Search results for: adipose tissue engineering
3803 Anatomical, Light and Scanning Electron Microscopical Study of Ostrich (Struthio camelus) Integument
Authors: Samir El-Gendy, Doaa Zaghloul
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The current study dealt with the gross and microscopic anatomy of the integument of male ostrich in addition to the histological features of different areas of skin by light and SEM. The ostrich skin is characterized by prominent feather follicles and bristles. The number of feather follicles was determined per cm2 in different regions. The integument of ostrich had many modifications which appeared as callosities and scales, nail and toe pads. They were sternal, pubic and Achilles tendon callosities. The vacuolated epidermal cells were seen mainly in the skin of legs and to a lesser extent in the skin of back and Achilles areas. Higher lipogenic potential was expressed by epidermis from glabrous areas of ostrich skin. The dermal papillae were found in the skin of feathered area of neck and back and this was not a common finding in bird's skin which may give resistance against shearing forces in these regions of ostrich skin. The thickness of the keratin layer of ostrich varied, being thick and characteristically loose in the skin at legs, very thin and wavy at neck, while at Achilles skin area, scale and toe pad were thick and more compact, with the thickest very dense and wavy keratin layer at the nail. The dermis consisted of superficial layer of dense irregular connective tissue characterized by presence of many vacuoles of different sizes just under the basal lamina of the epithelium of epidermis and deep layer of dense regular connective tissue. This result suggested presence of fat droplets in this layer which may be to overcome the lack of good barrier of cutaneous water loss in epidermis.Keywords: ostrich, light microscopy, scanning electron microscopy, integument, skin modifications
Procedia PDF Downloads 2333802 Coagulase Negative Staphylococci: Phenotypic Characterization and Antimicrobial Susceptibility Pattern
Authors: Lok Bahadur Shrestha, Narayan Raj Bhattarai, Basudha Khanal
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Introduction: Coagulase-negative staphylococci (CoNS) are the normal commensal of human skin and mucous membranes. The study was carried out to study the prevalence of CoNS among clinical isolates, to characterize them up to species level and to compare the three conventional methods for detection of biofilm formation. Objectives: to characterize the clinically significant coagulase-negative staphylococci up to species level, to compare the three phenotypic methods for the detection of biofilm formation and to study the antimicrobial susceptibility pattern of the isolates. Methods: CoNS isolates were obtained from various clinical samples during the period of 1 year. Characterization up to species level was done using biochemical test and study of biofilm formation was done by tube adherence, congo red agar, and tissue culture plate method. Results: Among 71 CoNS isolates, seven species were identified. S. epidermidis was the most common species followed by S. saprophyticus, S. haemolyticus. Antimicrobial susceptibility pattern of CoNS documented resistance of 90% to ampicillin. Resistance to cefoxitin and ceftriaxone was observed in 55% of the isolates. We detected biofilm formation in 71.8% of isolates. The sensitivity of tube adherence method was 82% while that of congo red agar method was 78%. Conclusion: Among 71 CoNS isolated, S. epidermidis was the most common isolates followed by S. saprophyticus and S. haemolyticus. Biofilm formation was detected in 71.8% of the isolates. All of the methods were effective at detecting biofilm-producing CoNS strains. Biofilm former strains are more resistant to antibiotics as compared to biofilm non-formers.Keywords: CoNS, congo red agar, bloodstream infections, foreign body-related infections, tissue culture plate
Procedia PDF Downloads 1923801 Biomechanical Performance of the Synovial Capsule of the Glenohumeral Joint with a BANKART Lesion through Finite Element Analysis
Authors: Duvert A. Puentes T., Javier A. Maldonado E., Ivan Quintero., Diego F. Villegas
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Mechanical Computation is a great tool to study the performance of complex models. An example of it is the study of the human body structure. This paper took advantage of different types of software to make a 3D model of the glenohumeral joint and apply a finite element analysis. The main objective was to study the change in the biomechanical properties of the joint when it presents an injury. Specifically, a BANKART lesion, which consists in the detachment of the anteroinferior labrum from the glenoid. Stress and strain distribution of the soft tissues were the focus of this study. First, a 3D model was made of a joint without any pathology, as a control sample, using segmentation software for the bones with the support of medical imagery and a cadaveric model to represent the soft tissue. The joint was built to simulate a compression and external rotation test using CAD to prepare the model in the adequate position. When the healthy model was finished, it was submitted to a finite element analysis and the results were validated with experimental model data. With the validated model, it was sensitized to obtain the best mesh measurement. Finally, the geometry of the 3D model was changed to imitate a BANKART lesion. Then, the contact zone of the glenoid with the labrum was slightly separated simulating a tissue detachment. With this new geometry, the finite element analysis was applied again, and the results were compared with the control sample created initially. With the data gathered, this study can be used to improve understanding of the labrum tears. Nevertheless, it is important to remember that the computational analysis are approximations and the initial data was taken from an in vitro assay.Keywords: biomechanics, computational model, finite elements, glenohumeral joint, bankart lesion, labrum
Procedia PDF Downloads 1573800 Developing Customizable Scaffolds With Antimicrobial Properties for Vascular Tissue Regeneration Using Low Temperature Plasma
Authors: Komal Vig, Syamala Soumyakrishnan, Yadav Baral
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Bypass surgery, using the autologous vein has been one of the most effective treatments for cardiovascular diseases (CVD). More recently tissue engineering including engineered vascular grafts to synthesize blood vessels is gaining usage. Dacron and ePTFE has been employed for vascular grafts, however, these does not work well for small diameter grafts (<6 mm) due to intimal hyperplasia and thrombosis. In the present study PTFE was treated with LTP to improve the endothelialization of intimal surface of graft. Scaffolds were also modified with polyvinylpyrrolidone coated silver nanoparticles (Ag-PVP) and the antimicrobial peptides, p753 and p359. Human umbilical vein endothelial cells (HUVEC) were plated on the developed scaffolds and cell proliferation was determined by the MTT assay. Cells attachment on scaffolds was visualized by microscopy. mRNA expressions levels of different cell markers were investigated using quantitative real-time PCR (qPCR). X ray photoelectron spectroscopic confirmed the introduction of oxygenated functionalities from LTP air plasma. Microscopic and MTT assays indicated increase in cell viability in LTP treated scaffolds. Gene expression studies shows enhanced expression of cell adhesion marker Integrin- α 5 gene after LTP treatment. The KB test displayed a zone of inhibition for Ag-PVP, p753 and p359 of 19mm, 14mm, and 12mm respectively. To determine toxicity of antimicrobial agents to cells, MTT Assay was performed using HEK293 cells. MTT Assay exhibited that Ag-PVP and the peptides were non-toxic to cells at 100μg/mL and 50μg/mL, respectively. Live/dead analysis and plate count of treated bacteria exhibited bacterial inhibition on develop scaffold compared to non-treated scaffold. SEM was performed to analyze the structural changes of bacteria after treatment with antimicrobial agents. Gene expression studies were conducted on RNA from bacteria treated with Ag-PVP and peptides using qRT-PCR. Based on our initial results, more scaffolds alternatives will be developed and investigated for cell growth and vascularization studies.Keywords: low temperature plasma, vascular graft, HUVEC cells, antimicrobial
Procedia PDF Downloads 2343799 Treatment of Porphyromonas gingivalis Induced Gingivitis in Albino Rats with Tetracycline-Loaded Nanochitosan, an Immunohistochemical Analysis
Authors: Rania Hanafi Said, Rasha Mohamed Taha
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Background: By using nanoparticles as drug delivery, it may be possible to avoid the drawbacks of systemic antibiotic dosing, including bacterial antibiotic resistance. The goal of this study was to see how well tetracycline loaded on nanochitosan worked to treat gingival inflammation in albino rats caused by Porphyromonas gingivalis. The study analyzed immunohistochemically the localization of the pro-inflammatory cytokine Interleukin-1beta (IL-1β). Material and methods: In this study, fifty mature male albino rats weighing 150 to 180 grams each were used. They were randomly divided into five groups. We checked for weight changes in rats. Ten male albino rats were included in Group I, which served as a negative control group. Ten rats were included in Group II, where they were exposed once to Porphyromonas. Group III contained ten rats, which were treated the same as Group II plus daily injections of diluted tetracycline powder at the infection sites. Ten rats in Group IV received the same procedure as those in Group II before receiving daily injections of nanochitosan at the injection sites. Finally, Group V, which had ten rats. Following the same protocol as Group II, they received localized injections of tetracycline loaded on nanochitosan once daily. Rats' gingivae were extracted and prepared after they were anesthetized. The biopsies were examined histologically and immunohistochemically by light microscopy. Results: Groups I and V had a nearly normal histological appearance of gingival tissue. In Groups II, III, and IV, degeneration was seen because the epithelial cells were bigger, collagen fibers were pulling away from the lamina propria connective tissue, and the basement membranes had come to an end. There was no discernible difference between groups V and I when they were examined immunohistochemically. Conclusion: The use of nano chitosan as a tetracycline carrier is a novel technique to overcome the drug's rising level of resistance.Keywords: Immunohistochemistry, Nanochitosan, porphyromonas gingivitis, Tetracycline
Procedia PDF Downloads 773798 Non-Cytotoxic Natural Sourced Inorganic Hydroxyapatite (HAp) Scaffold Facilitate Bone-like Mechanical Support and Cell Proliferation
Authors: Sudip Mondal, Biswanath Mondal, Sudit S. Mukhopadhyay, Apurba Dey
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Bioactive materials improve devices for a long lifespan but have mechanical limitations. Mechanical characterization is one of the very important characteristics to evaluate the life span and functionality of the scaffold material. After implantation of scaffold material the primary stage rejection of scaffold occurs due to non biocompatible effect of host body system. The second major problems occur due to the effect of mechanical failure. The mechanical and biocompatibility failure of the scaffold materials can be overcome by the prior evaluation of the scaffold materials. In this study chemically treated Labeo rohita scale is used for synthesizing hydroxyapatite (HAp) biomaterial. Thermo-gravimetric and differential thermal analysis (TG-DTA) is carried out to ensure thermal stability. The chemical composition and bond structures of wet ball-milled calcined HAp powder is characterized by Fourier Transform Infrared spectroscopy (FTIR), X-ray Diffraction (XRD), Field Emission Scanning Electron Microscopy (FE-SEM), Transmission Electron Microscopy (TEM), Energy Dispersive X-ray (EDX) analysis. Fish scale derived apatite materials consists of nano-sized particles with Ca/P ratio of 1.71. The biocompatibility through cytotoxicity evaluation and MTT assay are carried out in MG63 osteoblast cell lines. In the cell attachment study, the cells are tightly attached with HAp scaffolds developed in the laboratory. The result clearly suggests that HAp material synthesized in this study do not have any cytotoxic effect, as well as it has a natural binding affinity for mammalian cell lines. The synthesized HAp powder further successfully used to develop porous scaffold material with suitable mechanical property of ~0.8GPa compressive stress, ~1.10 GPa a hardness and ~ 30-35% porosity which is acceptable for implantation in trauma region for animal model. The histological analysis also supports the bio-affinity of processed HAp biomaterials in Wistar rat model for investigating the contact reaction and stability at the artificial or natural prosthesis interface for biomedical function. This study suggests the natural sourced fish scale-derived HAp material could be used as a suitable alternative biomaterial for tissue engineering application in near future.Keywords: biomaterials, hydroxyapatite, scaffold, mechanical property, tissue engineering
Procedia PDF Downloads 4523797 A Histopathological Study on Leech (Hirudo medicinalis) Application in the Management of Vicarcikā (Eczema)
Authors: K. M. Pratap Shankar, Dattatreya Rao, Sai Prasad
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Background: Skin diseases are among the most common health problems worldwide and are associated with a considerable burden. Eczema is such a skin ailment which cause psychological, social and financial burden on the patient and their families. Management of eczema with antibiotics, antihistamines, steroids etc., are available but even after their use relapses, recurrences and other complications are very common. Aim: The aim of this study was to assess the efficacy of leech application in the management of vicarcikā (Eczema) with Histopathological study. Methods: For the present study 10 patients having the classical symptoms of Vicarcikā, were randomly selected as per the inclusion and exclusion criteria from O.P.D. & I.P.D. sections of Śalya department, S.V. Āyurvedic Hospital, Tirupati. Minimum 4 sittings of Leech application was carried out with seven days interval. Total duration of treatment was 6 weeks. Biopsy samples were collected from the lesion site before and after treatment. Histopathological examination was done by the pathologist. Results: In eczema (dermatitis) the leech application therapy gives excellent response by reducing the inflammatory component, hyperkeratosis, spongiosis, irregular acanthosis and by evoking a granulation tissue response in the dermis and in most of the cases with complete recovery from the lesion. Most of the cases in the study were chronic dermatitis and sebhoric keratosis, almost all local/focal pigmented lesions is totally relieved by leech therapy especially in cases of sebhoric keratosis. Conclusion: In the present study it was found that, leech application evokes significant changes at histological level specifically in reduction of inflammatory component, hyperkeratosis, spongiosis and irregular acanthosis. It was also found that there was a considerable formation of granulation tissue, which helps in formation of healthy new tissues.Keywords: acanthosis, eczema, hyperkeratosis, leech application, spongiosis
Procedia PDF Downloads 2943796 CT Images Based Dense Facial Soft Tissue Thickness Measurement by Open-source Tools in Chinese Population
Authors: Ye Xue, Zhenhua Deng
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Objectives: Facial soft tissue thickness (FSTT) data could be obtained from CT scans by measuring the face-to-skull distances at sparsely distributed anatomical landmarks by manually located on face and skull. However, automated measurement using 3D facial and skull models by dense points using open-source software has become a viable option due to the development of computed assisted imaging technologies. By utilizing dense FSTT information, it becomes feasible to generate plausible automated facial approximations. Therefore, establishing a comprehensive and detailed, densely calculated FSTT database is crucial in enhancing the accuracy of facial approximation. Materials and methods: This study utilized head CT scans from 250 Chinese adults of Han ethnicity, with 170 participants originally born and residing in northern China and 80 participants in southern China. The age of the participants ranged from 14 to 82 years, and all samples were divided into five non-overlapping age groups. Additionally, samples were also divided into three categories based on BMI information. The 3D Slicer software was utilized to segment bone and soft tissue based on different Hounsfield Unit (HU) thresholds, and surface models of the face and skull were reconstructed for all samples from CT data. Following procedures were performed unsing MeshLab, including converting the face models into hollowed cropped surface models amd automatically measuring the Hausdorff Distance (referred to as FSTT) between the skull and face models. Hausdorff point clouds were colorized based on depth value and exported as PLY files. A histogram of the depth distributions could be view and subdivided into smaller increments. All PLY files were visualized of Hausdorff distance value of each vertex. Basic descriptive statistics (i.e., mean, maximum, minimum and standard deviation etc.) and distribution of FSTT were analysis considering the sex, age, BMI and birthplace. Statistical methods employed included Multiple Regression Analysis, ANOVA, principal component analysis (PCA). Results: The distribution of FSTT is mainly influenced by BMI and sex, as further supported by the results of the PCA analysis. Additionally, FSTT values exceeding 30mm were found to be more sensitive to sex. Birthplace-related differences were observed in regions such as the forehead, orbital, mandibular, and zygoma. Specifically, there are distribution variances in the depth range of 20-30mm, particularly in the mandibular region. Northern males exhibit thinner FSTT in the frontal region of the forehead compared to southern males, while females shows fewer distribution differences between the northern and southern, except for the zygoma region. The observed distribution variance in the orbital region could be attributed to differences in orbital size and shape. Discussion: This study provides a database of Chinese individuals distribution of FSTT and suggested opening source tool shows fine function for FSTT measurement. By incorporating birthplace as an influential factor in the distribution of FSTT, a greater level of detail can be achieved in facial approximation.Keywords: forensic anthropology, forensic imaging, cranial facial reconstruction, facial soft tissue thickness, CT, open-source tool
Procedia PDF Downloads 563795 Assessment of Knowledge, Awareness about Hemorrhoids Causes and Stages among the General Public of Saudi Arabia
Authors: Asaiel Mubark Al Hadi
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Background: A frequent anorectal condition known as hemorrhoids, sometimes known as piles, is characterized by a weakening of the anal cushion and the supporting tissue as well as spasms of the internal sphincter. Hemorrhoids are most frequently identified by painless bright red bleeding, prolapse, annoying grape-like tissue prolapse, itching, or a combination of symptoms. digital rectal examination (DRE) and anoscope are used to diagnose it. Constipation, a low-fiber diet, a high body- mass index (BMI), pregnancy, and a reduced physical activity are among the factors that are typically thought to increase the risk of hemorrhoids. Golighers is the most commonly used hemorrhoid classification scheme It is 4 degrees, which determines the degree of the event. The purpose of this study is to assess knowledge and awareness level of the causes and stages of Hemorrhoids in the public of Saudi Arabia. Method: This cross-sectional study was conducted in the Saudi Arabia between Oct 2022- Dec 2022. The study group included at least 384 aged above 18 years. The outcomes of this study were analyzed using the SPSS program using a pre-tested questionnaire. Results: The study included 1410 participants, 69.9% of them were females and 30.1% were males. 53.7% of participants aged 20- 30 years old. 17% of participants had hemorrhoids and 42% had a relative who had hemorrhoids. 42.8% of participants could identify stage 1 of hemorrhoids correctly, 44.7% identified stage 2 correctly, 46.7% identified stage 3 correctly and 58.1% identified stage 4 correctly. Only 28.9% of participants had high level of knowledge about hemorrhoids, 62.7% had moderate knowledge and 8.4% had low knowledge. Conclusion: In conclusion, Saudi general population has poor knowledge of hemorrhoids, their causes and their management approach. There was a significant association between knowledge scores of hemorrhoids with age, gender, residence area and employment.Keywords: hemorrhoids, external hemorrhoid, internal hemorrhoid, anal fissure, hemorrhoid stages, prolapse, rectal bleeding
Procedia PDF Downloads 853794 Acoustic Radiation Force Impulse Elastography of the Hepatic Tissue of Canine Brachycephalic Patients
Authors: A. C. Facin, M. C. Maronezi , M. P. Menezes, G. L. Montanhim, L. Pavan, M. A. R. Feliciano, R. P. Nociti, R. A. R. Uscategui, P. C. Moraes
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The incidence of brachycephalic syndrome (BS) in the clinical routine of small animals has increased significantly giving the higher proportion of brachycephalic pets in the last years and has been considered as an animal welfare problem. The treatment of BS is surgical and the clinical signs related can be considerably attenuated. Nevertheless, the systemic effects of the BS are still poorly reported and little is known about these when the surgical correction is not performed early. Affected dogs are more likely to develop cardiopulmonary, gastrointestinal and sleep disorders in which the chronic hypoxemia plays a major role. This syndrome is compared with the obstructive sleep apnea (OSA) in humans, both considered as causes of systemic and metabolic dysfunction. Among the several consequences of the BS little is known if the syndrome also affects the hepatic tissue of brachycephalic patients. Elastography is a promising ultrasound technique that evaluates tissue elasticity and has been recently used with the purpose of diagnosis of liver fibrosis. In medicine, it is a growing concern regarding the hepatic injury of patients affected by OSA. This prospective study hypothesizes if there is any consequence of BS in the hepatic parenchyma of brachycephalic dogs that don’t receive any surgical treatment. This study was conducted following the approval of the Animal Ethics and Welfare Committee of the Faculdade de Ciências Agrárias e Veterinárias, UNESP, Campus Jaboticabal, Brazil (protocol no 17944/2017) and funded by Sao Paulo Research Foundation (FAPESP, process no 2017/24809-4). The methodology was based in ARFI elastography using the ACUSON S2000/SIEMENS device, with convex multifrequential transducer and specific software as well as clinical evaluation of the syndrome, in order to determine if they can be used as a prognostic non-invasive tool. On quantitative elastography, it was collected three measures of shear wave velocity (meters per second) and depth in centimeters in the left lateral, left medial, right lateral, right medial and caudate lobe of the liver. The brachycephalic patients, 16 pugs and 30 french bulldogs, were classified using a previously established 4-point functional grading system based on clinical evaluation before and after a 3-minute exercise tolerance test already established and validated. The control group was based on the same features collected in 22 beagles. The software R version 3.3.0 was used for the analysis and the significance level was set at 0.05. The data were analysed for normality of residuals and homogeneity of variances by Shapiro-Wilks test. Comparisons of parametric continuous variables between breeds were performed by using ANOVA with a post hoc test for pair wise comparison. The preliminary results show significant statistic differences between the brachycephalic groups and the control group in all lobes analysed (p ≤ 0,05), with higher values of shear wave velocities in the hepatic tissue of brachycephalic dogs. In this context, the results obtained in this study contributes to the understanding of BS as well as its consequences in our patients, reflecting in evidence that one more systemic consequence of the syndrome may occur in brachycephalic patients, which was not related in the veterinary literature yet.Keywords: airway obstruction, brachycephalic airway obstructive syndrome, hepatic injury, obstructive sleep apnea
Procedia PDF Downloads 1153793 Expression of Selected miRNAs in Placenta of the Intrauterine Restricted Growth Fetuses in Cattle
Authors: Karolina Rutkowska, Hubert Pausch, Jolanta Oprzadek, Krzysztof Flisikowski
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The placenta is one of the most important organs that plays a crucial role in the fetal growth and development. Placenta dysfunction is one of the primary cause of the intrauterine growth restriction (IUGR). Cattle have the cotyledonary placenta which consists of two anatomical parts: fetal and maternal. In the case of cattle during the first months of pregnancy, it is very easy to separate maternal caruncle from fetal cotyledon tissue, easier in fact than removing an ordinary glove from one's hand. Which in fact make easier to conduct tissue-specific molecular studies. Typically, animal models for the study of IUGR are created using surgical methods and malnutrition of the pregnant mother or in the case of mice by genetic modifications. However, proposed cattle model with MIMT1Del/WT deletion is unique because it was created without any surgical methods what significantly distinguish it from the other animal models. The primary objective of the study was to identify differential expression of selected miRNAs in the placenta from normal and intrauterine growth restricted fetuses. There was examined the expression of miRNA in the fetal and maternal part of the placenta from 24 fetuses (12 samples from the fetal part of the placenta and 12 samples from maternal part of the placenta). In the study, there was done miRNAs sequencing in the placenta of MIMT1Del/WT fetuses and MIMT1WT/WT fetuses. Then, there were selected miRNAs that are involved in fetal growth and development. Analysis of miRNAs expression was conducted on ABI7500 machine. miRNAs expression was analyzed by reverse-transcription polymerase chain reaction (RT-PCR). As the reference gene was used SNORD47. The results were expressed as 2ΔΔCt: ΔΔCt = (Ctij − CtSNORD47j) − (Cti1 − CtSNORD471). Where Ctij and CtSNORD47j are the Ct values for gene i and for SNORD47 in a sample (named j); Cti1 and CtSNORD471 are the Ct values in sample 1. Differences between groups were evaluated with analysis of variance by using One-Way ANOVA. Bonferroni’s tests were used for interpretation of the data. All normalised miRNA expression values are expressed on a value of natural logarithm. The data were expressed as least squares mean with standard errors. Significance was declared when P < 0.05. The study shows that miRNAs expression depends on the part of the placenta where they origin (fetal or maternal) and on the genotype of the animal. miRNAs offer a particularly new approach to study IUGR. Corresponding tissue samples were collected according to the standard veterinary protocols according to the European Union Normative for Care and Use of Experimental Animals. All animal experiments were approved by the Animal Ethics Committee of the State Provincial Office of Southern Finland (ESAVI-2010-08583/YM-23).Keywords: placenta, intrauterine growth restriction, miRNA, cattle
Procedia PDF Downloads 3113792 The Effect of Elapsed Time on the Cardiac Troponin-T Degradation and Its Utility as a Time Since Death Marker in Cases of Death Due to Burn
Authors: Sachil Kumar, Anoop K.Verma, Uma Shankar Singh
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It’s extremely important to study postmortem interval in different causes of death since it assists in a great way in making an opinion on the exact cause of death following such incident often times. With diligent knowledge of the interval one could really say as an expert that the cause of death is not feigned hence there is a great need in evaluating such death to have been at the CRIME SCENE before performing an autopsy on such body. The approach described here is based on analyzing the degradation or proteolysis of a cardiac protein in cases of deaths due to burn as a marker of time since death. Cardiac tissue samples were collected from (n=6) medico-legal autopsies, (Department of Forensic Medicine and Toxicology), King George’s Medical University, Lucknow India, after informed consent from the relatives and studied post-mortem degradation by incubation of the cardiac tissue at room temperature (20±2 OC) for different time periods (~7.30, 18.20, 30.30, 41.20, 41.40, 54.30, 65.20, and 88.40 Hours). The cases included were the subjects of burn without any prior history of disease who died in the hospital and their exact time of death was known. The analysis involved extraction of the protein, separation by denaturing gel electrophoresis (SDS-PAGE) and visualization by Western blot using cTnT specific monoclonal antibodies. The area of the bands within a lane was quantified by scanning and digitizing the image using Gel Doc. As time postmortem progresses the intact cTnT band degrades to fragments that are easily detected by the monoclonal antibodies. A decreasing trend in the level of cTnT (% of intact) was found as the PM hours increased. A significant difference was observed between <15 h and other PM hours (p<0.01). Significant difference in cTnT level (% of intact) was also observed between 16-25 h and 56-65 h & >75 h (p<0.01). Western blot data clearly showed the intact protein at 42 kDa, three major (28 kDa, 30kDa, 10kDa) fragments, three additional minor fragments (12 kDa, 14kDa, and 15 kDa) and formation of low molecular weight fragments. Overall, both PMI and cardiac tissue of burned corpse had a statistically significant effect where the greatest amount of protein breakdown was observed within the first 41.40 Hrs and after it intact protein slowly disappears. If the percent intact cTnT is calculated from the total area integrated within a Western blot lane, then the percent intact cTnT shows a pseudo-first order relationship when plotted against the time postmortem. A strong significant positive correlation was found between cTnT and PM hours (r=0.87, p=0.0001). The regression analysis showed a good variability explained (R2=0.768) The post-mortem Troponin-T fragmentation observed in this study reveals a sequential, time-dependent process with the potential for use as a predictor of PMI in cases of burning.Keywords: burn, degradation, postmortem interval, troponin-T
Procedia PDF Downloads 4443791 Methylation Analysis of PHF20L1 and DACT2 Gene Promoters in Women with Breast Cancer
Authors: Marta E. Hernandez-Caballero, Veronica Borgonio-Cuadra, Antonio Miranda-Duarte, Xochitl Rojas-Toledo, Normand Garcia-Hernandez, Maura Cardenas-Garcia, Teresa Abad-Camacho
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Breast cancer (BC) is the most common tumor in women over the world. DNA methylation is an epigenetic modification critical in CpG sites, aberrant methylation of CpG islands in promoters is a hallmark of cancer. So, gene expression can be regulated by alterations in DNA methylation. In cell lines DACT2 gene reduces the growth and migration of tumor cells by its participation in the suppression of TGFb/SMAD2/3. PHF20L1 is involved in histone acetylation therefore, it regulates transcription. Our aim was to analyze the methylation status of the DACT2 and PHF20L1 promoter regions in tumoral and healthy mammary tissue from women with BC in different progression states. The study included 77 patients from Centro Medico Nacional La Raza in Mexico City. After identifying a CpG island in DACT2 and PHF20L1 promoters, DNA methylation status was analyzed through sodium bisulfite with subsequent amplification using methylation-specific PCR. Results revealed no changes in methylation status of PHF20L1 and cancer stages (II y III) or in comparison to healthy tissues, it was demethylated. DACT2 promoter methylation was no significant between tumoral stages (II, P = 0.37; III, P = 0.17) or with healthy tissue. Previous data reported DACT2 methylated in nasopharyngeal carcinoma but in this study promoter methylation was not observed. PHF20L1 protein contains N-terminal Tudor and C-terminal plant homeodomain domains, it has been suggested that can stabilize DNMT1 regulating DNA methylation, therefore, was associated with poor prognostic in BC. We found no evidence of methylation in patients and controls in PHF20L1 promoter, so its association with BC may have no direct relation with promoter methylation. More studies including other methylation sites in these genes in BC are necessary.Keywords: bisulfite conversion, breast cancer, DACT2, DNA methylation, PHF20L1, tumoral status
Procedia PDF Downloads 2943790 Possible Modulation of FAS and PTP-1B Signaling in Ameliorative Potential of Bombax ceiba against High Fat Diet Induced Obesity
Authors: Paras Gupta, Rohit Goyal, Yamini Chauhan, Pyare Lal Sharma
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Background: Bombax ceiba Linn., commonly called as Semal, is used in various gastro-intestinal disturbances. It contains lupeol which inhibits PTP-1B, adipogenesis, TG synthesis and accumulation of lipids in adipocytes and adipokines whereas the flavonoids isolated from B. ceiba has FAS inhibitory activity. The present study was aimed to investigate ameliorative potential of Bombax ceiba to experimental obesity in Wistar rats, and its possible mechanism of action. Methods: Male Wistar albino rats weighing 180–220 g were employed in present study. Experimental obesity was induced by feeding high fat diet for 10 weeks. Methanolic extract of B. ceiba extract 100, 200 and 400 mg/kg and Gemfibrozil 50 mg/kg as standard drug were given orally from 7th to 10th week. Results: Induction with HFD for 10 weeks caused significant (p < 0.05) increase in % body wt, BMI, LEE indices; serum glucose, triglyceride, LDL, VLDL, cholesterol, free fatty acid, ALT, AST; tissue TBARS, nitrate/nitrite levels; different fat pads and relative liver weight; and significant decrease in food intake (g and kcal), serum HDL and tissue glutathione levels in HFD control rats. Treatment with B. ceiba extract and Gemfibrozil significantly attenuated these HFD induced changes, as compared to HFD control. The effect of B. ceiba 200 and 400 mg/kg was more pronounced in comparison to Gemfibrozil. Conclusion: On the basis of results obtained, it may be concluded that the methanolic extract of stem bark of Bombax ceiba has significant ameliorative potential against HFD induced obesity in rats, possibly through modulation of FAS and PTP-1B signaling due to the presence of flavonoids and lupeol.Keywords: obesity, Bombax ceiba, free fatty acid, protein tyrosine phosphatase-1B, fatty acid synthase
Procedia PDF Downloads 3933789 The Effect of Hesperidin on Troponin's Serum Level Changes as a Heart Tissue Damage Biomarker Due to Gamma Irradiation of Rat's Mediastinum
Authors: G. H. Haddadi, S. Sajadi, R. Fardid, Z. Haddadi
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The heart is a radiosensitive organ, and its damage is a dose-limiting factor in radiotherapy. Different side effects including vascular plaque and heart fibrosis occur in patients with thorax irradiation. The present study aimed to evaluate the radioprotective efficacy of Hesperidin (HES), a naturally occurring citrus flavanoglycone, against γ-radiation induced tissue damage in the heart of male rats. Sixty-eight rats were divided into four groups. The rats in group 1 received PBS, and those in group 2 received HES. Also, the rats in group 3 received PBS and underwent γ-irradiation, and those in group 4 received HES and underwent γ-irradiation. They were exposed to 20 Gy γ-radiation using a single fraction cobalt-60 unit, and the dose of Hesperidin was (100 mg/kg/d, orally) for 7 days prior irradiation. Each group was divided into two subgroups. Samplings of rats in subgroup A was done 4-6 hours after irradiation. The samples were sent to laboratory for determination of Troponin’s I (TnI) serum level changes as a cardiac biomarker. The remaining animals (subgroups B) were sacrificed 8 weeks after radiotherapy for histopathological evaluation. In group 3, TnI obviously increased in comparison with group 1 (p < 0.05). The comparison of groups 1 and 4 showed no significant difference. Evaluation of histopathological parameters in subgroup B showed significant differences between groups 1 and 3 in some of the cases. Inflammation (p=0.008), pericardial effusion (p=0.001) and vascular plaque (p=0.001) increased in the rats exposed to 20 Gy γ-irradiation. Using oral administration of HES significantly decreased all the above factors when compared to group 4 (P > 0.016). Administration of 100 mg/kg/day Hesperidin for 7 days resulted in decreased Troponin I and radiation heart injury. This agent may have protective effects against radiation-induced heart damage.Keywords: hesperidin, radioprotector, troponin I, cardiac inflammation, vascular plaque
Procedia PDF Downloads 2463788 Branched Chain Amino Acid Kinesio PVP Gel Tape from Extract of Pea (Pisum sativum L.) Based on Ultrasound-Assisted Extraction Technology
Authors: Doni Dermawan
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Modern sports competition as a consequence of the increase in the value of the business and entertainment in the field of sport has been demanding athletes to always have excellent physical endurance performance. Physical exercise is done in a long time, and intensive may pose a risk of muscle tissue damage caused by the increase of the enzyme creatine kinase. Branched Chain Amino Acids (BCAA) is an essential amino acid that is composed of leucine, isoleucine, and valine which serves to maintain muscle tissue, keeping the immune system, and prevent further loss of coordination and muscle pain. Pea (Pisum sativum L.) is a kind of leguminous plants that are rich in Branched Chain Amino Acids (BCAA) where every one gram of protein pea contains 82.7 mg of leucine; 56.3 mg isoleucine; and 56.0 mg of valine. This research aims to develop Branched Chain Amino Acids (BCAA) from pea extract is applied in dosage forms Gel PVP Kinesio Tape technology using Ultrasound-assisted Extraction. The method used in the writing of this paper is the Cochrane Collaboration Review that includes literature studies, testing the quality of the study, the characteristics of the data collection, analysis, interpretation of results, and clinical trials as well as recommendations for further research. Extraction of BCAA in pea done using ultrasound-assisted extraction technology with optimization variables includes the type of solvent extraction (NaOH 0.1%), temperature (20-250C), time (15-30 minutes) power (80 watt) and ultrasonic frequency (35 KHz). The advantages of this extraction method are the level of penetration of the solvent into the membrane of the cell is high and can increase the transfer period so that the BCAA substance separation process more efficient. BCAA extraction results are then applied to the polymer PVP (Polyvinylpyrrolidone) Gel powder composed of PVP K30 and K100 HPMC dissolved in 10 mL of water-methanol (1: 1) v / v. Preparations Kinesio Tape Gel PVP is the BCAA in the gel are absorbed into the muscle tissue, and joints through tensile force then provides stimulation to the muscle circulation with variable pressure so that the muscle can increase the biomechanical movement and prevent damage to the muscle enzyme creatine kinase. Analysis and evaluation of test preparation include interaction, thickness, weight uniformity, humidity, water vapor permeability, the levels of the active substance, content uniformity, percentage elongation, stability testing, release profile, permeation in vitro and in vivo skin irritation testing.Keywords: branched chain amino acid, BCAA, Kinesio tape, pea, PVP gel, ultrasound-assisted extraction
Procedia PDF Downloads 2833787 Preliminary Assessment for Protective Effect of Rhodiola rosea in Chemically Induced Ulcerative Colitis
Authors: Santram Lodhi, Alok Pal Jain, Awesh K. Yadav, Gopal Rai
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Rhodiola rosea L. (Crassulaceae) is commonly known as golden root or rose root. It is a perennial herbaceous plant and most investigated species of the genus Rhodiola. Rhodiola rosea contains flavonoids, terpenoids, phenylpropanoid glycosides and phenylethanol derivatives in the roots of the plant. The objective of present study was to investigate the protective effect of hydroalcoholic extract from Rhodiola rosea roots in DSS induced colitis in mice. The ulcerative colitis was induced by DSS (3%, w/v) in mice and estimated weight loss and stool consistency. Various parameters including Colon length, spleen weights and ulcer index were also measured. The histological observations were observed by H&E staining. Effect of hydroalcoholic extract on various antioxidant parameter of rat colon such as tissue myeloperoxidase (MPO), reduced GSH, SOD concentrations and lipid peroxidation were determined. Pro-inflammatory mediators, such as tumour necrosis factor-α (TNF-α) and nitric oxide (NO) were determined by ELISA. In DSS induced group, mice body weight decreased gradually as compared to the control group. Redness and edema were observed in the colons intensely and scores representing inflammation in this group. The extract treated showed with tissue levels of TNF-α, IL-6 and MPO activity were significantly (p<0.05) increased. The mice treated with higher doses of hydroalcoholic extract (300 mg/kg) significantly reduced the activity compared with standard drug sulfasalazine (100 mg/kg. B.wt). Conclusion: Results of this study were suggested that the efficacy of hydroalcoholic extract, especially at the higher dose, was similar to that of standard drug, which concerned its potential application as a natural medicine for the treatment of ulcerative colitis.Keywords: phenylpropanoid, Rhodiola rosea, sulfasalazin, ulcerative colitis
Procedia PDF Downloads 2383786 A Multiple Freezing/Thawing Cycles Influence Internal Structure and Mechanical Properties of Achilles Tendon
Authors: Martyna Ekiert, Natalia Grzechnik, Joanna Karbowniczek, Urszula Stachewicz, Andrzej Mlyniec
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Tendon grafting is a common procedure performed to treat tendon rupture. Before the surgical procedure, tissues intended for grafts (i.e., Achilles tendon) are stored in ultra-low temperatures for a long time and also may be subjected to unfavorable conditions, such as repetitive freezing (F) and thawing (T). Such storage protocols may highly influence the graft mechanical properties, decrease its functionality and thus increase the risk of complications during the transplant procedure. The literature reports on the influence of multiple F/T cycles on internal structure and mechanical properties of tendons stay inconclusive, confirming and denying the negative influence of multiple F/T at the same time. An inconsistent research methodology and lack of clear limit of F/T cycles, which disqualifies tissue for surgical graft purposes, encouraged us to investigate the issue of multiple F/T cycles by the mean of biomechanical tensile tests supported with Scanning Electron Microscope (SEM) imaging. The study was conducted on male bovine Achilles tendon-derived from the local abattoir. Fresh tendons were cleaned of excessive membranes and then sectioned to obtained fascicle bundles. Collected samples were randomly assigned to 6 groups subjected to 1, 2, 4, 6, 8 and 12 cycles of freezing-thawing (F/T), respectively. Each F/T cycle included deep freezing at -80°C temperature, followed by thawing at room temperature. After final thawing, thin slices of the side part of samples subjected to 1, 4, 8 and 12 F/T cycles were collected for SEM imaging. Then, the width and thickness of all samples were measured to calculate the cross-sectional area. Biomechanical tests were performed using the universal testing machine (model Instron 8872, INSTRON®, Norwood, Massachusetts, USA) using a load cell with a maximum capacity of 250 kN and standard atmospheric conditions. Both ends of each fascicle bundle were manually clamped in grasping clamps using abrasive paper and wet cellulose wadding swabs to prevent tissue slipping while clamping and testing. Samples were subjected to the testing procedure including pre-loading, pre-cycling, loading, holding and unloading steps to obtain stress-strain curves for representing tendon stretching and relaxation. The stiffness of AT fascicles bundle samples was evaluated in terms of modulus of elasticity (Young’s modulus), calculated from the slope of the linear region of stress-strain curves. SEM imaging was preceded by chemical sample preparation including 24hr fixation in 3% glutaraldehyde buffered with 0.1 M phosphate buffer, washing with 0.1 M phosphate buffer solution and dehydration in a graded ethanol solution. SEM images (Merlin Gemini II microscope, ZEISS®) were taken using 30 000x mag, which allowed measuring a diameter of collagen fibrils. The results confirm a decrease in fascicle bundles Young’s modulus as well as a decrease in the diameter of collagen fibrils. These results confirm the negative influence of multiple F/T cycles on the mechanical properties of tendon tissue.Keywords: biomechanics, collagen, fascicle bundles, soft tissue
Procedia PDF Downloads 1213785 Process Improvement and Redesign of the Immuno Histology (IHC) Lab at MSKCC: A Lean and Ergonomic Study
Authors: Samantha Meyerholz
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MSKCC offers patients cutting edge cancer care with the highest quality standards. However, many patients and industry members do not realize that the operations of the Immunology Histology Lab (IHC) are the backbone for carrying out this mission. The IHC lab manufactures blocks and slides containing critical tissue samples that will be read by a Pathologist to diagnose and dictate a patient’s treatment course. The lab processes 200 requests daily, leading to the generation of approximately 2,000 slides and 1,100 blocks each day. Lab material is transported through labeling, cutting, staining and sorting manufacturing stations, while being managed by multiple techs throughout the space. The quality of the stain as well as wait times associated with processing requests, is directly associated with patients receiving rapid treatments and having a wider range of care options. This project aims to improve slide request turnaround time for rush and non-rush cases, while increasing the quality of each request filled (no missing slides or poorly stained items). Rush cases are to be filled in less than 24 hours, while standard cases are allotted a 48 hour time period. Reducing turnaround times enable patients to communicate sooner with their clinical team regarding their diagnosis, ultimately leading faster treatments and potentially better outcomes. Additional project goals included streamlining tech and material workflow, while reducing waste and increasing efficiency. This project followed a DMAIC structure with emphasis on lean and ergonomic principles that could be integrated into an evolving lab culture. Load times and batching processes were analyzed using process mapping, FMEA analysis, waste analysis, engineering observation, 5S and spaghetti diagramming. Reduction of lab technician movement as well as their body position at each workstation was of top concern to pathology leadership. With new equipment being brought into the lab to carry out workflow improvements, screen and tool placement was discussed with the techs in focus groups, to reduce variation and increase comfort throughout the workspace. 5S analysis was completed in two phases in the IHC lab, helping to drive solutions that reduced rework and tech motion. The IHC lab plans to continue utilizing these techniques to further reduce the time gap between tissue analysis and cancer care.Keywords: engineering, ergonomics, healthcare, lean
Procedia PDF Downloads 2213784 Simulation of GAG-Analogue Biomimetics for Intervertebral Disc Repair
Authors: Dafna Knani, Sarit S. Sivan
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Aggrecan, one of the main components of the intervertebral disc (IVD), belongs to the family of proteoglycans (PGs) that are composed of glycosaminoglycan (GAG) chains covalently attached to a core protein. Its primary function is to maintain tissue hydration and hence disc height under the high loads imposed by muscle activity and body weight. Significant PG loss is one of the first indications of disc degeneration. A possible solution to recover disc functions is by injecting a synthetic hydrogel into the joint cavity, hence mimicking the role of PGs. One of the hydrogels proposed is GAG-analogues, based on sulfate-containing polymers, which are responsible for hydration in disc tissue. In the present work, we used molecular dynamics (MD) to study the effect of the hydrogel crosslinking (type and degree) on the swelling behavior of the suggested GAG-analogue biomimetics by calculation of cohesive energy density (CED), solubility parameter, enthalpy of mixing (ΔEmix) and the interactions between the molecules at the pure form and as a mixture with water. The simulation results showed that hydrophobicity plays an important role in the swelling of the hydrogel, as indicated by the linear correlation observed between solubility parameter values of the copolymers and crosslinker weight ratio (w/w); this correlation was found useful in predicting the amount of PEGDA needed for the desirable hydration behavior of (CS)₄-peptide. Enthalpy of mixing calculations showed that all the GAG analogs, (CS)₄ and (CS)₄-peptide are water-soluble; radial distribution function analysis revealed that they form interactions with water molecules, which is important for the hydration process. To conclude, our simulation results, beyond supporting the experimental data, can be used as a useful predictive tool in the future development of biomaterials, such as disc replacement.Keywords: molecular dynamics, proteoglycans, enthalpy of mixing, swelling
Procedia PDF Downloads 683783 Synthesis and Characterization of Silver/Graphene Oxide Co-Decorated TiO2 Nanotubular Arrays for Biomedical Applications
Authors: Alireza Rafieerad, Bushroa Abd Razak, Bahman Nasiri Tabrizi, Jamunarani Vadivelu
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Recently, reports on the fabrication of nanotubular arrays have generated considerable scientific interest, owing to the broad range of applications of the oxide nanotubes in solar cells, orthopedic and dental implants, photocatalytic devices as well as lithium-ion batteries. A more attractive approach for the fabrication of oxide nanotubes with controllable morphology is the electrochemical anodization of substrate in a fluoride-containing electrolyte. Consequently, titanium dioxide nanotubes (TiO2 NTs) have been highly considered as an applicable material particularly in the district of artificial implants. In addition, regarding long-term efficacy and reasons of failing and infection after surgery of currently used dental implants required to enhance the cytocompatibility properties of Ti-based bone-like tissue. As well, graphene oxide (GO) with relevant biocompatibility features in tissue sites, osseointegration and drug delivery functionalization was fully understood. Besides, the boasting antibacterial ability of silver (Ag) remarkably provided for implantable devices without infection symptoms. Here, surface modification of Ti–6Al–7Nb implants (Ti67IMP) by the development of Ag/GO co-decorated TiO2 NTs was examined. Initially, the anodic TiO2 nanotubes obtained at a constant potential of 60 V were annealed at 600 degree centigrade for 2 h to improve the adhesion of the coating. Afterward, the Ag/GO co-decorated TiO2 NTs were developed by spin coating on Ti67IM. The microstructural features, phase composition and wettability behavior of the nanostructured coating were characterized comparably. In a nutshell, the results of the present study may contribute to the development of the nanostructured Ti67IMP with improved surface properties.Keywords: anodic tio2 nanotube, biomedical applications, graphene oxide, silver, spin coating
Procedia PDF Downloads 3193782 In vivo Mechanical Characterization of Facial Skin Combining Digital Image Correlation and Finite Element
Authors: Huixin Wei, Shibin Wang, Linan Li, Lei Zhou, Xinhao Tu
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Facial skin is a biomedical material with complex mechanical properties of anisotropy, viscoelasticity, and hyperelasticity. The mechanical properties of facial skin are crucial for a number of applications including facial plastic surgery, animation, dermatology, cosmetic industry, and impact biomechanics. Skin is a complex multi-layered material which can be broadly divided into three main layers, the epidermis, the dermis, and the hypodermis. Collagen fibers account for 75% of the dry weight of dermal tissue, and it is these fibers which are responsible for the mechanical properties of skin. Many research on the anisotropic mechanical properties are mainly concentrated on in vitro, but there is a great difference between in vivo and in vitro for mechanical properties of the skin. In this study, we presented a method to measure the mechanical properties of facial skin in vivo. Digital image correlation (DIC) and indentation tests were used to obtain the experiment data, including the deformation of facial surface and indentation force-displacement curve. Then, the experiment was simulated using a finite element (FE) model. Application of Computed Tomography (CT) and reconstruction techniques obtained the real tissue geometry. A three-dimensional FE model of facial skin, including a bi-layer system, was obtained. As the epidermis is relatively thin, the epidermis and dermis were regarded as one layer and below it was hypodermis in this study. The upper layer was modeled as a Gasser-Ogden-Holzapfel (GOH) model to describe hyperelastic and anisotropic behaviors of the dermis. The under layer was modeled as a linear elastic model. In conclusion, the material properties of two-layer were determined by minimizing the error between the FE data and experimental data.Keywords: facial skin, indentation test, finite element, digital image correlation, computed tomography
Procedia PDF Downloads 1053781 Fibroblast Compatibility of Core-Shell Coaxially Electrospun Hybrid Poly(ε-Caprolactone)/Chitosan Scaffolds
Authors: Hilal Turkoglu Sasmazel, Ozan Ozkan, Seda Surucu
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Tissue engineering is the field of treating defects caused by injuries, trauma or acute/chronic diseases by using artificial scaffolds that mimic the extracellular matrix (ECM), the natural biological support for the tissues and cells within the body. The main aspects of a successful artificial scaffold are (i) large surface area in order to provide multiple anchorage points for cells to attach, (ii) suitable porosity in order to achieve 3 dimensional growth of the cells within the scaffold as well as proper transport of nutrition, biosignals and waste and (iii) physical, chemical and biological compatibility of the material in order to obtain viability throughout the healing process. By hybrid scaffolds where two or more different materials were combined with advanced fabrication techniques into complex structures, it is possible to combine the advantages of individual materials into one single structure while eliminating the disadvantages of each. Adding this to the complex structure provided by advanced fabrication techniques enables obtaining the desired aspects of a successful artificial tissue scaffold. In this study, fibroblast compatibility of poly(ε-caprolactone) (PCL)/chitosan core-shell electrospun hybrid scaffolds with proper mechanical, chemical and physical properties successfully developed in our previous study was investigated. Standard 7-day cell culture was carried out with L929 fibroblast cell line. The viability of the cells cultured with the scaffolds was monitored with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) viability assay for every 48 h starting with 24 h after the initial seeding. In this assay, blank commercial tissue culture polystyrene (TCPS) Petri dishes, single electrospun PCL and single electrospun chitosan mats were used as control in order to compare and contrast the performance of the hybrid scaffolds. The adhesion, proliferation, spread and growth of the cells on/within the scaffolds were observed visually on the 3rd and the 7th days of the culture period with confocal laser scanning microscopy (CSLM) and scanning electron microscopy (SEM). The viability assay showed that the hybrid scaffolds caused no toxicity for fibroblast cells and provided a steady increase in cell viability, effectively doubling the cell density for every 48 h for the course of 7 days, as compared to TCPS, single electrospun PCL or chitosan mats. The cell viability on the hybrid scaffold was ~2 fold better compared to TCPS because of its 3D ECM-like structure compared to 2D flat surface of commercially cell compatible TCPS, and the performance was ~2 fold and ~10 fold better compared to single PCL and single chitosan mats, respectively, even though both fabricated similarly with electrospinning as non-woven fibrous structures, because single PCL and chitosan mats were either too hydrophobic or too hydrophilic to maintain cell attachment points. The viability results were verified with visual images obtained with CSLM and SEM, in which cells found to achieve characteristic spindle-like fibroblast shape and spread on the surface as well within the pores successfully at high densities.Keywords: chitosan, core-shell, fibroblast, electrospinning, PCL
Procedia PDF Downloads 1723780 Effect of Z-VAD-FMK on in Vitro Viability of Dog Follicles
Authors: Leda Maria Costa Pereira, Maria Denise Lopes, Nucharin Songsasen
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Mammalian ovaries contain thousands of follicles that eventually degenerate or die after culture in vitro. Caspase-3 is a key enzyme that regulating cell death. Our objective was to examine the influence of anti-apoptotic drug Z-VAD-FMK (pan-caspase inhibitor) on in vitro viability of dog follicles within the ovarian cortex. Ovaries were obtained from prepubertal (age, 2.5–6 months) and adult (age, 8 months to 2 years) bitches and ovarian cortical fragments were recovered. The cortices were then incubated on 1.5% (w/v) agarose gel blocks within a 24-wells culture plate (three cortical pieces/well) containing Minimum Essential Medium Eagle - Alpha Modification (Alpha MEM) supplemented with 4.2 µg/ml insulin, 3.8 µg/ml transferrin, 5 ng/ml selenium, 2 mM L-glutamine, 100 µg/mL of penicillin G sodium, 100 µg/mL of streptomycin sulfate, 0.05 mM ascorbic acid, 10 ng/mL of FSH and 0.1% (w/v) polyvinyl alcohol in humidified atmosphere of 5% CO2 and 5% O2. The cortices were divided in six treatment groups: 1) 10 ng/mL EGF (EGF V0); 2) 10 ng/mL of EGF plus 1 mM Z-VAD-FMK (EGF V1); 3) 10 ng/mL of EGF and 10 mM Z-VAD-FMK (EGF V10); 4) 1 mM Z-VAD-FMK; 5) 10 mM Z-VAD-FMK and (6) no EGF and Z-VAD-FMK supplementation. Ovarian follicles within the tissues were processed for histology and assessed for follicle density, viability (based on morphology) and diameter immediately after collection (Control) or after 3 or 7 days of in vitro incubation. Comparison among fresh and culture treatment group was performed using ANOVA test. There were no differences (P > 0.05) in follicle density and viability among different culture treatments. However, there were differences in this parameter between culture days. Specifically, culturing tissue for 7 days resulted in significant reduction in follicle viability and density, regardless of treatments. We found a difference in size between culture days when these follicles were cultured using 10 mM Z-VAD-FMK or 10 ng/mL EGF (EGF V0). In sum, the finding demonstrated that Z-VAD-FMK at the dosage used in the present study does not provide the protective effect to ovarian tissue during in vitro culture. Future studies should explore different Z-VAD-FMK dosages or other anti-apoptotic agent, such as surviving in protecting ovarian follicles against cell death.Keywords: anti apoptotic drug, bitches, follicles, Z-VAD-FMK
Procedia PDF Downloads 3543779 Probing Mechanical Mechanism of Three-Hinge Formation on a Growing Brain: A Numerical and Experimental Study
Authors: Mir Jalil Razavi, Tianming Liu, Xianqiao Wang
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Cortical folding, characterized by convex gyri and concave sulci, has an intrinsic relationship to the brain’s functional organization. Understanding the mechanism of the brain’s convoluted patterns can provide useful clues into normal and pathological brain function. During the development, the cerebral cortex experiences a noticeable expansion in volume and surface area accompanied by tremendous tissue folding which may be attributed to many possible factors. Despite decades of endeavors, the fundamental mechanism and key regulators of this crucial process remain incompletely understood. Therefore, to taking even a small role in unraveling of brain folding mystery, we present a mechanical model to find mechanism of 3-hinges formation in a growing brain that it has not been addressed before. A 3-hinge is defined as a gyral region where three gyral crests (hinge-lines) join. The reasons that how and why brain prefers to develop 3-hinges have not been answered very well. Therefore, we offer a theoretical and computational explanation to mechanism of 3-hinges formation in a growing brain and validate it by experimental observations. In theoretical approach, the dynamic behavior of brain tissue is examined and described with the aid of a large strain and nonlinear constitutive model. Derived constitute model is used in the computational model to define material behavior. Since the theoretical approach cannot predict the evolution of cortical complex convolution after instability, non-linear finite element models are employed to study the 3-hinges formation and secondary morphological folds of the developing brain. Three-dimensional (3D) finite element analyses on a multi-layer soft tissue model which mimics a small piece of the brain are performed to investigate the fundamental mechanism of consistent hinge formation in the cortical folding. Results show that after certain amount growth of cortex, mechanical model starts to be unstable and then by formation of creases enters to a new configuration with lower strain energy. By further growth of the model, formed shallow creases start to form convoluted patterns and then develop 3-hinge patterns. Simulation results related to 3-hinges in models show good agreement with experimental observations from macaque, chimpanzee and human brain images. These results have great potential to reveal fundamental principles of brain architecture and to produce a unified theoretical framework that convincingly explains the intrinsic relationship between cortical folding and 3-hinges formation. This achieved fundamental understanding of the intrinsic relationship between cortical folding and 3-hinges formation would potentially shed new insights into the diagnosis of many brain disorders such as schizophrenia, autism, lissencephaly and polymicrogyria.Keywords: brain, cortical folding, finite element, three hinge
Procedia PDF Downloads 2313778 Biomechanical Prediction of Veins and Soft Tissues beneath Compression Stockings Using Fluid-Solid Interaction Model
Authors: Chongyang Ye, Rong Liu
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Elastic compression stockings (ECSs) have been widely applied in prophylaxis and treatment of chronic venous insufficiency of lower extremities. The medical function of ECS is to improve venous return and increase muscular pumping action to facilitate blood circulation, which is largely determined by the complex interaction between the ECS and lower limb tissues. Understanding the mechanical transmission of ECS along the skin surface, deeper tissues, and vascular system is essential to assess the effectiveness of the ECSs. In this study, a three-dimensional (3D) finite element (FE) model of the leg-ECS system integrated with a 3D fluid-solid interaction (FSI) model of the leg-vein system was constructed to analyze the biomechanical properties of veins and soft tissues under different ECS compression. The Magnetic Resonance Imaging (MRI) of the human leg was divided into three regions, including soft tissues, bones (tibia and fibula) and veins (peroneal vein, great saphenous vein, and small saphenous vein). The ECSs with pressure ranges from 15 to 26 mmHg (Classes I and II) were adopted in the developed FE-FSI model. The soft tissue was assumed as a Neo-Hookean hyperelastic model with the fixed bones, and the ECSs were regarded as an orthotropic elastic shell. The interfacial pressure and stress transmission were simulated by the FE model, and venous hemodynamics properties were simulated by the FSI model. The experimental validation indicated that the simulated interfacial pressure distributions were in accordance with the pressure measurement results. The developed model can be used to predict interfacial pressure, stress transmission, and venous hemodynamics exerted by ECSs and optimize the structure and materials properties of ECSs design, thus improving the efficiency of compression therapy.Keywords: elastic compression stockings, fluid-solid interaction, tissue and vein properties, prediction
Procedia PDF Downloads 1093777 Biomechanical Assessment of Esophageal Elongation
Authors: Marta Kozuń, Krystian Toczewski, Sylwester Gerus, Justyna Wolicka, Kamila Boberek, Jarosław Filipiak, Dariusz Patkowski
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Long gap esophageal atresia is a congenital defect and is a challenge for pediatric surgeons all over the world. There are different surgical techniques in use to treat atresia. One of them is esophageal elongation but the optimal suture placement technique to achieve maximum elongation with low-risk complications is still unknown. The aim of the study was to characterize the process of esophageal elongation from the biomechanical point of view. Esophagi of white Pekin Duck was used as a model based on the size of this animal which is similar to a newborn (2.5-4kg). The specimens were divided into two groups: the control group (CG) and the group with sutures (SG). The esophagi of the control group were mounted in the grips of the MTS Tytron 250 testing machine and tensile test until rupture was performed. The loading speed during the test was 10mm/min. Then the SG group was tested. Each esophagus was cut into two equal parts and that were fused together using surgical sutures. The distance between both esophagus parts was 20mm. Ten both ends were mounted on the same testing machine and the tensile test with the same parameters was conducted. For all specimens, force and elongation were recorded. The biomechanical properties, i.e., the maximal force and maximal elongation, were determined on the basis of force-elongation curves. The maximal elongation was determined at the point of maximal force. The force achieved with the suture group was 10.1N±1.9N and 50.3N±11.6N for the control group. The highest elongation was also obtained for the control group: 18mm±3mm vs. 13.5mm ±2.4mm for the suture group. The presented study expands the knowledge of elongation of esophagi. It is worth emphasizing that the duck esophagus differs from the esophagus of a newborn, i.e., its wall lacks striated muscle cells. This is why the parts of animal esophagi used in the research are may characterized by different biomechanical properties in comparison with newborn tissue.Keywords: long gap atresia treatment, esophageal elongation, biomechanical properties, soft tissue
Procedia PDF Downloads 943776 Nanowire Substrate to Control Differentiation of Mesenchymal Stem Cells
Authors: Ainur Sharip, Jose E. Perez, Nouf Alsharif, Aldo I. M. Bandeas, Enzo D. Fabrizio, Timothy Ravasi, Jasmeen S. Merzaban, Jürgen Kosel
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Bone marrow-derived human mesenchymal stem cells (MSCs) are attractive candidates for tissue engineering and regenerative medicine, due to their ability to differentiate into osteoblasts, chondrocytes or adipocytes. Differentiation is influenced by biochemical and biophysical stimuli provided by the microenvironment of the cell. Thus, altering the mechanical characteristics of a cell culture scaffold can directly influence a cell’s microenvironment and lead to stem cell differentiation. Mesenchymal stem cells were cultured on densely packed, vertically aligned magnetic iron nanowires (NWs) and the effect of NWs on the cell cytoskeleton rearrangement and differentiation were studied. An electrochemical deposition method was employed to fabricate NWs into nanoporous alumina templates, followed by a partial release to reveal the NW array. This created a cell growth substrate with free-standing NWs. The Fe NWs possessed a length of 2-3 µm, with each NW having a diameter of 33 nm on average. Mechanical stimuli generated by the physical movement of these iron NWs, in response to a magnetic field, can stimulate osteogenic differentiation. Induction of osteogenesis was estimated using an osteogenic marker, osteopontin, and a reduction of stem cell markers, CD73 and CD105. MSCs were grown on the NWs, and fluorescent microscopy was employed to monitor the expression of markers. A magnetic field with an intensity of 250 mT and a frequency of 0.1 Hz was applied for 12 hours/day over a period of one week and two weeks. The magnetically activated substrate enhanced the osteogenic differentiation of the MSCs compared to the culture conditions without magnetic field. Quantification of the osteopontin signal revealed approximately a seven-fold increase in the expression of this protein after two weeks of culture. Immunostaining staining against CD73 and CD105 revealed the expression of antibodies at the earlier time point (two days) and a considerable reduction after one-week exposure to a magnetic field. Overall, these results demonstrate the application of a magnetic NW substrate in stimulating the osteogenic differentiation of MSCs. This method significantly decreases the time needed to induce osteogenic differentiation compared to commercial biochemical methods, such as osteogenic differentiation kits, that usually require more than two weeks. Contact-free stimulation of MSC differentiation using a magnetic field has potential uses in tissue engineering, regenerative medicine, and bone formation therapies.Keywords: cell substrate, magnetic nanowire, mesenchymal stem cell, stem cell differentiation
Procedia PDF Downloads 1883775 Cosmetic Dermatology Procedures: Survey Results of American Society for Dermatologic Surgery
Authors: Marina S. Basta, Kirollos S. Basta
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Cosmetic dermatology procedures have witnessed exponential growth and diversification over the last 10 years. Thus, the purpose of this study was to collect data about the latest trends for cosmetic procedures reported by dermatologists during the year 2018. This study was performed by American Society for Dermatologic Surgery (ASDS) in 2018 through sending survey invitations to 3,358 practicing dermatologists in the U.S. containing streamline questions as well as statistical questions targeted to specific analysis of cosmetic dermatology trends. Out of the targeted physicians, only 596 dermatologists reply to the survey invitation (15% overall response rate). It was noted that data collected from that survey was generalized to represent all ASDS members. Results show that there is an increase in cosmetic dermatology procedures since 12.5 million procedures were reported for 2018 compared to only 7.8 million for 2012. Injectable neuromodulators and soft tissue fillers have topped the list with a 3.7 million procedure count. Body sculpting, chemical peeling, hair transplantation, and microneedling procedures were reported to be 1.57 million cases combined. Also, the top two procedures using laser were represented in wrinkle treatment as well as sun damage correction, while the lowest two trends for laser usage were for treatments of tattoos and birthmarks. Cryolipolysis was found to be at the head of body sculpting procedures with 287,435 cases, while tumescent liposuction was reported as the least performed body sculpting procedure (18,286 cases). In conclusion, comparing the procedural trends for the last 7 years has indicated that there has been a 78% increase in soft tissue filler treatment compared to 2012. In addition, it was further noted that laser procedures scored 74% increase in the last 7 years while body contouring procedures have had four folds increase in general compared to 2012.Keywords: cosmetic dermatology, ASDS procedure survey, laser, body sculpting
Procedia PDF Downloads 1203774 Genetic Association and Functional Significance of Matrix Metalloproteinase-14 Promoter Variants rs1004030 and rs1003349 in Gallbladder Cancer Pathogenesis
Authors: J. Vinay , Kusumbati Besra, Niharika Pattnaik, Shivaram Prasad Singh, Manjusha Dixit
Abstract:
Gallbladder cancer (GBC) is rare but highly malignant cancer; its prevalence is more in certain geographical regions and ethnic groups, which include the Northern and Eastern states of India. Previous studies in India have reported genetic predisposition as one of the risk factors in GBC pathogenesis. Although the matrix metalloproteinase-14 (MMP14) is a well-known modulator of the tumor microenvironment and tumorigenesis and TCGA data also suggests its upregulation yet, its role in the genetic predisposition for GBC is completely unknown. We elucidated the role of MMP14 promoter variants as genetic risk factors and their implications in expression modulation. We screened MMP14 promoter variants association with GBC using Sanger’s sequencing in approximately 300 GBC and 300 control subjects and 26 GBC tissue samples of Indian ethnicity. The immunohistochemistry was used to check the MMP14 protein expression in GBC tissue samples. The role of promoter variants on expression levels was elucidated using a luciferase reporter assay. The variants rs1004030 (p-value = 0.0001) and rs1003349 (p-value = 0.0008) were significantly associated with gallbladder cancer. The luciferase assay in two different cell lines, HEK-293 (p = 0.0006) and TGBC1TKB (p = 0.0036) showed a significant increase in relative luciferase activity in the presence of risk alleles for both the single nucleotide polymorphisms (SNPs). Similarly, genotype-phenotype correlation in patients samples confirmed that the presence of risk alleles at rs1004030 and rs1003349 increased MMP14 expression. Overall, this study unravels the genetic association of MMP14 promoter variants with gallbladder cancer, which may contribute to pathogenesis by increasing its expression.Keywords: gallbladder cancer, matrix metalloproteinase-14, single nucleotide polymorphism, case control study, genetic association study
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