Search results for: bone tissue engineering

Authors: I. Ramirez D. Edgar, J. Angeles H. José, Ruiz C. Osvaldo, H. Jacobo A. Victor, Ortiz P. Armando

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Knowledge of bone mechanical properties is important for bone substitutes design and fabrication, and more efficient prostheses development. The aim of this study is to characterize the viscoelastic behavior of bone specimens, through stress relaxation and fatigue tests performed to trabecular bone samples from bovine femoral heads. Relaxation tests consisted on preloading the samples at five different magnitudes and evaluate them for 1020 seconds, adjusting the results to a KWW mathematical model. Fatigue tests consisted of 700 load cycles and analyze their status at the end of the tests. As a conclusion we have that between relaxation stress and each preload there is linear relation and for samples with initial Young´s modulus greater than 1.5 GPa showed no effects due fatigue test loading cycles.

Keywords: bone viscoelasticity, fatigue test, stress relaxation test, trabecular bone properties

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Authors: Mehieddine Bouatrous

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Recently bioactive ceramic materials have been applied in the biomedical field as bulk, granular, or coating materials for more than half a century. More recently, bone tissue engineering scaffolds made of highly porous bioactive ceramic, glass-ceramic, and composite materials have also been created. As a result, recent bioactive ceramic structures have a high bioactivity rate, an open pores network, and good mechanical characteristics simulating cortical bone. Cyclowollastonite frameworks are also suggested for use as a graft material. As a porogenous agent, various amounts of the polymethyl methacrylate (PMMA) powders were used in this study successfully to synthesize a highly interrelated, nanostructured porous cyclowollastonite with a large specific surface area where the morphology and porosity were investigated. Porous cyclowollastonite bioactive ceramics were synthesized with a cost-effective and eco-friendly wet chemical method. The synthesized biomaterial is bioactive according to in vitro tests and can be used for bone tissue engineering scaffolds where cyclowollastonite sintered dense discs were submerged in simulated body fluid (S.B.F.) for various periods of time (1-4 weeks), resulting in the formation of a dense and consistent layer of hydroxyapatite on the surface of the ceramics, indicating its good in vitro bioactivity. Therefore, the cyclowollastonite framework exhibits good in vitro bioactivity due to its highly interconnecting porous structure and open macropores. The results demonstrate that even after soaking for several days, the surface of cyclowollastonite ceramic can generate a dense and consistent layer of hydroxyapatite. The results showed that cyclowollastonite framework exhibits good in vitro bioactivity due to highly interconnecting porous structure and open macropores.

Keywords: porous, bioactive, biomaterials, S.B.F, cyclowollastonite, biodegradability

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Authors: Pakize Neslihan Taslı, Alev Cumbul, Gul Merve Yalcın, Fikrettin Sahin

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A key experimental trial in the regeneration of large oral and craniofacial defects is the neogenesis of osseous and ligamentous interfacial structures. Currently, oral regenerative medicine strategies are unpredictable for repair of tooth supporting tissues destroyed as a consequence of trauma, chronic infection or surgical resection. A different approach combining the gel-casting method with Hydroxy Apatite HA-based scaffold and different cell lineages as a hybrid system leads to successively mimic the early stage of tooth development, in vitro. HA is widely accepted as a bioactive material for guided bone and tooth regeneration. In this study, it was reported that, HA porous scaffold preparation, characterization and evaluation of structural and chemical properties. HA is the main factor that exists in tooth and it is in harmony with structural, biological, and mechanical characteristics. Here, this study shows mimicking immature tooth at the late bell stage design and construction of HA scaffolds for cell transplantation of human Adipose Stem Cells (hASCs), human Bone Marrow Stem Cells (hBMSCs) and Gingival Epitelial cells for the formation of human tooth dentin-pulp-enamel complexes in vitro. Scaffold characterization was demonstrated by SEM, FTIR and pore size and density measurements. The biological contraction of dental tissues against each other was demonstrated by mRNA gene expressions, histopatologic observations and protein release profile by ELISA tecnique. The tooth shaped constructs with a pore size ranging from 150 to 300 µm arranged by gathering right amounts of materials provide interconnected macro-porous structure. The newly formed tissue like structures that grow and integrate within the HA designed constructs forming tooth cementum like tissue, pulp and bone structures. These findings are important as they emphasize the potential biological effect of the hybrid scaffold system. In conclusion, this in vitro study clearly demonstrates that designed 3D scaffolds shaped as a immature tooth at the late bell stage were essential to form enamel-dentin-pulp interfaces with an appropriate cell and biodegradable material combination. The biomimetic architecture achieved here is providing a promising platform for dental tissue engineering.

Keywords: tooth regeneration, tissue engineering, adipose stem cells, hydroxyapatite tooth engineering, porous scaffold

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Authors: C. Gallottini, W. Di Mari, A. Amaddeo, K. Barbaro, A. Dolci, G. Dolci, L. Gallottini, G. Barraco, S. Eramo

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A group of 10 dogs (group A) with Periodontal Disease in the third stage, were subjected to regenerative therapy of periodontal tissues, by use of nano hydroxy apatite (NHA). These animals induced by general anesthesia, where treated by ultrasonic scaling, root planning, and at the end by a mucogingival flap in which it was applied NHA. The flap was closed and sutured with simple steps. Another group of 10 dogs (group B), control group, was treated only by scaling and root planning. No patient was subjected to antibiotic therapy. After three months, a check was made by inspection of the oral cavity, radiography and bone biopsy at the alveolar level. Group A showed a total restitutio ad integrum of the periodontal structures, and in group B still mild gingivitis in 70% of cases and 30% of the state remains unchanged. Numerous experimental studies both in animals and humans have documented that the grafts of porous hydroxyapatite are rapidly invaded by fibrovascular tissue which is subsequently converted into mature lamellar bone tissue by activating osteoblast. Since we acted on the removal of necrotic cementum and rehabilitating the root tissue by polishing without intervention in the ligament but only on anatomical functional interface of cement-blasts, we can connect the positive evolution of the clinical-only component of the cement that could represent this perspective, the only reason that Periodontal Disease become a Cement Disease, while all other clinical elements as nothing more than a clinical pathological accompanying.

Keywords: nanoidroxiaphatite, parodontal disease, cement disease, regenerative therapy

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Authors: Ashok K. Gupta

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Over the past few decades, since the bio-engineering revolution, autologous cell therapy (ACT) has become a rapidly evolving field. Currently, this form of therapy has broad applications in modern medicine and plastic surgery, ranging from the treatment/improvement of wound healing to life-saving operations. A study was conducted on 50 patients having to disfigure, and deform post burn scars and was treated by injection of extracted, refined adipose tissue grafts with their unique stem cell properties. To compare the outcome, a control of 20 such patients was treated with conventional skin or soft-tissue flaps or skin grafting, and a control of 10 was treated with more advanced microsurgical techniques such as Pre-fabricated flaps/pre laminated flaps / free flaps. Assessment of fat volume and survival post- follow up period was done by radiological aid, using MRI and clinically (Survival of the autograft and objective parameters for scar elasticity were evaluated skin elasticity parameters 3 to 9 months postoperatively). Recently, an enzyme that is involved in collagen crosslinking in fibrotic tissue, lysyl hydroxylase (LH2), was identified. This enzyme is normally active in bone and cartilage but hardly in the skin. It has been found that this enzyme is highly expressed in scar tissue and subcutaneous fat; this is in contrast to the dermis, where the enzyme is hardly expressed. Adipose tissue-derived stem cell injections are an effective method in the treatment of various extensive post-burn scar deformities that makes it possible to re-create the lost sub-dermal tissue for improvement in the function of involved joint movements.

Keywords: adipose tissue-derived stem cell injections, treatment of various extensive post-burn scar deformities, re-create the lost sub-dermal tissue, improvement in function of involved joint movements

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

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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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Authors: Sadek A. Makhlouf, Hassan M. Nouh

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Bone hydatidosis is an infection in worldwide distribution. Although there is no evidence in literature on Bone Hydatid disease in Libya, we tried to present the first epidemiological study of this disease in Eastern Libya through retrospective study from 1995 to 2013. Our data were collected from 3 hospitals in Eastern Libya particularly the sheep-raising areas with total number of musculoskeletal infection cases of two thousand one hundred ninety-four (2,194). There were five (5) five cases of bone infection, four (4) of it have been diagnosed after more than three (3) months. Our study is comparable to other international study but this type of bone infection need further studies for effective control strategies for all dogs to avoid serious complications that might happened from the delay in diagnosing this type of disease.

Keywords: bone infection, hydatidosis, Eastern Libya, sheep-raising areas

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Authors: Zeynep Busra Velioglu, Deniz Pulat, Beril Demirbakan, Burak Ozcan, Ece Bayrak, Cevat Erisken

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Bone tissue has the ability to perform a wide array of functions including providing posture, load-bearing capacity, protection for the internal organs, initiating hematopoiesis, and maintaining the homeostasis of key electrolytes via calcium/phosphate ion storage. The most common cause for bone defects is extensive trauma and subsequent infection. Bone tissue has the self-healing capability without a scar tissue formation for the majority of the injuries. However, some may result with delayed union or fracture non-union. Such cases include reconstruction of large bone defects or cases of compromised regenerative process as a result of avascular necrosis and osteoporosis. Several surgical methods exist to treat bone defects, including Ilizarov method, Masquelete technique, growth factor stimulation, and bone replacement. Unfortunately, these are technically demanding and come with noteworthy disadvantages such as lengthy treatment duration, adverse effects on the patient’s psychology, repeated surgical procedures, and often long hospitalization times. These limitations associated with surgical techniques make bone substitutes an attractive alternative. Here, it was hypothesized that a 3D printed scaffold will mimic trabecular bone in terms of biomechanical properties and that such scaffolds will support cell attachment and survival. To test this hypothesis, this study aimed at fabricating poly(lactic acid), PLA, structures using 3D printing technology for trabecular bone defects, characterizing the scaffolds and comparing with bovine trabecular bone. Capacity of scaffolds on human bone marrow stem cell (hBMSC) attachment and survival was also evaluated. Cubes with a volume of 1 cm³ having pore sizes of 0.50, 1.00 and 1.25 mm were printed. The scaffolds/grafts were characterized in terms of porosity, contact angle, compressive mechanical properties as well cell response. Porosities of the 3D printed scaffolds were calculated based on apparent densities. For contact angles, 50 µl distilled water was dropped over the surface of scaffolds, and contact angles were measured using ‘Image J’ software. Mechanical characterization under compression was performed on scaffolds and native trabecular bone (bovine, 15 months) specimens using a universal testing machine at a rate of 0.5mm/min. hBMSCs were seeded onto the 3D printed scaffolds. After 3 days of incubation with fully supplemented Dulbecco’s modified Eagle’s medium, the cells were fixed using 2% formaldehyde and glutaraldehyde mixture. The specimens were then imaged under scanning electron microscopy. Cell proliferation was determined by using EZQuant dsDNA Quantitation kit. Fluorescence was measured using microplate reader Spectramax M2 at the excitation and emission wavelengths of 485nm and 535nm, respectively. Findings suggested that porosity of scaffolds with pore dimensions of 0.5mm, 1.0mm and 1.25mm were not affected by pore size, while contact angle and compressive modulus decreased with increasing pore size. Biomechanical characterization of trabecular bone yielded higher modulus values as compared to scaffolds with all pore sizes studied. Cells attached and survived in all surfaces, demonstrating higher proliferation on scaffolds with 1.25mm pores as compared with those of 1mm. Collectively, given lower mechanical properties of scaffolds as compared to native bone, and biocompatibility of the scaffolds, the 3D printed PLA scaffolds of this study appear as candidate substitutes for bone repair and regeneration.

Keywords: 3D printing, biomechanics, bone repair, stem cell

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Authors: Janis Zarins, Kristaps Blums, Oskars Radzins, Renars Deksnis, Atis Svare, Santa Salaka

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Wide tumor resection remains the first choice method for tumors of the oral cavity. Nevertheless, remained tissue defect impacts patients functional and aesthetical outcome, which could be improved using microvascular tissue transfers. Mandibular reconstruction is challenging due to the complexity of composite tissue defects and occlusal relationships for normal eating, chewing, and pain free jaw motions. Individual 3-D virtual planning would provide better symmetry and functional outcome. The main goal of preoperative planning is to develop a customized surgical approach with patient specific cutting guides of the mandible, osteotomy guides of the fibula, pre-bended osteosynthesis plates to perform more precise reconstruction, to decrease the surgery time and reach the best outcome. Our study is based on the analysis of 32 patients operated on between 2019 to 2021. All patients underwent mandible reconstruction with vascularized fibula flaps. Patients characteristics, surgery profile, survival, functional outcome, and quality of life was evaluated. Preoperative planning provided a significant decrease of surgery time and the best arrangement of bone closely similar as before the surgery. In cases of bone asymmetry, deformity and malposition, a new mandible was created using 3D planning to restore the appearance of lower jaw anatomy and functionality.

Keywords: mandibular, 3D planning, cutting guides, fibula flap, reconstruction

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Authors: Özgür Öztürk, Cumhur Sipahi, Hande Yeşil

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Implant retained restorations commonly consist of a metal-framework veneered with ceramic or composite facings. The increasing and expanding use of indirect resin composites in dentistry is a result of innovations in materials and processing techniques. Of special interest to the implant restorative field is the possibility that composites present significantly lower peak vertical and transverse forces transmitted at the peri-implant level compared to metal-ceramic supra structures in implant-supported restorations. A 43-year-old male patient referred to the department of prosthodontics for an implant retained fixed prosthesis. The clinical and radiographic examination of the patient demonstrated the presence of an implant in the right mandibular first molar tooth region. A considerable amount of marginal bone loss around the implant was detected in radiographic examinations combined with a remarkable peri-implant soft tissue deficiency. To minimize the chewing loads transmitted to the implant-bone interface it was decided to fabricate an indirect composite resin veneered single metal crown over a screw-retained abutment. At the end of the treatment, the functional and aesthetic deficiencies were fully compensated. After a 6 months clinical and radiographic follow-up period the not any additional pathologic invasion was detected in the implant-bone interface and implant retained restoration did not reveal any vehement complication.

Keywords: dental implant, fixed partial dentures, indirect composite resin, peri-implant defects

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Authors: Gulsharat A. Baigonakova, Ekaterina S. Marchenko, Venera R. Luchsheva

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The preferred materials for bone grafting are titanium-nickel alloys. They have a porous, permeable structure similar to that of bone tissue, can withstand long-term physiological stress in the body, and retain the scaffolding function for bone tissue ingrowth. Despite the excellent functional properties of these alloys, there is a possibility of post-operative infectious complications that prevent the newly formed bone tissue from filling the spaces created in a timely manner and prolong the rehabilitation period of patients. In order to minimise such consequences, it is necessary to use biocompatible materials capable of simultaneously fulfilling the function of a long-term functioning implant and an osteoreplacement carrier saturated with drugs. Methods to modify the surface by saturation with bioactive substances, in particular macrocyclic compounds, for the controlled release of drugs, biologically active substances, and cells are becoming increasingly important. This work is dedicated to the functionalisation of the surface of porous titanium nickelide by the deposition of macrocyclic compounds in order to provide titanium nickelide with antibacterial activity and accelerated osteogenesis. The paper evaluates the effect of macrocyclic compound deposition methods on the continuity, structure, and cytocompatibility of the surface properties of porous titanium nickelide. Macrocyclic compounds were deposited on the porous surface of titanium nickelide under the influence of various physical effects. Structural research methods have allowed the evaluation of the surface morphology of titanium nickelide and the nature of the distribution of these compounds. The method of surface functionalisation of titanium nickelide influences the size of the deposited bioactive molecules and the nature of their distribution. The surface functionalisation method developed has enabled titanium nickelide to be deposited uniformly on the inner and outer surfaces of the pores, which will subsequently enable the material to be uniformly saturated with various drugs, including antibiotics and inhibitors. The surface-modified porous titanium nickelide showed high biocompatibility and low cytotoxicity in in vitro studies. The research was carried out with financial support from the Russian Science Foundation under Grant No. 22-72-10037.

Keywords: biocompatibility, NiTi, surface, porous structure

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

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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

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Authors: Farhad Forouharmajd, Hossein Ebrahimi, Siamak Pourabdian

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Introduction: Prevalent use of cell phones (mobile phones) has led to increasing worries about the effect of radiofrequency waves on the physiology of human body. This study was done to determine different reactions of the temperatures in different depths of brain tissue in confronting with radiofrequency waves of cell phones. Methodology: This study was an empirical research. A cow's brain tissue was placed in a compartment and the effects of radiofrequency waves of the cell phone was analyzed during confrontation and after confrontation, in three different depths of 2, 12, and 22 mm of the tissue, in 4 mm and 4 cm distances of the tissue to a cell phone, for 15 min. Lutron thermometer was used to measure the tissue temperatures. Data analysis was done by Lutron software. Findings: The rate of increasing the temperature at the depth of 22 mm was higher than 2 mm and 12mm depths, during confrontation of the brain tissue at the distance of 4 mm with the cell phone, such that the tissue temperatures at 2, 12, and 22 mm depths increased by 0.29 ˚C, 0.31 ˚C, and 0.37 ˚C, respectively, relative to the base temperature (tissue temperature before confrontation). Moreover, the temperature of brain tissue at the distance of 4 cm by increasing the tissue depth was more than other depths. Increasing the tissue temperature also existed by increasing the brain tissue depth after the confrontation with the cell phone. The temperature of the 22 mm depth increased with higher speed at the time confrontation. Conclusion: Not only radiofrequency waves of cell phones increased the tissue temperature in all the depths of the brain tissue, but also the temperature due to radiofrequency waves of the cell phone was more at the depths higher than 22 mm of the tissue. In fact, the thermal effect of radiofrequency waves was higher in higher depths.

Keywords: mobile phone, radio frequency waves, brain tissue, temperature

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Authors: Mikaela Kutrolli, Noah S. Pereira, Vanessa Scanlon, Mohamadmahdi Samandari, Ali Tamayol

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Bone tissue engineering has drawn significant attention and various biomaterials have been tested. Polymers such as polycaprolactone (PCL) offer excellent biocompatibility, reasonable mechanical properties, and biodegradability. However, PCL scaffolds suffer a critical drawback: a lack of micro/mesoporosity, affecting cell attachment, tissue integration, and mineralization. It also results in a slow degradation rate. While 3D-printing has addressed the issue of macroporosity through CAD-guided fabrication, PCL scaffolds still exhibit poor smaller-scale porosity. To overcome this, we generated composites of PCL, hydroxyapatite (HA), and powdered sucrose (PS). The latter serves as a sacrificial material to generate porous particles after sucrose dissolution. Additionally, we have incorporated dexamethasone (DEX) to boost the PCL osteogenic properties. The resulting scaffolds maintain controlled macroporosity from the lattice print structure but also develop micro/mesoporosity within PCL fibers when exposed to aqueous environments. The study involved mixing PS into solvent-dissolved PCL in different weight ratios of PS to PCL (70:30, 50:50, and 30:70 wt%). The resulting composite was used for 3D printing of scaffolds at room temperature. Printability was optimized by adjusting pressure, speed, and layer height through filament collapse and fusion test. Enzymatic degradation, porogen leaching, and DEX release profiles were characterized. Physical properties were assessed using wettability, SEM, and micro-CT to quantify the porosity (percentage, pore size, and interconnectivity). Raman spectroscopy was used to verify the absence of sugar after leaching. Mechanical characteristics were evaluated via compression testing before and after porogen leaching. Bone marrow stromal cells (BMSCs) behavior in the printed scaffolds was studied by assessing viability, metabolic activity, osteo-differentiation, and mineralization. The scaffolds with a 70% sugar concentration exhibited superior printability and reached the highest porosity of 80%, but performed poorly during mechanical testing. A 50% PS concentration demonstrated a 70% porosity, with an average pore size of 25 µm, favoring cell attachment. No trace of sucrose was found in Raman after leaching the sugar for 8 hours. Water contact angle results show improved hydrophilicity as the sugar concentration increased, making the scaffolds more conductive to cell adhesion. The behavior of bone marrow stromal cells (BMSCs) showed positive viability and proliferation results with an increasing trend of mineralization and osteo-differentiation as the sucrose concentration increased. The addition of HA and DEX also promoted mineralization and osteo-differentiation in the cultures. The integration of PS as porogen at a concentration of 50%wt within PCL scaffolds presents a promising approach to address the poor cell attachment and tissue integration issues of PCL in bone tissue engineering. The method allows for the fabrication of scaffolds with tunable porosity and mechanical properties, suitable for various applications. The addition of HA and DEX further enhanced the scaffolds. Future studies will apply the scaffolds in an in-vivo model to thoroughly investigate their performance.

Keywords: bone, PCL, 3D printing, tissue engineering

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Authors: Ramandeep Kaur, Makula Ajitha

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Fluvastatin has been reported for increasing bone mineral density in osteoporosis since last decade. Systemically administered drug undergoes extensive hepatic first-pass metabolism, thus very small amount of drug reaches the bone tissue which is highly insignificant. The present study aims to deliver fluvastatin in the form of nanoemulsion (NE) gel directly to the bone tissue through transdermal route thereby bypassing hepatic first pass metabolism. The NE formulation consisted of isopropyl myristate as oil, tween 80 as surfactant, transcutol as co-surfactant and water as the aqueous phase. Pseudoternary phase diagrams were constructed using aqueous titration method and NE’s obtained were subjected to thermodynamic-kinetic stability studies. The stable NE formulations were evaluated for their droplet size, zeta potential, and transmission electron microscopy (TEM). The nano-sized formulations were incorporated into 0.5% carbopol 934 gel matrix. Ex-vivo permeation behaviour of selected formulations through rat skin was investigated and compared with the conventional formulations (suspension and emulsion). Further, in-vivo pharmacokinetic study was carried using male Wistar rats. The optimized NE formulations mean droplet size was 11.66±3.2 nm with polydispersity index of 0.117. Permeation flux of NE gel formulations was found significantly higher than the conventional formulations i.e. suspension and emulsion. In vivo pharmacokinetic study showed significant increase in bioavailability (1.25 fold) of fluvastatin than oral formulation. Thus, it can be concluded that NE gel was successfully developed for transdermal delivery of fluvastatin for the treatment of osteoporosis.

Keywords: fluvastatin, nanoemulsion gel, osteoporosis, transdermal

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Authors: Z. Karahaliloğlu, E. Yalçın, M. Demirbilek, E.B. Denkbaş

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Critical-sized bone defects caused by trauma, bone diseases, prosthetic implant revision or tumor excision cannot be repaired by physiological regenerative processes. Current orthopedic applications for critical-sized bone defects are to use autologous bone grafts, bone allografts, or synthetic graft materials. However, these strategies are unable to solve completely the problem, and motivate the development of novel effective biological scaffolds for tissue engineering applications and regenerative medicine applications. In particular, scaffolds combined with a variety of bio-agents as fundamental tools emerge to provide the regeneration of damaged bone tissues due to their ability to promote cell growth and function. In this study, a magnetic silk fibroin (SF) hydrogel scaffold was prepared by electrogelation process of the concentrated Bombxy mori silk fibroin (8 %wt) aqueous solution. For enhancement of osteoblast-like cells (SaOS-2) growth and adhesion, basal fibroblast growth factor (bFGF) were conjugated physically to the HSA-coated magnetic nanoparticles (Fe3O4) and magnetic SF e-gel scaffolds were prepared by incorporation of Fe3O4, HSA (human serum albumin)=Fe3O4 and HSA=Fe3O4-bFGF nanoparticles. HSA=Fe3O4, HSA=Fe3O4-bFGF loaded and bare SF e-gels scaffolds were characterized using scanning electron microscopy (SEM.) For cell studies, human osteoblast-like cell line (SaOS-2) was used and an MTT assay was used to assess the cytotoxicity of magnetic silk fibroin e-gel scaffolds and cell density on these surfaces. For the evaluation osteogenic activation, ALP (alkaline phosphatase), the amount of mineralized calcium, total protein and collagen were studied. Fe3O4 nanoparticles were successfully synthesized and bFGF was conjugated to HSA=Fe3O4 nanoparticles with %97.5 of binding yield which has a particle size of 71.52±2.3 nm. Electron microscopy images of the prepared HSA and bFGF incorporated SF e-gel scaffolds showed a 3D porous morphology. In terms of water uptake results, bFGF conjugated HSA=Fe3O4 nanoparticles has the best water absorbability behavior among all groups. In the in-vitro cell culture studies realized using SaOS-2 cell line, the coating of Fe3O4 nanoparticles surface with a protein enhance the cell viability and HSA coating and bFGF conjugation, the both have an inductive effect in the cell proliferation. One of the markers of bone formation and osteoblast differentiation, according to the ALP activity and total protein results, HSA=Fe3O4-bFGF loaded SF e-gels had significantly enhanced ALP activity. Osteoblast cultured HSA=Fe3O4-bFGF loaded SF e-gels deposited more calcium compared with SF e-gel. The proposed magnetic scaffolds seem to be promising for bone tissue regeneration and used in future work for various applications.

Keywords: basic fibroblast growth factor (bFGF), e-gel, iron oxide nanoparticles, silk fibroin

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Authors: R. Janyaphadungpong, A. Pimkhaokham

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This prospective clinical study determined the insertion torque (IT) value and monitored the changes in implant stability quotient (ISQ) values during the 12 weeks healing period from implant placement without bone graft (control group) and with bone graft using the guided bone regeneration (GBR) technique (study group). The relationship between the IT and ISQ values of the implants was also assessed. The control and study groups each consisted of 6 patients with 8 implants per group. The ASTRA TECH Implant System™ EV 4.2 mm in diameter was placed in the posterior mandibular region. In the control group, implants were placed in bone without bone graft, whereas in the study group implants were placed simultaneously with the GBR technique at favorable bone defect. IT (Ncm) of each implant was recorded when fully inserted. ISQ values were obtained from the Osstell® ISQ at the time of implant placement, and at 2, 4, 8, and 12 weeks. No difference in IT was found between groups (P = 0.320). The ISQ values in the control group were significantly higher than in the study group at the time of implant placement and at 4 weeks. There was no significant association between IT and ISQ values either at baseline or after the 12 weeks. At 12 weeks of healing, the control and study groups displayed different trends. Mean ISQ values for the control group decreased over the first 2 weeks and then started to increase. ISQ value increases were statistically significant at 8 weeks and later, whereas mean ISQ values in the study group decreased over the first 4 weeks and then started to increase, with statistical significance after 12 weeks. At 12 weeks, all implants achieved osseointegration with mean ISQ values over the threshold value (ISQ>70). These results indicated that implants, in which guided bone regeneration technique was performed during implant placement for treating favorable bone defects, were as predictable as implants placed without bone graft. However, loading in implants placed with the GBR technique for correcting favorable bone defects should be performed after 12 weeks of healing to ensure implant stability and osseointegration.

Keywords: dental implant, favorable bone defect, guided bone regeneration technique, implant stability

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Authors: Djamel Remache, Marie Semaan, Cécile Baron, Martine Pithioux, Patrick Chabrand, Jean-Marie Rossi, Jean-Louis Milan

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Cortical bone is a complex multi-scale structure. Even though several works have contributed significantly to understanding its mechanical behavior, this behavior remains poorly understood. Nanoindentation testing is one of the primary testing techniques for the mechanical characterization of bone at small scales. The purpose of this study was to provide new nanoindentation data of cortical bovine bone in different directions and at different bone microstructures (osteonal, interstitial and laminar bone), and then to identify anisotropic properties of samples with FEM (finite element method) based inverse method. Experimentally and numerical results were compared. Experimental and numerical results were compared. The results compared were in good agreement.

Keywords: mechanical behavior of bone, nanoindentation, finite element analysis, inverse optimization approach

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Authors: Ibrahima Farikou, Kolontchang Gatchou Alberic Lionel, Tsiagadgui Jean Gustave, Ngo Yamben Marie-Ange, Handy Eone Daniel

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Introduction: It was in 1956 that Ilizarov pioneered the concept of osteogenesis in distraction by the device that bears his name to help produce bone and soft tissue regeneration and bone consolidation. This technique is not widely used in Africa where, however, its applications are numerous (loss of bone substances, congenital or acquired malformations). Our goal is to bring the indications of Ilizarov's device back to our practice conditions. Methods: Our study was conducted in 2 hospitals over a period of 20 years. For the retrospective phase, this study included all complete usable records of patients operated on in the Ilizarov external fixator department, and for the prospective phase, all patients operated on in the departments with complete usable records. Our sample was consecutive and not exhaustive. Data were analyzed by SPSS software version 23.0. Results: A total of 52 patients were reviewed. The average age of our patients was 14.7 years. The sex ratio was 1.6 in favor of men. The lower limb was the most affected (49), with a predominance of the tibia (62.4%). The average elongation was 6.4 cm. Traumatic acquired pathologies (delayed union, malunion) represented 60.6%. The mean time to union was seven months. Correction of the limb length discrepancy or filling of loss of bone substance was obtained in 75% of cases. Functionally, 80.8% of the patients treated had regained autonomy at the end of treatment, but in 17.3% of the patients, pain and limping persisted. Conclusion: This technique should be popularized in Africa because the benefit that would accrue to patients is invaluable and would be an attractive alternative to many amputations sometimes carried out in Africa by despair.

Keywords: ilizarov, external fixator, limb lengthening, bone regeneration, africa

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Authors: Jui-Ting Hsu, Heng-Li Huang, Ming-Tzu Tsai, Kuo-Chih Su, Lih-Jyh Fuh

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The objectives of this study were to measure the initial stability of the dental implant (ISQ and PTV) in the artificial foam bone block with three different quality levels. In addition, the 3D bone to implant contact percentage (BIC%) was measured based on the micro-computed tomography images. Furthermore, the relation between the initial stability of dental implant (ISQ and PTV) and BIC% were calculated. The experimental results indicated that enhanced the material property of the artificial foam bone increased the initial stability of the dental implant. The Pearson’s correlation coefficient between the BIC% and the two approaches (ISQ and PTV) were 0.652 and 0.745.

Keywords: dental implant, implant stability quotient, peak insertion torque, bone-implant contact, micro-computed tomography

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Authors: Changsoo Chon, Sangkuy Han, Donghyun Seo, Jihyung Park, Bokku Kang, Hansung Kim, Keyoungjin Chun, Cheolwoong Ko

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The purpose of the study is to develop a finite element model based on 3D bone structural images of Micro-CT and to analyze the stress distribution for the osteoporosis mouse femora. In this study, results of finite element analysis show that the early osteoporosis of mouse model decreased a bone density in trabecular region; however, the bone density in cortical region increased.

Keywords: micro-CT, finite element analysis, osteoporosis, bone strength

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Authors: Syazwani Ramli, Norhidayu Muhamad Zain

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Nowadays, the use of gelatin as biomaterials in tissue engineering are evolving especially in skin graft and wound dressing applications. Towards year 2018, Malaysia is in the way of planning to get the halal certification for biomedical device in order to cater the needs of Muslims and non-Muslims in Malaysia. However, the use of gelatins in tissue engineering are mostly derived from non-halal sources. Currently, gelatin production mostly comes from mammalian gelatin sources. Moreover, within these past years, just a few studies of the uses of gelatin in tissue engineering from halal perspective has been studied. Thus, this paper aims to give overview of the use of gelatin from different sources from halal perspectives. This review also discussing the current status of halal for the emerging biomedical devices. In addition, the different sources of gelatin used in tissue engineering are being identified and provides better alternatives for halal gelatin. Cold- water fish skin gelatin could be an effective alternative to substitute the mammalian sources. Therefore, this review is important because the information about the halal biomedical devices will delighted Muslim consumers and give better insight of halal gelatin in tissue engineering application.

Keywords: biomedical device, gelatin, halal, skin graft, tissue engineering

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Authors: Getenet Aseged Zeleke

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The aim of this project was to study the synthesis of activated carbon from low-cost animal beef and the characterization of the product obtained. The bone was carbonized in an inert atmosphere at three different temperatures (500°C, 700oC and 900°C) in an electric furnace, followed by activation with hydrochloric acid. The activated animal bone charcoals obtained were characterized by using scanning electron microscopy (SEM)to observe the effect of activation compared to the unactivated bone charcoal. The following parameters were also determined: ash content, moisture content, volatile content, fixed carbon, pH, pore volume and bulk (apparent) density. The characterization result showed that the activated bone charcoal has good properties and is compared favorably with other reference activated carbons.

Keywords: bones, carbonization, activation, characterization, activated carbon

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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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Authors: Jung Park, Anca Mazare, Klaus Von Der Mark, Patrik Schmuki

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In clinical application of TiO2 implants on tooth and hip replacement, migration, adhesion and differentiation of neighboring mesenchymal stem cells onto implant surfaces are critical steps for successful bone regeneration. In a recent decade, accumulated attention has been paid on nanoscale electrochemical surface modifications on TiO2 layer for improving bone-TiO2 surface integration. We generated, on titanium surfaces, self-assembled layers of vertically oriented TiO2 nanotubes with defined diameters between 15 and 100 nm and here we show that mesenchymal stem cells finely sense TiO2 nanotubular geometry and quickly decide their cell fate either to differentiation into osteoblasts or to programmed cell death (apoptosis) on TiO2 nanotube layers. These cell fate decisions are critically dependent on nanotube size differences (15-100nm in diameters) of TiO2 nanotubes sensing by integrin clustering. We further demonstrate that nanoscale topography-sensing is feasible not only in mesenchymal stem cells but rather seems as generalized nanoscale microenvironment-cell interaction mechanism in several cell types composing bone tissue network including osteoblasts, osteoclast, endothelial cells and hematopoietic stem cells. Additionally we discuss the synergistic effect of simultaneous stimulation by nanotube-bound growth factor and nanoscale topographic cues on enhanced bone regeneration.

Keywords: TiO2 nanotube, stem cell fate decision, nano-scale microenvironment, bone regeneration

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Authors: D. Ficai, A. Ficai, D. Gudovan, I. A. Gudovan, I. Ardelean, R. Trusca, E. Andronescu, V. Mitran, A. Cimpean

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In the last years, scientists struggled hardly to mimic bone structures to develop implants and biostructures which present higher biocompatibility and reduced rejection rate. One way to obtain this goal is to use similar materials as that of bone, namely collagen/hydroxyapatite composite materials. However, it is very important to tailor both compositions but also the microstructure of the bone that would ensure both the optimal osteointegartion and the mechanical properties required by the application. In this study, new collagen/hydroxyapatites composite materials doped with Cu, Li, Mn, Zn were successfully prepared. The synthesis method is described below: weight the Ca(OH)₂ mass, i.e., 7,3067g, and ZnCl₂ (0.134g), CuSO₄ (0.159g), LiCO₃ (0.133g), MnCl₂.4H₂O (0.1971g), and suspend in 100ml distilled water under magnetic stirring. The solution thus obtained is added a solution of NaH₂PO₄*H2O (8.247g dissolved in 50ml distilled water) under slow dropping of 1 ml/min followed by adjusting the pH to 9.5 with HCl and finally filter and wash until neutral pH. The as-obtained slurry was dried in the oven at 80°C and then calcined at 600°C in order to ensure a proper purification of the final product of organic phases, also inducing a proper sterilization of the mixture before insertion into the collagen matrix. The collagen/hydroxyapatite composite materials are tailored from morphological point of view to optimize their biocompatibility and bio-integration against mechanical properties whereas the addition of the dopants is aimed to improve the biological activity of the samples. The addition of transitional metals can improve the biocompatibility and especially the osteoblasts adhesion (Mn²⁺) or to induce slightly better osteoblast differentiation of the osteoblast, Zn²⁺ being a cofactor for many enzymes including those responsible for cell differentiation. If the amount is too high, the final material can become toxic and lose all of its biocompatibility. In order to achieve a good biocompatibility and not reach the cytotoxic effect, the amount of transitional metals added has to be maintained at low levels (0.5% molar). The amount of transitional metals entering into the elemental cell of HA will be verified using inductively-coupled plasma mass spectrometric system. This highly sensitive technique is necessary, because, at such low levels of transitional metals, the difference between biocompatible and cytotoxic is a very thin line, thus requiring proper and thorough investigation using a precise technique. In order to determine the structure and morphology of the obtained composite materials, IR spectroscopy, X-Ray diffraction (XRD), scanning electron microscopy (SEM), and Energy Dispersive X-Ray Spectrometry (EDS) were used. Acknowledgment: The present work was possible due to the EU-funding grant POSCCE-A2O2.2.1-2013-1, Project No. 638/12.03.2014, code SMIS-CSNR 48652. The financial contribution received from the national project “Biomimetic porous structures obtained by 3D printing developed for bone tissue engineering (BIOGRAFTPRINT), No. 127PED/2017 is also highly acknowledged.

Keywords: collagen, composite materials, hydroxyapatite, bone tissue engineering

Procedia PDF Downloads 169

Authors: K. Mardaleishvili, G. Loladze, G. Shatirishivili, D. Chakhunashvili, A. Vishnevskaya, Z. Kakabadze

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Benign osteoblastoma is a benign tumor of the bone, usually affecting the vertebrae and long tubular bones. It is a rarely seen tumor of the facial bones. The authors present a case of a 28-year-old male patient with a tumor in mandibular body. The lesion was radically resected and histological analysis of the specimen demonstrated features typical of a benign osteoblastoma. The defect of the jaw was reconstructed with titanium implants and decellularized and lyophilized cattle bone matrix with mesenchymal bone marrow stem cells transplantation. This presentation describes the procedures for rehabilitating a patient with decellularized bone scaffold in the region of the face, recovering the facial contours and esthetics of the patient.

Keywords: facial bones, osteoblastoma, stem cells, transplantation

Procedia PDF Downloads 394

Authors: I. O. Oladele, J. A. Omotoyinbo, A. M. Okoro, A. G. Okikiola, J. L. Olajide

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This work was carried out to investigate comparatively the effects of modified and unmodified cow bone particles on the mechanical properties of polyester matrix composites in order to investigate the suitability of the materials as biomaterial. Cow bones were procured from an abattoir, sun dried for 4 weeks and crushed. The crushed bones were divided into two, where one part was turned to ash while the other part was pulverized with laboratory ball mill before the two grades were sieved using 75 µm sieve size. Bone ash and bone particle reinforced tensile and flexural composite samples were developed from pre-determined proportions of 2, 4, 6, and 8 %. The samples after curing were stripped from the moulds and were allowed to further cure for 3 weeks before tensile and flexural tests were performed on them. The tensile test result showed that, 8 wt % bone particle reinforced polyester composites has higher tensile properties except for modulus of elasticity where 8 wt % bone ash particle reinforced composites has higher value while for flexural test, bone ash particle reinforced composites demonstrate the best flexural properties. The results show that these materials are structurally compatible.

Keywords: biomedical, composites, cow bone, mechanical properties, polyester, reinforcement

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Authors: Ajay R, Rammohan B, Sridhar K S S, Gurusharan N

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The objective of the work is to develop a numerical method to solve the inverse mode shape problem by determining the cross-sectional area of a structure for the desired mode shape via the vibration response study of the humerus bone, which is in the form of a cantilever beam with anisotropic material properties. The humerus bone is the long bone in the arm that connects the shoulder to the elbow. The mode shape is assumed to be a higher-order polynomial satisfying a prescribed set of boundary conditions to converge the numerical algorithm. The natural frequency and the mode shapes are calculated for different boundary conditions to find the cross-sectional area of humerus bone from Eigenmode shape with the aid of the inverse mode shape algorithm. The cross-sectional area of humerus bone validates the mode shapes of specific boundary conditions. The numerical method to solve the inverse mode shape problem is validated in the biomedical application by finding the cross-sectional area of a humerus bone in the human arm.

Keywords: Cross-sectional area, Humerus bone, Inverse mode shape problem, Mode shape

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Authors: Yu Luan

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The bone-conducted objective occlusion effect (OE) is characterized by a discomforting sensation of fullness experienced in an occluded ear. This phenomenon arises from various external stimuli, such as human speech, chewing, and walking, which generate vibrations transmitted through the body to the ear canal walls. The bone-conducted OE occurs due to the pressure build-up inside the occluded ear caused by sound radiating into the ear canal cavity from its walls. In the hearing aid industry, artificial ears are utilized as a tool for developing hearing aids. However, the currently available commercial artificial ears primarily focus on pure acoustics measurements, neglecting the bone-conducted vibration aspect. This research endeavors to develop an artificial ear specifically designed for bone-conducted occlusion measurements. Finite element analysis (FEA) modeling has been employed to gain insights into the behavior of the artificial ear.

Keywords: artificial ear, bone conducted vibration, occlusion measurement, finite element modeling

Procedia PDF Downloads 52