Search results for: myeloma bone disease

Authors: K. Jankowski, M. Pawlikowski, A. Makuch, K. Skalski

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Depth-sensing indentation (DSI) or nanoindentation is becoming a more and more popular method of measuring mechanical properties of various materials and tissues at a micro-scale. This technique allows measurements without complicated sample preparation procedures which makes this method very useful. As a result of measurement force and displacement of the intender are obtained. It is also possible to determine three measures of hardness i.e. Martens hardness (HM), nanohardness (HIT), Vickers hardness (HV) and Young modulus EIT. In this work trabecular bone mechanical properties were investigated. The bone samples were harvested from human femoral heads during hip replacement surgery. Patients were of different age, sexes and stages of tissue degeneration caused by osteoarthritis. The specimens were divided into three groups. Each group contained samples harvested from patients of different range of age. All samples were investigated with the same measurement conditions. The maximum load was Pmax=500 mN and the loading rate was 500 mN/min. The tests were held without hold at the peak force. The tests were conducted with indenter Vickers tip and spherical tip of the diameter 0.2 mm. Each trabecular bone sample was tested 7 times in a close area of the same trabecula. The measured loading P as a function of indentation depth allowed to obtain hysteresis loop and HM, HIT, HV, EIT. Results for arbitrarily chosen sample are HM=289.95 ± 42.31 MPa, HIT=430.75 ± 45.37 MPa, HV=40.66 ± 4.28 Vickers, EIT=7.37 ± 1.84 GPa for Vickers tip and HM=115.19 ± 15.03 MPa, HIT=165.80 ± 19.30 MPa, HV=16.90 ± 1.97 Vickers, EIT=5.30 ± 1.31 GPa for spherical tip. Results of nanoindentation tests show that this method is very useful and is perfect for obtaining mechanical properties of trabecular bone. Estimated values of elastic modulus are similar. The differences between hardness are significant but it is a result of using two different types of tips. However, it has to be emphasised that the differences in the values of elastic modulus and hardness result from different testing protocols, anisotropy and asymmetry of the micro-samples and the hydration of bone.

Keywords: human bone, mechanical properties, nano hardness nanoindentation, trabecular bone

Procedia PDF Downloads 255

Authors: M. G. R. S. Perera, B. S. Weerakoon, L. P. G. Sherminie, M. L. Jayatilake, R. D. Jayasinghe, W. Huang

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The aim of this study was to measure and evaluate the Longitudinal Relaxation Times (T1) in bone marrow of an Acute Myeloid Leukaemia (AML) patient in order to explore the potential for a prognostic biomarker using Magnetic Resonance Imaging (MRI) which will be a non-invasive prognostic approach to AML. MR image data were collected in the DICOM format and MATLAB Simulink software was used in the image processing and data analysis. For quantitative MRI data analysis, Region of Interests (ROI) on multiple image slices were drawn encompassing vertebral bodies of L3, L4, and L5. T1 was evaluated using the T1 maps obtained. The estimated bone marrow mean value of T1 was 790.1 (ms) at 3T. However, the reported T1 value of healthy subjects is significantly (946.0 ms) higher than the present finding. This suggests that the T1 for bone marrow can be considered as a potential prognostic biomarker for AML patients.

Keywords: acute myeloid leukaemia, longitudinal relaxation time, magnetic resonance imaging, prognostic biomarker.

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Authors: Mahmoud M. Zakaria, Omnia F. Elmoursi, Mahmoud M. Gabr, Camelia A. AbdelMalak, Mohamed A. Ghoneim

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Many protocols were publicized for differentiation of human mesenchymal stem cells (MSCS) into insulin-producing cells (IPCs) in order to excrete insulin hormone ingoing to treat diabetes disease. Our aim is to evaluate relative gene expression for each independent protocol. Human bone marrow cells were derived from three volunteers that suffer diabetes disease. After expansion of mesenchymal stem cells, differentiation of these cells was done by three different protocols (the one-step protocol was used conophylline protein, the two steps protocol was depending on trichostatin-A, and the three-step protocol was started by beta-mercaptoethanol). Evaluation of gene expression was carried out by real-time PCR: Pancreatic endocrine genes, transcription factors, glucose transporter, precursor markers, pancreatic enzymes, proteolytic cleavage, extracellular matrix and cell surface protein. Quantitation of insulin secretion was detected by immunofluorescence technique in 24-well plate. Most of the genes studied were up-regulated in the in vitro differentiated cells, and also insulin production was observed in the three independent protocols. There were some slight increases in expression of endocrine mRNA of two-step protocol and its insulin production. So, the two-step protocol was showed a more efficient in expressing of pancreatic endocrine genes and its insulin production than the other two protocols.

Keywords: mesenchymal stem cells, insulin producing cells, conophylline protein, trichostatin-A, beta-mercaptoethanol, gene expression, immunofluorescence technique

Procedia PDF Downloads 190

Authors: Rozeena Baig, R. Rehana Rehman, Rifat Ahmed

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Background: Osteoporosis, characterized by low bone mineral density, poses a global health concern. Diagnosis increases the likelihood of developing osteoporosis, a multifactorial disorder marked by low bone mass, elevating the risk of fractures in the lumbar spine, femoral neck, hip, vertebras, and distal forearm, particularly in postmenopausal women due to bone loss influenced by various pathophysiological factors. Objectives: The aim is to investigate the association of serum cytokine, bone turnover marker, bone mineral density and TLR4 gene polymorphism in pre and post-menopausal women and to find if any of these can be the potential predictor of osteoporosis in postmenopausal women. Material and methods: The study participants consisted of Group A (n=91) healthy pre-menopausal women and Group B (n=102) healthy postmenopausal women having ≥ 5 years’ history of menopause. ELISA was performed for cytokine (TNFα) and bone turnover markers (carboxytelopeptides), respectively. Bone Mineral Density (BMD)was measured through a dual X-ray absorptiometry (DEXA) scan. Toll-like Receptors 4 (TLR4) gene polymorphisms (A896G; Asp299Gly) and (C1196T; Thr399Ile) were investigated by PCR and Sanger sequencing. Results: Statistical analysis reveals a positive correlation of age and BMI with T scores in the premenopausal group, whereas in post-menopausal group found a significant negative correlation between age and T-score at hip (r = - 0.352**), spine (r = - .306**), and femoral neck (r = - 0.344**) and a significant negative correlation of BMI with TNF-α (- 0.316**). No association and significant differences were observed for TLR4 genotype and allele frequencies among studied groups However, both SNPs exhibited significant association with each other. Conclusions: This study concludes that BMI, BMD and TNF-α are the potential predictors of osteoporosis in post-menopausal women. However, CTX and TLR4 gene polymorphism did not appear as potential predictors of bone loss in this study and apparently cannot help in predicting bone loss in post-menopausal women.

Keywords: osteoporosis, post-menopausal, pre-menopausal woemn, genetics mutaiont, TLR4 genepolymorphsum

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Authors: Manal M. Shehata, Kawther M. Abdel-Hamid, Nashwa A. Mohamed, Marwa H. Bakr, Maged S. Mahmoud, Hala M. Elbadre

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Introduction: The worldwide rapid increase in diabetes poses a significant challenge to current therapeutic approaches. Therapeutic utility of bone marrow transplantation in diabetes is an attractive approach. However, the oxidative stress generated by hyperglycemia may hinder β-cell regeneration. Curcumin, is a dietary spice with antioxidant activity. Aim of work: The present study was undertaken to investigate the therapeutic potential of curcumin, bone marrow transplantation, and their combined effects in the reversal of experimental diabetes. Material and Methods: Fifty adult male healthy albino rats were included in the present study.They were divided into two groups: Group І: (control group) included 10 rats. Group П: (diabetic group): included 40 rats. Diabetes was induced by single intraperitoneal injection of streptozotocin (STZ). Group II will be further subdivided into four groups (10 rats for each): Group II-a (diabetic control). Group II-b: rats were received single intraperitoneal injection of bone marrow suspension (un-fractionated bone marrow cells) prepared from rats of the same family. Group II-c: rats were treated with curcumin orally by gastric intubation for 6 weeks. Group II-d: rats were received a combination of single bone marrow transplantation and curcumin for 6 weeks. After 6 weeks, blood glucose, insulin levels were measured and the pancreas from all rats were processed for Histological, Immunohistochemical and morphometric examination. Results: Diabetic group, showed progressive histological changes in the pancreatic islets. Treatment with either curcumin or bone marrow transplantation improved the structure of the islets and reversed streptozotocin-induced hyperglycemia and hypoinsulinemia. Combination of curcumin and bone marrow transplantation elicited more profound alleviation of streptozotocin-induced changes including islet regeneration and insulin secretion. Conclusion: The use of natural antioxidants combined with bone marrow transplantation to induce pancreatic regeneration is a promising strategy in the management of diabetes.

Keywords: diabtes, panceatic islets, bone marrow transplantation, curcumin

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Authors: K. Yadamma, K. Rudrama Devi

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The protective effect of Ginger Extract against cyclophosphamide induced cytotoxicity was evaluated in in vivo animal model using analysis of chromosomal aberrations in somatic cells of mice. Three doses of Ginger Extract (150mg/kg, 200mg/kg, and 250mg/kg body weight) were selected for modulation and given to animals after priming. The animals were sacrificed 24, 48, 72 hrs after the treatment and slides were prepared for the incidence of chromosomal aberrations in bone marrow cells of mice. When animals were treated with cyclophosphamide 50mg/kg, showed cytogenetic damage in somatic cells. However, a significant decrease was observed in the percentage of chromosomal aberrations when animals were primed with various doses of Ginger Extract. The present results clearly indicate the protective nature of Ginger Extract against cyclophosphamide induced genetic damage in mouse bone marrow cells.

Keywords: ginger extract, protection, bone marrow cells, swiss albino mice

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Authors: Asim Abbasi, Amer Habib, Sajid Hussain, Muhammad Sufyan, Iqra, Hasnain Sajjad

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Except for few scattered cases, powdery mildew disease was not a big problem for watermelon in the past but with the outbreaks of its pathotypes, races 1W and 2W, this disease becomes a serious issue all around the globe. The severe outbreak of this disease also increased the rate of fungicide application for its proper management. Twelve varieties of watermelon were screened in Research Area of Department of Plant pathology, University of Agriculture, Faisalabad to check the incidence of powdery mildew disease. Disease inoculum was prepared and applied with the help of foliar spray method. Fungicides and plants extracts were also applied after the disease incidence. Percentage leaf surface area diseased was assessed visually with a modified Horsfall-Barratt scale. The results of the experiment revealed that among all varieties, WT2257 and Zcugma F1 were highly resistant showing less than 5% disease incidence while Anar Kali and Sugar baby were highly susceptible with disease incidence of more than 65%. Among botanicals neem extract gave best results with disease incidence of less than 20%. Besides neem, all other botanicals also gave significant control of powdery mildew disease than the untreated check. In case of fungicides, Gemstar showed least disease incidence i.e. < 10%, however besides control maximum disease incidence was observed in Curzate (> 30%).

Keywords: botanicals, fungicides, pathotypes, powdery mildew

Procedia PDF Downloads 263

Authors: Aura Maria Lopera Echavarria, Angela Maria Lema Perez, Daniela Medrano David, Pedronel Araque Marin, Marta Elena Londoño Lopez

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Bone regeneration is the process by which the formation of new bone is stimulated. Bone fractures can originate at any time due to trauma, infections, tumors, congenital malformations or skeletal diseases. Currently there are different strategies to treat bone defects that in some cases, regeneration does not occur on its own. That is why they are treated with bone substitutes, which provide a necessary environment for the cells to synthesize new bone. The Demineralized Bone Matrix (DBM) is widely used as a bone implant due to its good properties, such as osteoinduction and bioactivity. However, the use of DBM is limited, because its presentation is powder, which is difficult to implant with precision and is susceptible to migrating to other sites through blood flow. That is why the DBM is commonly incorporated into a variety of vehicles or carriers. The objective of this project is to evaluate the bioactive and confinement properties of a bone substitute based on demineralized bone matrix (DBM). Also, structural and morphological properties were evaluated. Bone substitute was obtained from EIA Biomaterials Laboratory of EIA University and the DBM was facilitated by Tissue Bank Foundation. Morphological and structural properties were evaluated by scanning electron microscopy (SEM), X-ray diffraction (DRX) and Fourier transform infrared spectroscopy with total attenuated reflection (FTIR-ATR). Water absorption capacity and degradation were also evaluated during three months. The cytotoxicity was evaluated by the MTT test. The bioactivity of the bone substitute was evaluated through immersion of the samples in simulated body fluid during four weeks. Confinement tests were performed on tibial fragments of a human donor with bone defects of determined size, to ensure that the substitute remains in the defect despite the continuous flow of fluid. According of the knowledge of the authors, the methodology for evaluating samples in a confined environment has not been evaluated before in real human bones. The morphology of the samples showed irregular surface and presented some porosity. DRX confirmed a semi-crystalline structure. The FTIR-ATR determined the organic and inorganic phase of the sample. The degradation and absorption measurements stablished a loss of 3% and 150% in one month respectively. The MTT showed that the system is not cytotoxic. Apatite clusters formed from the first week were visualized by SEM and confirmed by EDS. These calcium phosphates are necessary to stimulate bone regeneration and thanks to the porosity of the developed material, osteinduction and osteoconduction are possible. The results of the in vitro evaluation of the confinement of the material showed that the migration of the bone filling to other sites is negligible, although the samples were subjected to the passage of simulated body fluid. The bone substitute, putty type, showed stability, is bioactive, non-cytotoxic and has handling properties for specialists at the time of implantation. The obtained system allows to maintain the osteoinductive properties of DBM and it can fill completely fractures in any way; however, it does not provide a structural support, that is, it should only be used to treat fractures without requiring a mechanical load.

Keywords: bone regeneration, cytotoxicity, demineralized bone matrix, hydrogel

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Authors: Kumaran Letchmanan, Shou-Cang Shen, Wai Kiong Ng

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Postoperative implant-associated infections in soft tissues and bones remain a serious complication in orthopaedic surgery, which leads to impaired healing, re-implantation, prolong hospital stay and increase cost. Drug-loaded implants with sustained release of antibiotics at the local site are current research interest to reduce the risk of post-operative infections and osteomyelitis, thus, minimize the need for follow-up care and increase patient comfort. However, the improved drug release of the drug-loaded bone cements is usually accompanied by a loss in mechanical strength, which is critical for weight-bearing bone cement. Recently, more attempts have been undertaken to develop techniques to enhance the antibiotic elution as well as preserve the mechanical properties of the bone cements. The present study investigates the potential influence of addition of mesoporous silica nanoparticles (MSN) on the in vitro drug release kinetics of gentamicin (GTMC), along with the mechanical properties of bone cements. Simplex P was formulated with MSN and loaded with GTMC by direct impregnation. Meanwhile, Simplex P with water soluble poragen (xylitol) and high loading of GTMC as well as commercial bone cement CMW Smartset GHV were used as controls. MSN-formulated bone cements are able to increase the drug release of GTMC by 3-fold with a cumulative release of more than 46% as compared with other control groups. Furthermore, a sustained release could be achieved for two months. The loaded nano-sized MSN with uniform pore channels significantly build up an effective nano-network path in the bone cement facilitates the diffusion and extended release of GTMC. Compared with formulations using xylitol and high GTMC loading, incorporation of MSN shows no detrimental effect on biomechanical properties of the bone cements as no significant changes in the mechanical properties as compared with original bone cement. After drug release for two months, the bending modulus of MSN-formulated bone cements is 4.49 ± 0.75 GPa and the compression strength is 92.7 ± 2.1 MPa (similar to the compression strength of Simplex-P: 93.0 ± 1.2 MPa). The unaffected mechanical properties of MSN-formulated bone cements was due to the unchanged microstructures of bone cement, whereby more than 98% of MSN remains in the matrix and supports the bone cement structures. In contrast, the large portions of extra voids can be observed for the formulations using xylitol and high drug loading after the drug release study, thus caused compressive strength below the ASTM F541 and ISO 5833 minimum of 70 MPa. These results demonstrate the potential applicability of MSN-functionalized poly(methyl methacrylate)-based bone cement as a highly efficient, sustained and local drug delivery system with good mechanical properties.

Keywords: antibiotics, biomechanical properties, bone cement, sustained release

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Authors: Syed Dawood M. Taimur

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Background: Peripheral arterial disease (PAD) is under diagnosed in primary care practices, yet the extent of unrecognized PAD in patients with coronary artery disease (CAD) is unknown. Objective: To assess the prevalence of previously unrecognized PAD in patients undergoing coronary angiogram and to determine the relationship between the presence of PAD and severity of CAD. Material & Methods: This five years retrospective study was conducted at an invasive lab of the department of Cardiology, Ibrahim Cardiac Hospital & Research Institute from January 2010 to December 2014. Total 77 patients were included in this study. Study variables were age, sex, risk factors like hypertension, diabetes mellitus, dyslipidaemia, smoking habit and positive family history for ischemic heart disease, coronary artery and peripheral artery profile. Results: Mean age was 56.83±13.64 years, Male mean age was 53.98±15.08 years and female mean age was 54.5±1.73years. Hypertension was detected in 55.8%, diabetes in 87%, dyslipidaemia in 81.8%, smoking habits in 79.2% and 58.4% had a positive family history. After catheterization 88.3% had peripheral arterial disease and 71.4% had coronary artery disease. Out of 77 patients, 52 had both coronary and peripheral arterial disease which was statistically significant (p < .014). Coronary angiogram revealed 28.6% (22) patients had triple vessel disease, 23.3% (18) had single vessel disease, 19.5% (15) had double vessel disease and 28.6% (22) were normal coronary arteries. The peripheral angiogram revealed 54.5% had superficial femoral artery disease, 26% had anterior tibial artery disease, 27.3% had posterior tibial artery disease, 20.8% had common iliac artery disease, 15.6% had common femoral artery disease and 2.6% had renal artery disease. Conclusion: There is a strong and definite correlation between coronary and peripheral arterial disease. We found that cardiovascular risk factors were in fact risk factors for both PAD and CAD.

Keywords: coronary artery disease (CAD), peripheral artery disease(PVD), risk, factors, correlation, cathetarization

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Authors: Caighley Logan

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The outcome of fire related fatalities, along with other research, has found fires can have a detrimental effect to the mineral and crystalline structures within bone. This study focused on the mineral and crystalline structures within porcine bone samples to analyse the changes caused, with the intent of effectively ‘reverse engineering’ the data collected from burned bone samples to discover what may have happened. Using Fourier Transform Infrared (FT-IR), and X-Ray Fluorescence (XRF), the data collected from a controlled source of intense heat (muffle furnace) and an open fire, based in a living room setting in a standard size shipping container (8.5ft x 8ft) of a similar temperature with a known ignition source, a gasoline lighter. This approach is to analyse the changes to the samples and how the changes differ depending on the heat source. Results have found significant differences in the levels of remaining minerals for each type of heat/burning (p=<0.001), particularly Phosphorus and Calcium, this also includes notable additions of absorbed elements and minerals from the surrounding materials, i.e., Cerium (Ce), Bromine (Br) and Neodymium (Ne). The analysis techniques included provide validated results in conjunction with previous studies.

Keywords: forensic anthropology, thermal alterations, porcine bone, FTIR, XRF

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Authors: Mohammad Shakir, Reshma Jolly, Mohammad Shoeb Khan, Noor-E-Iram

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A nanocomposite system incorporating dextrin into nano-hydroxyapatite/chitin matrix (n-HA/DX/CT) has been successfully synthesized via co-precipitation route at room temperature for the application in bone tissue engineering by investigating biocompatibility, cytotoxicity and mechanical properties. The FTIR spectra of n-HA/DX/CT nanocomposite indicated a considerable intermolecular interaction between the various components of the system. The results of XRD, TEM and TGA/DTA revealed that the crystallinity, size and thermal stability of the n-HA/DX/CT scaffold has decreased and increased respectively. The result of SEM image of the n-HA/DX/CT scaffold indicated that the incorporation of dextrin affected the surface morphology while considerable in-vitro bioactivity has been observed in n-HA/DX/CT based on SBF study, referring a step towards possibility of making direct bond to living bone if implanted. Moreover, MTT assay suggested the non-toxic nature of n-HA/DX/CT to murine fibroblast L929 cells. The swelling study of n-HA/DX/CT scaffold indicated the low swelling rate for n-HADX/CT. All these results have paved the way for n-HA/DX/CT to be used as a competent material for bone tissue engineering.

Keywords: autograft, chitin, dextrin, nanocomposite

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Authors: Xianjia Meng, Chuanyong Qu

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Compact bone produces fatigue damage under the inevitable physiological load. The accumulation of fatigue damage can change the bone’s micro-structure at different scales and cause the catastrophic failure eventually. However, most tests were limited to the macroscopic modulus of bone and there is a need to assess the microscopic modulus during fatigue progress. In this paper, nano-identation was used to investigate the bone specimen subjected to four point bending. The microscopic modulus of the same area were measured at different degrees of damage including fracture. So microscopic damage can be divided into three stages: first, the modulus decreased rapidly and then They fell slowly, before fracture the decline became fast again. After fracture, the average modulus decreased by 20%. The results of inner and outer planes explained the influence of compressive and tensile loads on modulus. Both the compressive and tensile moduli decreased with the accumulation of damage. They reached the minimum at ending and increased after fracture. The modulus evolution under different strains were revealed by the side. They all fell slowly and then fast with the accumulation of damage. The fractured results indicated that the elastic modulus decreased obviously at the high strain while decreased less at the low strain. During the fatigue progress, there was a significant difference in modulus at low degree of damage. However, the dispersed modulus tended to be similar at high degree of damage, but they became different again after the failure.

Keywords: fatigue damage, fracture, microscopic modulus, bone, nano-identation

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Authors: F. Ruiz-Navarro, M. Matzner, G. Kobinia

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Background: Cerebral palsy (CP) encompasses the largest group of childhood movement disorders, the patterns and severity varies widely. Today, the management focuses only on a rehabilitation therapy that tries to secure the functions remained and prevents complications. However the treatments are not aimed to cure the disease. Stem cells (SCs) transplant via intrathecal is a new approach to the disease. Method: Our aim was to performed a pilot study under the condition of unproven treatment on clinical practice to assessed the safety and efficacy of Neuron Point-of-care Stem cell Therapy (N-POCST), an ambulatory procedure of autologous bone marrow derived SCs (BM-SCs) harvested from the posterior superior iliac crest undergo an on-site cell separation for intrathecal infusion via lumbar puncture. Results: 82 patients were treated in a period of 28 months, with a follow-up after 6 months. They had a mean age of 6,2 years old and male predominance (65,9%). Our preliminary results show that: A. No patient had any major side effects, B. Only 20% presented mild headache due to LP, C. 53% of the patients had an improvement in spasticity, D. 61% improved the coordination abilities, 23% improved the motor function, 15% improved the speech, 23% reduced the number of convulsive events with the same doses or less doses of anti-convulsive medication and 94% of the patients report a subjective general improvement. Conclusions: These results support previous worldwide publications that described the safety and effectiveness of autologous BM-SCs transplant for patients wit CP.

Keywords: autologous transplant, cerebral palsy, point of care, childhood movement disorders

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Authors: Kadir Gök

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In this study, a vacuum system was developed for orthopedic drilling processes, and the most efficient processing parameters were determined using statistical analysis of temperature rise. A reverse engineering technique was used to obtain a 3D model of the chip vacuum system, and the obtained point cloud data was transferred to Solidworks software in STL format. An experimental design method was performed by selecting different parameters and their levels, such as RPM, feed rate, and drill bit diameter, to determine the most efficient processing parameters in temperature rise using ANOVA. Additionally, the bone chip-vacuum device was developed and performed successfully to collect the whole chips and fragments in the bone drilling experimental tests, and the chip-collecting device was found to be useful in removing overheating from the drilling zone. The effects of processing parameters on the temperature levels during the chip-vacuuming were determined, and it was found that bone chips and fractures can be used as autograft and allograft for tissue engineering. Overall, this study provides significant insights into the development of a vacuum system for orthopedic drilling processes and the use of bone chips and fractures in tissue engineering applications.

Keywords: vacuum system, orthopedic drilling, temperature rise, bone chips

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Authors: Marzieh Jafarzadeh, Fatemeh Rezaee

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Introduction: The effect of ionizing radiation on human health can be effective for genomic integrity and cell viability. It also increases the risk of cancer and malignancy. Therefore, X-ray behavior and absorption dose calculation are considered. One of the applicable tools for calculating and evaluating the absorption dose in human tissues is Monte Carlo simulation. Monte Carlo offers a straightforward way to simulate and integrate, and because it is simple and straightforward, Monte Carlo is easy to use. The Monte Carlo BEAMnrc code is one of the most common diagnostic X-ray simulation codes used in this study. Method: In one of the understudy hospitals, a certain number of CT scan images of patients who had previously been imaged were extracted from the hospital database. BEAMnrc software was used for simulation. The simulation of the head of the device with the energy of 0.09 MeV with 500 million particles was performed, and the output data obtained from the simulation was applied for phantom construction using CT CREATE software. The percentage of depth dose (PDD) was calculated using STATE DOSE was then compared with international standard values. Results and Discussion: The ratio of surface dose to depth dose (D/Ds) in the measured energy was estimated to be about 4% to 8% for bone and 3% to 7% for bone marrow. Conclusion: MC simulation is an efficient and accurate method for simulating bone marrow and calculating the absorbed dose.

Keywords: Monte Carlo, absorption dose, BEAMnrc, bone marrow

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Authors: Anuj Kumar, Kummara M. Rao, Sung S. Han

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Bone tissue engineering has emerged as a potentially alternative method for localized bone defects or diseases, congenital deformation, and surgical reconstruction. The designing and the fabrication of the ideal scaffold is a great challenge, in restoring of the damaged bone tissues via cell attachment, proliferation, and differentiation under three-dimensional (3D) biological micro-/nano-environment. In this case, hydrogel system composed of high hydrophilic 3D polymeric-network that is able to mimic some of the functional physical and chemical properties of the extracellular matrix (ECM) and possibly may provide a suitable 3D micro-/nano-environment (i.e., resemblance of native bone tissues). Thus, this proposed hydrogel system is highly permeable and facilitates the transport of the nutrients and metabolites. However, the use of hydrogels in bone tissue engineering is limited because of their low mechanical properties (toughness and stiffness) that continue to posing challenges in designing and fabrication of tough and stiff hydrogels along with improved bioactive properties. For this purpose, in our lab, polyacrylamide-based hybrid hydrogels were synthesized by involving sodium alginate, cellulose nanocrystals and silica-based glass using one-step free-radical polymerization. The results showed good in vitro apatite-forming ability (biomineralization) and improved mechanical properties (under compression in the form of strength and stiffness in both wet and dry conditions), and in vitro osteoblastic (MC3T3-E1 cells) cytocompatibility. For in vitro cytocompatibility assessment, both qualitative (attachment and spreading of cells using FESEM) and quantitative (cell viability and proliferation using MTT assay) analyses were performed. The obtained hybrid hydrogels may potentially be used in bone tissue engineering applications after establishment of in vivo characterization.

Keywords: bone tissue engineering, cellulose nanocrystals, hydrogels, polyacrylamide, sodium alginate

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Authors: M. Benlebna, B. Serier, B. Bachir Bouiadjra, S. Khalkhal

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This study focuses on the mechanical behavior of a dental prosthesis subjected to dynamic loads chewing. It covers a three-dimensional analysis by the finite element method, the level of distribution of equivalent stresses induced in the bone between the implants (depending on the number of implants). The studied structure, consisting of a braced, implant and mandibular bone is subjected to dynamic loading of variable amplitude in three directions corrono-apical, mesial-distal and bucco-lingual. These efforts simulate those of mastication. We show that compared to the implantation of a single implant, implantology using two implants promotes the weakening of the bones. This weakness is all the more likely that the implants are located in close proximity to one another.

Keywords: stress, bone, dental implant, distribution, stress levels, dynamic, effort, interaction, prosthesis

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Authors: Irena Baranowska-Bosiacka, Mateusz Bosiacki, Patrycja Kupnicka, Anna Lubkowska, Dariusz Chlubek

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Physical activity and a balanced diet are among the key factors of "healthy ageing". Physical effort, including swimming in cold water (including bathing in natural water reservoirs), is widely recognized as a hardening factor, with a positive effect on the mental and physical health. At the same time, there is little scientific evidence to verify this hypothesis. In the literature to date, it is possible to obtain data on the impact of these factors on selected physiological and biochemical parameters of the blood, at the same time there are no results of research on the effect of immersing in cold water on mineral metabolism, especially bones, hence it seems important to perform such an analysis in relation to the key elements such as calcium (Ca), magnesium (Mg) and phosphorus (P). Taking the above into account, a hypothesis was put forward about the possibility of a positive effect of exercise in cold water on mineral metabolism and bone density in aging rats. The aim of the study was to evaluate the effect of an 8-week swimming training on mineral metabolism and bone density in aging rats in response to exercise in cold water (5oC) in comparison to swimming in thermal comfort (36oC) and sedentary (control) rats of both sexes. The examination of the concentration of the examined elements in the bones was carried out using inductively coupled plasma atomic emission spectrometry (ICP-OES). The mineral density of the femurs of the rats was measured using the Hologic Horizon DEXA System® densitometer. The results of our study showed that swimming in cold water affects bone mineral metabolism in aging rats by changing the Ca, Mg, P concentration and at the same time increasing their bone density. In males, a decrease in Mg concentration and no changes in bone density were observed. In the light of the research results, it seems that swimming in cold water may be a factor that positively modifies the bone aging process by improving the mechanisms affecting their density.

Keywords: swimming in cold water, adaptation to cold water, bone mineral metabolism, aging

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Authors: W. Soufi, F. Boukli Hacene, S. Ghalem

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Alzheimer's disease (AD) is a neurodegenerative disease that leads to a progressive and permanent deterioration of nerve cells. This disease is progressively accompanied by an intellectual deterioration leading to psychological manifestations and behavioral disorders that lead to a loss of autonomy. It is the most frequent of degenerative dementia. Alzheimer's disease (AD), which affects a growing number of people, has become a major public health problem in a few years. In the context of the study of the mechanisms governing the evolution of AD disease, we have found that natural flavonoids are good acetylcholinesterase inhibitors that reduce the rate of ßA secretion in neurons. This work is to study the inhibition of acetylcholinesterase (AChE) which is an enzyme involved in Alzheimer's disease, by methods of molecular modeling. These results will probably help in the development of an effective therapeutic tool in the fight against the development of Alzheimer's disease. Our goal of the research is to study the inhibition of acetylcholinesterase (AChE) by molecular modeling methods.

Keywords: Alzheimer's disease, acetylcholinesterase, flavonoids, molecular modeling

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Authors: Alaa T. Ali, Nevine H. Kheir El Din, Ehab S. Abdelhamid, Ahmed E. Amr

Abstract:

Objective: Severe alveolar bone resorption is usually associated with a deficient amount of soft tissues. soft tissue expansion is introduced to provide an adequate amount of soft tissue over the grafted area. This study aimed to assess the efficacy of sub-periosteal self-inflating osmotic tissue expanders used as preparatory surgery before horizontal alveolar ridge augmentation using autogenous onlay block bone graft. Methods: A prospective randomized controlled clinical trial was performed. Sixteen partially edentulous patients demanding horizontal bone augmentation in the anterior maxilla were randomly assigned to horizontal ridge augmentation with autogenous bone block grafts harvested from the mandibular symphysis. For the test group, soft tissue expanders were placed sub-periosteally before horizontal ridge augmentation. Impressions were taken before and after STE, and the cast models were optically scanned and superimposed to be used for volumetric analysis. Horizontal ridge augmentation was carried out after STE completion. For the control group, a periosteal releasing incision was performed during bone augmentation procedures. Implants were placed in both groups at re-entry surgery after six months period. A core biopsy was taken. Histomorphometric assessment for newly formed bone surface area, mature collagen area fraction, the osteoblasts count, and blood vessel count were performed. The change in alveolar ridge width was evaluated through bone caliper and CBCT. Results: Soft tissue expander successfully provides a Surplus amount of soft tissues in 5 out of 8 patients in the test group. Complications during the expansion period were perforation through oral mucosa occurred in two patients. Infection occurred in one patient. The mean soft tissue volume gain was 393.9 ± 322mm. After 6 months. The mean horizontal bone gains for the test and control groups were 3.14 mm and 3.69 mm, respectively. Conclusion: STE with a sub-periosteal approach is an applicable method to achieve an additional soft tissue and to reduce bone block graft exposure and wound dehiscence.

Keywords: soft tissue expander, ridge augmentation, block graft, symphysis bone block

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Authors: Somaiya Mateen, Shagufta Moin, Mohammad Owais, Abdul Khan, Atif Zafar

Abstract:

In rheumatoid arthritis (RA), impaired oxidative metabolism and imbalance between pro-and anti-inflammatory cytokines are responsible for causing inflammation and the degradation of cartilage and bone. The present study was done to evaluate the level and hence the role of reactive oxygen species (ROS) and inflammatory cytokines in the pathogenesis of RA. The present study was performed in the blood of 80 RA patients and 55 age and sex-matched healthy controls. The level of ROS (in 5% hematocrit) and the plasma level of pro-inflammatory cytokines [TNF-α, interleukin-6 (IL-6), IL-22] and anti-inflammatory cytokines (IL-4 and IL-5) were monitored in healthy subjects and RA patients. For evaluating the role of rheumatoid factor (RF) in the pathogenesis of RA, patients were sub-divided on the basis of presence or absence of RF. Reactive species and inflammatory cytokines were correlated with disease activity measure-Disease Activity Score for 28 joints (DAS28). The level of ROS, TNF-α, IL-6 and IL-22 were found to be significantly higher in RA patients as compared to the healthy controls, with the increase being more significant in patients positive for rheumatoid factor and those having high disease severity. On the other hand, a significant decrease in the level of IL-4 and IL-10 were observed in RA patients compared with healthy controls, with the decrease being more prominent in severe cases of RA. Higher ROS (indicative of impaired anti-oxidant defence system) and pro-inflammatory cytokines level in RA patients may lead to the damage of biomolecules which in turn contributes to tissue damage and hence to the development of more severe RA. The imbalance between pro-and anti-inflammatory cytokines may lead to the development of multi-system immune complications. ROS and inflammatory cytokines may also serve as a potential biomarker for assessing the disease severity.

Keywords: rheumatoid arthritis, reactive oxygen species, pro-inflammatory cytokines, anti-inflammatory cytokines

Procedia PDF Downloads 287

Authors: N. Haj Salem, M. Belhadj, S. Ben Jomâa, R. Dhouieb, S. Saadi, M. A. Mesrati, A. Chadly

Abstract:

Introduction: The hyoid bone is considered as one of many bones used to identify a missed person. There is a specificity of each population group in human identifications. Objective: To analyze the relationship between age, sex and metric parameters of hyoid bone in Tunisian population sample, using CT-scan. Materials and Methods: A prospective study was conducted in the Department of Forensic Medicine of FattoumaBourguiba Hospital of Monastir-Tunisia during 4 years. A total of 240 samples of hyoid bone were studied. The age of cases ranged from 18 days to 81 years. The specimens were collected only from the deceased of known age. Once dried, each hyoid bone was scanned using CT scan. For each specimen, 10 measurements were taken using a computer program. The measurements consisted of 6 lengths and 4 widths. A regression analysis was used to estimate the relationship between age, sex, and different measurements. For age estimation, a multiple logistic regression was carried out for samples ≤ 35 years. For sex determination, ROC curve was performed. Discriminant value finally retained was based on the best specificity with the best sensitivity. Results: The correlation between real age and estimated age was good (r²=0.72) for samples aged 35 years or less. The unstandardised canonical function equation was estimated using three variables: maximum length of the right greater cornua, length from the middle of the left joint space to the middle of the right joint space and perpendicular length from the centre point of a line between the distal ends of the right and left greater cornua to the centre point of the anterior view of the body of the hyoid bone. For sex determination, the ROC curve analysis reveals that the area under curve was at 81.8%. Discriminant value was 0.451 with a specificity of 73% and sensibility of 79%. The equation function was estimated based on two variables: maximum length of the greater cornua and maximum length of the hyoid bone. Conclusion: The findings of the current study suggest that metric analysis of the hyoid bone may predict the age ≤ 35 years. Sex estimation seems to be more reliable. Further studies dealing with the fusion of the hyoid bone and the current study could help to achieve more accurate age estimation rates.

Keywords: anthropology, age estimation, CT scan, sex determination, Tunisia

Procedia PDF Downloads 148

Authors: Laura Canagueral-Pellice, Antonio Munar-Frau, Adaia Valls-Ontanon, Joao Carames, Federico Hernandez-Alfaro, Jordi Caballe-Serrano

Abstract:

Objective: Guided bone regeneration (GBR) aims to replace the missing bone with a new structure to achieve long-term stability of rehabilitations. The aim of the present systematic review and meta-analysis is to determine the effect of barrier membranes on histological outcomes after GBR procedures. Moreover, the effect of the grafting material and tissue gain were analyzed. Materials & methods: Two independent reviewers performed an electronic search in Pubmed and Scopus, identifying all eligible publications up to March 2020. Only randomized controlled trials (RCTs) assessing a histological analysis of augmented areas were included. Results: A total of 6 publications were included for the present systematic review. A total of 110 biopsied sites were analysed; 10 corresponded to vertical bone augmentation procedures, whereas 100 analysed horizontal regeneration procedures. A mean tissue gain of 3 ± 1.48mm was obtained for horizontal defects. Histological assessment of new bone formation, residual particle and sub-epithelial connective tissue (SCT) was reported. The four main barrier membranes used were natural collagen membranes, e-PTFE, polylactic resorbable membranes and acellular dermal matrix membranes (AMDG). The analysis demonstrated that resorbable membranes result in higher values of new bone formation and lower values of residual particles and SCT. Xenograft resulted in lower new bone formation compared to allograft; however, no statistically significant differences were observed regarding residual particle and SCT. Overall, regeneration procedures adding autogenous bone, plasma derivate or growth factors achieved in general greater new bone formation and tissue gain. Conclusions: There is limited evidence favoring the effect of a certain type of barrier membrane in GBR. Data needs to be evaluated carefully; however, resorbable membranes are correlated with greater new bone formation values, especially when combined with allograft materials and/or the addition of autogenous bone, platelet reach plasma (PRP) or growth factors in the regeneration area. More studies assessing the histological outcomes of different GBR protocols and procedures testing different biomaterials are needed to maximize the clinical and histological outcomes in bone regeneration science.

Keywords: barrier membrane, graft material, guided bone regeneration, implant surgery, histology

Procedia PDF Downloads 124

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

Abstract:

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

Procedia PDF Downloads 159

Authors: Moetazza M. Al-Shafei, Hala Abdel Karim, Eitedal M. Daoud, Hassan Zaki Hassuna

Abstract:

Epilepsy is a common neurological disorder affecting all age groups. It is one of the world's most prevalent non-communicable diseases. Increased evidence suggesting that long term usage of anti-epileptic drugs can have adverse effects on bone mineralization and bone molding .Aiming to study these effects and to give guide lines to support bone health through early intervention. From Neurology Out-Patient Clinic kaser Elaini University Hospital, 60 Patients were enrolled, 40 patients on antiepileptic drugs for at least two years and 20 controls matched with age and sex, epileptic but before starting treatment both chosen under specific criteria. Patients were divided into four groups, three groups with monotherapy treated with either Phynetoin, Valporic acid or Carbamazipine and fourth group treated with both Valporic acid and Carbamazipine. Estimation of serum Carboxy-Terminal Telopeptide of Type I- Collagen(ICTP) bone resorption marker, serum 25(OH )vit D3, calcium ,magnesium and phosphorus were done .Results showed that all patients on AED had significant low levels of 25(OH) vit D3 (p<0.001) ,with significant elevation of ICTP (P<0.05) versus controls. In group treated with Phynotoin highly significant elevation of (ICTP) marker and decrease of both serum 25(OH) vit D3 (P<0, 0001) and serum calcium(P<0.05)versus control. Double drug group showed significant decrease of serum 25(OH) vit D3 (P<0.0001) and decrease in Phosphorus (P<0.05) versus controls. Serum magnesium showed no significant differences between studied groups. We concluded that Anti- epileptic drugs appears to be an aggravating factor on bone mineralization ,so therapeutically it can be worth wile to supplement calcium and vitamin D even before initiation of antiepileptic therapy. ICTP marker can be used to evaluate change in bone resorption before and during AED therapy.

Keywords: antiepileptic drugs, bone minerals, carboxy teminal telopeptidetype-1-collagen bone resorption marker, vitamin D

Procedia PDF Downloads 473

Authors: Rajendra Kumar Kanojia

Abstract:

Background; Massive bone gaps are common following road side accidents and injury to the tibia, specially open grade three fractures. It has been seen that the diaphyseal fractures in the tibia are prone to non-union, there are certain reasons known very well, like less soft tissues around the lower third tibia, less vascularity, less options of fixation of the fractures after trauma and prolonged surgical time, operation theatre time and special surgical means. Aim of study; To know the suitability of the ilizarov ring fixators in staged treatment of the fracture of the both bones leg, including tibia, we wish to see the role of ilizarov in management of open grade three fractures which have been operated and debrided, for getting the length use of ilizaorv ring in a tertiary canter is the aim of the study.

Keywords: open fracture, staged management, ilizarov, bone grafting, lengthening

Procedia PDF Downloads 284

Authors: Suraj Muke, Vikas Mankumare, Sadhana Sathaye

Abstract:

Osteoporosis is the most common metabolic bone disease which affects an estimated 25 million people worldwide, leading to 1 million fractures, 40,000 annual deaths and health costs of billions of dollars. It is estimated that about 80% of total osteoporosis patients are women, amongst which majority are above the age of 45 years. Postmenopausal osteoporosis is associated with lack of intestinal calcium absorption, increasing pro-oxidant and inflammatory mediators. Lawsonia inermis is a biennial dicotyledonous herbaceous shrub is reported to possess a high flavonoid, high phenolic and Inhibitors of osteoclastogenesis like Daphneside and Daphnorin. The present study aimed to screen osteoprotective effect of methanolic extract of Lawsonia inermis (LIM) in rat model along with its antioxidant activity. LIM shows phenolic content 146.3Milligrams of Gallic acid equivalent present per gram of extract and 19.8 Milligrams of rutin per gram of extract of Total flavonoid content with IC50 value 42.99μg/ml. bilateral ovariectomized rat model in which Healthy female wistar rats were used for screening. Treatment with LIM was carried out using graded doses of 25mg/kg, 50mg/kg and 100mg/kg for period of 28 days. The negative control group comprised of ovariectomized rats along with saline treatment for four weeks whereas sham operated rats were used as positive control.LIM showed a decrease in bone turnover by preventing loss of urinary calcium and phosphorous moreover it decreased the alkaline phosphatase levels and loss of bone density is prevented by LIM suggesting decrease in osteoclast activity.

Keywords: antioxidant, osteoclast, osteoporosis, ovariectomized

Procedia PDF Downloads 384

Authors: Hong Je Kang, Young Chae Choi, Jin Sung Park, Isac Kim

Abstract:

Purpose: Osteoporotic fractures are an important among postmenopausal women. When osteoporotic distal radial fractures occur, osteoporosis must be treated to prevent the hip and spine fractures. Intravenous injection of Zoledronic acid is expected to improve preventing osteoporotic fractures. Many articles reported the effect of intravenous Zoledronic acid to BMD of the hip and spine fracture or non-fracture patients with low BMD. However, that with distal radial fractures has rarely been reported. Therefore, the authors decided to study the effect of Zoledronic acid in BMD score, bone union, and bone turnover markers in the patients who underwent volar plating due to osteoporotic distal radial fractures. Materials: From April 2018 to May 2022, postmenopausal women aged 55 years or older who had osteoporotic distal radial fractures and who underwent surgical treatment using volar plate fixation were included. Zoledronic acid (5mg) was injected intravenously between 3 and 5 days after surgery. BMD scores after 1 year of operation were compared with the initial scores. Bone turnover markers were measured before surgery, after 3 months, and after 1 year. Radiological follow-up was performed every 2 weeks until the bone union and at 1 year postoperatively. Clinical outcome indicators were measured one year after surgery, and the occurrence of side effects was observed. Result: Total of 23 patients were included, with a lumbar BMD T score of -2.89±0.2 before surgery to -2.27±0.3 one year after surgery (p=0.012) and a femoral neck BMD T score of -2.45±0.3 before surgery to -2.36±0.3 (p=0.041) after one year, and all were statistically significant. Measured as bone resorption markers, serum CTX-1 was 337.43±10.4 pg/mL before surgery, 160.86±8.7 pg/mL (p=0.022) after three months, and 250.12±12.7 pg/mL (p=0.031) after one year. Urinary NTX-1 was 39.24±2.2 ng/mL before surgery, 24.46±1.2 ng/mL (p=0.014) after three months and 30.35±1.6 ng/mL (p=0.042) after one year. Measured as bone formation markers, serum osteocalcin was 13.04±1.1 ng/mL before surgery, 8.84±0.7 ng/mL (p=0.037) after 3 months and 11.1±0.4 ng/mL (p=0.026) after one year. Serum bone-specific ALP was 11.24±0.9 IU/L before surgery, 8.25±0.9 IU/L (p=0.036) after three months, and 10.2±0.9 IU/L (p=0.027) after one year. All were statistically significant. All cases showed bone union within an average of 6.91±0.3 weeks without any signs of failure. Complications were found in 5 out of 23 cases (21.7%), such as headache, nausea, muscle pain, and fever. Conclusion: When Zoledronic acid was used, BMD was improved in both the spine and femoral neck. This may reduce the likelihood and subsequent morbidity of additional osteoporotic fractures. This study is meaningful in that there was no difference in the duration of bone union and radiological characteristics in patients with distal radial fractures administrated with intravenous BP early after the fractures, and improvement in BMD and bone turnover indicators was measured.

Keywords: zeoldreonic acid, BMD, osteoporosis, distal radius

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Authors: Naveen Kumar B. T., Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, Shanthanagouda A. H., Orawan Boodde, Shankar K. M., Prakash Patil, Shubhkaramjeet Kaur

Abstract:

Since 2009, Acute Hepatopancreatic Necrosis Disease (AHPND) outbreaks have increased rapidly, and these have led to the major economic losses to the global shrimp industry. In comparison to other treatments, passive immunity and monoclonal antibody (MAb) based farmer level kit have proved their importance in controlling and treating the diseases in the shrimp industry. In the present study, MAbs were produced against the recombinant PirB protein Vibrio parahaemolyticus strain causing AHPND. Briefly, Balb/C mice were immunized with rPirB at 15 days interval, and antibody titer was determined by ELISA. Spleen cells from mice showing high antibody titer were fused with SP2O myeloma cells for hybridoma production. Among 130 hybridomas, four showed high antibody titer and positive reactivity in an immunoblot assay. In Western blot assay, three out of four MAbs (4C4, 2C2 and 4G3) showed reactivity to rPirB protein. However, in the natural host, only Mab clone 4G3 show strong reactivity (with a strain of V. parahemolyticus causing EMS/AHPND). These clones also showed reactivity with less than 20 kDa proteins in AHPND free V. parahaemolyticus (Thailand stain). Further, on from MAb 4G3 clone, four panels of single cell MAbs clones (G3F5, G3B8, G3H2, and G3D6) were produced of which three showed strong positive reactivity to rPirB protein in the Western blot. These MAbs have potential for controlling and prevention of the AHPND through passive immunity and development of filed level rapid diagnostic kits.

Keywords: shrimp, economic loss, AHPND, MAb

Procedia PDF Downloads 228