Search results for: bone scaffold

Authors: S. A. Hashemvarzi, A. Heidarianpour, Z. Fallahmohammadi, M. Pourghasem, M. Kaviani

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Introduction: Parkinson’s disease (PD) is a progressive neurodegenerative in the central nervous system characterized by the loss of dopaminergic neurons in the substantia nigra resulting in loss of dopamine release in the striatum. Non-drug treatment options such as Stem cell transplantation and exercise have been considered for treatment of Parkinson's disease. Purpose: The purpose of this study was to evaluate the effect of aerobic exercise after bone marrow stem cells transplantation on recovery of dopaminergic neurons and promotion of angiogenesis markers in the striatum of parkinsonian rats. Materials and Methods: 42 male Wistar rats were divided randomly into six groups: Normal (N), Sham (S), Parkinson’s (P), Stem cells transplanted Parkinson’s (SP), Exercised Parkinson’s (EP) and Stem cells transplanted + Exercised Parkinson’s (SEP). To create a model of Parkinson's, the striatum was destroyed by injection of 6-hydroxy-dopamine into the striatum through stereotaxic apparatus. Stem cells were derived from the bone marrow of femur and tibia of male rats with 6-8 weeks old. After cultivation, approximately 5×105 cells in 5 microliter of medium were injected into the striatum of rats through the channel. Aerobic exercise was included 8 weeks of running on the treadmill with a speed of 15 meters per minute. At the end, all subjects were decapitated and striatum tissues were separately isolated for measurement of vascular endothelial growth factor (VEGF), dopamine (DA) and tyrosine hydroxylase (TH) levels. Results: VEGF, DA and TH levels in the striatum of parkinsonian rats significantly increased in treatment groups (SP, EP and SEP), especially in SEP group compared to P group after treatment (P<0.05). Conclusion: The findings implicate that the BMSCs transplantation in combination with exercise would have synergistic effects leading to functional recovery, dopaminergic neurons recovery and promotion of angiogenesis marker in the striatum of parkinsonian rats.

Keywords: stem cells, treadmill training, neurotrophic factors, Parkinson

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Authors: Udoaka A. I., Didia B. C.

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Background: The maturation of a child’s bones as it grows to adulthood can be viewed radiologically. The skeletal age (bone age) is the average age at which a particular stage of bone maturation is achieved. The Greulich and Pyle standard is the commonest method used to assess the skeletal age using the hand and wrist radiograph throughout the world. This atlas was compiled solely from Caucasian children and made use of the orderly sequence of carpal ossification to determine the skeletal age. Several authors have faulted this atlas for not being suitable for other races. Aim: The aim of this study is to determine if the Greulich and Pyle Atlas is applicable to Nigerian children when compared to their chronological ages. Methods: The total number of 78 normal radiographs of the hand and wrist of Nigerian children obtained from several hospitals were used for this study . These radiographs were compared with the atlas and their skeletal ages noted form the atlas. The child’s chronological age in each case was also recorded. Results: The result shows a mean increase of two months in the skeletal ages of the Nigerian children compared to the atlas. This difference, however, was not significant. The skeletal age (in months) was greater in 77% of the children than the expected age in the atlas. Conclusion: The mean skeletal age of Nigerian children, though more than the standard in the atlas, is not statistically significant; as a result the study finds the radiographic atlas of Greulich and Pyle atlas applicable to Nigerian children.

Keywords: Greulich and Pyle Atlas, radiograph, skeletal age

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

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Background: The lunate bone dislocation can lead to the compression of the median nerve and subsequent carpal tunnel syndrome. The dislocation can interrupt the vasculature and would cause avascular necrosis. The objective of the present study was to study the morphology and number of the nutrient foramina in the cadaveric dried lunate bones of the Indian population. Methods: The present study included 28 lunate bones (13 right sided and 15 left sided) which were obtained from the gross anatomy laboratory of our institution. The bones were macroscopically observed for the nutrient foramina and the data was collected with respect to their number. The tabulation of the data and analysis were done. Results: All of our specimens (100%) exhibited the nutrient foramina over the non-articular surfaces. The foramina were observed only over the palmar and dorsal surfaces of the lunate bones. The foramen ranged between 2 and 10. The foramina were more in number over the dorsal surface (average number 3.3) in comparison to the palmar surface (average number 2.4). Conclusion: We believe that the present study has provided important data about the nutrient foramina of the lunate bones. The data is enlightening to the orthopedic surgeon and would help in the hand surgeries. The morphological knowledge of the vasculature, their foramina of entry and their number is required to understand the concepts in the lunatomalacia and Kienbock’s disease.

Keywords: avascular necrosis, foramen, lunate, nutrient

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Authors: Ching-Yuan Chen, Lung-Kwang Pan

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Objective: Back pain is a major health problem costing billions of health budgets annually in Taiwan. Thousands of back pain surgeries are performed annually with up to 40% of patients complaining of back pain at time of post-surgery causing failed back surgery syndrome (FBSS), although diagnosis in these patients may be complex. The aim of study is to assess the feasibility of using bone SPECT-CT imaging to localize the active lesions causing persistent, recurrent or new backache after spine surgery. Materials and Methods: Bone SPECT-CT imaging was performed after the intravenous injection of 20 mCi of 99mTc-MDP for all the patients with diagnosis of FBSS. Patients were evaluated using status of subjectively pain relief, functional improvement and degree of satisfaction by reviewing the medical records and questionnaires in a 2 more years’ follow-up. Results: We enrolled a total of 16 patients were surveyed in our hospital from Jan. 2015 to Dec. 2016. Four people on SPEC/CT imaging ensured significant lesions were undergone a revised surgery (surgical treatment group). The mean visual analogue scale (VAS) decreased 5.3 points and mean Oswestry disability index (ODI) improved 38 points in the surgical group. The remaining 12 on SPECT/CT imaging were diagnosed as no significant lesions then received drug treatment (medical treatment group). The mean VAS only decreased 2 .1 point and mean ODI improved 12.6 points in the medical treatment group. In the posttherapeutic evaluation, the pain of the surgical treatment group showed a satisfactory improvement. In the medical treatment group, 10 of the 12 were also satisfied with the symptom relief while the other 2 did not improve significantly. Conclusions: Findings on SPECT-CT imaging appears to be easily explained the patients' pain. We recommended that SPECT/CT imaging was a feasible and useful clinical tool to improve diagnostic confidence or specificity when evaluating patients with FBSS.

Keywords: failed back surgery syndrome, oswestry disability index, SPECT-CT imaging, 99mTc-MDP, visual analogue scale

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Authors: Pármenas Costa Macedo do Nascimento

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Periodontal disease is the most common disease in the oral cavity of felines. It starts with the accumulation of bacteria on the tooth surface supporting the tissues of the periodontal tissue, namely gums, alveolar bone, cementum, and periodontal ligament. The main clinical symptom observed by the owner is bad breath, which may lead to local and systemic consequences depending on the stage of periodontal disease, such as bleeding and bone loss. Therefore, the study is important to educate tutors to take better care of the felines oral health in order to try to prevent the disease. For this epidemiological study, the target population has been felines, located on the outskirts of Manaus, in the state of Amazonas, with a geographic area of 155.68 km², with no defined breed, from October 1st to 10th, 2021, whose samples has been randomly selected, with a detailed profile. The variables of interest for this study have been: absence or presence of periodontal disease, gender, age (delimited by age group), and condition (domiciled or homeless). Using a sample of 40 felines from 4 districts of the east side of Manaus chosen at random, an oral exam has been made to identify the studied disease. The animal's apparent age, condition, sex, and presence or absence of periodontal disease has been noted. It has been observed that 70% (28/40) of them had periodontal disease, mostly females, aged between 0 and 5 years and domiciled, totaling 30% (12/40).

Keywords: felines, oral cavity, oral exam, periodontal disease

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Authors: Yuncang Li, Cuie Wen

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Biodegradable metallic materials such as magnesium (Mg)-based alloys have attracted extensive interest for use as bone implant materials. However, the high biodegradation rate of existing Mg alloys in the physiological environment of human body leads to losing mechanical integrity before adequate bone healing and producing a large volume of hydrogen gas. Therefore, slowing down the biodegradation rate of Mg alloys is a critical task in developing new biodegradable Mg alloy implant materials. One of the most effective approaches to achieve this is to strategically design new Mg alloys with low biodegradation rate, excellent biocompatibility, and enhanced mechanical properties. Our research selected biocompatible and biofunctional alloying elements such as zirconium (Zr), strontium (Sr), and rare earth elements (REEs) to alloy Mg and has developed a new series of Mg-Zr-Sr-REEs alloys for biodegradable implant applications. Research results indicated that Sr and Zr additions could refine the grain size, decrease the biodegradation rate, and enhance the biological behaviors of the Mg alloys. The REE addition, such as holmium (Ho) and dysprosium (Dy) to Mg-Zr-Sr alloys resulted in enhanced mechanical strength and decreased biodegradation rate. In addition, Ho and Dy additions (≤ 5 wt.%) to Mg-Zr-Sr alloys led to enhancement of cell adhesion and proliferation of osteoblast cells on the Mg-Zr-Sr-Ho/Dy alloys.

Keywords: biocompatibility, magnesium, mechanical and biodegrade properties, rare earth elements

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Authors: Muhammad Saleem

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

Keywords: 1. health and physical education risk factors male athlete associated with the male athlete traid in young elite athlete from pakistan., 2. sports sciences pakistan, 3. risk factors sports sciences pakistan, 4. triad and young elite athlete from pakistan

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Authors: Harry Burton, Alexios Dimitrios Iliadis, Neil Jones, Aaron Saini, Nicola Bystrzonowski, Alexandros Vris, Georgios Pafitanis

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This article portrays the authors’ experience with a complex lower limb bone and soft tissue defect, following chronic osteomyelitis and pathological fracture, which was managed by the multidisciplinary orthoplastic team. The decision for functional amputation versus limb salvage was deemed necessary, enhanced by the principles of “spares parts” in reconstructive microsurgery. This case describes a successful use of the osteocutaneous distal tibia turn-over fillet flap that allowed ‘lowering the level of the amputation’ from a through knee to the conventional level of a below-knee amputation to preserve the knee joint function. This case demonstrates the value of ‘spare-parts’ surgery principles and how these concepts refine complex orthoplastic approaches when limb salvage is not possible to enhance function. The osteocutaneous distal tibia turn-over fillet flap is a robust technique for modified BKA reconstructions that provides sufficient bone length to achieve a tough, sensate stump and functional knee joint.

Keywords: osteocutaneous flap, fillet flap, spare-parts surgery, Below knee amputation

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Authors: Awf A. Al-Khan, Michael J. Day, Judith Nimmo, Mourad Tayebi, Stewart D. Ryan, Samantha J. Richardson, Janine A. Danks

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Osteosarcoma (OS) is the most common type of malignant primary bone tumour in dogs. In addition to their critical roles in bone formation and remodeling, parathyroid hormone-related protein (PTHrP) and its receptor (PTHR1) are involved in progression and metastasis of many types of tumours in humans. The aims of this study were to determine the localisation and expression levels of PTHrP and PTHR1 in canine OS tissues using immunohistochemistry and to investigate if this expression is correlated with survival time. Formalin-fixed, paraffin-embedded tissue samples from 44 dogs with known survival time that had been diagnosed with primary osteosarcoma were analysed for localisation of PTHrP and PTHR1. Findings showed that both PTHrP and PTHR1 were present in all OS samples. The dogs with high level of PTHR1 protein (16%) had decreased survival time (P<0.05) compared to dogs with less PTHR1 protein. PTHrP levels did not correlate with survival time (P>0.05). The results of this study indicate that the PTHR1 is expressed differently in canine OS tissues and this may be correlated with poor prognosis. This may mean that PTHR1 may be useful as a prognostic indicator in canine OS and could represent a good therapeutic target in OS.

Keywords: dog, expression, osteosarcoma, parathyroid hormone receptor 1 (PTHR1), parathyroid hormone-related protein (PTHrP), survival

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Authors: Yung-Shan Lu, Chia-Fone Lee, Shang-Hsuan Li, Chien-Hao Liu

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Recently, various bioelectronic devices have been developed for neurologic disease treatments via electro-stimulations such as cochlear implants and retinal prosthesis. Since the electric signal needs electrodes to be transmitted to an organism, electrodes play an important role of stimulations. The materials of stimulation electrodes affect the efficiency of the delivered currents. The higher the efficiency of the electrodes, the lower the threshold current can be used to stimulate the organism which minimizes the potential damages to the adjacent tissues. In this study, we proposed a biocompatible composite electrode composed of high-charge-capacity iridium oxide (IrOₓ) film for a bone-guide extra-cochlear implant. IrOₓ was exploited to decrease the threshold current due to its high capacitance and low impedance. The IrOₓ electrode was fabricated via microelectromechanical systems (MEMS) photolithography and examined with in-vivo tests with guinea pigs. Based on the measured responses of brain waves to sound, the results demonstrated that IrOₓ electrodes have a lower threshold current compared with the Platinum (Pt) electrodes. The research results are expected to be beneficial for implantable and biocompatible electrodes for electrical stimulations.

Keywords: cochlear implants, electrode, electrical stimulation, iridium oxide

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Authors: H. Yousefnia, S. Zolghadri

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The measurement of organ radiation exposure dose is one of the most important steps to be taken initially, for developing a new radiopharmaceutical. In this study, the dosimetric studies of a novel agent for SPECT-imaging of the bone metastasis, 111In-1,4,7,10-tetraazacyclododecane-1,4,7,10 tetraethylene phosphonic acid (111In-DOTMP) complex, have been carried out to estimate the dose in human organs based on the data derived from rats. The radiolabeled complex was prepared with high radiochemical purity in the optimal conditions. Biodistribution studies of the complex was investigated in the male Syrian rats at selected times after injection (2, 4, 24 and 48 h). The human absorbed dose estimation of the complex was made based on data derived from the rats by the radiation absorbed dose assessment resource (RADAR) method. 111In-DOTMP complex was prepared with high radiochemical purity of >99% (ITLC). Total body effective absorbed dose for 111In-DOTMP was 0.061 mSv/MBq. This value is comparable to the other 111In clinically used complexes. The results show that the dose with respect to the critical organs is satisfactory within the acceptable range for diagnostic nuclear medicine procedures. Generally, 111In-DOTMP has interesting characteristics and can be considered as a viable agent for SPECT-imaging of the bone metastasis in the near future.

Keywords: In-111, DOTMP, Internal Dosimetry, RADAR

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Authors: Maria Angelica Trak-Fellermeier, Alison K. Macchi, Rodolfo Galvan, Yolangel Hernandez, Thresia Gambon, Rebeca Martinez, Cristina Palacios

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Poor lifestyle habits, vitamin D deficiency, and inadequate calcium intake, particularly during the COVID-19 pandemic, may contribute to severe osteopenia in childhood, increasing future fractures and osteoporosis risk. We here present a case of osteopenia in a 13-year-old white, Hispanic, premenarchal girl who completed the baseline visit of the MetA-Bone Trial during the COVID-19 pandemic. The premenarchal girl has a family history of osteoporosis (maternal grandfather) but no previous fractures; moderate outdoor activity was <1 hour/day 3 times/week with 8 hours/day of sleep. Consumption of dairy products and vegetables was <1 serving/day. Lab blood tests confirmed vitamin D deficiency (serum 25(OH)D: 9 ng/L) and hyperphosphatemia (5.2 mg/dL); other tests were normal. DXA scan Z‐score was ‐2.2 SD (indicative of osteopenia by age and sex). The premenarchal girl was referred to a pediatrician, who confirmed the results, and prescribed a daily supplement with 2000 IU of vitamin D and 1000 mg of calcium. Seclusion during the COVID-19 pandemic may have contributed to the severity of the findings. Therefore, we recommend screening children undergoing growth spurts for vitamin D, calcium, and poor lifestyle habits during and after the pandemic.

Keywords: bone mass, vitamin D, puberty, Hispanic

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Authors: Krzysztof J. Czarnocki, E. Czarnocka, K. Szaniawska

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Main goal of the research project is Scaffold Use Risk Assessment Model (SURAM) formulation, developed for the assessment of risk levels as a various construction process stages with various work trades. Finally, in 2016, the project received financing by the National Center for Research and development according to PBS3/A2/19/2015–Research Grant. The presented data, calculations and analyzes discussed in this paper were created as a result of the completion on the first and second phase of the PBS3/A2/19/2015 project. Method: One of the arms of the research project is the assessment of worker visual concentration on the sight zones as well as risky visual point inadequate observation. In this part of research, the mobile eye-tracker was used to monitor the worker observation zones. SMI Eye Tracking Glasses is a tool, which allows us to analyze in real time and place where our eyesight is concentrated on and consequently build the map of worker's eyesight concentration during a shift. While the project is still running, currently 64 construction sites have been examined, and more than 600 workers took part in the experiment including monitoring of typical parameters of the work regimen, workload, microclimate, sound vibration, etc. Full equipment can also be useful in more advanced analyses. Because of that technology we have verified not only main focus of workers eyes during work on or next to scaffolding, but we have also examined which changes in the surrounding environment during their shift influenced their concentration. In the result of this study it has been proven that only up to 45.75% of the shift time, workers’ eye concentration was on one of three work-related areas. Workers seem to be distracted by noisy vehicles or people nearby. In opposite to our initial assumptions and other authors’ findings, we observed that the reflective parts of the scaffoldings were not more recognized by workers in their direct workplaces. We have noticed that the red curbs were the only well recognized part on a very few scaffoldings. Surprisingly on numbers of samples, we have not recognized any significant number of concentrations on those curbs. Conclusion: We have found the eye-tracking method useful for the construction of the SURAM model in the risk perception and worker’s behavior sub-modules. We also have found that the initial worker's stress and work visual conditions seem to be more predictive for assessment of the risky developing situation or an accident than other parameters relating to a work environment.

Keywords: accident assessment model, eye tracking, occupational safety, scaffolding

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Authors: Neeraj Malhotra

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Background: Bioreactors in tissue engineering are used as devices that apply mechanical means to influence biological processes. They are commonly employed for stem cell culturing, growth and expansion as well as in 3D tissue culture. Contemporarily there use is well established and is tested extensively in the medical sciences, for tissue-regeneration and tissue engineering of organs like bone, cartilage, blood vessels, skin grafts, cardiac muscle etc. Methodology: Literature search, both electronic and hand search, was done using the following MeSH and keywords: bioreactors, bioreactors and dentistry, bioreactors & dental tissue engineering, bioreactors and regenerative dentistry. Articles published only in English language were included for review. Results: Bioreactors like, spinner flask-, rotating wall-, flow perfusion-, and micro-bioreactors and in-vivo bioreactor have been employed and tested for the regeneration of dental and like-tissues. These include gingival tissue, periodontal ligament, alveolar bone, mucosa, cementum and blood vessels. Based on their working dynamics they can be customized in future for regeneration of pulp tissue and whole tooth regeneration. Apart from this, they have been successfully used in testing the clinical efficacy and biological safety of dental biomaterials. Conclusion: Bioreactors have potential use in testing dental biomaterials and tissue engineering approaches aimed at regenerative dentistry.

Keywords: bioreactors, biological process, mechanical stimulation, regenerative dentistry, stem cells

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Authors: Azieva A. M., Yastremsky E. V., Kirillova D. A., Patsaev T. D., Sharikov R. V., Kamyshinsky R. A., Lukanina K. I., Sharikova N. A., Grigoriev T. E., Vasiliev A. L.

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Adhesive properties of scaffolds, which predominantly depend on the chemical and structural features of their surface, play the most important role in tissue engineering. The basic requirements for such scaffolds are biocompatibility, biodegradation, high cell adhesion, which promotes cell proliferation and differentiation. In many cases, synthetic polymers scaffolds have proven advantageous because they are easy to shape, they are tough, and they have high tensile properties. The regeneration of nerve tissue still remains a big challenge for medicine, and neural stem cells provide promising therapeutic potential for cell replacement therapy. However, experiments with stem cells have their limitations, such as low level of cell viability and poor control of cell differentiation. Whereas the study of already differentiated neuronal cell culture obtained from newborn mouse brain is limited only to cell adhesion. The growth and implantation of neuronal culture requires proper scaffolds. Moreover, the polymer scaffolds implants with neuronal cells could demand specific morphology. To date, it has been proposed to use numerous synthetic polymers for these purposes, including polystyrene, polylactic acid (PLA), polyglycolic acid, and polylactide-glycolic acid. Tissue regeneration experiments demonstrated good biocompatibility of PLA scaffolds, despite the hydrophobic nature of the compound. Problem with poor wettability of the PLA scaffold surface could be overcome in several ways: the surface can be pre-treated by poly-D-lysine or polyethyleneimine peptides; roughness and hydrophilicity of PLA surface could be increased by plasma treatment, or PLA could be combined with natural fibers, such as collagen or chitosan. This work presents a study of adhesion of both induced pluripotent stem cells (iPSCs) and mouse primary neuronal cell culture on the polylactide scaffolds of various types: oriented and non-oriented fibrous nonwoven materials and sponges – with and without the effect of plasma treatment and composites with collagen and chitosan. To evaluate the effect of different types of PLA scaffolds on the neuronal differentiation of iPSCs, we assess the expression of NeuN in differentiated cells through immunostaining. iPSCs more effectively differentiate into neurons on PLA scaffolds with high adhesive properties for primary neuronal cells.

Keywords: PLA scaffold, neurons, neuronal differentiation, stem cells, polylactid

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Authors: Zhongyang Sun, Shu Zhang, Manjiang Xie

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Emerging evidence indicates that microRNAs (miRNAs) play important roles in modulating osteoblast function and bone formation. However, the influence of miRNA on osteoblast proliferation and the possible mechanisms underlying remain to be defined. In this study, we aimed to investigate whether miR-103 regulates osteoblast proliferation under simulated microgravity condition through regulating Cav1.2, the primary subunit of L-type voltage sensitive calcium channels (LTCCs). We first investigated the effect of simulated microgravity on osteoblast proliferation and the outcomes clearly demonstrated that the mechanical unloading inhibits MC3T3-E1 osteoblast-like cells proliferation. Using quantitative Real-Time PCR (qRT-PCR), we provided data showing that miR-103 was up-regulated in response to simulated microgravity. In addition, we observed that up-regulation of miR-103 inhibited and down-regulation of miR-103 promoted osteoblast proliferation under simulated microgravity condition. Furthermore, knocking-down or over-expressing miR-103, respectively, up- or down-regulated the level of Cav1.2 expression and LTCCs currents, suggesting that miR-103 acts as an endogenous attenuator of Cav1.2 in osteoblasts under the condition of simulated microgravity. More importantly, we showed that the effect of miR-103 on osteoblast proliferation was diminished in simulated microgravity, when co-transfecting miR-103 mimic or inhibitor with Cav1.2 siRNA. Taken together, our data suggest that miR-103 inhibits osteoblast proliferation mainly through suppression of Cav1.2 expression under simulated microgravity condition. This work may provide a novel mechanism of microgravity-induced detrimental effects on osteoblast, identifying miR-103 as a novel possible therapeutic target in bone remodeling disorders in this mechanical unloading.

Keywords: microRNA, osteoblasts, cell proliferation, Cav1.2, simulated microgravity

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Authors: Yang Yang, Luciana Magalhães Rebelo Alencar, Martha Sahylí Ortega Pijeira, Beatriz da Silva Batista, Alefe Roger Silva França, Erick Rafael Dias Rates, Ruana Cardoso Lima, Sara Gemini-Piperni, Ralph Santos-Oliveira

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Radium-²²³ dichloride ([²²³Rₐ]RₐCl₂) is an alpha particle-emitting radiopharmaceutical currently approved for the treatment of patients with castration-resistant prostate cancer, symptomatic bone metastases, and no known visceral metastatic disease. [²²³Rₐ]RₐCl₂ is bone-seeking calcium mimetic that bonds into the newly formed bone stroma, especially osteoblastic or sclerotic metastases, killing the tumor cells by inducing DNA breaks in a potent and localized manner. Nonetheless, the successful therapy of osteosarcoma as primary bone tumors is still a challenge. Nanomicelles are colloidal nanosystems widely used in drug development to improve blood circulation time, bioavailability, and specificity of therapeutic agents, among other applications. In addition, the enhanced permeability and retention effect of the nanosystems, and the renal excretion of the nanomicelles reported in most cases so far, are very attractive to achieve selective and increased accumulation in tumor site as well as to increase the safety of [²²³Rₐ]RₐCl₂ in the clinical routine. In the present work, [²²³Rₐ]RₐCl₂ nanomicelles were produced, characterized, in vitro evaluated, and compared with pure [²²³Rₐ]RₐCl2 solution using SAOS2 osteosarcoma cells. The [²²³Rₐ]RₐCl₂ nanomicelles were prepared using the amphiphilic copolymer Pluronic F127. The dynamic light scattering analysis of freshly produced [²²³Rₐ]RₐCl₂ nanomicelles demonstrated a mean size of 129.4 nm with a polydispersity index (PDI) of 0.303. After one week stored in the refrigerator, the mean size of the [²²³Rₐ]RₐCl₂ nanomicelles increased to 169.4 with a PDI of 0.381. Atomic force microscopy analysis of [223Rₐ]RₐCl₂ nanomicelles exhibited spherical structures whose heights reach 1 µm, suggesting the filling of 127-Pluronic nanomicelles with [²²³Rₐ]RₐCl₂. The viability assay with [²²³Rₐ]RₐCl₂ nanomicelles displayed a dose-dependent response as it was observed using pure [²²³Rₐ]RₐCl2. However, at the same dose, [²²³Rₐ]RₐCl₂ nanomicelles were 20% higher efficient in killing SAOS2 cells when compared with pure [²²³Rₐ]RₐCl₂. These findings demonstrated the effectiveness of the nanosystem validating the application of nanotechnology in targeted alpha therapy with [²²³Ra]RₐCl₂. In addition, the [²²³Rₐ]RaCl₂nanomicelles may be decorated and incorporated with a great variety of agents and compounds (e.g., monoclonal antibodies, aptamers, peptides) to overcome the limited use of [²²³Ra]RₐCl₂.

Keywords: nanomicelles, osteosarcoma, radium dichloride, targeted alpha therapy

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Authors: H. Pegram, R. Stevens, L. De Girolamo

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Aligned electrospun nanofibres act as effective neuronal and glial cell scaffolds that can be layered to contain multiple sheets harboring different cell populations. This allows personalised biofunctional prostheses to be manufactured with both acellular and cellularised layers for the treatment of spinal cord injury. Additionally, the manufacturing route may be configured to produce in-vitro 3D cell based model of spinal cord injury to aid drug development and enhance prosthesis performance. The goal of this investigation was to optimise the multi-layer scaffold design parameters for prosthesis manufacture, to enable the development of multi-layer patient specific implant therapies. The work has also focused on the fabricating aligned nanofibre scaffolds that promote in-vitro neuronal and glial cell population growth, cell-to-cell interaction and long-term survival following trauma to mimic an in-vivo spinal cord lesion. The approach has established reproducible lesions and has identified markers of trauma and regeneration marked by effective neuronal migration across the lesion with glial support. The investigation has advanced the development of an in-vitro model of traumatic spinal cord injury and has identified a route to manufacture prostheses which target the repair spinal cord injury. Evidence collated to investigate the multi-layer concept suggests that physical cues provided by nanofibres provide both a natural extra-cellular matrix (ECM) like environment and controls cell proliferation and migration. Specifically, aligned nanofibre layers act as a guidance system for migrating and elongating neurons. On a larger scale, material type in multi-layer systems also has an influence in inter-layer migration as cell types favour different material types. Results have shown that layering nanofibre membranes create a multi-level scaffold system which can enhance or prohibit cell migration between layers. It is hypothesised that modifying nanofibre layer material permits control over neuronal/glial cell migration. Using this concept, layering of neuronal and glial cells has become possible, in the context of tissue engineering and also modelling in-vitro induced lesions.

Keywords: electrospinning, layering, lesion, modeling, nanofibre

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Authors: Mian Huang, Chi Ma, Junyu Lin, William Lu

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Introduction: Bone health attracts more attention recently and an intelligent question and answer (QA) chatbot for osteoporosis is helpful for science popularization. With Generative Pretrained Transformer (GPT) technology developing, we build an osteoporosis corpus dataset and then fine-tune LLaMA, a famous open-source GPT foundation large language model(LLM), on our self-constructed osteoporosis corpus. Evaluated by clinical orthopedic experts, our fine-tuned model outperforms vanilla LLaMA on osteoporosis QA task in Chinese. Three-dimensional quantitative computed tomography (QCT) measured bone mineral density (BMD) is considered as more accurate than DXA for BMD measurement in recent years. We develop an automatic Phantom-less QCT(PL-QCT) that is more efficient for BMD measurement since no need of an external phantom for calibration. Combined with LLM on osteoporosis, our PL-QCT provides efficient and accurate BMD measurement for our chatbot users. Material and Methods: We build an osteoporosis corpus containing about 30,000 Chinese literatures whose titles are related to osteoporosis. The whole process is done automatically, including crawling literatures in .pdf format, localizing text/figure/table region by layout segmentation algorithm and recognizing text by OCR algorithm. We train our model by continuous pre-training with Low-rank Adaptation (LoRA, rank=10) technology to adapt LLaMA-7B model to osteoporosis domain, whose basic principle is to mask the next word in the text and make the model predict that word. The loss function is defined as cross-entropy between the predicted and ground-truth word. Experiment is implemented on single NVIDIA A800 GPU for 15 days. Our automatic PL-QCT BMD measurement adopt AI-associated region-of-interest (ROI) generation algorithm for localizing vertebrae-parallel cylinder in cancellous bone. Due to no phantom for BMD calibration, we calculate ROI BMD by CT-BMD of personal muscle and fat. Results & Discussion: Clinical orthopaedic experts are invited to design 5 osteoporosis questions in Chinese, evaluating performance of vanilla LLaMA and our fine-tuned model. Our model outperforms LLaMA on over 80% of these questions, understanding ‘Expert Consensus on Osteoporosis’, ‘QCT for osteoporosis diagnosis’ and ‘Effect of age on osteoporosis’. Detailed results are shown in appendix. Future work may be done by training a larger LLM on the whole orthopaedics with more high-quality domain data, or a multi-modal GPT combining and understanding X-ray and medical text for orthopaedic computer-aided-diagnosis. However, GPT model gives unexpected outputs sometimes, such as repetitive text or seemingly normal but wrong answer (called ‘hallucination’). Even though GPT give correct answers, it cannot be considered as valid clinical diagnoses instead of clinical doctors. The PL-QCT BMD system provided by Bone’s QCT(Bone’s Technology(Shenzhen) Limited) achieves 0.1448mg/cm2(spine) and 0.0002 mg/cm2(hip) mean absolute error(MAE) and linear correlation coefficient R2=0.9970(spine) and R2=0.9991(hip)(compared to QCT-Pro(Mindways)) on 155 patients in three-center clinical trial in Guangzhou, China. Conclusion: This study builds a Chinese osteoporosis corpus and develops a fine-tuned and domain-adapted LLM as well as a PL-QCT BMD measurement system. Our fine-tuned GPT model shows better capability than LLaMA model on most testing questions on osteoporosis. Combined with our PL-QCT BMD system, we are looking forward to providing science popularization and early morning screening for potential osteoporotic patients.

Keywords: GPT, phantom-less QCT, large language model, osteoporosis

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Authors: Adae Amoako, Daniel Montero, Peter Murray, George Pujalte

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We present a case of a 42-year-old, right-handed Caucasian male who presented to a medical orthopedics clinic with left wrist pain. The patient indicated that the pain started two months prior to the visit. He could only remember helping a friend move furniture prior to the onset of pain. Examination of the left wrist showed limited extension compared to the right. There was clicking with flexion and extension of the wrist on the dorsal aspect. Mild tenderness was noticed over the distal radioulnar joint. There was ulnar and radial deviation on provocation. Initial 4-view x-rays of the left wrist showed mild radiocarpal and scapho-trapezium-trapezoid (ST-T) osteoarthritis, with subchondral cysts seen in the lunate and scaphoid, with no obvious fractures. The patient was initially put in a wrist brace and diclofenac topical gel was prescribed for pain control, as a patient could not take non-steroidal anti-inflammatory drugs (NSAIDs) due to gastritis. Despite diclofenac topical gel use and bracing, symptoms remained, and a steroid injection with 1 mL of lidocaine with 10 mg of triamcinolone acetonide was performed under fluoroscopy. He obtained some relief but after 3 months, the injection had to be repeated. On 2-month follow up after the initial evaluation, symptoms persisted. Magnetic resonance imaging (MRI) was obtained which showed an abnormal T1 hypodense signal involving the proximal pole of the scaphoid and articular collapse proximally of the scaphoid, with marked irregularity of the overlying cartilage, suggesting a remote injury, findings consistent with avascular necrosis of the proximal pole of the scaphoid. A month after that, the patient had the left proximal pole of the scaphoid debrided and an intercompartmental supraretinacular artery vascularized. Pedicle bone graft reconstruction of the proximal pole of the left scaphoid was done. A non-vascularized autograft from the left radius was also applied. He was put in a thumb spica cast with the interphalangeal joint free for 6 weeks. On 6-week follow-up after surgery, the patient was healing well and could make a composite fist with his left hand. The diagnosis of Preiser’s disease is primarily based on radiological findings. Due to the fact that necrosis happens over a period of time, most AVNs are diagnosed at the late stages of the disease. There appear to be no specific guidelines on the management AVN of the scaphoid. In the past, immobilization and arthroscopic debridement had been used. Radial osteotomy has also been tried. Vascularized bone grafts have also been used to treat Preiser’s disease. In our patient, we used three of these treatment modalities, starting with conservative management with topical NSAIDS and immobilization, then debridement with vascularized bone grafts.

Keywords: wrist pain, avascular necrosis of the scaphoid, Preiser’s disease, vascularized bone grafts

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Authors: Massimiliano Panella, Sara Bortoluzzi, Sophia Russotto, Daniele Nicolini, Carmela Rinaldi

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Osteoporosis affects more than 200 million people worldwide. About 75% of osteoporosis cases are undiagnosed or diagnosed only when a bone fracture occurs. Since osteoporosis related fractures are significant determinants of the burden of disease and health and social costs of aging populations, we believe that this is the early identification and treatment of high-risk patients should be a priority in actual healthcare systems. Screening for osteoporosis by dual energy x-ray absorptiometry (DEXA) is not cost-effective for general population. An alternative is pulse-echo ultrasound (PEUS) because of the minor costs. To this end, we developed an early detection program for osteoporosis with PEUS, and we evaluated is possible impact and sustainability. We conducted a cross-sectional study including 1,050 people in Italy. Subjects with >1 major or >2 minor risk factors for osteoporosis were invited to PEUS bone mass density (BMD) measurement at the proximal tibia. Based on BMD values, subjects were classified as healthy subjects (BMD>0.783 g/cm²) and pathological including subjects with suspected osteopenia (0.783≤BMD>0.719 g/cm²) or osteoporosis (BMD ≤ 0.719 g/cm²). The responder rate was 60.4% (634/1050). According to the risk, PEUS scan was recommended to 436 people, of whom 300 (mean age 45.2, 81% women) accepted to participate. We identified 240 (80%) healthy and 60 (20%) pathological subjects (47 osteopenic and 13 osteoporotic). We observed a significant association between high risk people and reduced bone density (p=0.043) with increased risks for female gender, older ages, and menopause (p<0.01). The yearly cost of the screening program was 8,242 euros. With actual Italian fracture incidence rates in osteoporotic patients, we can reasonably expect in 20 years that at least 6 fractures will occur in our sample. If we consider that the mean costs per fracture in Italy is today 16,785 euros, we can estimate a theoretical cost of 100,710 euros. According to literature, we can assume that the early treatment of osteoporosis could avoid 24,170 euros of such costs. If we add the actual yearly cost of the treatments to the cost of our program and we compare this final amount of 11,682 euros to the avoidable costs of fractures (24,170 euros) we can measure a possible positive benefits/costs ratio of 2.07. As a major outcome, our study let us to early identify 60 people with a significant bone loss that were not aware of their condition. This diagnostic anticipation constitutes an important element of value for the project, both for the patients, for the preventable negative outcomes caused by the fractures, and for the society in general, because of the related avoidable costs. Therefore, based on our finding, we believe that the PEUS based screening performed could be a cost-effective approach to early identify osteoporosis. However, our study has some major limitations. In fact, in our study the economic analysis is based on theoretical scenarios, thus specific studies are needed for a better estimation of the possible benefits and costs of our program.

Keywords: osteoporosis, prevention, public health, screening

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Authors: Aftab Ahmed, Farhan Saeed, Muhammad Afzaal, Shinawar Waseem Ali, Ali Imran, Sadaf Munir

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Malnutrition is a globally increasing public health concern among children; it affects number of school children influencing their growth, development and academic performance. The tenet of the current cross sectional study was to assess the nutritional biomarkers of school going children of age 12-15 years resulting in stunting, underweight, overweight, bone deformities and other health disparities in nutritionally deprived urban and rural populations of Pakistan. A sample size comprising of 180 school going children was stipulated from the targeted urban and rural populations. The fallouts of investigation unveiled that both rural and urban populations were experiencing nutritional challenges however; on account of awareness paucity the rustic population was nutritionally more compromised. Hematological tests elucidated 16.7% and 7.8% cases for high glucose level, 35.6% and 27.8% cases for low hemoglobin levels, 14.4% and 15.6% cases for low calcium indices, 12.2% and 4.4% high white blood cell count (WBC), 20% and 14.4% low red blood cell count, 76.7% and 74.4% low hematocrit (HCT) values, among the rural and urban populations respectively. The above mentioned outcomes can serve as a way forward for policy and law maker institutions to curb the possible barricades in the way of healthy nutritional status in these areas

Keywords: malnutrition, hematological study, child nutrition, bone mineral density, calcium, RBC

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Authors: Carolina Pereira Lobato Costa, Cristina Pascual-González, Monica Echeverry, Javier LLorca, Carlos Gonzáléz, Juan Pedro Fernández-Bláquez

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Although the potential of PLA self-reinforced composites for bone applications, not much literature addresses optimal manufacturing conditions. In this regard, this paper describes the woven self-reinforced PLA composites manufacturing processes: the commingling of yarns, weaving, and hot pressing and characterizes the manufactured laminates. Different structures and properties can be achieved by varying the hot compaction process parameters (pressure, holding time, and temperature). The specimens manufactured were characterized in terms of thermal properties (DSC), microstructure (C-scan optical microscope and SEM), strength (tensile test), and biocompatibility (MTT assays). Considering the final device, 155 ℃ for 10 min at 2 MPa act as the more appropriate hot pressing parameters. The laminate produced with these conditions has few voids/porosity, a tensile strength of 30.39 ± 1.21 MPa, and a modulus of 4.09 ± 0.24 GPa. Subsequently to the tensile testing was possible to observe fiber pullout from the fracture surfaces, confirming that this material behaves as a composite. From the results, no single laminate can fulfill all the requirements, being necessary to compromise in function of the priority property. Further investigation is required to improve materials' mechanical performance. Subsequently, process parameters and materials configuration can be adjusted depending on the place and type of implant to suit its function.

Keywords: woven fabric, self-reinforced polymer composite, poly(lactic acid), biodegradable

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Authors: Manoj Kumar, Sujata Mohanty, D. N. Rao, Arul Selvi, Sanjeev K. Bhoi

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Background: Hematopoietic progenitor cells (HPC) mobilized from bone marrow to peripheral blood has been observed in severe trauma and hemorrhagic shock patients. Granulocyte-colony stimulating factor (G-CSF) is a potent stimulator that mobilized HPC from bone marrow to peripheral blood. Objective: Our aim of the study was to investigate the serum G-CSF levels and correlate with HPC and outcome. Methods: Peripheral blood sample from 50 hemorrhagic shock patients was collected on arrival for determination of G-CSF and peripheral blood HPC (PBHPC) and compared with healthy control (n=15). Determination of serum levels of G-CSF by sandwich ELISA and PBHPC by Sysmex XE-2100. Data were categorized by age, sex, Injury Severity Score (ISS), and laboratory data was prospectively collected. Data are expressed as mean±SD and median (min, max). Results: Significantly increased the serum level of G-CSF (264.8 vs. 79.1 pg/ml) and peripheral blood HPC (0.1 vs. 0.01 %) in the T/HS patients when compared with control group. Conclusions: Our studies suggest serum G-CSF elevated in T/HS patients. The elevated in G-CSF was also associated with mobilization of HPC from BM to peripheral blood HPC. Increased the levels of G-CSF in T/HS may play a significant role in the alteration of the hematopoietic compartment.

Keywords: granulocyte colony stimulating factor, G-CSF, hematopoietic progenitor cells, HPC, trauma hemorrhagic shock, T/HS, outcome

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Authors: Samah Oda, Asmaa Khafaga, Mohammed Hashim, Asmaa Khamis

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Toxicity of hydroxyurea (HU), a treatment for certain tumors, polycythemia, and thrombocytosis, was evaluated in rats in one-month toxicity study. Sixty male albino rats were equally classified into four groups. Rats received daily oral gavage of HU in 0, 250, 500, and 750 mg/kg b.wt. Chemical and histopathological assessment of liver, lung, spleen, and bone marrow was performed at 10, 20, and 30 days of the experiment. No significant change was reported in alanine aminotransferase (ALT), aspartate aminotransferase (AST), globulin, and albumin/ globulin ratio during the experiment. Significant decreases in alkaline phosphatase (ALP) and total albumin were reported in rats received 500 and 750 mg/kg b.wt of HU. In addition, total cholesterol level increased significantly after 10 days; however, it significantly decreased after 20 and 30 days of the experiment. Moreover, hepatocytic vacuolation and necrosis with portal inflammatory infiltrates were reported along experimental periods. Pulmonary congestion, hemorrhage, interstitial mononuclear infiltration, peribronchitis, and bronchial epithelial necrosis were also reported. Severe lymphocytic necrosis in spleen and severe loss of hematopoietic cells and replacement with corresponding adipose tissue in bone marrow tissues was demonstrated. In conclusion, HU could be able to induce severe dose and time-dependent hepato-pulmonary toxicity and lymphoid depression in rats.

Keywords: hydroxyurea, hepato-pulmonary toxicity, lymphoid depression, histopathology

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Authors: Doran C. Kalmin

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The descending genicular artery (DGA) perforator free flap provides an alternative to free flap reconstruction based on a review of the literature detailing both anatomical and clinical studies. The descending genicular artery (DGA) supplies skin, muscle, tendon, and bone located around the medial aspect of the knee that has been used in several pioneering reports in reconstructing defects located in various areas throughout the body. After the success of the medial femoral condyle flap in early studies, a small number of studies have been published detailing the use of the DGA in free flap reconstruction. Despite early success in the use of the DGA flap, acceptance within the Plastic and Reconstructive Surgical community has been limited due primarily to anatomical variations of the pedicle. This literature review is aimed at detailing the progression of the DGA perforator free flap and its variations as an alternative and reliable free flap for reconstruction of composite defects with an exploration into both anatomical and clinical studies. A literature review was undertaken, and the progression of the DGA flap is explored from the early review by Acland et al. pioneering the saphenous free flap to exploring modern changes and studies of the anatomy of the DGA. An extensive review of the literature was undertaken that details the anatomy and its variations, approaches to harvesting the flap, the advantages, and disadvantages of the DGA perforator free flap as well as flap outcomes. There are 15 published clinical series of DGA perforator free flaps that incorporate cutaneous, osteoperiosteal, cartilage, osteocutaneous, osteoperiosteal and muscle, osteoperiosteal and subcutaneous and tendocutatenous. The commonest indication for using a DGA free flap was for non-union of bone, particularly that of the scaphoid whereby the medial femoral condyle could be used. In the case series, a success rate of over 90% was established, showing that these early studies have had good success with a wide range of tissue transfers. The greatest limitation is the anatomical variation of the DGA and therefore, the challenges associated with raising the flap. Despite the variation in anatomy and around 10-15% absence of the DGA, the saphenous artery can be used as well as the superior medial genicular artery if the vascular bone is required as part of the flap. Despite only a handful of anatomical and clinical studies describing the DGA perforator free flap, it ultimately provides a reliable flap that can include a variety of composite structure used for reconstruction in almost any area throughout the body. Although it has limitations, it provides a reliable option for free flap reconstruction that can routinely be performed as a single-stage procedure.

Keywords: anatomical study, clinical study, descending genicular artery, literature review, perforator free flap reconstruction

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

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Traction therapy has been used in the treatment of spinal pain for decades. However, a case study reported the occurrence of large disc protrusion during motorized traction therapy. In this study, we hypothesized that additional local decompression with a global axial traction could be helpful for risk reduction of intervertebral disc damage. A validated three dimensional finite element model of the lumbar spine was used. Two-step traction therapy using the axial global traction (the first step) with 1/3 body weight and the additional local decompression (the second step) with 7 mm translation of L4 spinal bone was determined for the traction therapy. During two-step traction therapy, the sacrum was constrained in all translational directions. Reduced lordosis angle by the global axial traction recovered with the additional local decompression. Stress on fibers of the annulus fibrosus by the axial global traction decreased with the local decompression by 17%~96% in the posterior region of intervertebral disc. Stresses on ligaments except anterior longitudinal ligaments in all motion segments decreased till 4.9 mm~5.6 mm translation of L4 spinal bone. The results of this study showed that the additional local decompression is very useful for reducing risk of damage in the intervertebral disc and ligaments caused by the global axial traction force. Moreover, the local decompression could be used to enhance reduction of intradiscal pressure.

Keywords: lumbar spine, traction-therapy, biomechanics, finite element analysis

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Authors: Amin Davoudi

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Aim: New approaches have been introduced to improve soft tissue indices of the dental implants. This systematic review aimed to investigate the effect of computer-aided design and computer-assisted manufacture (CAD/CAM) zirconia (Zr) implant abutments on periodontal aspects. Materials and Methods: Five electronic databases were searched thoroughly based on prior defined MeSH and non-MeSH keywords. Clinical studies were collected via hand searches in English language journals up to September 2020. Interproximal papilla stability, papilla recession, pink and white esthetic score (PES, WES), bone and gingival margin levels, color, and contour of soft tissue were reviewed. Results: The initial literature search yielded 412 articles. After the evaluation of abstracts and full texts, six studies were eligible to be screened. The study design of the included studies was a prospective cohort (n=3) and randomized clinical trial (n=3). The outcome was found to be significantly better for Zr than titanium abutments, however, the studies did not show significant differences between stock and CAD/CAM abutments. Conclusion: Papilla fill, WES, PES, and the distance from the contact point to dental crest bone of adjacent tooth and inter-tooth–implant distance were not significantly different between Zr CAD/CAM and Zr stock abutments. However, soft tissue stability and recession index were better in Zr CAD/CAM abutments.

Keywords: zirconia, CADCAM, periodental, implant

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Authors: Adrian Ionut Nicoara, Denisa Ionela Ene, Alina Maria Holban, Cristina Busuioc

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At present, the development of materials with biomedical applications is a domain of interest that will produce a full series of benefits in engineering and medicine. In this sense, it is required to use a natural material, and this paper is focused on the development of a composite material based on bacterial cellulose – hydroxyapatite and silver nanoparticles with applications in hard tissue. Bacterial cellulose own features like biocompatibility, non-toxicity character and flexibility. Moreover, the bacterial cellulose can be conjugated with different forms of active silver to possess antimicrobial activity. Hydroxyapatite is well known that can mimic at a significant level the activity of the initial bone. The material was synthesized by using an ultrasound probe and finally characterized by several methods. Thereby, the morphological properties were analyzed by using Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). Because the synthesized material has medical application in restore the tissue and to fight against microbial invasion, the samples were tested from the biological point of view by evaluating the biodegradability in phosphate-buffered saline (PBS) and simulated body fluid (SBF) and moreover the antimicrobial effect was performed on Gram-positive bacterium Staphylococcus aureus, Gram-negative bacterium Escherichia coli, and fungi Candida albicans. The results reveal that the obtained material has specific characteristics for bone regeneration.

Keywords: bacterial cellulose, biomaterials, hydroxyapatite, scaffolds materials

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Authors: Dinara Ikramova, Karin A. Hing, Simon C. F. Rawlinson

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Silicate-substituted hydroxyapatite (SiHA) has been shown to enhance bone regeneration in vivo compared with phase pure stoichiometric hydroxyapatite. Evidence suggests that substrate chemistry dependent formation of a permissive protein layer on the surface of synthetic bone graft substitute materials is key for bioactivity and cell attachment. However, little information is available on whether the substrate chemistry may affect cell migration and recruitment. The aim of this study is to investigate whether or not human Mesenchymal Stem Cells (hMSCs) exhibit a chemotactic response to SiHA porous granules and if it can be linked to either the ion exchange or protein sequestering and enrichment on the surface of the material. 150mg of SiHA granules with 80% total porosity and 20% strut porosity were incubated in 1ml of either Serum Free Media (SFM) or 10% Serum Containing Media (SCM) under static cell culture conditions (37°C, 5% CO2) in absence of cells. Protein sequestering and exchange of calcium, phosphate and silicate ions were analysed at 0.5, 1, 2, 4, 8, 16 and 24 hours with n=12 per time point. Migration of hMSCs in the presence of 150mg of SiHA granules was assessed over 24 hours using a modified transwell migration system in either SFM or SCM (n=6) with 30% serum containing media acting as a positive control. At 24 hours protein sequestering and ionic exchange were analysed, and the number of cells was quantified using a high throughput confocal microscope (IN Cell Analyser 6000). In acellular condition, both calcium and phosphate ion concentrations in media showed a decrease at 24 hours which was greater in SFM than in SCM. This suggests possible formation and precipitation of a bone like apatite on the surface of SiHA. Reduction in this activity observed in SCM indicates that the presence of serum proteins is interfering with the ion exchange at the material and media interface. Adsorbed protein levels showed fluctuation over time followed by sharp decrease at 24 hours, suggesting a possible protein rearrangement on the surface of the material. The ion analysis performed on SFM and SCM after 24-hour incubation with cells in the presence of granules showed a greater reduction in phosphate concentration in both SFM and SCM compared to phosphate levels in acellular condition. Silicate concentration in SCM increased from 1.6mM (absence of cells) to 5.1mM (presence of cells). This indicates that the cells are promoting the uptake of phosphate and release of silicate ions. No significant change was seen in levels of adsorbed proteins in the presence and absence of cells. Further analysis is required to determine whether the species of these proteins change over time. The analysis of cell migration after 24-hour incubation showed more cells migrating towards the granules, 12.7% in SFM and 8.3% in SCM, than in positive control, 4.5% in SFM and 3.6% in SCM respectively. These results suggest that SiHA has a chemotactic activity independent of serum proteins. A property which has not previously been demonstrated for a synthetic bone graft material.

Keywords: cell migration, hMSCs, SiHA, transwell migration system

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