Search results for: bone mimic
Commenced in January 2007
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Edition: International
Paper Count: 961

Search results for: bone mimic

781 Forced Swim Stress Does Not Induce Structural Chromosomal Aberrations in Rat Bone Marrow

Authors: Mohammad Y. Alfaifi

Abstract:

Anything that poses a challenge or a threat to our well-being is a stress. Understanding the genetic material and cellular response of rats threatened with Repeated swimming stress provides insights that can influence human health. The aim of the present study was to assess the genetical damage and cytological changes caused by exposure of the test organism (Rattus rattus) to forced swimming stress. For this purpose, animals have been submerged in water path 15 minutes daily for 2 weeks. Following that, we performed a micronuclei (MN) test using MNNCE (Micronucleated 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 slightly but not significant increase in the frequency of micronucleated as well as in cytological parameters in bone marrow cells.

Keywords: submergence stress, micronucleus, NDI, NDCI, toxicity, chromosomal aberrations

Procedia PDF Downloads 372
780 Immunomodulatory Effect of Deer Antler Extract

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

Abstract:

Velvet antler (VA), the immature antlers of male deer, is traditionally used for thousands of years in Asian countries, such as Korea, China, Taiwan, and Mongolia. It has been considered to improve immune system and physical strength. The goal of this study was to investigate the immunomodulatory effect of deer antler velvet using in vitro system. In the first step, the effects of VA (70% ethanol extract) on the proliferation of splenocytes, bone marrow cell, and macrophages were determined. Next, the effect of VA on the production of nitric oxide and phagocytic activity in macrophage were measured. The results showed that VA treatment increased concanavalin-A stimulated splenocyte, bone marrow cells, and macrophage proliferation in a dose dependent manner. VA at 50 and 100 ug/mL concentrations significantly enhanced the concanavalin-A stimulated splenocyte proliferation by 8.8% and 18.5%, respectively. The proliferation of bone marrow cells, isolated from 5wk-old ICR mice, were increased by 25.2% and 46.5% by 50 and 100 ug/mL VA treatment. RAW 264.7 cell proliferation reached peak value at 50 ug/mL of VA treatment exhibiting 108% of the basal value. Nitric oxide production by RAW 264.7 macrophage cells was slightly reduced by VA treatment but was not statistically significant. Moreover, the phagocytic activity of macrophages was enhanced by VA treatment. These results indicate that VA is effective in immune system.

Keywords: deer antler, splenocyte, bone marrow cells, macrophage proliferation, phagocytosis

Procedia PDF Downloads 249
779 Synthesis and Characterization of Hydroxyapatite from Biowaste for Potential Medical Application

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

Abstract:

Over the period of time, several approaches have been undertaken to mitigate the challenges associated with bone regeneration. This includes but not limited to xenografts, allografts, autografts as well as artificial substitutions like bioceramics, synthetic cements and metals. The former three techniques often come along with peculiar limitation and problems such as morbidity, availability, disease transmission, collateral site damage or absolute rejection by the body as the case may be. Synthetic routes remain the only feasible alternative option for treatment of bone defects. Hydroxyapatite (HA) is very compatible and suitable for this application. However, most of the common methods for HA synthesis are either expensive, complicated or environmentally unfriendly. Interestingly, extraction of HA from bio-wastes have been perceived not only to be cost effective, but also environment friendly. In this research, HA was synthesized from bio-waste: namely bovine bones through three different methods which are hydrothermal chemical processes, ultrasound assisted synthesis and ordinary calcination techniques. Structure and property analysis of the HA was carried out through different characterization techniques such as TGA, FTIR, and XRD. All the methods applied were able to produce HA with similar compositional properties to biomaterials found in human calcified tissues. Calcination process was however observed to be more efficient as it eliminated all the organic components from the produced HA. The HA synthesized is unique for its minimal cost and environmental friendliness. It is also perceived to be suitable for tissue and bone engineering applications.

Keywords: hydroxyapatite, bone, calcination, biowaste

Procedia PDF Downloads 222
778 Evaluation of Some Serum Proteins as Markers for Myeloma Bone Disease

Authors: V. T. Gerov, D. I. Gerova, I. D. Micheva, N. F. Nazifova-Tasinova, M. N. Nikolova, M. G. Pasheva, B. T. Galunska

Abstract:

Multiple myeloma (MM) is the most frequent plasma cell (PC) dyscrasia that involves the skeleton. Myeloma bone disease (MBD) is characterized by osteolytic bone lesions as a result of increased osteoclasts activity not followed by reactive bone formation due to osteoblasts suppression. Skeletal complications cause significant adverse effects on quality of life and lead to increased morbidity and mortality. Last decade studies revealed the implication of different proteins in osteoclast activation and osteoblast inhibition. The aim of the present study was to determine serum levels of periostin, sRANKL and osteopontin and to evaluate their role as bone markers in MBD. Materials and methods. Thirty-two newly diagnosed MM patients (mean age: 62.2 ± 10.7 years) and 33 healthy controls (mean age: 58.9 ± 7.5 years) were enrolled in the study. According to IMWG criteria 28 patients were with symptomatic MM and 4 with monoclonal gammopathy of undetermined significance (MGUS). In respect to their bone involvement all symptomatic patients were divided into two groups (G): 9 patients with 0-3 osteolytic lesions (G1) and 19 patients with >3 osteolytic lesions and/or pathologic fractures (G2). Blood samples were drawn for routine laboratory analysis and for measurement of periostin, sRANKL and osteopontin serum levels by ELISA kits (Shanghai Sunred Biological Technology, China). Descriptive analysis, Mann-Whitney test for assessment the differences between groups and non-parametric correlation analysis were performed using GraphPad Prism v8.01. Results. The median serum levels of periostin, sRANKL and osteopontin of ММ patients were significantly higher compared to controls (554.7pg/ml (IQR=424.0-720.6) vs 396.9pg/ml (IQR=308.6-471.9), p=0.0001; 8.9pg/ml (IQR=7.1-10.5) vs 5.6pg/ml (IQR=5.1-6.4, p<0.0001 and 514.0ng/ml (IQR=469.3-754.0) vs 387.0ng/ml (IQR=335.9-441.9), p<0.0001, respectively). for assessment of differences between groups and non-parametric correlation analysis were performed using GraphPad Prism v8.01. Statistical significance was found for all tested bone markers between symptomatic MM patients and controls: G1 vs controls (p<0.03), G2 vs controls (p<0.0001) for periostin; G1 vs controls (p<0.0001), G2 vs controls (p<0.0001) for sRANKL; G1 vs controls (p=0.002), G2 vs controls (p<0.0001) for osteopontin, as well between symptomatic MM patients and MGUS patients: G1 vs MGUS (p<0.003), G2 vs MGUS (p=0.003) for periostin; G1 vs MGUS (p<0.05), G2 vs MGUS (p<0.001) for sRANKL; G1 vs MGUS (p=0.011), G2 vs MGUS (p=0.0001) for osteopontin. No differences were detected between MGUS and controls and between patients in G1 and G2 groups. Spearman correlation analysis revealed moderate positive correlation between periostin and beta-2-microglobulin (r=0.416, p=0.018), percentage bone marrow myeloma PC (r=0.432, p=0.014), and serum total protein (r=0.427, p=0.015). Osteopontin levels were also positively related to beta-2-microglobulin (r=0.540, p=0.0014), percentage bone marrow myeloma PC (r=0.423, p=0.016), and serum total protein (r=0.413, p=0.019). Serum sRANKL was only related to beta-2-microglobulin levels (r=0.398, p=0.024). Conclusion: In the present study, serum levels of periostin, sRANKL and osteopontin in newly diagnosed MM patients were evaluated. They gradually increased from MGUS to more advanced stages of MM reflecting the severity of bone destruction. These results support the idea that some new protein markers could be used in monitoring the MBD as a most severe complication of MM.

Keywords: myeloma bone disease, periostin, sRANKL, osteopontin

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777 Finite Element Analysis of Mini-Plate Stabilization of Mandible Fracture

Authors: Piotr Wadolowski, Grzegorz Krzesinski, Piotr Gutowski

Abstract:

The aim of the presented investigation is to recognize the possible mechanical issues of mini-plate connection used to treat mandible fractures and to check the impact of different factors for the stresses and displacements within the bone-stabilizer system. The mini-plate osteosynthesis technique is a common type of internal fixation using metal plates connected to the fractured bone parts by a set of screws. The selected two types of plate application methodology used by maxillofacial surgeons were investigated in the work. Those patterns differ in location and number of plates. The bone geometry was modeled on the base of computed tomography scans of hospitalized patient done just after mini-plate application. The solid volume geometry consisting of cortical and cancellous bone was created based on gained cloud of points. Temporomandibular joint and muscle system were simulated to imitate the real masticatory system behavior. Finite elements mesh and analysis were performed by ANSYS software. To simulate realistic connection behavior nonlinear contact conditions were used between the connecting elements and bones. The influence of the initial compression of the connected bone parts or the gap between them was analyzed. Nonlinear material properties of the bone tissues and elastic-plastic model of titanium alloy were used. The three cases of loading assuming the force of magnitude of 100N acting on the left molars, the right molars and the incisors were investigated. Stress distribution within connecting plate shows that the compression of the bone parts in the connection results in high stress concentration in the plate and the screws, however the maximum stress levels do not exceed material (titanium) yield limit. There are no significant differences between negative offset (gap) and no-offset conditions. The location of the external force influences the magnitude of stresses around both the plate and bone parts. Two-plate system gives generally lower von Misses stress under the same loading than the one-plating approach. Von Mises stress distribution within the cortical bone shows reduction of high stress field for the cases without the compression (neutral initial contact). For the initial prestressing there is a visible significant stress increase around the fixing holes at the bottom mini-plate due to the assembly stress. The local stress concentration may be the reason of bone destruction in those regions. The performed calculations prove that the bone-mini-plate system is able to properly stabilize the fractured mandible bone. There is visible strong dependency between the mini-plate location and stress distribution within the stabilizer structure and the surrounding bone tissue. The results (stresses within the bone tissues and within the devices, relative displacements of the bone parts at the interface) corresponding to different models of the connection provide a basis for the mechanical optimization of the mini-plate connections. The results of the performed numerical simulations were compared to clinical observation. They provide information helpful for better understanding of the load transfer in the mandible with the stabilizer and for improving stabilization techniques.

Keywords: finite element modeling, mandible fracture, mini-plate connection, osteosynthesis

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776 Association of Clostridium difficile Infection and Bone Cancer

Authors: Daniela Prado, Lexi Frankel, Amalia Ardeljan, Lokesh Manjani, Matthew Cardeiro, Omar Rashid

Abstract:

Background: Clostridium difficile (C. diff) is a gram-positive bacterium that is known to cause life-threatening diarrhea and severe inflammation of the colon. It originates as an alteration of the gut microbiome and can be transmitted through spores. Recent studies have shown a high association between the development of C. diff in cancer patients due to extensive hospitalization. However, research is lacking regarding C. diff’s association in the causation or prevention of cancer. The objective of this study was to therefore assess the correlation between Clostridium difficile infection (CDI) and the incidence of bone cancer. Methods: This retrospective analysis used data provided by a Health Insurance Portability and Accountability Act (HIPAA) compliant national database to evaluate the patients infected versus patients not infected with C. diff using ICD-10 and ICD-9 codes. Access to the database was granted by the Holy Cross Health, Fort Lauderdale, for the purpose of academic research. Standard statistical methods were used. Results: Between January 2010 and December 2019, the query was analyzed and resulted in 78863 patients in both the infected and control group, respectively. The two groups were matched by age range and CCI score. The incidence of bone cancer was 659 patients (0.835%) in the C. diff group compared to 1941 patients (2.461%) in the control group. The difference was statistically significant by a P-value < 2.2x10^-16 with an odds ratio (OR)= 0.33 (0.31-0.37) with a 95% confidence interval (CI). Treatment for CDI was analyzed for both C. diff infected and noninfected populations. 91 out of 16,676 (0.55%) patients with a prior C. diff infection and treated with antibiotics were compared to the control group were 275 out of 16,676 (1.65%) patients with no history of CDI and received antibiotic treatment. Results remained statistically significant by P-value <2.2x10-16 with an OR= 0.42 (0.37, 0.48). and a 95% CI. Conclusion: The study shows a statistically significant correlation between C. diff and a reduced incidence of bone cancer. Further evaluation is recommended to assess the potential of C. difficile in reducing bone cancer incidence.

Keywords: bone cancer, colitis, clostridium difficile, microbiome

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775 Piled Critical Size Bone-Biomimetic and Biominerizable Nanocomposites: Formation of Bioreactor-Induced Stem Cell Gradients under Perfusion and Compression

Authors: W. Baumgartner, M. Welti, N. Hild, S. C. Hess, W. J. Stark, G. Meier Bürgisser, P. Giovanoli, J. Buschmann

Abstract:

Perfusion bioreactors are used to solve problems in tissue engineering in terms of sufficient nutrient and oxygen supply. Such problems especially occur in critical size grafts because vascularization is often too slow after implantation ending up in necrotic cores. Biominerizable and biocompatible nanocomposite materials are attractive and suitable scaffold materials for bone tissue engineering because they offer mineral components in organic carriers – mimicking natural bone tissue. In addition, human adipose derived stem cells (ASCs) can potentially be used to increase bone healing as they are capable of differentiating towards osteoblasts or endothelial cells among others. In the present study, electrospun nanocomposite disks of poly-lactic-co-glycolic acid and amorphous calcium phosphate nanoparticles (PLGA/a-CaP) were seeded with human ASCs and eight disks were stacked in a bioreactor running with normal culture medium (no differentiation supplements). Under continuous perfusion and uniaxial cyclic compression, load-displacement curves as a function of time were assessed. Stiffness and energy dissipation were recorded. Moreover, stem cell densities in the layers of the piled scaffold were determined as well as their morphologies and differentiation status (endothelial cell differentiation, chondrogenesis and osteogenesis). While the stiffness of the cell free constructs increased over time caused by the transformation of the a-CaP nanoparticles into flake-like apatite, ASC-seeded constructs showed a constant stiffness. Stem cell density gradients were histologically determined with a linear increase in the flow direction from the bottom to the top of the 3.5 mm high pile (r2 > 0.95). Cell morphology was influenced by the flow rate, with stem cells getting more roundish at higher flow rates. Less than 1 % osteogenesis was found upon osteopontin immunostaining at the end of the experiment (9 days), while no endothelial cell differentiation and no chondrogenesis was triggered under these conditions. All ASCs had mainly remained in their original pluripotent status within this time frame. In summary, we have fabricated a critical size bone graft based on a biominerizable bone-biomimetic nanocomposite with preserved stiffness when seeded with human ASCs. The special feature of this bone graft was that ASC densities inside the piled construct varied with a linear gradient, which is a good starting point for tissue engineering interfaces such as bone-cartilage where the bone tissue is cell rich while the cartilage exhibits low cell densities. As such, this tissue-engineered graft may act as a bone-cartilage interface after the corresponding differentiation of the ASCs.

Keywords: bioreactor, bone, cartilage, nanocomposite, stem cell gradient

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774 Bone Mineral Density in Long-Living Patients with Coronary Artery Disease

Authors: Svetlana V. Topolyanskaya, Tatyana A. Eliseeva, Olga N. Vakulenko, Leonid I. Dvoretski

Abstract:

Introduction: Limited data are available on osteoporosis in centenarians. Therefore, we evaluated bone mineral density in long-living patients with coronary artery disease (CAD). Methods: 202 patients hospitalized with CAD were enrolled in this cross-sectional study. The patients' age ranged from 90 to 101 years. The majority of study participants (64.4%) were women. The main exclusion criteria were any disease or medication that can lead to secondary osteoporosis. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry. Results: Normal lumbar spine BMD was observed in 40.9%, osteoporosis – in 26.9%, osteopenia – in 32.2% of patients. Normal proximal femur BMD values were observed in 21.3%, osteoporosis – in 39.9%, and osteopenia – in 38.8% of patients. Normal femoral neck BMD was registered only in 10.4% of patients, osteoporosis was observed in 60.4%, osteopenia in 29.2%. Significant positive correlation was found between all BMD values and body mass index of patients (p < 0.001). Positive correlation was registered between BMD values and serum uric acid (p=0.0005). The likelihood of normal BMD values with hyperuricemia increased 3.8 times, compared to patients with normal uric acid, who often have osteoporosis (Odds Ratio=3.84; p = 0.009). Positive correlation was registered between all BMD values and body mass index (p < 0.001). Positive correlation between triglycerides levels and T-score (p=0.02), but negative correlation between BMD and HDL-cholesterol (p=0.02) were revealed. Negative correlation between frailty severity and BMD values (p=0.01) was found. Positive correlation between BMD values and functional abilities of patients assessed using Barthel index (r=0,44; p=0,000002) and IADL scale (r=0,36; p=0,00008) was registered. Fractures in history were observed in 27.6% of patients. Conclusions: The study results indicate some features of BMD in long-livers. In the study group, significant relationships were found between bone mineral density on the one hand, and patients' functional abilities on the other. It is advisable to further study the state of bone tissue in long-livers involving a large sample of patients.

Keywords: osteoporosis, bone mineral density, centenarians, coronary artery disease

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773 The Role of Bone Marrow Stem Cells Transplantation in the Repair of Damaged Inner Ear in Albino Rats

Authors: Ahmed Gaber Abdel Raheem, Nashwa Ahmed Mohamed

Abstract:

Introduction: Sensorineural hearing loss (SNHL) is largely caused by the degeneration of the cochlea. Therapeutic options for SNHL are limited to hearing aids and cochlear implants. The cell transplantation approach to the regeneration of hair cells has gained considerable attention because stem cells are believed to accumulate in the damaged sites and have the potential for the repair of damaged tissues. The aim of the work: was to assess the use of bone marrow transplantation in repair of damaged inner ear hair cells in rats after the damage had been inflicted by Amikacin injection. Material and Methods: Thirty albino rats were used in this study. They were divided into three groups. Each group ten rats. Group I: used as control. Group II: Were given Amikacin- intratympanic injection till complete loss of hearing function. This could be assessed by Distortion product Otoacoustic Emission (DPOAEs) and / or auditory brain stem evoked potential (ABR). GroupIII: were given intra-peritoneal injection of bone marrow stem cell after complete loss of hearing caused by Amikacin. Clinical assessment was done using DPOAEs and / or auditory brain stem evoked potential (ABR), before and after bone marrow injection. Histological assessment of the inner ear was done by light and electron microscope. Also, Detection of stem cells in the inner ear by immunohistochemistry. Results: Histological examination of the specimens showed promising improvement in the structure of cochlea that may be responsible for the improvement of hearing function in rats detected by DPOAEs and / or ABR. Conclusion: Bone marrow stem cells transplantation might be useful for the treatment of SNHL.

Keywords: amikacin, hair cells, sensorineural hearing loss, stem cells

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772 Understanding the Information in Principal Component Analysis of Raman Spectroscopic Data during Healing of Subcritical Calvarial Defects

Authors: Rafay Ahmed, Condon Lau

Abstract:

Bone healing is a complex and sequential process involving changes at the molecular level. Raman spectroscopy is a promising technique to study bone mineral and matrix environments simultaneously. In this study, subcritical calvarial defects are used to study bone composition during healing without discomposing the fracture. The model allowed to monitor the natural healing of bone avoiding mechanical harm to the callus. Calvarial defects were created using 1mm burr drill in the parietal bones of Sprague-Dawley rats (n=8) that served in vivo defects. After 7 days, their skulls were harvested after euthanizing. One additional defect per sample was created on the opposite parietal bone using same calvarial defect procedure to serve as control defect. Raman spectroscopy (785 nm) was established to investigate bone parameters of three different skull surfaces; in vivo defects, control defects and normal surface. Principal component analysis (PCA) was utilized for the data analysis and interpretation of Raman spectra and helped in the classification of groups. PCA was able to distinguish in vivo defects from normal surface and control defects. PC1 shows that the major variation at 958 cm⁻¹, which corresponds to ʋ1 phosphate mineral band. PC2 shows the major variation at 1448 cm⁻¹ which is the characteristic band of CH2 deformation and corresponds to collagens. Raman parameters, namely, mineral to matrix ratio and crystallinity was found significantly decreased in the in vivo defects compared to surface and controls. Scanning electron microscope and optical microscope images show the formation of newly generated matrix by means of bony bridges of collagens. Optical profiler shows that surface roughness increased by 30% from controls to in vivo defects after 7 days. These results agree with Raman assessment parameters and confirm the new collagen formation during healing.

Keywords: Raman spectroscopy, principal component analysis, calvarial defects, tissue characterization

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771 Influence of Ligature Tightening on Bone Fracture Risk in Interspinous Process Surgery

Authors: Dae Kyung Choi, Won Man Park, Kyungsoo Kim, Yoon Hyuk Kim

Abstract:

The interspinous process devices have been recently used due to its advantages such as minimal invasiveness and less subsidence of the implant to the osteoporotic bone. In this paper, we have analyzed the influences of ligature tightening of several interspinous process devices using finite element analysis. Four types of interspinous process implants were inserted to the L3-4 spinal motion segment based on their surgical protocols. Inferior plane of L4 vertebra was fixed and 7.5 Nm of extension moment were applied on superior plane of L3 vertebra with 400N of compressive load along follower load direction and pretension of the ligature. The stability of the spinal unit was high enough than that of intact model. The higher value of pretension in the ligature led the decrease of dynamic stabilization effect in cases of the WallisTM, DiamTM, Viking, and Spear®. The results of present study could be used to evaluate surgical option and validate the biomechanical characteristics of the spinal implants.

Keywords: interspinous process device, bone fracture risk, lumbar spine, finite element analysis

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770 Effects of Li2O Doping on Mechanical and Electrical Properties of Bovine Hydroxyapatite Composites (BHA)

Authors: Sibel Daglilar, Isil Kerti, Murat Karagoz, Fatih Dumludag, Oguzhan Gunduz, Faik Nuzhet Oktar

Abstract:

Hydroxyapatite (HA) materials have common use in bone repairing due to its ability to accelerate the bone growth around the implant. In spite of being a biocompatible and bioactive material, HA has a limited usage as an implant material because of its weak mechanical properties. HA based composites are required to improve the strength and toughness properties of the implant materials without compromising of biocompatibility. The excellent mechanical properties and higher biocompatibilities are expected from each of biomedical composites. In this study, HA composites were synthesized by using bovine bone reinforced doped with different amount of (wt.%) Li2O. The pressed pellets were sintered at various sintering temperatures between 1000ºC and 1300°C, and mechanical, electrical properties of the obtained products were characterized. In addition to that, in vitro stimulated body fluid (SBF) tests for these samples were conducted. The most suitable composite composition for biomedical applications was discussed among the composites studied.

Keywords: biocomposites, sintering temperature, biocompatibility, electrical property, conductivity, mechanical property

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769 Osteitis in the Diabetic Foot in Algeria

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

Abstract:

— 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: diabetic foot, bone biopsy, osteitis, algeria

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768 Tc-99m MIBI Scintigraphy to Differentiate Malignant from Benign Lesions, Detected on Planar Bone Scan

Authors: Aniqa Jabeen

Abstract:

The aim of this study was to evaluate the effectiveness of Tc-99m MIBI (Technetium 99-methoxy-iso-butyl-isonitrile) scintigraphy to differentiate malignancies from benign lesions, which were detected on planar bone scans. Materials and Methods: 59 patients with bone lesions were enrolled in the study. The scintigraphic findings were compared with the clinical, radiological and the histological findings. Each patient initially underwent a three-phase bone scan with Tc-99m MDP (Methylene Diphosphonate) and if evidence of lesion found, the patient then underwent a dynamic and static MIBI scintigraphy after three to four days. The MDP and MIBI scans were evaluated visually and quantitatively. For quantitative analysis count ratios of lesions and contralateral normal side (L/C) were taken by region of interests drawn on scans. The Student T test was applied to assess the significant difference between benign and malignant lesions p-value < 0.05 was considered significant. Result: The MDP scans showed the increase tracer uptake, but there was no significant difference between benign and malignant uptake of the radiotracer. However significant difference (p-value 0.015), in uptake was seen in malignant (L/C = 3.51 ± 1.02) and benign lesion (L/C = 2.50±0.42) on MIBI scan. Three of thirty benign lesions did not show significant MIBI uptake. Seven malignant appeared as false negatives. Specificity of the scan was 86.66%, and its Negative Predictive Value (NPV) was 81.25% whereas the sensitivity of scan was 79.31%. In excluding the axial metastasis from the lesions, the sensitivity of MIBI scan increased to 91.66% and the NPV also increased to 92.85%. Conclusion: MIBI scintigraphy provides its usefulness by distinguishing malignant from benign lesions. MIBI also correctly identifies metastatic lesions. The negative predictive value of the scan points towards its ability to accurately diagnose the normal (benign) cases. However, biopsy remains the gold standard and a definitive diagnostic modality in musculoskeletal tumors. MIBI scan provides useful information in preoperative assessment and in distinguishing between malignant and benign lesions.

Keywords: benign, malignancies, MDP bone scan, MIBI scintigraphy

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767 Physiopathology of Osteitis in the Diabetic Foot

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

Abstract:

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 therapy, bone biopsy, diabetic foot

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766 Diagnostic Physiopathology of Osteitis in the Diabetic Foot

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

Abstract:

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 therapy, bone biopsy, diabetic foot

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765 Advancement in Adhesion and Osteogenesis of Stem Cells with Histatin Coated 3D-Printed Bio-Ceramics

Authors: Haiyan Wang, Dongyun Wang, Yongyong Yan, Richard T. Jaspers, Gang Wu

Abstract:

Mesenchymal stem cell and 3D printing-based bone tissue engineering present a promising technique to repair large-volume bone defects. Its success is highly dependent on cell attachment, spreading, osteogenic differentiation, and in vivo survival of stem cells on 3D-printed scaffolds. In this study, human salivary histatin-1 (Hst1) was utilized to enhance the interactions between human adipose-derived stem cells (hASCs) and 3D-printed β-tricalcium phosphate (β-TCP) bioceramic scaffolds. Fluorescent images showed that Hst1 significantly enhanced the adhesion of hASCs to both bioinert glass and 3D-printed β-TCP scaffold. In addition, Hst1 was associated with significantly higher proliferation and osteogenic differentiation of hASCs on 3D-printed β-TCP scaffolds. Moreover, coating 3D-printed β-TCP scaffolds with histatin significantly promotes the survival of hASCs in vivo. The ERK and p38 but not JNK signaling was found to be involved in the superior adhesion of hASCs to β-TCP scaffolds with the aid of Hst1. In conclusion, Hst1 could significantly promote the adhesion, spreading, osteogenic differentiation, and in vivo survival of hASCs on 3D-printed β-TCP scaffolds, bearing a promising application in stem cell/3D printing-based constructs for bone tissue engineering.

Keywords: 3d printing, adipose-derived stem cells, bone tissue engineering, histatin-1, osteogenesis

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764 Oncological and Antiresorptive Treatment of Breast Cancer: Dental Assessment and Risk of MRONJ Development

Authors: Magdalena Korytowska, Gunnar Lengstrand, Cecilia Larsson Wexell

Abstract:

Background: Breast cancer (BC) is the most common cancer among women worldwide, and cases are continuing to increase in Sweden. Bone is the most common metastatic site in breast cancer patients, where > 65-75% of women with advanced breast cancer develop bone metastases during their disease. To prevent the skeletal-related events of metastases (e.g., pathological fractures, bone loss, cancer-induced bone pain, and hypercalcemia bone), two different classes of antiresorptive medications (AR), bisphosphonate and denosumab are typically administered every 3 to 4 weeks. Since 2015, adjuvant bisphosphonate treatment has been used every six months for three to five years in postmenopausal women for the prevention of skeletal metastases and improved survival. Methods: A case-control study was conducted to test the hypotheses that patients treated with high-dose AR are at higher risk of developing MRONJ than breast cancer patients with adjuvant bisphosphonate treatment at a lower dose. Medical and odontological data was collected between 2015-2020. Assessment of oral health and dental care before and during oncological treatment took place at the specialist clinic for Orofacial medicine linked to the specific hospital. Results: In total, 220 patients were included, 101 patients in the high-dose group and 119 patients in the adjuvant BP-treatment group. MRONJ was diagnosed in 13 patients (14%) in the high-dose group. The mandible was affected in most of the cases (84.6%), with a mean duration of high-dose treatment of 19.7 months. In 46.2% of cases, no dental cause of MRONJ could be identified. Overall, estrogen receptor-positive (ER+) BC was the most representative type in 172 patients (78.2%). However, this was 83.9% in the high-dose cases group. The most used drug was denosumab. Twenty-five patients (26.9%) switched their medication from ZOL to denosumab during their oncological treatment. Patients with ER+ breast cancer were reported in 88 patients (87.8%) in the adjuvant group that was treated with ZOL. Conclusions: MRONJ was diagnosed only in the high-dose AR group. Dental assessment and care of patients in the adjuvant group should be considered, with a recommendation to potentially prolong ZOL treatment from 3 to 5 years, with concomitant use of hormonal therapy in patients diagnosed with ER+ breast cancer to prevent bone loss induced by oncological treatment. A new referral for dental assessment is very important in the case of bone metastases when treatment with high dose AR will be required since it is associated with a higher risk of MRONJ.

Keywords: antiresorptive therapy, breast cancer, dental care, MRONJ

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763 A Radiographic Superimposition in Orthognathic Surgery of Class III Skeletal Malocclusion

Authors: Albert Suryaprawira

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Patients requiring correction of severe Class III skeletal discrepancy historically has been among the most challenging treatments for orthodontists. Correction of an aesthetic and functional problem is crucially important. This is a case report of an adult male aged 18 years who complained of difficulty in chewing and speaking. Patient has a prominent profile with mandibular excess. The pre-treatment cephalometric radiograph was taken to analyse the skeletal problem and to measure the amount of bone movement and the prediction soft tissue response. The panoramic radiograph was also taken to analyse bone quality, bone abnormality, third molar impaction, etc. Before the surgery, the pre-surgical cephalometric radiograph was taken to re-evaluate the plan and to settle the final amount of bone cut. After the surgery, the post-surgical cephalometric radiograph was taken to confirm the result with the plan. The superimposition between those radiographs was performed to analyse the outcome. It includes the superimposition of the cranial base, maxilla, and mandible. Superimposition is important to describe the amount of hard and soft tissue movement. It is also important to predict the possibility of relapse after the surgery. The patient needs to understand all the surgical plan, outcome and relapse prevention. The surgery included mandibular set back by bilateral sagittal split osteotomies. Although the discrepancy was severe using this combination of treatment and the use of radiographic superimposition, an aesthetically pleasing and stable result was achieved.

Keywords: cephalometric, mandibular set back, orthognathic, superimposition

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762 Poly(propylene fumarate) Copolymers with Phosphonic Acid-based Monomers Designed as Bone Tissue Engineering Scaffolds

Authors: Görkem Cemali̇, Avram Aruh, Gamze Torun Köse, Erde Can ŞAfak

Abstract:

In order to heal bone disorders, the conventional methods which involve the use of autologous and allogenous bone grafts or permanent implants have certain disadvantages such as limited supply, disease transmission, or adverse immune response. A biodegradable material that acts as structural support to the damaged bone area and serves as a scaffold that enhances bone regeneration and guides bone formation is one desirable solution. Poly(propylene fumarate) (PPF) which is an unsaturated polyester that can be copolymerized with appropriate vinyl monomers to give biodegradable network structures, is a promising candidate polymer to prepare bone tissue engineering scaffolds. In this study, hydroxyl-terminated PPF was synthesized and thermally cured with vinyl phosphonic acid (VPA) and diethyl vinyl phosphonate (VPES) in the presence of radical initiator benzoyl peroxide (BP), with changing co-monomer weight ratios (10-40wt%). In addition, the synthesized PPF was cured with VPES comonomer at body temperature (37oC) in the presence of BP initiator, N, N-Dimethyl para-toluidine catalyst and varying amounts of Beta-tricalcium phosphate (0-20 wt% ß-TCP) as filler via radical polymerization to prepare composite materials that can be used in injectable forms. Thermomechanical properties, compressive properties, hydrophilicity and biodegradability of the PPF/VPA and PPF/VPES copolymers were determined and analyzed with respect to the copolymer composition. Biocompatibility of the resulting polymers and their composites was determined by the MTS assay and osteoblast activity was explored with von kossa, alkaline phosphatase and osteocalcin activity analysis and the effects of VPA and VPES comonomer composition on these properties were investigated. Thermally cured PPF/VPA and PPF/VPES copolymers with different compositions exhibited compressive modulus and strength values in the wide range of 10–836 MPa and 14–119 MPa, respectively. MTS assay studies showed that the majority of the tested compositions were biocompatible and the overall results indicated that PPF/VPA and PPF/VPES network polymers show significant potential for applications as bone tissue engineering scaffolds where varying PPF and co-monomer ratio provides adjustable and controllable properties of the end product. The body temperature cured PPF/VPES/ß-TCP composites exhibited significantly lower compressive modulus and strength values than the thermal cured PPF/VPES copolymers and were therefore found to be useful as scaffolds for cartilage tissue engineering applications.

Keywords: biodegradable, bone tissue, copolymer, poly(propylene fumarate), scaffold

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761 Effect of Operative Stabilization on Rib Fracture Healing in Porcine Experimental Model: A Pilot Study

Authors: Maria Stepankova, Lucie Vistejnova, Pavel Klein, Tereza Blassova, Marketa Slajerova, Radek Sedlacek, Martin Bartos, Jaroslav Chlupac

Abstract:

Background: Clinical outcome benefits of the segment rib fracture surgical therapy are well known and follow from better stabilization of the chest wall. Despite this, some authors still incline to conservative therapy and point out to possible rib fracture healing failure in connection with the bone vascular supply disturbance caused by metal plate implantation. This suggestion met neither experimental nor clinical verification and remains the object of discussion. In our pilot study we investigated the titanium plate fixation effect on the rib fracture healing in porcine model and its histological, biomechanical and radiological aspects. Materials and Method: Two porcine models (experimental group) underwent the operative chest wall stabilization with a titanium plate implantation after osteotomy. Two other porcine models (control group) were treated conservatively after osteotomy. Three weeks after surgery, all animals were sacrificed, treated ribs were explanted and the histological analysis, µCT imaging and biomechanical testing of the calluses tissue were performed. Results: In µCT imaging, experimental group showed a higher cortical bone volume compared to the control group. Histological analysis using the non-decalcified bone tissue blocks demonstrated more maturated callus with higher newly-formed osseous tissue ratio in experimental group in comparison to controls. In contrast, no significant differences in bone blood vessels supply in both groups were observed. This finding suggests that the bone blood supply in experimental group was not impaired. Biomechanical analysis using 3-point bending test demonstrated significantly higher bending stiffness and the maximum force in experimental group. Conclusion: Based on our observation, it could be concluded, that the titanium plate fixation of the rib fractures leads to faster bone callus maturation whereas does not cause the vascular supply impairment after 3 weeks and thus has a beneficial effect on the rib fracture healing.

Keywords: bone vascular supply, chest wall stabilization, fracture healing, histological analysis, titanium plate implantation

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760 Severe Bone Marrow Edema on Sacroiliac Joint MRI Increases the Risk of Low BMD in Patients with Axial Spondyloarthritis

Authors: Kwi Young Kang

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Objective: To determine the association between inflammatory and structural lesions on sacroiliac joint (SIJ) MRI and BMD and to identify risk factors for low BMD in patients with axial spondyloarthritis (axSpA). Methods: Seventy-six patients who fulfilled the ASAS axSpA criteria were enrolled. All underwent SIJ MRI and BMD measurement at the lumbar spine, femoral neck, and total hip. Inflammatory and structural lesions on SIJ MRI were scored. Laboratory tests and assessment of radiographic and disease activity were performed at the time of MRI. The association between SIJ MRI findings and BMD was evaluated. Results: Among the 76 patients, 14 (18%) had low BMD. Patients with low BMD showed significantly higher bone marrow edema (BME) and deep BME scores on MRI than those with normal BMD (p<0.047 and 0.007, respectively). Inflammatory lesions on SIJ MRI correlated with BMD at the femoral neck and total hip. Multivariate analysis identified the presence of deep BME on SIJ MRI, increased CRP, and sacroiliitis on X-ray as risk factors for low BMD (OR: 5.6, 14.6, and 2.5, respectively). Conclusion: The presence of deep BME on SIJ MRI, increased CRP levels, and severity of sacroiliitis on X-ray were independent risk factors for low BMD.

Keywords: axial spondyloarthritis, sacroiliac joint MRI, bone mineral density, sacroiliitis

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759 The Effect of Calcium Phosphate Composite Scaffolds on the Osteogenic Differentiation of Rabbit Dental Pulp Stem Cells

Authors: Ling-Ling E, Lin Feng, Hong-Chen Liu, Dong-Sheng Wang, Zhanping Shi, Juncheng Wang, Wei Luo, Yan Lv

Abstract:

The objective of this study was to compare the effects of the two calcium phosphate composite scaffolds on the attachment, proliferation and osteogenic differentiation of rabbit dental pulp stem cells (DPSCs). One nano-hydroxyapatite/collagen/poly (L-lactide) (nHAC/PLA), imitating the composition and the micro-structure characteristics of the natural bone, was made by Beijing Allgens Medical Science & Technology Co., Ltd. (China). The other beta-tricalcium phosphate (β-TCP), being fully interoperability globular pore structure, was provided by Shanghai Bio-lu Biomaterials Co, Ltd. (China). We compared the absorption water rate and the protein adsorption rate of two scaffolds and the characterization of DPSCs cultured on the culture plate and both scaffolds under osteogenic differentiation media (ODM) treatment. The constructs were then implanted subcutaneously into the back of severe combined immunodeficient (SCID) mice for 8 and 12 weeks to compare their bone formation capacity. The results showed that the ODM-treated DPSCs expressed osteocalcin (OCN), bone sialoprotein (BSP), type I collagen (COLI) and osteopontin (OPN) by immunofluorescence staining. Positive alkaline phosphatase (ALP) staining, calcium deposition and calcium nodules were also observed on the ODM-treated DPSCs. The nHAC/PLA had significantly higher absorption water rate and protein adsorption rate than ß-TCP. The initial attachment of DPSCs seeded onto nHAC/PLA was significantly higher than that onto ß-TCP; and the proliferation rate of the cells was significantly higher than that of ß-TCP on 1, 3 and 7 days of cell culture. DPSCs+ß-TCP had significantly higher ALP activity, calcium/phosphorus content and mineral formation than DPSCs+nHAC/PLA. When implanted into the back of SCID mice, nHAC/PLA alone had no new bone formation, newly formed mature bone and osteoid were only observed in β-TCP alone, DPSCs+nHAC/PLA and DPSCs+β-TCP, and this three groups displayed increased bone formation over the 12-week period. The percentage of total bone formation area had no difference between DPSCs+β-TCP and DPSCs+nHAC/PLA at each time point,but the percentage of mature bone formation area of DPSCs+β-TCP was significantly higher than that of DPSCs+nHAC/PLA. Our results demonstrated that the DPSCs on nHAC/PLA had a better proliferation and that the DPSCs on β-TCP had a more mineralization in vitro, much more newly formed mature bones in vivo were presented in DPSCs+β-TCP group. These findings have provided a further knowledge that scaffold architecture has a different influence on the attachment, proliferation and differentiation of cells. This study may provide insight into the clinical periodontal bone tissue repair with DPSCs+β-TCP construct.

Keywords: dental pulp stem cells, nano-hydroxyapatite/collagen/poly(L-lactide), beta-tricalcium phosphate, periodontal tissue engineering, bone regeneration

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758 The MicroRNA-2110 Suppressed Cell Proliferation and Migration Capacity in Hepatocellular Carcinoma Cells

Authors: Pelin Balcik Ercin

Abstract:

Introduction: ZEB transcription factor family member ZEB2, has a role in epithelial to mesenchymal transition during development and metastasis. The altered circulating extracellular miRNAs expression is observed in diseases, and extracellular miRNAs have an important role in cancer cell microenvironment. In ChIP-Seq study, the expression of miR-2110 was found to be regulated by ZEB2. In this study, the effects of miR2110 on cell proliferation and migration of hepatocellular carcinoma (HCC) cells were examined. Material and Methods: SNU398 cells transfected with mimic miR2110 (20nM) (HMI0375, Sigma-Aldrich) and negative control miR (HMC0002, Sigma-Aldrich). MicroRNA isolation was accomplished with miRVANA isolation kit according to manufacturer instructions. cDNA synthesis was performed expression, respectively, and calibrated with Ct of controls. The real-time quantitative PCR (RT-qPCR) reaction was performed using the TaqMan Fast Advanced Master Mix (Thermo Sci.). Ct values of miR2110 were normalized to miR-186-5p and miR16-5p for the intracellular gene. Cell proliferation analysis was analyzed with the xCELLigence RTCA System. Wound healing assay was analyzed with the ImageJ program and relative fold change calculated. Results: The mimic-miR-2110 transfected SNU398 cells nearly nine-fold (log2) more miR-2110 expressed compared to negative control transfected cells. The mimic-miR-2110 transfected HCC cell proliferation significantly inhibited compared to the negative control cells. Furthermore, miR-2110-SNU398 cell migration capacity was relatively four-fold decreased compared to negative control-miR-SNU398 cells. Conclusion: Our results suggest the miR-2110 inhibited cell proliferation and also miR-2110 negatively affect cell migration compared to control groups in HCC cells. These data suggest the complexity of microRNA EMT transcription factors regulation. These initial results are pointed out the predictive biomarker capacity of miR-2110 in HCC.

Keywords: epithelial to mesenchymal transition, EMT, hepatocellular carcinoma cells, micro-RNA-2110, ZEB2

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757 Estimated Human Absorbed Dose of 111 In-BPAMD as a New Bone-Seeking Spect-Imaging Agent

Authors: H. Yousefnia, S. Zolghadri

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An early diagnosis of bone metastases is very important for providing a profound decision on a subsequent therapy. A prerequisite for the clinical application of new diagnostic radiopharmaceutical is the measurement of organ radiation exposure dose from biodistribution data in animals. In this study, the dosimetric studies of a novel agent for SPECT-imaging of bone methastases, 111In-(4-{[(bis(phosphonomethyl))carbamoyl]methyl}-7,10-bis(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl) acetic acid (111In-BPAMD) complex, have been estimated in human organs based on mice data. The radiolabeled complex was prepared with high radiochemical purity at the optimal conditions. Biodistribution studies of the complex were investigated in male Syrian mice at selected times after injection (2, 4, 24 and 48 h). The human absorbed dose estimation of the complex was performed based on mice data by the radiation absorbed dose assessment resource (RADAR) method. 111In-BPAMD complex was prepared with high radiochemical purity >95% (ITLC) and specific activities of 2.85 TBq/mmol. Total body effective absorbed dose for 111In-BPAMD was 0.205 mSv/MBq. This value is comparable to the other 111In clinically used complexes. The results show that the dose to critical organs the complex is well within the acceptable considered range for diagnostic nuclear medicine procedures. Generally, 111In-BPAMD has interesting characteristics and can be considered as a viable agent for SPECT-imaging of the bone metastases in the near future.

Keywords: In-111, BPAMD, absorbed dose, RADAR

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756 Toward Automatic Chest CT Image Segmentation

Authors: Angely Sim Jia Wun, Sasa Arsovski

Abstract:

Numerous studies have been conducted on the segmentation of medical images. Segmenting the lungs is one of the common research topics in those studies. Our research stemmed from the lack of solutions for automatic bone, airway, and vessel segmentation, despite the existence of multiple lung segmentation techniques. Consequently, currently, available software tools used for medical image segmentation do not provide automatic lung, bone, airway, and vessel segmentation. This paper presents segmentation techniques along with an interactive software tool architecture for segmenting bone, lung, airway, and vessel tissues. Additionally, we propose a method for creating binary masks from automatically generated segments. The key contribution of our approach is the technique for automatic image thresholding using adjustable Hounsfield values and binary mask extraction. Generated binary masks can be successfully used as a training dataset for deep-learning solutions in medical image segmentation. In this paper, we also examine the current software tools used for medical image segmentation, discuss our approach, and identify its advantages.

Keywords: lung segmentation, binary masks, U-Net, medical software tools

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755 The Diagnostic Utility and Sensitivity of the Xpert® MTB/RIF Assay in Diagnosing Mycobacterium tuberculosis in Bone Marrow Aspirate Specimens

Authors: Nadhiya N. Subramony, Jenifer Vaughan, Lesley E. Scott

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In South Africa, the World Health Organisation estimated 454000 new cases of Mycobacterium tuberculosis (M.tb) infection (MTB) in 2015. Disseminated tuberculosis arises from the haematogenous spread and seeding of the bacilli in extrapulmonary sites. The gold standard for the detection of MTB in bone marrow is TB culture which has an average turnaround time of 6 weeks. Histological examinations of trephine biopsies to diagnose MTB also have a time delay owing mainly to the 5-7 day processing period prior to microscopic examination. Adding to the diagnostic delay is the non-specific nature of granulomatous inflammation which is the hallmark of MTB involvement of the bone marrow. A Ziehl-Neelson stain (which highlights acid-fast bacilli) is therefore mandatory to confirm the diagnosis but can take up to 3 days for processing and evaluation. Owing to this delay in diagnosis, many patients are lost to follow up or remain untreated whilst results are awaited, thus encouraging the spread of undiagnosed TB. The Xpert® MTB/RIF (Cepheid, Sunnyvale, CA) is the molecular test used in the South African national TB program as the initial diagnostic test for pulmonary TB. This study investigates the optimisation and performance of the Xpert® MTB/RIF on bone marrow aspirate specimens (BMA), a first since the introduction of the assay in the diagnosis of extrapulmonary TB. BMA received for immunophenotypic analysis as part of the investigation into disseminated MTB or in the evaluation of cytopenias in immunocompromised patients were used. Processing BMA on the Xpert® MTB/RIF was optimised to ensure bone marrow in EDTA and heparin did not inhibit the PCR reaction. Inactivated M.tb was spiked into the clinical bone marrow specimen and distilled water (as a control). A volume of 500mcl and an incubation time of 15 minutes with sample reagent were investigated as the processing protocol. A total of 135 BMA specimens had sufficient residual volume for Xpert® MTB/RIF testing however 22 specimens (16.3%) were not included in the final statistical analysis as an adequate trephine biopsy and/or TB culture was not available. Xpert® MTB/RIF testing was not affected by BMA material in the presence of heparin or EDTA, but the overall detection of MTB in BMA was low compared to histology and culture. Sensitivity of the Xpert® MTB/RIF compared to both histology and culture was 8.7% (95% confidence interval (CI): 1.07-28.04%) and sensitivity compared to histology only was 11.1% (95% CI: 1.38-34.7%). Specificity of the Xpert® MTB/RIF was 98.9% (95% CI: 93.9-99.7%). Although the Xpert® MTB/RIF generates a faster result than histology and TB culture and is less expensive than culture and drug susceptibility testing, the low sensitivity of the Xpert® MTB/RIF precludes its use for the diagnosis of MTB in bone marrow aspirate specimens and warrants alternative/additional testing to optimise the assay.

Keywords: bone marrow aspirate , extrapulmonary TB, low sensitivity, Xpert® MTB/RIF

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754 Conservative and Surgical Treatment of Antiresorptive Drug-Related Osteonecrosis of the Jaw with Ultrasonic Piezoelectric Bone Surgery under Polyvinylpyrrolidone Iodine Irrigation: A Case Series of 13 Treated Sites

Authors: Esra Yuce, Isil D. S. Yamaner, Murude Yazan

Abstract:

Aims and objective: Antiresorptive agents including bisphosphonates and denosumab as strong suppressors of osteoclasts are the most commonly used antiresorptive medications for the treatment of osteoporosis which counteract the negative quantitative alteration of trabecular and cortical bone by inhibition of bone turnover. Oral bisphosphonate therapy for the treatment of osteopenia, osteoporosis or Paget's disease is associated with the low-grade risk of osteonecrosis of the jaw, while higher-grade risk is associated with receiving intravenous bisphosphonates therapy in the treatment of multiple myeloma and bone metastases. On the other hand, there has been a remarkable increase in incidences of antiresorptive related osteonecrosis of the jaw (ARONJ) in oral bisphosphonate users. This clinical presentation will evaluate the healing outcomes via piezoelectric bone surgery under the irrigation of PVP-I solution irrigation in patients received bisphosphonate therapy. Material-Method: The study involved 8 female and 5 male patients that have been treated for ARONJ. Among 13 necrotic sites, 9 were in the mandible and 4 were in the maxilla. All of these 13 patients treated with surgical debridement via piezoelectric bone surgery under irrigation by solution with 3% PVP-I concentration in combination with long-term antibiotic therapy and 5 also underwent removal of mobile segments of bony sequestrum. All removable prosthesis in 8 patients were relined with soft liners during the healing periods in order to eliminate chronic minor traumas. Results: All patients were on oral bisphosphonate therapy for at least 2 years and 5 of which had received intravenous bisphosphonates up to 1 year before therapy with oral bisphosphonates was started. According to the AAOMS staging system, four cases were stage II, eight cases were stage I, and one case was stage III. The majority of lesions were identified at sites of dental prostheses (38%) and dental extractions (62%). All patients diagnosed with ARONJ stage I had used unadjusted removable prostheses. No recurrence of the symptoms was observed during the present follow-up (9–37 months). Conclusion: Despite their confirmed effectiveness, the prevention and treatment of osteonecrosis of the jaw secondary to oral bisphosphonate therapy remain major medical challenges. Treatment with piezoelectric bone surgery with irrigation of povidone-iodine solution was effective for management of bisphosphonate-related osteonecrosis of the jaw. Taking precautions for patients treated with oral bisphosphonates, especially also denture users, may allow for a reduction in the rate of developing osteonecrosis of the maxillofacial region.

Keywords: antiresorptive drug related osteonecrosis, bisphosphonate therapy, piezoelectric bone surgery, povidone iodine

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753 Biological Optimization following BM-MSC Seeding of Partially Demineralized and Partially Demineralized Laser-Perforated Structural Bone Allografts Implanted in Critical Femoral Defects

Authors: S. AliReza Mirghasemi, Zameer Hussain, Mohammad Saleh Sadeghi, Narges Rahimi Gabaran, Mohamadreza Baghaban Eslaminejad

Abstract:

Background: Despite promising results have shown by osteogenic cell-based demineralized bone matrix composites, they need to be optimized for grafts that act as structural frameworks in load-bearing defects. The purpose of this experiment is to determine the effect of bone-marrow-mesenchymal-stem-cells seeding on partially demineralized laser-perforated structural allografts that have been implanted in critical femoral defects. Materials and Methods: P3 stem cells were used for graft seeding. Laser perforation in four rows of three holes was achieved. Cell-seeded grafts were incubated for one hour until they were planted into the defect. We used four types of grafts: partially demineralized only (Donly), partially demineralized stem cell seeded (DST), partially demineralized laser-perforated (DLP), and partially demineralized laser-perforated stem cell seeded (DLPST). histologic and histomorphometric analysis were performed at 12 weeks. Results: Partially demineralized laser-perforated had the highest woven bone formation within graft limits, stem cell seeded demineralized laser-perforated remained intact, and the difference between partially demineralized only and partially demineralized stem cell seeded was insignificant. At interface, partially demineralized laser-perforated and partially demineralized only had comparable osteogenesis, but partially demineralized stem cell seeded was inferior. The interface in stem cell seeded demineralized laser-perforated was almost replaced by distinct endochondral osteogenesis with higher angiogenesis in the vicinity. Partially demineralized stem cell seeded and stem cell seeded demineralized laser-perforated graft surfaces had extra vessel-ingrowth-like porosities, a sign of delayed resorption. Conclusion: This demonstrates that simple cell-based composites are not optimal and necessitates the supplementation of synergistic stipulations and surface changes.

Keywords: structural bone allograft, partial demineralization, laser perforation, mesenchymal stem cell

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752 Viscoelastic Behavior of Human Bone Tissue under Nanoindentation Tests

Authors: Anna Makuch, Grzegorz Kokot, Konstanty Skalski, Jakub Banczorowski

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Cancellous bone is a porous composite of a hierarchical structure and anisotropic properties. The biological tissue is considered to be a viscoelastic material, but many studies based on a nanoindentation method have focused on their elasticity and microhardness. However, the response of many organic materials depends not only on the load magnitude, but also on its duration and time course. Depth Sensing Indentation (DSI) technique has been used for examination of creep in polymers, metals and composites. In the indentation tests on biological samples, the mechanical properties are most frequently determined for animal tissues (of an ox, a monkey, a pig, a rat, a mouse, a bovine). However, there are rare reports of studies of the bone viscoelastic properties on microstructural level. Various rheological models were used to describe the viscoelastic behaviours of bone, identified in the indentation process (e. g Burgers model, linear model, two-dashpot Kelvin model, Maxwell-Voigt model). The goal of the study was to determine the influence of creep effect on the mechanical properties of human cancellous bone in indentation tests. The aim of this research was also the assessment of the material properties of bone structures, having in mind the energy aspects of the curve (penetrator loading-depth) obtained in the loading/unloading cycle. There was considered how the different holding times affected the results within trabecular bone.As a result, indentation creep (CIT), hardness (HM, HIT, HV) and elasticity are obtained. Human trabecular bone samples (n=21; mean age 63±15yrs) from the femoral heads replaced during hip alloplasty were removed and drained from alcohol of 1h before the experiment. The indentation process was conducted using CSM Microhardness Tester equipped with Vickers indenter. Each sample was indented 35 times (7 times for 5 different hold times: t1=0.1s, t2=1s, t3=10s, t4=100s and t5=1000s). The indenter was advanced at a rate of 10mN/s to 500mN. There was used Oliver-Pharr method in calculation process. The increase of hold time is associated with the decrease of hardness parameters (HIT(t1)=418±34 MPa, HIT(t2)=390±50 MPa, HIT(t3)= 313±54 MPa, HIT(t4)=305±54 MPa, HIT(t5)=276±90 MPa) and elasticity (EIT(t1)=7.7±1.2 GPa, EIT(t2)=8.0±1.5 GPa, EIT(t3)=7.0±0.9 GPa, EIT(t4)=7.2±0.9 GPa, EIT(t5)=6.2±1.8 GPa) as well as with the increase of the elastic (Welastic(t1)=4.11∙10-7±4.2∙10-8Nm, Welastic(t2)= 4.12∙10-7±6.4∙10-8 Nm, Welastic(t3)=4.71∙10-7±6.0∙10-9 Nm, Welastic(t4)= 4.33∙10-7±5.5∙10-9Nm, Welastic(t5)=5.11∙10-7±7.4∙10-8Nm) and inelastic (Winelastic(t1)=1.05∙10-6±1.2∙10-7 Nm, Winelastic(t2) =1.07∙10-6±7.6∙10-8 Nm, Winelastic(t3)=1.26∙10-6±1.9∙10-7Nm, Winelastic(t4)=1.56∙10-6± 1.9∙10-7 Nm, Winelastic(t5)=1.67∙10-6±2.6∙10-7)) reaction of materials. The indentation creep increased logarithmically (R2=0.901) with increasing hold time: CIT(t1) = 0.08±0.01%, CIT(t2) = 0.7±0.1%, CIT(t3) = 3.7±0.3%, CIT(t4) = 12.2±1.5%, CIT(t5) = 13.5±3.8%. The pronounced impact of creep effect on the mechanical properties of human cancellous bone was observed in experimental studies. While the description elastic-inelastic, and thus the Oliver-Pharr method for data analysis, may apply in few limited cases, most biological tissues do not exhibit elastic-inelastic indentation responses. Viscoelastic properties of tissues may play a significant role in remodelling. The aspect is still under an analysis and numerical simulations. Acknowledgements: The presented results are part of the research project founded by National Science Centre (NCN), Poland, no.2014/15/B/ST7/03244.

Keywords: bone, creep, indentation, mechanical properties

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