Search results for: bovine bone

Authors: Sonia A. Stoica, Lexi Frankel, Amalia Ardeljan, Selena Rashid, Ali Yasback, Omar Rashid

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Introduction Enterococcus is a vast genus of lactic acid bacteria, gram-positivecocci species. They are common commensal organisms in the intestines of humans: E. faecalis (90–95%) and E. faecium (5–10%). Rare groups of infections can occur with other species, including E. casseliflavus, E. gallinarum, and E. raffinosus. The most common infections caused by Enterococcus include urinary tract infections, biliary tract infections, subacute endocarditis, diverticulitis, meningitis, septicemia, and spontaneous bacterial peritonitis. The treatment for sensitive strains of these bacteria includes ampicillin, penicillin, cephalosporins, or vancomycin, while the treatment for resistant strains includes daptomycin, linezolid, tygecycline, or streptogramine. Enterococcus faecalis CECT7121 is an encouraging nominee for being considered as a probiotic strain. E. faecalis CECT7121 enhances and skews the profile of cytokines to the Th1 phenotype in situations such as vaccination, anti-tumoral immunity, and allergic reactions. It also enhances the secretion of high levels of IL-12, IL-6, TNF alpha, and IL-10. Cytokines have been previously associated with the development of cancer. The intention of this study was to therefore evaluate the correlation between Enterococcus infections and incidence of bone carcinoma. Methods A retrospective cohort study (2010-2019) was conducted through a Health Insurance Portability and Accountability Act (HIPAA) compliant national database and conducted using International Classification of Disease (ICD) 9th and 10th codes for bone carcinoma diagnosis in a previously Enterococcus infected population. Patients were matched for age range and Charlson Comorbidity Index (CCI). Access to the database was granted by Holy Cross Health for academic research. Chi-squared test was used to assess statistical significance. Results A total number of 17,056 patients was obtained in Enterococcus infected group as well as in the control population (matched by Age range and CCI score). Subsequent bone carcinoma development was seen at a rate of 1.07% (184) in the Enterococcal infectious group and 3.42% (584) in the control group, respectively. The difference was statistically significant by p= 2.2x10-¹⁶, Odds Ratio = 0.355 (95% CI 0.311 - 0.404) Treatment for enterococcus infection was analyzed and controlled for in both enterococcus infected and noninfected populations. 78 out of 6,624 (1.17%) patients with a prior enterococcus infection and treated with antibiotics were compared to 202 out of 6,624 (3.04%) patients with no history of enterococcus infection (control) and received antibiotic treatment. Both populations subsequently developed bone carcinoma. Results remained statistically significant (p<2.2x10-), Odds Ratio=0.456 (95% CI 0.396-0.525). Conclusion This study shows a statistically significant correlation between Enterococcus infection and a decreased incidence of bone carcinoma. The immunologic response of the organism to Enterococcus infection may exert a protecting mechanism from developing bone carcinoma. Further exploration is needed to identify the potential mechanism of Enterococcus in reducing bone carcinoma incidence.

Keywords: anti-tumoral immunity, bone carcinoma, enterococcus, immunologic response

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Authors: Eva Filová, Jana Daňková, Věra Sovková, Matej Daniel

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Titanium alloys are biocompatible metals that are widely used in clinical practice as load bearing implants. The chemical modification may influence cell adhesion, proliferation, and differentiation as well as stiffness of the material. The aim of the study was to evaluate the adhesion, growth and differentiation of pig mesenchymal stem cells on the novel beta titanium alloy Ti36Nb6Ta compared to standard medical titanium alloy Ti6Al4V. Discs of Ti36Nb6Ta and Ti6Al4V alloy were sterilized by ethanol, put in 48-well plates, and seeded by pig mesenchymal stem cells at the density of 60×103/cm2 and cultured in Minimum essential medium (Sigma) supplemented with 10% fetal bovine serum and penicillin/streptomycin. Cell viability was evaluated using MTS assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay;Promega), cell proliferation using Quant-iT™ ds DNA Assay Kit (Life Technologies). Cells were stained immunohistochemically using monoclonal antibody beta-actin, and secondary antibody conjugated with AlexaFluor®488 and subsequently the spread area of cells was measured. Cell differentiation was evaluated by alkaline phosphatase assay using p-nitrophenyl phosphate (pNPP) as a substrate; the reaction was stopped by NaOH, and the absorbance was measured at 405 nm. Osteocalcin, specific bone marker was stained immunohistochemically and subsequently visualized using confocal microscopy; the fluorescence intensity was analyzed and quantified. Moreover, gene expression of osteogenic markers osteocalcin and type I collagen was evaluated by real-time reverse transcription-PCR (qRT-PCR). For statistical evaluation, One-way ANOVA followed by Student-Newman-Keuls Method was used. For qRT-PCR, the nonparametric Kruskal-Wallis Test and Dunn's Multiple Comparison Test were used. The absorbance in MTS assay was significantly higher on titanium alloy Ti6Al4V compared to beta titanium alloy Ti36Nb6Ta on days 7 and 14. Mesenchymal stem cells were well spread on both alloys, but no difference in spread area was found. No differences in alkaline phosphatase assay, fluorescence intensity of osteocalcin as well as the expression of type I collagen, and osteocalcin genes were observed. Higher expression of type I collagen compared to osteocalcin was observed for cells on both alloys. Both beta titanium alloy Ti36Nb6Ta and titanium alloy Ti6Al4V Ti36Nb6Ta supported mesenchymal stem cellsˈ adhesion, proliferation and osteogenic differentiation. Novel beta titanium alloys Ti36Nb6Ta is a promising material for bone implantation. The project was supported by the Czech Science Foundation: grant No. 16-14758S, the Grant Agency of the Charles University, grant No. 1246314 and by the Ministry of Education, Youth and Sports NPU I: LO1309.

Keywords: beta titanium, cell growth, mesenchymal stem cells, titanium alloy, implant

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Authors: Mohammed A. Alshehri

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Salvia officinalis is an aromatic plant member of the mint (Labiatae) family. It is popular kitchen herb. Not surprise to find that the name of this herb related to cure, in Latin language Salvia means to cure where officinalis means medicinal which answer why the sage has a top place in the list of medicinal plants. The aim of the present study was to assess the genetic damage and cytological changes caused by exposure of the test organism (Rattusrattus) to Salvia officinals. For this purpose, adult female rats, weighing 200–250 g, were used as donors. A total of 36 adult Wister male rats were randomly assigned to five groups: the experimental groups (rats were intraperitonealy injected with Salvia officinalis pure extract at (0.1, 0.2, 0.5, 0.1mg/kg body weight, the same dose was administered once a day. Control group (rats were injected intraperitonealy physiological saline. And positive control were injected with Cyclophosphamide. On the 21st days following Salvia officinalis pure extract exposure, rats were sacrificed, and samples of bone marrow were collected. Following that, we performed a micronuclei (MN) test using MNNCE (Micro-nucleated normocromatic erythrocytes) and MNPCE (Micronucleated polychromatic erythrocytes), NDI (Nuclear division index), and cytological parameters using NDCI (nuclear division cytotoxicity index), necrotic, and apoptotic cells in rat's bone marrow samples. Results showed that there was a no significant increase in the frequency of micro-nucleatedas well as in cytological parameters in bone marrow cells. In light of these results, if Salvia officinalis pure extract may considered to be safe from the stand point of genotoxicity and cytotoxicity effects.

Keywords: Salvia officinalis, micronucleus, NDI, NDCI, toxicity, chromosomal aberrations

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Authors: Abdulsalam A. Al-Tamimi, Chris Peach, Paulo Rui Fernandes, Paulo J. Bartolo

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Bone fixation implants currently used to treat traumatic fractured bones and to promote fracture healing are built with biocompatible metallic materials such as stainless steel, cobalt chromium and titanium and its alloys (e.g., CoCrMo and Ti6Al4V). The noticeable stiffness mismatch between current metallic implants and host bone associates with negative outcomes such as stress shielding which causes bone loss and implant loosening leading to deficient fracture treatment. This paper, part of a major research program to design the next generation of bone fixation implants, describes the combined use of three-dimensional (3D) topology optimization (TO) and additive manufacturing powder bed technology (Electron Beam Melting) to redesign and fabricate the plates based on the current standard one (i.e., locking compression plate). Topology optimization is applied with an objective function to maximize the stiffness and constraint by volume reductions (i.e., 25-75%) in order to obtain optimized implant designs with reduced stress shielding phenomenon, under different boundary conditions (i.e., tension, bending, torsion and combined loads). The stiffness of the original and optimised plates are assessed through a finite-element study. The TO results showed actual reduction in the stiffness for most of the plates due to the critical values of volume reduction. Additionally, the optimized plates fabricated using powder bed techniques proved that the integration between the TO and additive manufacturing presents the capability of producing stiff reduced plates with acceptable tolerances.

Keywords: additive manufacturing, locking compression plate, finite element, topology optimization

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Authors: Patricia Shirley Almeida Prado, Liz Brito, Selma Paixão Argollo, Gracie Moreira, Leticia Matos Sobrinho

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Diffuse idiopathic skeletal hyperostosis (DISH) is a degenerative bone disease also known as Forestier´s disease and ankylosing hyperostosis of the spine is characterized by a tendency toward ossification of half the anterior longitudinal spinal ligament without intervertebral disc disease. DISH is not considered to be osteoarthritis, although the two conditions commonly occur together. Diagnostic criteria include fusion of at least four vertebrae by bony bridges arising from the anterolateral aspect of the vertebral bodies. These vertebral bodies have a 'dripping candle wax' appearance, also can be seen periosteal new bone formation on the anterior surface of the vertebral bodies and there is no ankylosis at zygoapophyseal facet joint. Clinically, patients with DISH tend to be asymptomatic some patients mention moderate pain and stiffness in upper back. This disease is more common in man, uncommon in patients younger than 50 years and rare in patients under 40 years old. In modern populations, DISH is found in association with obesity, (type II) diabetes; abnormal vitamin A metabolism and also associated with higher levels of serum uric acid. There is also some association between the increase of risk of stroke or other cerebrovascular disease. The DISH condition can be confused with Heterotopic Ossification, what is the bone formation in the soft tissues as the result of trauma, wounding, surgery, burnings, prolonged immobility and some central nervous system disorder. All these conditions have been described extensively as myositis ossificans which can be confused with the fibrodysplasia (myositis) ossificans progressive. As in the DISH symptomatology it can be asymptomatic or extensive enough to impair joint function. A third confusion osteoarthritis disease that can bring confusion are the enthesopathies that occur in the entire skeleton being common on the ischial tuberosities, iliac crests, patellae, and calcaneus. Ankylosis of the sacroiliac joint by bony bridges may also be found. CASE 1: this case is skeletal remains presenting skull, some vertebrae and scapulae. This case remains unidentified and due to lack of bone remains. Sex, age and ancestry profile was compromised, however the DISH pathognomonic findings and diagnostic helps to estimate sex and age characteristics. Moreover to presenting DISH these skeletal remains also showed some bone alterations and non-metrics as fusion of the first vertebrae with occipital bone, maxillae and palatine torus and scapular foramen on the right scapulae. CASE 2: this skeleton remains shows an extensive bone heterotopic ossification on the great trochanter area of left femur, right fibula showed a healed fracture in its body however in its inteosseous crest there is an extensive bone growth, also in the Ilium at the region of inferior gluteal line can be observed some pronounced bone growth and the skull presented a pronounced mandibular, maxillary and palatine torus. Despite all these pronounced heterotopic ossification the whole skeleton presents moderate bone overgrowth that is not linked with aging, since the skeleton belongs to a young unidentified individual. The appropriate osteopathological diagnosis support the human identification process through medical reports and also assist with epidemiological data that can strengthen vulnerable anthropological estimates.

Keywords: bone disease, DISH, human identification, human remains

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Authors: Fakhrosadat Sajjadian

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The majority of deaths in cancer patients are caused by distant metastasis. Breast cancer shows a unique spread pattern, often affecting bone, liver, lung, and brain. Breast cancer can be categorized into various subtypes according to gene expression patterns, and these subtypes exhibit specific preferences for organs where metastasis occurs. Breast tumors with luminal characteristics have a preference for spreading to the bone, whereas basal-like breast cancer (BLBC) shows a tendency to metastasize to the lungs. Still, the mechanisms behind this particular pattern of metastasis in organs have yet to be fully understood. In this evaluation, we will outline the latest progress in molecular signaling pathways and treatment methods for breast cancer lung metastasis.

Keywords: lung cancer, liver cancer, diagnosis, BLBC, metastasis

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Authors: Nabeeha Hassan Abdel Jalil, Lulwa Saeed Al Badi, Mouza Ghafan Alkhyeli, Khaja Mohteshamuddin, Ahmad Al Aiyan, Mohamed Elfatih Hamad, Robert Barigye

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The present study was done to elucidate the prevalence of enzootic bovine leucosis (EBL) in the UAE, the seroprevalence rates of EBL in dairy herds from the Al Ain area, Abu Dhabi (AD) and indigenous cattle at the Al Ain livestock market (AALM) were assessed. Of the 949 sera tested by ELISA, 657 were from adult Holstein-Friesians from three farms and 292 from indigenous cattle at the AALM. The level of significance between the proportions of seropositive cattle were analyzed by the Marascuilo procedure and questionnaire data on husbandry and biosecurity practices evaluated. Overall, the aggregated farm and AALM data demonstrated a seroprevalence of 25.9%, compared to 37.0% for the study farms, and 1.0% for the indigenous cattle. Additionally, the seroprevalence rates at farms #1, #2 and #3 were 54.7%, 0.0%, and 26.3% respectively. Except for farm #2 and the AALM, statistically significant differences were noted between the proportions of seropositive cattle for farms #1 and #2 (Critical Range or CR=0.0803), farms #1 and #3 (p=0.1069), and farms #2 and #3 (CR=0.0707), farm #1 and the AALM (CR=0.0819), and farm #3 and the AALM (CR=0.0726). Also, the proportions of seropositive animals on farm #1 were 9.8%, 59.8%, 29.3%, and 1.2% in the 12-36, 37-72, 73-108, and 109-144-mo-old age groups respectively compared to 21.5%, 60.8%, 15.2%, and 2.5% in the respective age groups for farm #2. On both farms and the AALM, the 37-72-mo-old age group showed the highest EBL seroprevalence rate while all the 57 cattle on farm #2 were seronegative. Additionally, farms #1 and #3 had 3,130 and 2,828 intensively managed Holstein-Friesian cattle respectively, and all animals were routinely immunized against several diseases except EBL. On both farms #1 and #3, artificial breeding was practiced using semen sourced from the USA, and USA and Canada respectively, all farms routinely quarantined new stock, and farm #1 previously imported dairy cattle from an unspecified country, and farm #3 from the Netherlands, Australia and South Africa. While farm #1 provided no information on animal nutrition, farm #3 cited using hay, concentrates, and ad lib water. To the authors’ best knowledge, this is the first serological evidence of EBL in the UAE and as previously reported, the seroprevalence rates are comparatively higher in the intensively managed dairy herds than in indigenous cattle. As two of the study farms previously sourced cattle and semen from overseas, biosecurity protocols need to be revisited to avoid inadvertent EBL incursion and the possibility of regional transboundary disease spread also needs to be assessed. After the proposed molecular studies have adduced additional data, the relevant UAE animal health authorities may need to develop evidence-based EBL control policies and programs.

Keywords: cattle, enzootic bovine leukosis, seroprevalence, UAE

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Authors: Shantanu Vyas, Neerja Meena

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Facial infiltrating lipomatosis is a rare lipomatous lesion, first described by Slavin in 1983. It is a benign pseudotumor pathology. It corresponds to a non-encapsulated collection of mature adipocytes infiltrating the local tissue and hyperplasia of underlying bone leading to a craniofacial deformity. Very few cases have been reported in the literature. We report the case of a 19-year-old female patient, who was consulted for a swelling of the right hemiface progressively evolving since birth. Physical examination revealed facial asymmetry. On palpation, the mass was soft, painless, not compressible, not pulsatile, not fluctuating. In view of the asymptomatic nature and slow progression of the lesion, a lipomatous tumour, namely lipoma, was suggested. CT scan image shows a hyperplastic subcutaneous fat on the right hemiface. On the right jugal and temporal areas, there is a subcutaneous formation of fatty density, poorly limited, with no detectable peripheral capsule. It merges with the adjacent fat. In the bone window, there was a hyperplasia of underlying bone. Facial lipomatosis infiltration of the face is a benign pseudotumor pathology. As a result, it can be confused with other disorders, in particular, hemifacial hyperplasia. Combination of physical and radiological findings can establish the diagnosis. Surgical treatment is done for cosmetic purposes.

Keywords: cosmetic correction and facial assemetry, aesthetic results, facial infiltration, surgery

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Authors: Mohua Chatterjee, Rajib De

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Introduction: Bone Marrow Transplant (BMT) is often performed to treat patients with various types of leukemia. A majority of these patients develop complications like chronic musculoskeletal GVHD post-BMT where patients get scleroderma, pain and restricted range of motion of joints of hand. If not treated early, it may cause permanent deformity of hand. This study was done to find the effectiveness of physiotherapy in hand dysfunction caused due to chronic musculoskeletal GVHD of leukemia patients after BMT. Methodology: 23 patients diagnosed with leukemia and having musculoskeletal GVHD were treated with a set of exercises including active exercises and stretching. The outcome was measured by Cochin Hand Function Scale (CHFS) at baseline and after four weeks of intervention. Results: Two patients were not able to carry out exercises beyond two weeks due to relapse of disease and one patient defaulted. It was found that all the patients who received physiotherapy had significant improvement in hand function. Mean CHFS decreased from 63.67 to 27.43 (P value < 0.001) indicating improvement in hand function after four weeks of physiotherapy. Conclusion: Early intervention of physiotherapy is effective in reducing hand dysfunction of leukemia patients with musculoskeletal GVHD post-BMT.

Keywords: bone marrow transplant, hand dysfunction, leukemia, musculoskeletal graft versus host disease, physiotherapy

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Authors: Huseyin Apdik, Aysegul Dogan, Selami Demirci, Ezgi Avsar Apdik, Fikrettin Sahin

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Mesenchymal stem cells (MSCs) are crucial cell types for bone maintenance and growth along with resident bone progenitor cells providing bone tissue integrity during osteogenesis and skeletal growth. Any deficiency in this regulation would result in vital bone diseases. Of those, osteoporosis, characterized by a reduction in bone mass and mineral density, is a critical skeletal disease for especially elderly people. The commonly used drugs for the osteoporosis treatment are bisphosphonates (BPs). The most prominent role of BPs is to prevent bone resorption arisen from high osteoclast activity. However, administrations of bisphosphonates may also cause bisphosphonate-induced osteonecrosis of the jaw (BIONJ). Up to the present, the researchers have proposed several circumstances for BIONJ. However, effects of long-term and/or high dose usage of BPs on stem cell’s proliferation, survival, differentiation or maintenance capacity have not been evaluated yet. The present study will be held to; figure out BPs’ effects on MSCs in vitro in the aspect of cell proliferation and toxicity, migration, angiogenic activity, lineage specific gene and protein expression levels, mesenchymal stem cell properties and potential signaling pathways affected by BP treatment. Firstly, mesenchymal stem cell characteristics of Dental Pulp Stem Cells (DPSCs) and Periodontal Ligament Stem Cells (PDLSCs) were proved using flow cytometry analysis. Cell viability analysis was completed to determine the cytotoxic effects of BPs (Zoledronate (Zol), Alendronate (Ale) and Risedronate (Ris)) on DPSCs and PDLSCs by the 3-(4,5-di-methyl-thiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium (MTS) assay. Non-toxic concentrations of BPs were determined at 24 h under growth condition, and at 21 days under osteogenic differentiation condition for both cells. The scratch assay was performed to evaluate their migration capacity under the usage of determined of BPs concentrations at 24 h. The results revealed that while the scratch closure is 70% in the control group for DPSCs, it was 57%, 66% and 66% in Zol, Ale and Ris groups, respectively. For PDLSs, while wound closure is 71% in control group, it was 65%, 66% and 66% in Zol, Ale and Ris groups, respectively. As future experiments, tube formation assay and aortic ring assay will be done to determinate angiogenesis abilities of DPSCs and PDLSCs treated with BPs. Expression levels of osteogenic differentiation marker genes involved in bone development will be determined using real time-polymerase change reaction (RT-PCR) assay and expression profiles of important proteins involved in osteogenesis will be evaluated using western blotting assay for osteogenically differentiated MSCs treated with or without BPs. In addition to these, von Kossa staining will be performed to measure calcium mineralization status of MSCs.

Keywords: bisphosphonates, bisphosphonate-induced osteonecrosis of the jaw, mesenchymal stem cells, osteogenesis

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Authors: Khaled Harrar, Rachid Jennane

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The purpose of this study is the discrimination of 28 postmenopausal with osteoporotic femoral fractures from an age-matched control group of 28 women using texture analysis based on fractals. Two pre-processing approaches are applied on radiographic images; these techniques are compared to highlight the choice of the pre-processing method. Furthermore, the values of the fractal dimension are compared to those of the fractal signature in terms of the classification of the two populations. In a second analysis, the BMD measure at proximal femur was compared to the fractal analysis, the latter, which is a non-invasive technique, allowed a better discrimination; the results confirm that the fractal analysis of texture on calcaneus radiographs is able to discriminate osteoporotic patients with femoral fracture from controls. This discrimination was efficient compared to that obtained by BMD alone. It was also present in comparing subgroups with overlapping values of BMD.

Keywords: osteoporosis, fractal dimension, fractal signature, bone mineral density

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Authors: Sara Mercedes Barroso Pinzón, Álvaro Jesús Caicedo Castro, Antonio Javer Sánchez Herencia

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In recent years there has been increased academic and industrial research into the development of orthopaedic implants with structural properties and functionality similar to mechanical strength, osseointegration, thermal stability and antibacterial capacity similar to bone structure. Hydroxyapatite has been considered for decades as an ideal biomaterial for bone regeneration due to its chemical and crystallographic similarity to the mineral structure bioapatites. However, the lack of trace elements in the hydroxyapatite structure confers very low mechanical and biological properties. Under this scenario, the objective of the research is the synthesis of hydroxyapatite with Mg from the francolite mineral present in phosphate rock from the central-eastern region of Colombia, taking advantage of the extraction of mineral species as natural precursors of Ca, P and Mg. The minerals present were studied, fluorapatite as the mineral of interest associated with magnesium carbonates and quartz. The chemical and mineralogical composition was determined by X-ray fluorescence (XRF) and X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX); the optimum conditions were established using the acid leaching mechanism in the wet concentration process. From the products obtained and characterised by XRD, XRF, SEM, FTIR, RAMAN, HAp-Mg biocomposite scaffolds are fabricated and the influence of Mg on morphometric parameters, mechanical and biological properties in the formed materials is evaluated.

Keywords: phosphate rock, hydroxyapatite, magnesium, biomaterials

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Authors: Sean Hall

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Prior to COVID-19, the world was preoccupied with opioids, effectiveness versus risk, and specifically toxicity versus abuse. Historically underpinning opioid use was a concept of safety. As use over time and real-world data evolved, a pursuit for efficacy associated with non-opioid alternatives became mainstream. On January 8, 2021, the US signed back into the opioid problem, with these two fundamental questions still unresolved. The author will share the current progression of a lead non-opioid cancer bone pain candidate, NanaBis™. NanaBis™ represents two innovative factors: The active ingredients are from cannabinoids; these ingredients are in a proprietary sub-micron delivery platform, NanoCelle®. The author will offer an opinion piece, potentiating the future role of delivery platforms in medicine to increase both patient safety and compliance.

Keywords: NanaBis, nanoCelle, opioids, toxicity

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Authors: Alazar Nebyou, Sujata Pandit

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Lactic acid bacteria (LAB) are essential ingredients in probiotic foods, intestinal microflora, and dairy products that are capable of coping up with harsh gastrointestinal tract conditions and are available in a variety of environments. The objective of this study is to evaluate the probiotic property of LAB isolated from bovine milk. Milk samples were collected from local dairy farms. Samples were obtained using sterile test tubes and transported to a laboratory in the icebox for further biochemical characterization. Preliminary physiological and biochemical identification of LAB isolates was conducted by growing on MRS agar after ten-fold serial dilution. Seven of the best isolates were selected for the evaluation of the probiotic property. The LAB isolates were checked for resistance to antibiotics and their antimicrobial activity by disc diffusion assay and agar well diffusion assay respectively. Bile salt hydrolase activity of isolates was studied by growing isolates in a BSH medium with bile salt. Cell surface property of isolates was assayed by studying their autoaggregation and coaggregation percentage with S. aerues. All isolates were found BSH positive. In addition, BCM2 and BGM1 were susceptible to all antibiotic disks except BBM1 which was resistant to all antibiotic disks. BCM1 and BGM1 had the highest autoaggregation and coaggregation potential respectively. Since all LAB isolates showed gastrointestinal tolerance and good cell surface property they could be considered as good potential probiotic candidates for treatment and probiotic starter culture preparation.

Keywords: probiotic, aggregation, lactic acid bacteria, antimicrobial activity

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Authors: S. Salah, S. A. El-Masry, H. F. Sheba, R. A. El-Banna, W. Saad

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Background: Familial Mediterranean fever (FMF) has episodic or subclinical inflammation that may lead to a decrease in bone mineral density (BMD). Objective: To assess BMD in Egyptian children with FMF on genetic basis. Subjects and Methods: A cross sectional study included 45 FMF patients and 25 control children of both sexes, with age range between 3-16 years old. The patients were reclassified into 2 groups: Group I (A) 23 cases used colchicines for 1 month or less, and Group I (B) 22 cases used colchicines for more than 6 months. For both patients and control, MEFV mutations were defined using molecular genetics technique and BMD was measured by DXA at 2 sites: proximal femur and the lumber spines. Results: four frequent gene mutations were found in the patient group: E148Q (35.6%), V726A (33.3%), M680I (28.9.0%) and M694V (2.2%). There were also 4 heterozygous gene mutations in 40% of control children. Patients received colchicines treatment for less than 1 month had highly significant lower values of BMD at femur and lumber spines than control children (p<0.05). Patients received colchicines treatment for more than 6 months had improved values of BMD at femur compared to control, but there were still significant differences between them at lumbar spine (p>0.05). There are insignificant effect of type of gene mutation on BMD and the risk of osteopenia among the patients. Conclusion: FMF had significant effect on BMD. However, regular use of colchicines treatment improves this effect mainly at femur.

Keywords: familial mediterranean fever, bone mineral density, genes, children

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Authors: Temesgen Kassa Getahun, Gezahegn Mamo, Beksisa Urge

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A cross-sectional study was conducted from December 2019 to May 2020 with the aim of determining the seroprevalence of brucellosis in dairy cows and their owners in the central highland of Oromia, Ethiopia. A total of 352 blood samples from dairy cattle, 149 from animal owners, and 17 from farm workers were collected and initially screened using the Rose Bengal Plate test and confirmed by the Complement Fixation test. Overall seroprevalence was 0.6% (95% CI: 0.0016–0.0209) in bovines and 1.2% (95% CI: 0.0032–0.0427) in humans. Market-based stock replacement (OR=16.55, p=0.002), breeding by artificial insemination (OR=7.58, p=0.05), and parturition pen (OR = 11.511, p=0.027) were found to be significantly associated with the seropositivity for Brucella infection in dairy cattle. Human housing (OR=1.8, p=0.002), contact with an aborted fetus (OR=21.19, p=0.017), drinking raw milk from non-aborted (OR=24.99, p=0.012), aborted (OR=5.72, p=0.019) and retained fetal membrane (OR=4.22, p=0.029) cows had a significant influence on human brucellosis. A structured interview question was administered to 284 respondents. Accordingly, most respondents had no knowledge of brucellosis (93.3%), and in contrast, 90% of them consumed raw milk. In conclusion, the present seroprevalence study revealed that brucellosis was low among dairy cattle and exposed individuals in the study areas. However, since there were no control strategies implemented in the study areas, there is a potential risk of transmission of brucellosis in dairy cattle and the exposed human population in the study areas. Implementation of a test and slaughter strategy with compensation to farmers is recommended, while in the case of human brucellosis, continuous social training and implementing one health approach framework must be applied.

Keywords: abortion, bovine brucellosis, human brucellosis, risk factors, seroprevalence

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Authors: Carlos Alfredo Camargo Vila, Fredy Augusto Avellaneda Vargas, Debora Alcida Nabarlatz

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Due to the current need to implement environmentally friendly technologies, the possibility of using renewable raw materials to produce bioproducts such as biofuels, or in this case, to produce biolubricant bases, from residual oils (tallow), originating has been studied of the bovine industry. Therefore, it is hypothesized that through the study and control of the operating variables involved in the reverse transesterification method, a biolubricant base with high performance is obtained on a laboratory scale using animal fats from the bovine industry as raw materials, as an alternative for material recovery and environmental benefit. To implement this process, esterification of the crude tallow oil must be carried out in the first instance, which allows the acidity index to be decreased ( > 1 mg KOH/g oil), this by means of an acid catalysis with sulfuric acid and methanol, molar ratio 7.5:1 methanol: tallow, 1.75% w/w catalyst at 60°C for 150 minutes. Once the conditioning has been completed, the biodiesel is continued to be obtained from the improved sebum, for which an experimental design for the transesterification method is implemented, thus evaluating the effects of the variables involved in the process such as the methanol molar ratio: improved sebum and catalyst percentage (KOH) over methyl ester content (% FAME). Finding that the highest percentage of FAME (92.5%) is given with a 7.5:1 methanol: improved tallow ratio and 0.75% catalyst at 60°C for 120 minutes. And although the% FAME of the biodiesel produced does not make it suitable for commercialization, it does ( > 90%) for its use as a raw material in obtaining biolubricant bases. Finally, once the biodiesel is obtained, an experimental design is carried out to obtain biolubricant bases using the reverse transesterification method, which allows the study of the effects of the biodiesel: TMP (Trimethylolpropane) molar ratio and the percentage of catalyst on viscosity and yield as response variables. As a result, a biolubricant base is obtained that meets the requirements of ISO VG (Classification for industrial lubricants according to ASTM D 2422) 32 (viscosity and viscosity index) for commercial lubricant bases, using a 4:1 biodiesel molar ratio: TMP and 0.51% catalyst at 120°C, at a pressure of 50 mbar for 180 minutes. It is necessary to highlight that the product obtained consists of two phases, a liquid and a solid one, being the first object of study, and leaving the classification and possible application of the second one incognito. Therefore, it is recommended to carry out studies of the greater depth that allows characterizing both phases, as well as improving the method of obtaining by optimizing the variables involved in the process and thus achieving superior results.

Keywords: biolubricant base, bovine tallow, renewable resources, reverse transesterification

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Authors: Noha Mohammed Ismael Awad Eladal, Aala Shoukry Emara

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Background: Wound healing after dental implant surgery is critical to the procedure's success. The aim of this study was to explore the effects of rutin+vitamin C supplementation in wound healing following the placement of dental implants. Methodology: There were 20 participants in this randomized controlled clinical trial who needed dental implants to replace missing teeth. Patients were divided into two groups, and group A received dental implants. Group B received dental implants with vitamin C administration. Follow-up appointments were performed on day 3, day 7, and day 14 post-surgery, during which soft tissue healing and pain response scores were evaluated using the visual analog scale. Postoperative digital panoramas were taken immediately after surgery, 3 months and 6 months postoperatively. Changes in bone density along with the bone-implant interface at the mesial, distal and apical sides were assessed using the digora software. Results: An independent t-test was used to compare the means of variables between the two groups. At the same time, repeated measures were employed to compare the means of variables between two groups. ANOVA was used to compare bone density for the same group at different dates. Significant increased differences were observed at the mesial, distal and apical sides Surrounding the implants of both groups per time. However, the rate of increase was significantly higher in group B The mean difference at the mesial side after 6 months was 21.99 ± 5.48 in the group B and 14.21 ± 4.95 in group A, while it read 21.74 ± 3.56 in the group B and 10.78 ± 3.90 in group A at the distal side and was 18.90 ± 5.91 in the group B and 10.39 ± 3.49 group A at the apical side. Significance was recorded at P = 0.004, P = 0.0001, and 0.001 at the mesial, distal and apical sides respectively. The mean pain score and wound healing were significantly higher in group A as compared to group B, respectively. Conclusion: The rutin c + vitamin c group significantly promoted bone healing and speeded up the osseointegration process and improved soft tissue healing.

Keywords: osseointegration, soft tissue, rutin c, dental implant

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Authors: Latha V. Prabhu, B.V. Murlimanju, P.J. Jiji, Mangala M. Pai

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Background: The capitate is the largest among the carpal bones. There exists no literature about the vascular foramina of the capitate bone. The objective of the present study was to investigate the morphology and number of the nutrient foramina in the cadaveric dried capitate bones of the Indian population. Methods: The present study included 59 capitate bones (25 right sided and 34 left sided) which were obtained from the gross anatomy laboratory of our institution. The bones were macroscopically observed for the nutrient foramina and the data was collected with respect to their number. The tabulation of the data and analysis were done. Results: All of our specimens (100%) exhibited the nutrient foramina over the non-articular and articular surfaces. The foramina were observed at the medial, lateral, palmar and dorsal surfaces of the capitate bones. The foramina were ranged from 6 to 23 in each capitate bone. In the medial surface, the foramina ranged from 1 to 6, lateral surface from 0 to 7, the foramina ranged between 0 and 5 in the palmar surface. However most of the foramina were located at the dorsal surface which ranged from 3 to 11. Conclusion: We believe that the present study has provided additional data about the nutrient foramina of the capitate bones. The data is enlightening to the orthopedic surgeon and would help in the hand surgeries. The knowledge about the foramina is also important to the radiologists to prevent the misinterpretation of the findings in the x ray and computed tomogram scan films. The foramina may mimick like erosions and ossicles. The morphological knowledge of the vasculature, their foramina of entry and number is required to understand the concepts in the avascular necrosis of the capitate.

Keywords: avascular necrosis, capitate, morphology, nutrient foramen

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Authors: Mohamed Amine Adaour, Mohamed Sadek Bachene, Mosaab Fortassi, Wafaa Siouda

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Foot infections are responsible for a significant number of hospitalizations and amputations in diabetic patients. The objective of our study is to analyze and evaluate the management of diabetic foot in a surgical setting. A retrospective study was conducted based on a selected case of suspected diabetic foot infections of osteitis treated at the Mohamed Boudiaf hospital in Medea.The case was reiterated as a therapeutic charge, consisting of treating first the infection of the soft tissues, then the osteitis: biopsy after at least 15 days of cessation of antibiotic therapy. Successful treatment of osteitis was defined at the end of a follow-up period of complete wound healing, lack of bone resection/amputation surgery at the initial bone site during follow-up , Instead, biopsies are prescribed in the treatment of soft tissue infection. The mean duration of treatment for soft tissue infection was 2-3 weeks, the duration of the antibiotic-free window of therapy prior to bone biopsy was 2-4 weeks. This patient received medical management without surgical resection. The success rate for treating osteitis at one year was 73%, and healing at one year was 88%.It is often limited to a sausage of the foot at the cost of repeated amputations. The best management remains prevention, which necessarily involves setting up a specialized and adapted centre.

Keywords: osteitis, antibiotic, biopsy, diabetic foot

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Authors: T. N. Goh, M. Hashmi, O. Hussain

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Temporomandibular joint (TMJ) dysfunction (TMD) is a condition that may affect patients via restricted mouth opening, significant pain during normal functioning, and/or reproducible joint noise. TMD includes myofascial pain, TMJ functional derangements (internal derangement, dislocation), and TMJ degenerative/inflammatory joint disease. Internal derangement (ID) is the most common cause of TMD-related clicking and locking. These patients are managed in a stepwise approach, from patient education (homecare advice and analgesia), splint therapy, physiotherapy, botulinum toxin treatment, to arthrocentesis. Arthrotomy is offered when the aforementioned treatment options fail to alleviate symptoms and improve quality of life. The aim of this prospective study was to review the outcomes of jaw joint open surgery in TMD patients. Patients who presented from 2015-2022 at the Oral and Maxillofacial Surgery Department in the Doncaster NHS Foundation Trust, UK, with a Wilkes classification of III -V were included. These patients underwent either i) discopexy with bone-anchoring suture (9); ii) intrapositional temporalis flap (ITF) with bone-anchoring suture (3); iii) eminoplasty and discopexy with suturing to the capsule (3); iii) discectomy + ITF with bone-anchoring suture (1); iv) discoplasty + bone-anchoring suture (1); v) ITF (1). Maximum incisal opening (MIO) was assessed pre-operatively and at each follow-up. Pain score, determined via the visual analogue scale (VAS, with 0 being no pain and 10 being the worst pain), was also recorded. A total of 18 eligible patients were identified with a mean age of 45 (range 22 - 79), of which 16 were female. The patients were scored by Wilkes Classification as III (14), IV (1), or V (4). Twelve patients had anterior disc displacement without reduction (66%) and six had degenerative/arthritic changes (33%) to the TMJ. The open joint procedure resulted in an increase in MIO and reduction in pain VAS and for the majority of patients, across all Wilkes Classifications. Pre-procedural MIO was 22.9 ± 7.4 mm and VAS was 7.8 ± 1.5. At three months post-procedure there was an increase in MIO to 34.4 ± 10.4 mm (p < 0.01) and a decrease in the VAS to 1.5 ± 2.9 (p < 0.01). Three patients were lost to follow-up prior to six months. Six were discharged at six month review and five patients were discharged at 12 months review as they were asymptomatic with good mouth opening. Four patients are still attending for annual botulinum toxin treatment. Two patients (Wilkes III and V) subsequently underwent TMJ replacement (11%). One of these patients (Wilkes III) had improvement initially to MIO of 40 mm, but subsequently relapsed to less than 20 mm due to lack of compliance with jaw rehabilitation device post-operatively. Clinical improvements in 89% of patients within the study group were found, with a return to near normal MIO range and reduced pain score. Intraoperatively, the operator found bone-anchoring suture used for discopexy/discoplasty more secure than the soft tissue anchoring suturing technique.

Keywords: bone anchoring suture, open temporomandibular joint surgery, temporomandibular joint, temporomandibular joint dysfunction

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Authors: Y. S. Darmanto, P. H. Riyadi, S. Susanti

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Recently, approximately 9.1 million people of 237.56 million of Indonesian population suffer diabetes. Such condition was caused by high rice consumption of most Indonesian people. It has been known that rice contains low amylose, high calorie, and possesses hyperglycemic properties. Through this study, we tried to solve that problem by creating a super food in order to provide an alternative healthy and balanced diet. We formulated Taro and Seaweed flour based analog rice that fortified by various fishes bone collagens. Corms of Taro contain easily digestible starch and seaweed is rich in fiber, vitamin, and mineral. That mixture was fortified with collagen-containing unique amino acids such as glysine, lysine, alanine, arginine, proline, and hydroxyprolin. Subsequently, super analog rice was characterized about its nutritional composition such are proximate analyses, water, dietary fiber and amylose content. Furthermore, its morphological structure was analyzed by using scanning electron microscopy while the level of consumer preferences was performed by hedonic test. Results demonstrated that fortification by using various fishes bone collagen into analog rice were significantly different in nutritional composition, morphological structure as well as its preferences. Thus, this study was expected as new avenue in functional food discovery especially in the treatment and prevention of diabetic diseases.

Keywords: analogue rice, taro, seaweed, collagen

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Authors: Martyna Ekiert, Andrzej Mlyniec

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Tendons are the biological structures, which primary function is to transfer force generated by muscles to the bones. Unfortunately, damages of tendons are also one of the most common injuries of the human musculoskeletal system. For the most severe cases of tendon rupture, such as the tear of calcaneus tendon or anterior cruciate ligament of the knee, a surgical procedure is the only possible way of full recovery. Tendons used as biological grafts are usually subjected to the process of deep freezing and subsequent thawing. This, in particular for multiple freezing/thawing cycles, may result in changes of tendon internal structure causing deterioration of mechanical properties of the tissue. Therefore, studies on the influence of freezing on tendons biomechanics, including internal water content in soft tissue, seems to be greatly needed. An experimental study of the influence of freezing on mechanical properties of the tendon was performed on fascicles samples dissected form bovine flexor tendons. The preparation procedure was performed with the presence of 0.9% saline solution in order to prevent an excessive tissue drying. All prepared samples were subjected to the different number of freezing/thawing cycles. For freezing part of the protocol we used -80°C temperature while for slow thawing we used fridge temperature (4°C) combined with equalizing temperatures in the standard state (25°C). After final thawing, the mechanical properties of each sample was examined using cyclic loading test. Our results may contribute for better understanding of negative effects of soft tissues freezing, resulting from abnormal thermal expansion of water. This also may help to determine the limit of freezing/thawing cycles disqualifying tissue for surgical purposes and thus help optimize tissues storage conditions.

Keywords: freezing, soft tissue, tendon, bovine fascicles

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Authors: Shiyin He, Zheng Huang

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In this paper, deep learning methods are applied in bio-medical field to detect and count different types of cells in an automatic way instead of manual work in medical practice, specifically in bone marrow examination. The process is mainly composed of two steps, detection and recognition. Mask-Region-Convolutional Neural Networks (Mask-RCNN) was used for detection and image segmentation to extract cells and then Convolutional Neural Networks (CNN), as well as Deep Residual Network (ResNet) was used to classify. Result of cell detection network shows high efficiency to meet application requirements. For the cell recognition network, two networks are compared and the final system is fully applicable.

Keywords: cell detection, cell recognition, deep learning, Mask-RCNN, ResNet

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Authors: Jia-Yu Wu, Chih-Han Chang, Shu-Fen Chuang, Rong-Yang Lai

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Objective: This study was to examine the stress distribution of tooth with different class V restorations under different loading situations and geometry by 3D finite element (FE) analysis. `Methods: A series of FE models of mandibular premolars containing class V cavities were constructed using micro-CT. The class V cavities were assigned as the combinations of different cavity depths x occlusal -gingival heights: 1x2, 1x4, 2x2, and 2x4 mm. Three alveolar bone loss conditions were examined: 0, 1, and 2 mm. 200 N force was exerted on the buccal cusp tip under various directions (vertical, V; obliquely 30° angled, O; oblique and parallel the individual occlusal cavity wall, P). A 3-D FE analysis was performed and the von-Mises stress was used to summarize the data of stress distribution and maximum stress. Results: The maximal stress did not vary in different alveolar bone heights. For each geometry, the maximal stress was found at bilateral corners of the cavity. The peak stress of restorations was significantly higher under load P compared to those under loads V and O while the latter two were similar. 2x2mm cavity exhibited significantly increased (2.88 fold) stress under load P compared to that under load V, followed by 1x2mm (2.11 fold), 2x4mm (1.98 fold) and 1x4mm (1.1fold). Conclusion: Load direction causes the greatest impact on the results of stress, while the effect of alveolar bone loss is minor. Load direction parallel to the cavity wall may enhance the stress concentration especially in deep and narrow class cavities.

Keywords: class v restoration, finite element analysis, loading situation, stress

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Authors: Ayda Khosravanihaghighi, Pramod Koshy, Bill Walsh, Vedran Lovric, Charles Christopher Sorrell

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There is an increasing demand for orthopedic implants owing to the increasing numbers of the aging population. Titanium alloy (Ti6Al4V) is a common material used for orthopedic implants owing to its advantageous properties in terms of good corrosion resistance, minimal elastic modulus mismatch with bone, bio-inertness, and high mechanical strength. However, it is important to improve the bioactivity and osseointegration of the titanium alloy and this can be achieved by coating the implant surface with suitable ceramic materials. In the present work, pure and doped-ceria (CeO₂) coatings were deposited by spin coating on the titanium alloy surface in order to enhance the biological interactions between the surface of the implant and the surrounding tissue. In order to examine the bone-binding ability of an implant, simulated body fluid (SBF) tests were conducted in order to assess the capability of apatite layer formation on the surface and thus predict in vivo bone bioactivity. Characterization was done using scanning electron microscopy (SEM) and X-ray diffraction (XRD) analyses to determine the extent of apatite formation. Preliminary tests showed that the CeO₂ coatings were biocompatible and that the extent of apatite formation and its characteristics can be enhanced by doping with suitable metal ions.

Keywords: apatite layer, biocompatibility, ceria, orthopaedic implant, SBF, spin coater, Ti-implant

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Authors: Jisoo Kim, Min Su Lee, Sunmook Lee

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Durability assessment of nanocomposite-based bone fixation device was performed by flexural fatigue tests, for which the changes in the life cycles of nanocomposite samples synthesized by blending bioabsorbable polymer (PLGA) and ceramic nanoparticles (β-TCP) with different ratios were monitored. The nanocomposite samples were kept in a constant temperature/humidity chamber at 37°C/50%RH for varied incubation periods for the degradation of nanocomposite samples under the temperature/humidity stress. It was found that the life cycles were increasing as the incubation time in the chamber were increasing in the initial stage irrespective of sample compositions, which was due to the annealing effect of the polymer. However, the life cycle was getting shorter as the incubation time increased afterward, which was due to the overall degradation of nanocomposites. It was found that the life cycle of the nanocomposite sample with high ceramic content was shorter than the one with low ceramic content, which was attributed to the increased brittleness of the composite with high ceramic content. The changes in chemical properties were also monitored by FT-IR, which indicated that the degradation of the biodegradable polymer could be confirmed by the increased intensities of carboxyl groups and hydroxyl groups since the hydrolysis of ester bonds connecting two successive monomers yielded carboxyl end groups and hydroxyl groups.

Keywords: bioabsorbable polymer, bone fixation device, ceramic nanoparticles, durability assessment, fatigue test

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Authors: Moncayo Donoso Miguelangel, Guevara Morales Johana, Kalenia Flores Kalenia, Barrera Avellaneda Luis Alejandro, Garzon Alvarado Diego Alexander

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In mammals, long bones are formed by ossification of a cartilage mold during early embryonic development, forming structures called secondary ossification centers (SOCs), a primary ossification center (POC) and growth plates. This last structure is responsible for long bone growth. During the femur growth, the morphology of the growth plate and the SOCs may vary during different developmental stages. So far there are no detailed morphological studies of the development process from embryonic to adult stages. In this work, we carried out a morphological characterization of femur development from embryonic period to adulthood in mice. 15, 17 and 19 days old embryos and 1, 7, 14, 35, 46 and 52 days old mice were used. Samples were analyzed by a computational approach, using 3D images obtained by micro-CT imaging. Results obtained in this study showed that femur, its growth plates and SOCs undergo morphological changes during different stages of development, including changes in shape, position and thickness. These variations may be related with a response to mechanical loads imposed for muscle development surrounding the femur and a high activity during early stages necessary to support the high growth rates during first weeks and years of development. This study is important to improve our knowledge about the ossification patterns on every stage of bone development and characterize the morphological changes of important structures in bone growth like SOCs and growth plates.

Keywords: development, femur, growth plate, mice

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Authors: Itsuo Yokoyama, Rikako Kikuti, Naoko Watabe, Tosinori Asai, Sarai Tsuyoshi

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Introduction: Chronic kidney disease presents significant changes in mineral and bone metabolism, referred to as CKD-MBD. These changes lead to decreased bone mass, heightened bone fragility, fractures, and increased vascular and valvular calcification, ultimately impacting cardiovascular outcomes. Key contributors to these complications in dialysis patients include calcium, phosphate, parathyroid hormone (PTH), fibroblast growth factor 23 (FGF23), and the vitamin D hormonal system. Methods: In our outpatient dialysis clinic, we monitor the long-term effects of vascular calcifications by calculating the volume of calcified areas in the abdominal aorta based on CT scan data. The results revealed a progressive nature of vascular calcification. To extend our study, we measured the volume of calcification in bones (vertebrae and femur) corresponding to Hounsfield units of 200 and 300. The study aims to investigate changes in osteoporosis during a 5-year follow-up period and its relationship with extraosseous calcification. Results and Considerations: While extraosseous calcification demonstrated a generally progressive nature, often resistant to medical treatment, the degree of osteoporotic change varied among patients. The majority exhibited continuous osteoporotic changes, while some showed improvement or minimal changes in bone calcification. Variations in the distribution and magnitude of osteoporotic changes were observed between groups based on the timing of hemodialysis initiation during the study. The former group tended to display more osteoporotic changes, possibly attributed to differences in medication between the groups. Other contributing factors may include the patient's age, duration of dialysis, or causes of renal disease. In conclusion, we emphasize the importance of carefully monitoring calcium and phosphate levels and maintaining adequate dialysis therapy to prevent osteoporosis in dialysis patients.

Keywords: CKD-MBD, dialysis, calcification, kidney

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Authors: Muhammad Saleem

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Background: Inattentive food choices and engagement in excessive physical activities by male athletes can potentially lead to adverse health consequences. Objective: The aim was to ascertain the occurrence of risk factors associated with the Male Athlete Triad among young elite athletes in Pakistan. Methodology: In 2018, a cross-sectional study based on questionnaires was conducted at the Pakistan Sports Board. The study aimed to explore the risk factors related to the Male Athlete Triad in young elite athletes who were part of national training camps in major metropolitan areas. The study included proficient male elite athletes aged 18 to 25 years, capable of understanding the English questionnaire. The athletes completed a survey encompassing aspects like demographic information, educational background, Body Mass Index (BMI), sports involvement, and hours of participation. Additionally, they filled out the Eating Attitude Test-26 (EAT-26) and questionnaires assessing risks of amenorrhea and low bone mineral density. The prevalence of risk factors for each of the three components was individually evaluated. The collected data underwent analysis using SPSS-20, with descriptive statistics being applied. Results: The study comprised a sample of 90 elite athletes (mean age: 23.57 ± 2.37 years, mean BMI: 21.97 ± 1.90) engaged in various sports. The EAT-26 results indicated that 50% of athletes were at risk of developing an eating disorder. Moreover, 83.3% exhibited disordered eating behaviors that necessitated referral. Risks for amenorrhea were observed in 15% of the participants, and regarding low bone mineral density, notable risks were absent except for the consumption of caffeinated beverages, which was noted in 51.7% of participants. Conclusion: The study identified a significant prevalence of disordered eating risk among male elite athletes in Pakistan. However, the risks associated with amenorrhea and low bone mineral density were not a major concern in this particular group.

Keywords: Pakistan, osteoporosis, female athlete triad, bone mineral density, athlete, amenorrhea, eating disorders

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