu, Catherine Roxas

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Background: Bone injuries in babies are common conditions that arise during delivery. Fractures of the clavicle, humerus, femur, and skull are the most common neonatal bone injuries sustained from labor and delivery. During operative deliveries, zealous tractions, ineffective delivery techniques, improper uterine incision, and inadequate relaxation of the uterus can lead to bone fractures in the newborn. Neonatal anatomy is unique. Just as children are not mini-adults, newborns are not mini children. A newborn’s anatomy and physiology are significantly different from a pediatric patient's. In this paper, we describe common orthopedic trauma in newborn babies. We provide a comprehensive overview of the different types of bone injuries in newborns. We hypothesize that the rate of bone fractures sustained at birth is higher in cases of operative deliveries. Methods: Relevant literature was selected by using the PubMed database. Search terms included orthopedic conditions in newborns, neonatal anatomy, and bone fractures in neonates during operative deliveries. Inclusion criteria included age, gender, race, type of bone injury and progression of bone injury. Exclusion criteria were limited in the medical history of cases reviewed and comorbidities. Results: This review finds that a clavicle fracture is the most common type of neonatal orthopedic injury sustained at birth in both operative and non-operative deliveries. We confirm the hypothesis that infants born via operative deliveries have a significantly higher rate of bone fractures than non-cesarean section deliveries. Conclusion: Newborn babies born via operative deliveries have a higher rate of bone fractures of the clavicle, humerus, and femur. A clavicle bone fracture in newborns is most common during emergency operative deliveries in new mothers. We conclude that infants born via an operative delivery sustained more bone injuries than infants born via non-cesarean section deliveries.

Keywords: clavicle fracture, humerus fracture, neonates, newborn orthopedics, orthopedic surgery, pediatrics, orthopedic trauma, orthopedic trauma during delivery, cesarean section, obstetrics, neonatal anatomy, neonatal fractures, operative deliveries, labor and delivery, bone injuries in neonates

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Authors: Omnia M. Kandil, Noha M. F. Hassan, Doaa Sedky, Hatem A. Shalaby, Heba M. Ashry, Nadia M. T. Abu El Ezz, Sahar M. Kandeel, Mohamed S. Abdelfattah Ying L, Ebtesam M. Al-Olayan

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The aim of the present study was to determine the prevalence of bovine cysticercosis in both cattle and buffaloes in Egypt and to assess the cysticidal efficacy of Balanites aegyptiacafruits (B. aegyptiaca) and Moringa oleiferaseeds (M. oleifera) extracts in experimentally infected mice. The study detected the level of tumor necrosis factor (TNF-α) to monitor the immune and inflammatory responses of experimentally infected mice. Through meat inspection, a total number of 2125 male bovines, 2 to 5 years old (1125 cattle and 1000 buffaloes) were examined at official abattoirs in Cairo Governorate. The prevalence of the disease among bovine was 7.8%, (6.31% of cattle and 9.5% of buffaloes). A decrease in the number of cysticerci was found in all treated mice groups, and up to 88% reduction was achieved in the B. aegyptiaca-treated group; higher than that was recorded in both M. oleifera (72.23%) and albendazole-treated ones (80.56%). Postmortem findings proved that M. oleifera and B. aegyptiaca reduced cysticerci numbers comparable to a commercial anthelmintic. The study showed a significant decrease (P<0.001) in TNF-α levels after treatment with Balanites and Moringa extracts, compared with the untreated control and the albendazole-treated groups.

Keywords: prevalence, bovine cysticercossis, extracts, mice

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Authors: S. Subramaniam, J. S. A. Rao, P. Ramdas, K. R. Selvaduray, N. M. Han, M. K. Kutty, A. K. Radhakrishnan

Abstract:

Immune system is a complex system where the immune cells have the capability to respond against a wide range of immune challenges including cancer progression. However, in the event of cancer development, tumour cells trigger immunosuppressive environment via activation of myeloid-derived suppressor cells and T regulatory (Treg) cells. The Treg cells are subset of CD4+ T lymphocytes, known to have crucial roles in regulating immune homeostasis and promoting the establishment and maintenance of peripheral tolerance. Dysregulation of these mechanisms could lead to cancer progression and immune suppression. Recently, there are many studies reporting on the effects of natural bioactive compounds on immune responses against cancer. It was known that tocotrienol-rich-fraction consisting 70% tocotrienols and 30% α-tocopherol is able to exhibit immunomodulatory as well as anti-cancer properties. Hence, this study was designed to evaluate the effects of gamma-tocotrienol (G-T3) supplementation on T-reg cells in a syngeneic mouse model of breast cancer. In this study, female BALB/c mice were divided into two groups and fed with either soy oil (vehicle) or gamma-tocotrienol (G-T3) for two weeks followed by inoculation with tumour cells. All the mice continued to receive the same supplementation until day 49. The results showed a significant reduction in tumour volume and weight in G-T3 fed mice compared to vehicle-fed mice. Lung and liver histology showed reduced evidence of metastasis in tumour-bearing G-T3 fed mice. Besides that, flow cytometry analysis revealed T-helper cell population was increased, and T-regulatory cell population was suppressed following G-T3 supplementation. Moreover, immunohistochemistry analysis showed that there was a marked decrease in the expression of FOXP3 in the G-T3 fed tumour bearing mice. In conclusion, the G-T3 supplementation showed good prognosis towards breast cancer by enhancing the immune response in tumour-bearing mice. Therefore, gamma-T3 can be used as immunotherapy agent for the treatment of breast cancer.

Keywords: breast cancer, gamma tocotrienol, immune suppression, supplement

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Authors: I. Nur Hilwani, R. Nasibah, S. Nurdiana, M. J. Norashirene

Abstract:

Impaired fertility may be the result of indirect consumption of anti-fertility agents through food. Monosodium glutamate (MSG) has been widely used as food additive, flavour enhancer and included in vaccines. This study focuses in determining the gonadotoxic and cytotoxic effect of MSG on selected sperm parameters such as sperm viability, sperm membrane integrity and testes cytoarchitecture of male mice via histological examination to determine its effect on spermatogenesis. Twenty-four Mus musculus were randomly divided into 4 groups and given intraperitoneal injections (IP) daily for 14 days of different MSG concentrations at 250, 500 and 1000mg/kg MSG to body weight to induce obesity. Saline was given to control group. Mice were sacrificed and analysis revealed abnormalities in values for sperm parameters and damages to testes cytoarchitecture of male mice. The results recorded decreased viability (p<0.05) and integrity of sperm membrane (p>0.05) with degenerative structures in seminiferous tubule of testes. The results indicated various implications of MSG on male mice reproductive system which has consequences in fertility potential.

Keywords: sperm parameter, testes histology, sperm viability, sperm membrane integrity

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Authors: Oluwabori Emmanuel Olukoyejo, Ogra Victor Ogra, Bosede Amodu, Tewogbade Adeoye Adedeji

Abstract:

Background: Disorders of thyroid function are the second most common endocrine disorders worldwide, with a direct relationship with metabolic bone diseases. These metabolic bone complications are often subtle but manifest as bone pains and an increased risk of fractures. The gold standard for diagnosis, Dual Energy X-ray Absorptiometry (DEXA), is limited in this environment due to unavailability, cumbersomeness and cost. However, bone biomarkers have shown prospects in assessing alterations in bone remodeling, which has not been studied in this environment. Aim: This study evaluates serum levels of bone-specific alkaline phosphatase (bone-specific ALP), osteopontin and osteoprotegerin biomarkers of bone turnover in patients with disorders of thyroid function. Methods: This is a cross-sectional study carried out over a period of one and a half years. Forty patients with thyroid dysfunctions, aged 20 to 50 years, and thirty-eight age and sex-matched healthy euthyroid controls were included in this study. Patients were further stratified into hyperthyroid and hypothyroid groups. Bone-specific ALP, osteopontin, and osteoprotegerin, alongside serum total calcium, ionized calcium and inorganic phosphate, were assayed for all patients and controls. A self-administered questionnaire was used to obtain data on sociodemographic and medical history. Then, 5 ml of blood was collected in a plain bottle and serum was harvested following clotting and centrifugation. Serum samples were assayed for B-ALP, osteopontin, and osteoprotegerin using the ELISA technique. Total calcium and ionized calcium were assayed using an ion-selective electrode, while the inorganic phosphate was assayed with automated photometry. Results: The hyperthyroid and hypothyroid patient groups had significantly increased median serum B-ALP (30.40 and 26.50) ng/ml and significantly lower median OPG (0.80 and 0.80) ng/ml than the controls (10.81 and 1.30) ng/ml respectively, p < 0.05. However, serum osteopontin in the hyperthyroid group was significantly higher and significantly lower in the hypothyroid group when compared with the controls (11.00 and 2.10 vs 3.70) ng/ml, respectively, p < 0.05. Both hyperthyroid and hypothyroid groups had significantly higher mean serum total calcium, ionized calcium and inorganic phosphate than the controls (2.49 ± 0.28, 1.27 ± 0.14 and 1.33 ± 0.33) mmol/l and (2.41 ± 0.04, 1.20 ± 0.04 and 1.15 ± 0.16) mmol/l vs (2.27 ± 0.11, 1.17 ± 0.06 and 1.08 ± 0.16) mmol/l respectively, p < 0.05. Conclusion: Patients with disorders of thyroid function have metabolic imbalances of all the studied bone markers, suggesting a higher bone turnover. The routine bone markers will be an invaluable tool for monitoring bone health in patients with thyroid dysfunctions, while the less readily available markers can be introduced as supplementary tools. Moreover, bone-specific ALP, osteopontin and osteoprotegerin were found to be the strongest independent predictors of metabolic bone markers’ derangements in patients with thyroid dysfunctions.

Keywords: metabolic bone diseases, biomarker, bone turnover, hyperthyroid, hypothyroid, euthyroid

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Authors: Hye Kyung Kim, Myung-Gyou Kim, Kang-Hyun Leem

Abstract:

Postmenopausal osteoporosis is characterized by low bone density which leads to increased bone fragility and greater susceptibility to fracture. Current treatments for osteoporosis are dominated by drugs that inhibit bone resorption although they also suppress bone formation that may contribute to pathogenesis of osteonecrosis. To restore the extensive bone loss, there is a great need for anabolic treatments that induce osteoblasts to build new bone. Pre-osteoblastic cells produce proteins of the extra-cellular matrix, including type I collagen at first, and then to successively produce alkaline phosphatase (ALP) and osteocalcin during differentiation to osteoblasts. Finally, osteoblasts deposit calcium. Present study investigated the effects of egg yolk peptide (EYP) on osteogenic activities and bone matrix gene expressions in human osteoblastic MG-63 cells. The effects of EYP on cell proliferation, alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization were measured. The expression of osteogenic genes including COL1A1 (collagen, type I, alpha 1), ALP, BGLAP (osteocalcin), and SPP1 (secreted phosphoprotein 1, osteopontin) were measured by quantitative realtime PCR. EYP dose-dependently increased MG-63 cell proliferation, ALP activity, collagen synthesis, and calcium deposition. Furthermore, COL1A1, ALP, and SPP1 gene expressions were increased by EYP treatment. Present study suggested that EYP treatment enhanced osteogenic activities and increased bone matrix osteogenicgenes. These results could provide a mechanistic explanation for the bone-strengthening effects of EYP.

Keywords: egg yolk peptide, osteoblastic MG-63 cells, alkaline phosphatase, collagen synthesis, osteogenic genes, COL1A1, osteocalcin, osteopontin

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Authors: Ping-Lun Jiang, Hung-Jun Lin, Shen-Fu Lin, Mei-Yin Chien, Ting-Wei Li, Chun-Han Lin, Der-Zen Liu

Abstract:

Mucosal vaccine induces both mucosal (secretory IgA) and systemic immune responses and it is considered an ideal vaccination strategy for prevention of infectious diseases. One important point to be considered in mucosal vaccination is effective antigen delivery system which can manage effective delivery of antigen to antigen-presenting cells (APCs) of mucosal. In the present study, cationic gelatin nanoparticles were prepared as ideal carriers for more efficient antigen delivery. The average diameter of cationic gelatin nanoparticle was approximate 190 nm, and the zeta potential was about +45 mV, then ovalbumin (OVA) was physically absorbed onto cationic gelatin nanoparticle. The OVA absorption rate was near 95% the zeta potential was about +20 mV. We show that cationic gelatin nanoparticle effectively facilitated antigen uptake by mice bone marrow-derived dendritic cells (mBMDCs) and RAW264.7 cells and induced higher levels of pro-inflammatory cytokines. C57BL/6 mice twice immunized intranasally with OVA-absorbed cationic gelatin nanoparticle induced high levels of OVA-specific IgG in the serum and IgA in their in the nasal and lung wash fluid. These results indicate that nasal administration of cationic gelatin nanoparticles induced both mucosal and systemic immune responses and cationic gelatin nanoparticles might be a potential antigen delivery carrier for further clinical applications.

Keywords: antigen delivery, antigen-presenting cells, gelatin nanoparticle, mucosal vaccine

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Authors: Al-Nouri Doha Mostfa, Al-Khalifa Abdulrahman Salih

Abstract:

The effects of long-term supplementation with different dietary omega-6/omega-3 (ω-6/ω-3) polyunsaturated fatty acid (PUFAs) ratios on the bone marrow fatty acids level, plasma biomarkers of bone metabolism, and minerals content in bone were evaluated in rabbits. Weanling male and female New Zealand white rabbits were randomly assigned to five groups and fed ad libitum for 100 days on diets containing 70 g/kg different dietary oils which providing the following ω-6/ω-3 ratios: soy bean oil (SBO control, 8.68), sesame oil (SO, 21.75), fish oil (FO, 0.39), DHA algae oil (DHA, 0.63), and DHA and ARA algae oils (DHA/ARA, 0.68). The bone marrow arachidonic (ARA), eicosapentaenoic (EPA), and docosahexaenoic (DHA) fatty acid levels were significantly influenced by and reflected the dietary ω-6/ω-3 ratios fed to rabbits. Rabbits fed on the FO diet maintained a lower ω-6/ω-3 ratio and a higher EPA and DHA levels, those fed on the DHA/ARA diet maintained a lower ω-6/ω-3 ratio and a higher ARA level, while those fed on the SO diet maintained a higher ω-6/ω-3 ratio and a lower ARA level. Plasma alkaline phosphatase (ALP) activity was significantly higher in male and female rabbits fed the DHA/ARA diet compared with those fed the control, SO, FO, or DHA diets. There was a significant main effect of dietary treatment on femur calcium (Ca), phosphorous (P), magnesium (Mg), and zinc (Zn) contents in both genders. This study confirmed that different dietary oil sources with varying ω-6/ω-3 ratios significantly altered the fatty acids level of bone marrow. In addition, the significant elevation in minerals content and the maintenance of optimal Ca/P ratio in bone of DHA/ARA and DHA fed groups beside the significant elevation in ALP activity in the DHA/ARA fed group proved that marine algae oils may be promising dietary sources for promoting bone mineralization and formation, thus improving bone mass during the growth stage.

Keywords: arachidonic (ARA), docosahexaenoic (DHA), eicosapentaenoic (EPA), growing rabbits

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Authors: Sizhu Wang, Cuisong Tang, Lin Zhang, Guangyu Tang

Abstract:

Objective: We aimed to detect the composition of bone marrow fatty acids early after ovariectomized (OVX) surgery and explore the potential mechanism. Methods: Thirty-two female Sprague-Dawley (SD) rats (12 weeks) were randomly divided into OVX group and Sham group (N=16/group), and received ovariectomy or sham surgery respectively. After 3 and 28 days, eight rats in each group were sacrificed to detect the composition of bone marrow fatty acids by gas chromatography–mass spectrometry (GC–MS) and evaluate the trabecular bone microarchitecture by micro-CT. Significant different fatty acids in the early stage of post-menopausal osteoporosis were selected by OPLS-DA and t test. Then selected fatty acids were further studied in the process of osteogenic differentiation through RT-PCR and Alizarin Red S staining. Results: An apparent sample clustering and group separation were observed between OVX group and sham group three days after surgery, which suggested the role of bone marrow fatty acids in the early stage of postmenopausal osteoporosis. Specifically, myristate, palmitoleate and arachidonate were found to play an important role in classification between OVX group and sham group. We further investigated the effect of palmitoleate and arachidonate on osteogenic differentiation and found that palmitoleate promoted the osteogenic differentiation of MC3T3-E1 cells while arachidonate inhibited this process. Conclusion: Profound bone marrow fatty acids changes have taken place in the early stage of post-menopausal osteoporosis. Bone marrow fatty acids may begin to affect osteogenic differentiation shortly after deficiency of estrogen.

Keywords: bone marrow fatty acids, GC-MS, osteoblast, osteoporosis, post-menopausal

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Authors: Laila Montaser, Hala Gabr, Maha El-Bassuony, Gehan Tawfeek

Abstract:

Orthotropic liver transplantation (OLT) is the final procedure of both end stage and metabolic liver diseases. Hepatocyte transplantation is an alternative for OLT, but the sources of hepatocytes are limited. Bone marrow mesenchymal stem cells (BM-MSCs) can differentiate into hepatocyte-like cells and are a potential alternative source for hepatocytes. The MSCs from bone marrow are a promising target population as they are capable of differentiating along multiple lineages and, at least in vitro, have significant expansion capability. MSCs from bone marrow may have the potential to differentiate in vitro and in vivo into hepatocytes. Our study examined whether mesenchymal stem cells (MSCs), which are stem cells originated from human bone marrow, are able to differentiate into functional hepatocyte-like cells in vitro. Our aim was to investigate the differentiation potential of BM-MSCs into hepatocyte-like cells. Adult stem cell therapy could solve the problem of degenerative disorders, including liver disease.

Keywords: bone marrow, differentiation, hepatocyte, stem cells

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Authors: Daeung Jung

Abstract:

Background: In case of patients with missing teeth, multiple implant restoration has been widely used and is inevitable. To increase its survival rate, it is important to understand the influence of different implant designs on inter-implant crestal bone resorption. There are several implant systems designed to minimize loss of crestal bone, and the Astra Tech and Brånemark Implant are two of them. Aim/Hypothesis: The aim of this 10-year study was to compare the height of inter-implant bone crest in two implant systems; the Astra Tech and the Brånemark implant system. Material and Methods: In this retrospective study, 40 consecutively treated patients were utilized; 23 patients with 30 sites for Astra Tech system and 17 patients with 20 sites for Brånemark system. The implant restoration was comprised of splinted crown in partially edentulous patients. Radiographs were taken immediately after 1st surgery, at impression making, at prosthetics setting, and annually after loading. Lateral distance from implant to bone crest, inter-implant distance was gauged, and crestal bone height was measured from the implant shoulder to the first bone contact. Calibrations were performed with known length of thread pitch distance for vertical measurement, and known diameter of abutment or fixture for horizontal measurement using ImageJ. Results: After 10 years, patients treated with Astra Tech implant system demonstrated less inter-implant crestal bone resorption when implants had a distance of 3mm or less between them. In cases of implants that had a greater than 3 mm distance between them, however, there appeared to be no statistically significant difference in crestal bone loss between two systems. Conclusion and clinical implications: In the situation of partially edentulous patients planning to have more than two implants, the inter-implant distance is one of the most important factors to be considered. If it is impossible to make sure of having sufficient inter-implant distance, the implants with less micro gap in the fixture-abutment junction, less traumatic 2nd surgery approach, and the adequate surface topography would be choice of appropriate options to minimize inter-implant crestal bone resorption.

Keywords: implant design, crestal bone loss, inter-implant distance, 10-year retrospective study

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