Search results for: carboxy teminal telopeptidetype-1-collagen bone resorption marker

Authors: Katarzyna Drela, Miroslaw Wielgos, Mikolaj Wrobel, Barbara Lukomska

Abstract:

Mesenchymal stem cells (MSCs) have a great potential in regenerative medicine. Since the initial number of isolated MSCs is limited, in vitro propagation is often required to reach sufficient numbers of cells for therapeutic applications. During long-term culture MSCs may undergo genetic or epigenetic alterations that subsequently increase the probability of spontaneous malignant transformation. Thus, factors that influence genomic stability of MSCs following long-term expansions need to be clarified before cultured MSCs are employed for clinical application. The aim of our study was to investigate the potential for spontaneous transformation of human neonatal cord blood (HUCB-MSCs) and adult bone marrow (BM-MSCs) derived MSCs. Materials and Methods: HUCB-MSCs and BM-MSCs were isolated by standard Ficoll gradient centrifugations method. Isolated cells were initially plated in high density 106 cells per cm2. After 48 h medium were changed and non-adherent cells were removed. The malignant transformation of MSCs in vitro was evaluated by morphological changes, proliferation rate, ability to enter cell senescence, the telomerase expression and chromosomal abnormality. Proliferation of MSCs was analyzed with WST-1 reduction method and population doubling time (PDT) was calculated at different culture stages. Then the expression pattern of genes characteristic for mesenchymal or epithelial cells, as well as transcriptions factors were examined by RT-PCR. Concomitantly, immunocytochemical analysis of gene-related proteins was employed. Results: Our studies showed that MSCs from all bone marrow isolations ultimately entered senescence and did not undergo spontaneous malignant transformation. However, HUCB-MSCs from one of the 15 donors displayed an increased proliferation rate, failed to enter senescence, and exhibited an altered cell morphology. In this sample we observed two different cell phenotypes: one mesenchymal-like exhibited spindle shaped morphology and express specific mesenchymal surface markers (CD73, CD90, CD105, CD166) with low proliferation rate, and the second one with round, densely package epithelial-like cells with significantly increased proliferation rate. The PDT of epithelial-like populations was around 1day and 100% of cells were positive for proliferation marker Ki-67. Moreover, HUCB-MSCs showed a positive expression of human telomerase reverse transcriptase (hTERT), cMYC and exhibit increased number of CFU during the long-term culture in vitro. Furthermore, karyotype analysis revealed chromosomal abnormalities including duplications. Conclusions: Our studies demonstrate that HUCB-MSCs are susceptible to spontaneous malignant transformation during long-term culture. Spontaneous malignant transformation process following in vitro culture has enormous effect on the biosafety issues of future cell-based therapies and regenerative medicine regimens.

Keywords: mesenchymal stem cells, spontaneous, transformation, long-term culture

Procedia PDF Downloads 238

Authors: Wenliang Song, Yu Zhang, Hua Jin, Il Kim

Abstract:

The self-assembly of the polypeptide (PP) into well-defined structures at different length scales is both biomimetic relevant and fundamentally interesting. Although there are various reports of nanostructures fabricated by the self-assembly of various PPs, directed self-assembly of PP into three-dimensional (3D) hierarchical structure has proven to be difficult, despite their importance for biological applications. Herein, an efficient method has been developed through living polymerization of phenylalanine N-Carboxy anhydride (NCA) towards the linear and cyclic polyphenylalanine, and the new invented swelling methodology can form diverse hierarchical polypeptide crystals. The solvent-dependent self-assembly behaviors of these homopolymers were characterized by high-resolution imaging tools such as atomic force microscopy, transmission electron microscopy, scanning electron microscope. The linear and cyclic polypeptide formed 3D nano hierarchical shapes, such as a sphere, cubic, stratiform and hexagonal star in different solvents. Notably, a crystalline packing model was proposed to explain the formation of 3D nanostructures based on the various diffraction patterns, looking forward to give an insight for their dissimilar shape inflection during the self-assembly process.

Keywords: self-assembly, polypeptide, bio-polymer, crystalline polymer

Procedia PDF Downloads 211

Authors: Chowdhury Md. Navim Kabir

Abstract:

Renal impairment is one of the most severe non-communicable diseases around the world. Especially patients with diagnosed/newly diagnosed renal impairment who need surgery are more focused on preoperative and postoperative preparation. Serum creatinine is the prime biochemical marker for assessing renal function, and the level of impairment is widely measured by this marker as well as Glomerular Filtration Rate (GFR). Objective: Factors responsible for fluctuating serum creatinine during preoperative and postoperative periods and minimizing the process of serum creatinine is the ultimate goal of this study. Method: 37 patients participated in this cross-sectional study who were previously diagnosed/newly diagnosed. They were admitted to different tertiary-level hospitals for emergency or elective surgery. Fifteen patients were admitted in the renal function impairment stage and 22 were admitted as normal patients’. Values of creatinine at the pre-admission stage and 2nd/3rd post-admission follow-up were compared. Results: 0.41 was the average of 22 patients' creatinine between pre-admission and 2nd/3rd follow-up. The responsible factor like prolonged staying, immobilization, co-morbidities, different preoperative antibiotics and Non-Steroidal Anti Inflammatory Drugs (NSAIDs) were also inducers for creatinine elevation. After postoperative hemodialysis rapid decrease of creatinine is seen in normal patients, but this decrease is very much minor in Chronic Kidney Disease (CKD) diagnosed patients.

Keywords: CKD, Meropenam, NSAID, comorbidities, immobilized

Procedia PDF Downloads 50

Authors: Elizabeta Sivevska, Sunchica Petrovska, Vaska Antevska, Lidija Todorovska, Sanja Manchevska, Beti Dejanova, Ivanka Karagjozova, Jasmina Pluncevic Gligoroska

Abstract:

Introduction: Body composition plays an important role in the selection of young soccer players and it is associated with their successful performance. The most commonly used model of body composition divides the body into two compartments: fat components and fat-free mass (muscular and bone components). The aims of the study were to determine the body composition parameters of young male soccer players and to show the differences in age groups. Material and methods: A sample of 52 young male soccer players, with an age span from 9 to 14 years were divided into two groups according to the age (group 1 aged 9 to 12 years and group 2 aged 12 to 14 years). Anthropometric measurements were taken according to the method of Mateigka. The following measurements were made: body weight, body height, circumferences (arm, forearm, thigh and calf), diameters (elbow, knee, wrist, ankle) and skinfold thickness (biceps, triceps, thigh, leg, chest, abdomen). The measurements were used in Mateigka’s equations. Results: Body mass components were analyzed as absolute values (in kilograms) and as percentage values: the muscular component (MC kg and MC%), the bone component (BCkg and BC%) and the body fat (BFkg and BF%). The group up to 12 years showed the following mean values of the analyzed parameters: MM=21.5kg; MM%=46.3%; BC=8.1kg; BC%=19.1%; BF= 6.3kg; BF%= 15.7%. The second group aged 12-14 year had mean values of body composition parameters as follows: MM=25.6 kg; MM%=48.2%; BC = 11.4 kg; BC%=21.6%; BF= 8.5 kg; BF%= 14. 7%. Conclusions: The young soccer players aged 12 up to 14 years who are in the pre-pubertal phase of growth and development had higher bone component (p<0.05) compared to younger players. There is no significant difference in muscular and fat body component between the two groups of young soccer players.

Keywords: body composition, young soccer players, body fat, fat-free mass

Procedia PDF Downloads 437

Authors: Anja Kurz, Marc Flynn, Tobias Good, Marco Caversaccio, Martin Kompis

Abstract:

Bone Anchored Hearing Implants (BAHI) are routinely used in patients with conductive or mixed hearing loss, e.g. if conventional air conduction hearing aids cannot be used. New sound processors and new fitting software now allow the adjustment of parameters such as loudness compression ratios or maximum power output separately. Today it is unclear, how the choice of these parameters influences aided speech understanding in BAHI users. In this prospective experimental study, the effect of varying the compression ratio and lowering the maximum power output in a BAHI were investigated. Twelve experienced adult subjects with a mixed hearing loss participated in this study. Four different compression ratios (1.0; 1.3; 1.6; 2.0) were tested along with two different maximum power output settings, resulting in a total of eight different programs. Each participant tested each program during two weeks. A blinded Latin square design was used to minimize bias. For each of the eight programs, speech understanding in quiet and in noise was assessed. For speech in quiet, the Freiburg number test and the Freiburg monosyllabic word test at 50, 65, and 80 dB SPL were used. For speech in noise, the Oldenburg sentence test was administered. Speech understanding in quiet and in noise was improved significantly in the aided condition in any program, when compared to the unaided condition. However, no significant differences were found between any of the eight programs. In contrast, on a subjective level there was a significant preference for medium compression ratios of 1.3 to 1.6 and higher maximum power output.

Keywords: Bone Anchored Hearing Implant, baha, compression, maximum power output, speech understanding

Procedia PDF Downloads 367

Authors: Suraj Muke, Vikas Mankumare, Sadhana Sathaye

Abstract:

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

Keywords: antioxidant, osteoclast, osteoporosis, ovariectomized

Procedia PDF Downloads 385

Authors: Naso Isaiah Thanavisuth

Abstract:

Medical cannabis can be used for treatment including pain, multiple sclerosis, Parkinson's disease, and cancer. However, medical cannabis leads to adverse effects (AEs), which is delta-9-tetrahydrocannabinol (THC). In previous studies, the major of THC metabolism enzymes are CYP2C9. Especially, the variation of CYP2C9 gene consist of CYP2C9*2 on exon 3 and CYP2C9*3 on exon 7 to decrease enzyme activity. Notwithstanding, there is no data describing whether the variant of CYP2C9 genes are apharmacogenetics marker for the prediction of THC-induced AEs in Thai patients. We want to investigate the association between CYP2C9 gene and THC-induced AEs in Thai patients. We enrolled 39 Thai patients with medical cannabis treatment who were classified by clinical data. The CYP2C9*2 and *3 genotyping were conducted using the TaqMan real time PCR assay. All Thai patients who received the medical cannabis consist of twenty-four (61.54%) patients were female, and fifteen (38.46%) were male, with age range 27- 87 years. Moreover, the most AEs in Thai patients who were treated with medical cannabis between cases and controls were tachycardia, arrhythmia, dry mouth, and nausea. Particularly, thirteen (72.22%) medical cannabis-induced AEs were female and age range 33 – 69 years. In this study, none of the medical cannabis groups carried CYP2C9*2 variants in Thai patients. The CYP2C9*3 variants (*1/*3, intermediate metabolizer, IM) and (*3/*3, poor metabolizer, PM) were found, three of thirty-nine (7.69%) and one of thirty-nine (2.56%), respectively. Although, our results indicate that there is no found the CYP2C9*2. However, the variation of CYP2C9 allele might serve as a pharmacogenetics marker for screening before initiating the therapy with medical cannabis for the prevention of medical cannabis-induced AEs.

Keywords: CYP2C9, medical cannabis, adverse effects, THC, P450

Procedia PDF Downloads 97

Authors: Irina Alexandra Paun, Mona Mihailescu, Marian Zamfirescu, Catalin Romeo Luculescu, Adriana Maria Acasandrei, Cosmin Catalin Mustaciosu, Roxana Cristina Popescu, Maria Dinescu

Abstract:

A major concern in bone tissue engineering is to develop complex 3D architectures that mimic the natural cells environment, facilitate the cells growth in a defined manner and allow the flow transport of nutrients and metabolic waste. In particular, porous structures of controlled pore size and positioning are indispensable for growing human-like bone structures. Another concern is to monitor both the structures and the seeded cells with high spatial resolution and without interfering with the cells natural environment. The present approach relies on laser-based technologies employed for fabricating porous biomimetic structures that support the growth of osteoblast-like cells and for their non-invasive 3D imaging. Specifically, the porous structures were built by two photon polymerization –direct writing (2PP_DW) of the commercially available photoresists IL-L780, using the Photonic Professional 3D lithography system. The structures consist of vertical tubes with micrometer-sized heights and diameters, in a honeycomb-like spatial arrangement. These were fabricated by irradiating the IP-L780 photoresist with focused laser pulses with wavelength centered at 780 nm, 120 fs pulse duration and 80 MHz repetition rate. The samples were precisely scanned in 3D by piezo stages. The coarse positioning was done by XY motorized stages. The scanning path was programmed through a writing language (GWL) script developed by Nanoscribe. Following laser irradiation, the unexposed regions of the photoresist were washed out by immersing the samples in the Propylene Glycol Monomethyl Ether Acetate (PGMEA). The porous structures were seeded with osteoblast like MG-63 cells and their osteogenic potential was tested in vitro. The cell-seeded structures were analyzed in 3D using the digital holographic microscopy technique (DHM). DHM is a marker free and high spatial resolution imaging tool, where the hologram acquisition is performed non-invasively i.e. without interfering with the cells natural environment. Following hologram recording, a digital algorithm provided a 3D image of the sample, as well as information about its refractive index, which is correlated with the intracellular content. The axial resolution of the images went down to the nanoscale, while the temporal scales ranged from milliseconds up to hours. The hologram did not involve sample scanning and the whole image was available in one frame recorded going over 200μm field of view. The digital holograms processing provided 3D quantitative information on the porous structures and allowed a quantitative analysis of the cellular response in respect to the porous architectures. The cellular shape and dimensions were found to be influenced by the underlying micro relief. Furthermore, the intracellular content gave evidence on the beneficial role of the porous structures in promoting osteoblast differentiation. In all, the proposed laser-based protocol emerges as a promising tool for the fabrication and non-invasive imaging of porous constructs for bone tissue engineering. Acknowledgments: This work was supported by a grant of the Romanian Authority for Scientific Research and Innovation, CNCS-UEFISCDI, project PN-II-RU-TE-2014-4-2534 (contract 97 from 01/10/2015) and by UEFISCDI PN-II-PT-PCCA no. 6/2012. A part of this work was performed in the CETAL laser facility, supported by the National Program PN 16 47 - LAPLAS IV.

Keywords: biomimetic, holography, laser, osteoblast, two photon polymerization

Procedia PDF Downloads 254

Authors: Pinar Erturk, Sevde Altuntas, Fatih Buyukserin

Abstract:

In order to obtain an effective integration between an implant and a bone, implant surfaces should have similar properties to bone tissue surfaces. Especially mimicry of the chemical, mechanical and topographic properties of the implant to the bone is crucial for fast and effective osseointegration. Titanium-based biomaterials are more preferred in clinical use, and there are studies of coating these implants with oxide layers that have chemical/nanotopographic properties stimulating cell interactions for enhanced osseointegration. There are low success rates of current implantations, especially in craniofacial implant applications, which are large and vital zones, and the oxide layer coating increases bone-implant integration providing long-lasting implants without requiring revision surgery. Our aim in this study is to examine bone-cell behavior on titanium implants with an aluminum oxide layer (AAO) on effective osseointegration potential in the deformation of large zones with difficult spontaneous healing. In our study, aluminum layer coated titanium surfaces were anodized in sulfuric, phosphoric, and oxalic acid, which are the most common used AAO anodization electrolytes. After morphologic, chemical, and mechanical tests on AAO coated Ti substrates, viability, adhesion, and mineralization of adult bone cells on these substrates were analyzed. Besides with atomic layer deposition (ALD) as a sensitive and conformal technique, these surfaces were coated with pure alumina (5 nm); thus, cell studies were performed on ALD-coated nanoporous oxide layers with suppressed ionic content too. Lastly, in order to investigate the effect of the topography on the cell behavior, flat non-porous alumina layers on silicon wafers formed by ALD were compared with the porous ones. Cell viability ratio was similar between anodized surfaces, but pure alumina coated titanium and anodized surfaces showed a higher viability ratio compared to bare titanium and bare anodized ones. Alumina coated titanium surfaces, which anodized in phosphoric acid, showed significantly different mineralization ratios after 21 days over other bare titanium and titanium surfaces which anodized in other electrolytes. Bare titanium was the second surface that had the highest mineralization ratio. Otherwise, titanium, which is anodized in oxalic acid electrolyte, demonstrated the lowest mineralization. No significant difference was shown between bare titanium and anodized surfaces except AAO titanium surface anodized in phosphoric acid. Currently, osteogenic activities of these cells on the genetic level are investigated by quantitative real-time polymerase chain reaction (qRT-PCR) analysis results of RUNX-2, VEGF, OPG, and osteopontin genes. Also, as a result of the activities of the genes mentioned before, Western Blot will be used for protein detection. Acknowledgment: The project is supported by The Scientific and Technological Research Council of Turkey.

Keywords: alumina, craniofacial implant, MG-63 cell line, osseointegration, oxalic acid, phosphoric acid, sulphuric acid, titanium

Procedia PDF Downloads 112

Authors: J. T. Pitale, S. Ghassab, H. Ay, N. Berme

Abstract:

Biomechanics researchers commonly use marker-based optical motion capture (MoCap) systems to extract human body kinematic data. These systems use cameras to detect passive or active markers placed on the subject. The cameras use triangulation methods to form images of the markers, which typically require each marker to be visible by at least two cameras simultaneously. Cameras in a conventional optical MoCap system are mounted at a distance from the subject, typically on walls, ceiling as well as fixed or adjustable frame structures. To accommodate for space constraints and as portable force measurement systems are getting popular, there is a need for smaller and smaller capture volumes. When the efficacy of a MoCap system is investigated, it is important to consider the tradeoff amongst the camera distance from subject, pixel density, and the field of view (FOV). If cameras are mounted relatively close to a subject, the area corresponding to each pixel reduces, thus increasing the image resolution. However, the cross section of the capture volume also decreases, causing reduction of the visible area. Due to this reduction, additional cameras may be required in such applications. On the other hand, mounting cameras relatively far from the subject increases the visible area but reduces the image quality. The goal of this study was to develop a quantitative methodology to investigate marker occlusions and optimize camera placement for a given capture volume and subject postures using three-dimension computer-aided design (CAD) tools. We modeled a 4.9m x 3.7m x 2.4m (LxWxH) MoCap volume and designed a mounting structure for cameras using SOLIDWORKS (Dassault Systems, MA, USA). The FOV was used to generate the capture volume for each camera placed on the structure. A human body model with configurable posture was placed at the center of the capture volume on CAD environment. We studied three postures; initial contact, mid-stance, and early swing. The human body CAD model was adjusted for each posture based on the range of joint angles. Markers were attached to the model to enable a full body capture. The cameras were placed around the capture volume at a maximum distance of 2.7m from the subject. We used the Camera View feature in SOLIDWORKS to generate images of the subject as seen by each camera and the number of markers visible to each camera was tabulated. The approach presented in this study provides a quantitative method to investigate the efficacy and efficiency of a MoCap camera setup. This approach enables optimization of a camera setup through adjusting the position and orientation of cameras on the CAD environment and quantifying marker visibility. It is also possible to compare different camera setup options on the same quantitative basis. The flexibility of the CAD environment enables accurate representation of the capture volume, including any objects that may cause obstructions between the subject and the cameras. With this approach, it is possible to compare different camera placement options to each other, as well as optimize a given camera setup based on quantitative results.

Keywords: motion capture, cameras, biomechanics, gait analysis

Procedia PDF Downloads 294

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

Abstract:

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

Keywords: biocompatibility, NiTi, surface, porous structure

Procedia PDF Downloads 59

Authors: Zuzana Chaloupková, Zdeňka Marková, Václav Ranc, Radek Zbořil

Abstract:

Prostate cancer is now one of the most serious oncological diseases in men with an incidence higher than that of all other solid tumors combined. Diagnosis of prostate cancer usually involves detection of related genes or detection of marker proteins, such as PSA. One of the new potential markers is PSMA (prostate specific membrane antigen). PSMA is a unique membrane bound glycoprotein, which is considerably overexpressed on prostate cancer as well as neovasculature of most of the solid tumors. Commonly applied methods for a detection of proteins include techniques based on immunochemical approaches, including ELISA and RIA. Magnetically assisted surface enhanced Raman spectroscopy (MA-SERS) can be considered as an interesting alternative to generally accepted approaches. This work describes a utilization of MA-SERS in a detection of PSMA in human blood. This analytical platform is based on magnetic nanocomposites Fe3O4@Ag, functionalized by a low-molecular selector labeled as JB303. The system allows isolating the marker from the complex sample using application of magnetic force. Detection of PSMA is than performed by SERS effect given by a presence of silver nanoparticles. This system allowed us to analyze PSMA in clinical samples with limits of detection lower than 1 ng/mL.

Keywords: diagnosis, cancer, PSMA, MA-SERS, Ag nanoparticles

Procedia PDF Downloads 205

Authors: Mehieddine Bouatrous

Abstract:

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

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

Procedia PDF Downloads 57

Authors: Hsiao-Chien Tsai, Yu-Pin Chen, Ruei-Ming Chen

Abstract:

Introduction: Osteoarthritis (OA) is characterized by joint destruction and causes chronic disability. One of the prominent symptoms is pain. Alleviating the pain is necessary and urgent for the therapy of OA patients. However, currently, understanding the mechanisms that drive OA-induced pain remains challenging, which hampers the optimistic management of pain in OA patients. Semaphorin 4D (Sema4D) participates in axon guidance pathway and bone remodeling, thus, may play a role in the regulation of pain in OA. In this study, we have established a rat model of OA to find out the mechanisms of OA-induced pain and to deliberate the roles of Sema4D. Methods: Behavioral changes and the pro-inflammatory cytokines (IL-1β, TNF-α, and IL-17) associated with pain were measured during the development of OA. Sema4D expression in cartilage and synovial membrane at 1, 4, and 12 weeks after inducing OA was analyzed. To assess if Sema4D is related to the neurogenesis in OA as an axon repellant, we analyzed the expression of PGP9.5 as well. Results: Synovitis and cartilage degradation were evident histologically during the development of OA. Mechanical hyperalgesia was most severe at week 1, then persisted thereafter. It was associated with stress coping strategies. Similar to the pain behavioral results, levels of IL-1β and TNF-α in synovial lavage fluid were significantly elevated in the OA group at weeks 1 and 4, respectively. Sema4D expression in cartilage and the synovial membrane was also enhanced in the OA group and was correlated with pain and pro-inflammatory cytokines. The marker of neurogenesis, PGP9.5, was also enhanced during the development of OA. Discussion: OA induced mechanical hyperalgesia, which might be through upregulating IL-1β/TNF-α-mediated Sema4D expressions. If anti-Sema4D treatment could reduce OA-induced mechanical hyperalgesia and prevent the subsequent progression of OA needs to be further investigated. Significance: OA can induce mechanical hyperalgesia through upregulation of IL-1β/TNF-α-mediated Sema4D and PGP9.5 expressions. And the upregulation of Sema4D may indicate the severity or active status of OA and OA-induced pain.

Keywords: traumatic osteoarthritis, mechanical hyperalgesia, Sema4D, inflammatory cytokines

Procedia PDF Downloads 59

Authors: Seyedmahdi Mousavihashemi

Abstract:

Natural Akermanite (NAM) has been successfully prepared by a modified sol-gel method. Optimization in calcination temperature and mechanical ball milling resulted in a pure and nano-sized powder which characterized by means of scanning electron microscopy (SEM), X-ray diffraction (XRD), transmission electron microscopy (TEM) and Fourier transform infrared Spectroscopy (FT–IR). We hypothesized that nano-sized Akermanite (AM) would mimic more efficiently the nanocrystal structure and function of natural bone apatite, owing to the higher surface area, compare to conventional micron-size Akermanite (AM). Accordingly, we used the unique advantage of nanotechnology to improve novel nano akermanite particles as a potential candidate for bone tissue regeneration whether as a per implant filling powder or in combination with other biomaterials as a composite scaffold. Pure Akermanite (PAM) powders were successfully obtained via a simple sol-gel method followed by calcination at 1250 °C. Mechanical grinding in a ceramic ball mill for 7 hours resulted in akermanite (AM) nanoparticles in the range of about 30- 45 nm.

Keywords: biomedical engineering, nano composite, SEM, TEM

Procedia PDF Downloads 220

Authors: G. Voicu, C. D. Ghitulica, A. Cucuruz, C. Busuioc

Abstract:

In the field of tissue engineering, the compositional and microstructural features of the employed materials play an important role, with implications on the mechanical and biological behaviour of the medical devices. In this context, the development of calcium phosphate-natural biopolymer composites represents a choice of many scientific groups. Thus, tricalcium phosphate powders were synthesized by a wet method, namely co-precipitation, starting from high purity reagents. Moreover, the substitution of calcium with magnesium have been approached, in the 5-10 wt.% range. Afterwards, the phosphate powders were integrated into two types of composites with chitosan, different from morphological point of view. First, 3D porous scaffolds were obtained by a freeze-drying procedure. Second, uniform compact films were achieved by film casting. The influence of chitosan molecular weight (low, medium and high), as well as phosphate powder to polymer ratio (1:1 and 1:2) on the morphological properties, were analysed in detail. In conclusion, the reported biocomposites, prepared by a straightforward route are suitable for bone substitution or repairing applications.

Keywords: bone reconstruction, chitosan, composite scaffolds, tricalcium phosphate

Procedia PDF Downloads 224

Authors: Cecília M. Antão, Paulo Jorge Nogueira

Abstract:

Breast cancer (BC) is the first cause of cancer mortality among Portuguese women. This retrospective observational study aimed at identifying comorbidities associated with BC female patients admitted to Portuguese public hospitals (2010-2018), investigating the effect of comorbidities on BC mortality rate, and building a predictive model using logistic regression. Results showed that the BC mortality in Portugal decreased in this period and reached 4.37% in 2018. Adjusted odds ratio indicated that secondary malignant neoplasms of liver, of bone and bone marrow, congestive heart failure, and diabetes were associated with an increased chance of dying from breast cancer. Although the Lisbon district (the most populated area) accounted for the largest percentage of BC patients, the logistic regression model showed that, besides patient’s age, being resident in Bragança, Castelo Branco, or Porto districts was directly associated with an increase of the mortality rate.

Keywords: breast cancer, comorbidities, logistic regression, adjusted odds ratio

Procedia PDF Downloads 66

Authors: Alsulami H., Alghamdi N., Alasker A., Almohen N., Shome D.

Abstract:

Most conventional chemotherapeutic agents which are used for the treatment of cancers not only eradicate cancer cells but also affect normal hematopoietic Stem cells (HSCs) that leads to severe pancytopenia during treatment. Therefore, a need exists for novel approaches to treat cancer without or with minimum effect on normal HSCs. Parthenolide (PTL), a herbal product occurring naturally in the plant Feverfew, is a potential new chemotherapeutic agent for the treatment of many cancers such as acute myeloid leukemia (AML) and chronic lymphocytic leukemia (CLL). In this study we investigated the effect of different PTL concentrations on the viability of normal HSCs and also on the ability of these cells to form colonies after they have been treated with PTL in vitro. Methods: In this study, 24 samples of bone marrow and cord blood were collected with consent, and mononuclear cells were separated using density gradient separation. These cells were then exposed to various concentrations of PTL for 24 hours. Cell viability after culture was determined using 7ADD in a flow cytometry test. Additionally, the impact of PTL on hematopoietic stem cells (HSCs) was evaluated using a colony forming unit assay (CFU). Furthermore, the levels of NFҝB expression were assessed by using a PE-labelled anti-pNFκBP65 antibody. Results: this study showed that there was no statistically significant difference in the percentage of cell death between untreated and PTL treated cells with 5 μM PTL (p = 0.7), 10 μM PTL (p = 0.4) and 25 μM (p = 0.09) respectively. However, at higher doses, PTL caused significant increase in the percentage of cell death. These results were significant when compared to untreated control (p < 0.001). The response of cord blood cells (n=4) on the other hand was slightly different from that for bone marrow cells in that the percentage of cell death was significant at 100 μM PTL. Therefore, cord blood cells seemed more resistant than bone marrow cells. Discussion &Conclusion: At concentrations ≤25 μM PTL has a minimum or no effect on HSCs in vitro. Cord blood HSCs are more resistant to PTL compared to bone marrow HSCs. This could be due to the higher percentage of T-lymphocytes, which are resistant to PTL, in CB samples (85% in CB vs. 56% in BM. Additionally, CB samples contained a higher proportion of CD34+ cells, with 14.5% of brightly CD34+ cells compared to only 1% in normal BM. These bright CD34+ cells in CB were mostly negative for early-stage stem cell maturation antigens, making them young and resilient to oxidative stress and high concentrations of PTL.

Keywords: stem cell, parthenolide, NFKB, CLL

Procedia PDF Downloads 24

Authors: Trad Khodja Rafik, Mahtout Amine, Ghoul Rachid, Benbouali Amine, Boulahlib Amine, Hariza Abdelmalik

Abstract:

Cranioplasty with custom implant realized using 3D printing technology. Cranioplasty is a surgical act that aims restoring cranial bone losses in order to protect the brain from external aggressions and to improve the patient aesthetic appearance. This objective can be achieved with taking advantage of the current technological development in computer science and biomechanics. The objective of this paper it to present an approach for the realization of high precision biocompatible cranial implants using new 3D printing technologies at the lowest cost. The proposed method is to reproduce the missing part of the skull by referring to its healthy contralateral part. Once the model is validated by the neurosurgeons, a mold is 3D printed for the production of a biocompatible implant in Poly-Methyl-Methacrylate (PMMA) acrylic cement. Using this procedure four patients underwent this procedure with excellent aesthetic results.

Keywords: cranioplasty, cranial bone loss, 3D printing technology, custom-made implants, PMMA

Procedia PDF Downloads 87

Authors: Predrag Kavaric, Vladimir L. Jubic, Maxim Cadenovic

Abstract:

The patient was transferred from his dentist to our tertiary medical institution. In anamnesis, we got information that his dental intervention was two years ago when he got dental implants but because of the coronavirus pandemic event, he didn’t finish the whole procedure. After two years, he decided that he will continue his work at his dentist, then his dentist noticed that there is no earlier inserted implant in the upper jaw on the right side. They do Panoramic X-ray and find that the implant is all in the maxillary sinus cavity. The flour of the maxilla was intact without any fistula on the place where the implant was inserted in the maxilla bone, After that initial diagnostic they sent the patient to maxillofacial surgery and otorhinolaryngology. We asked for a CT scan of paranasal sinuses, which confirmed the foreign body in the right maxillary sinus. The plan was that in general anesthesia we do FESS and try to find a foreign body in the maxillary sinus or in case of failure to do Caldwel Luc on that side. After preoperative preparation in GA, we do FESS. In inspection, we find small polyps and chronically changed mucosa of osteomeatal complex and right maxillary sinus. After removing polyps we did uncinectomy and medial maxillectomy. With Heuweiser Antrum grasping forceps after several attempts we managed to extract a foreign body from the bottom of the right maxillary sinus. On the first postoperative day we did detamponade, and then we discharge the patient from hospital. The Covid pandemic has contributed to the postponement of a large number of planned operations, which has resulted in various complications in the treatment of a number of patients. In this case, it happened that the implant was most likely rejected by the bone but in the direction of the maxillary sinus, which is not a common cause. On the other hand, the success was that less traumatic intervention was able to remove the foreign body from the maxillary sinus in which it was located. Since the sinus floor is free of bone defects, it can be continued relatively quickly with dental procedures.

Keywords: x-ray, surgery, maxillar sinus, complication, fees

Procedia PDF Downloads 133

Authors: Ane Escobar, Paula Angelomé, Mihaela Delcea, Marek Grzelczak, Sergio Enrique Moya

Abstract:

The antibacterial capacity of bone-anchoring implants can be improved by the use of antibiotics that can be delivered to the media after the surgery. Mesoporous films have shown great potential in drug delivery for orthopedic applications, since pore size and thickness can be tuned to produce different surface area and free volume inside the material. This work shows the synthesis of mesoporous titania films (MTF) by sol-gel chemistry and evaporation-induced self-assembly (EISA) on top of glass substrates. Pores with a diameter of 12nm were observed by Transmission Electron Microscopy (TEM). A film thickness of 100 nm was measured by Scanning Electron Microscopy (SEM). Gentamicin was used to study the antibiotic delivery from the film by means of High-performance liquid chromatography (HPLC). The Staphilococcus aureus strand was used to evaluate the effectiveness of the penicillin loaded films toward inhibiting bacterial colonization. MC3T3-E1 pre-osteoblast cell proliferation experiments proved that MTFs have a good biocompatibility and are a suitable surface for MC3T3-E1 cell proliferation. Moreover, images taken by Confocal Fluorescence Microscopy using labeled vinculin, showed good adhesion of the MC3T3-E1 cells to the MTFs, as well as complex actin filaments arrangement. In order to improve cell proliferation Bone Morphogenetic Protein-2 (BMP-2) was adsorbed on top of the mesoporous film. The deposition of the protein was proved by measurements in the contact angle, showing an increment in the hydrophobicity while the protein concentration is higher. By measuring the dehydrogenase activity in MC3T3-E1 cells cultured in dually functionalized mesoporous titatina films with gentamicin and BMP-2 is possible to find an improvement in cell proliferation. For this purpose, the absorption of a yellow-color formazan dye, product of a water-soluble salt (WST-8) reduction by the dehydrogenases, is measured. In summary, this study proves that by means of the surface modification of MTFs with proteins and loading of gentamicin is possible to achieve an antibacterial effect and a cell growth improvement.

Keywords: antibacterial, biocompatibility, bone morphogenetic protein-2, cell proliferation, gentamicin, implants, mesoporous titania films, osteoblasts

Procedia PDF Downloads 144

Authors: H. Maachou, A. Chagnes, G. Cote

Abstract:

The present work reports the properties of chitosan/hydroxyapatite (Cs/HA: 100/00, 70/30 and 30/70) composite microspheres obtained by emulsification processing route. The morphology of chitosane microspheres was observed by a scanning electron microscope (SEM) which shows an aggregate of spherical microspheres with a particle size, determined by optical microscope, ranged from 4 to 10 µm. Thereafter, a biomimetic approach was used to study the in vitro biomineralization of these composites. It concerns the composites immersion in simulated body fluid (SBF) for different times. The deposited calcium phosphate was studied using X-ray diffraction analysis (XRD), FTIR spectroscopy and ICP analysis of phosphorus. In fact, the mineral formed on Cs/HA microspheres was a mixture of carbonated HA and β-TCP as showed by FTIR peaks at 1419,5 and 871,8 cm-1 and XRD peak at 29,5°. This formation was induced by the presence of HA in chitosan microspheres. These results are confirmed by SEM micrographs which chow the Ca-P crystals growth in form of cauliflowers. So, these materials are of great interest for bone regeneration applications due to their ability to nucleate calcium phosphates in presence of simulated body fluid (SBF).

Keywords: hydroxyapatite, chitosan, microsphere, composite, bone regeneration

Procedia PDF Downloads 318

Authors: Norimichi Nagano, Yoshio Hirano, Tsutomu Yasukawa

Abstract:

Mesenchymal stem cells are thought to confer neuroprotection, facilitate tissue regeneration and exert their effects on retinal degenerative diseases, however, adverse events such as proliferative vitreoretinopathy and preretinal membrane disease associated with cell suspension transplantation have also been reported. We have recently developed human (allogeneic) adipose tissue-derived mesenchymal stem cell (adMSC) sheets through our proprietary sheet transformation technique, which could potentially mitigate these adverse events. To clarify the properties of our adMSC sheets named PAL-222, we performed in vitro studies such as viability testing, cytokine secretions by ELISA, immunohistochemical study, and migration assay. The viability of the cells exceeded 70%. Vascular Endothelial Growth Factor (VEGF) and Pigment Epithelium-Derived Factor (PEDF), which are quite important cytokines for the retinal area, were observed. PAL-222 expressed type I collagen, a strength marker, type IV collagen, a marker of the basement membrane, and elastin, an elasticity marker. Finally, the migration assay was performed and showed negative, which means that PAL-222 is stably kept in the topical area and does not come to pieces. Next, to evaluate the efficacy in vivo, we transplanted PAL-222 into the subretinal space of the eye of Royal College of Surgeons rats with congenital retinal degeneration and assessed it for three weeks after transplantation. We confirmed that PAL-222 suppressed the decrease in the thickness of the outer nuclear layer, which means that the photoreceptor protective effect treated with PAL-222 was significantly higher than that in the sham group. (p < 0.01). This finding demonstrates that PAL-222 showed their retinoprotective effect in a model of congenital retinal degeneration. As the study suggested the efficacy of PAL-222 in both in vitro and in vivo studies, we are presently engaged in clinical trials of PAL-222 for myopic chorioretinal atrophy, which is one of the retinal degenerative diseases, for the purpose of regenerative medicine.

Keywords: cell sheet, clinical trial, mesenchymal stem cell, myopic chorioretinal atrophy

Procedia PDF Downloads 62

Authors: Aulanni’am Aulanni’am

Abstract:

Incidence of Diabetes Mellitus (DM) progressively increasing it became a serious problem in Indonesia and it is a disease that government is priority to be addressed. The longer a person is suffering from diabetes the more likely to develop complications particularly diabetic patients who are not well maintained. Therefore, Incidence of Diabetes Mellitus needs to be done in the early diagnosis of pre-phase of the disease. In this pre-phase disease, already happening destruction of pancreatic beta cells and declining in beta cell function and the sign autoimmunity reactions associated with beta cell destruction. Type 1 DM is a multifactorial disease triggered by genetic and environmental factors, which leads to the destruction of pancreatic beta cells. Early marker of "beta cell autoreactivity" is the synthesis of autoantibodies against 65-kDa protein, which can be a molecule that can be detected early in the disease pathomechanism. The importance of early diagnosis of diabetic patients held in the phase of pre-disease is to determine the progression towards the onset of pancreatic beta cell destruction and take precautions. However, the price for this examination is very expensive ($ 150/ test), the anti-GAD65 abs examination cannot be carried out routinely in most or even in all laboratories in Indonesia. Therefore, production-based Rapid Test Recombinant Human Protein GAD65 with "Reverse Flow Immunchromatography Technique" in Indonesia is believed to reduce costs and improve the quality of care of patients with diabetes in Indonesia. Rapid Test Product innovation is very simple and suitable for screening and routine inspection of GAD65 autoantibodies. In the blood serum of patients with diabetes caused by autoimmunity, autoantibody-GAD65 is a major serologic marker to detect autoimmune reaction because their concentration level of stability.GAD65 autoantibodies can be found 10 years before clinical symptoms of diabetes. Early diagnosis is more focused to detect the presence autontibodi-GAD65 given specification and high sensitivity. Autoantibodies- GAD65 that circulates in the blood is a major indicator of the destruction of the islet cells of the pancreas. Results of research in collaboration with Biofarma has produced GAD65 autoantibodies based Rapid Test had conducted the soft launch of products and has been tested with the results of a sensitivity of 100 percent and a specificity between 90 and 96% compared with the gold standard (import product) which worked based on ELISA method.

Keywords: diabetes mellitus, GAD65 autoantibodies, rapid test, sensitivity, specificity

Procedia PDF Downloads 252

Authors: Ahmad Nazrun Shuid, Isa Naina Mohamed, Nurul Izzah Ibrahim, Norazlina Mohamed

Abstract:

Studies have shown that oral tocotrienol, a potent vitamin E, promoted fracture healing of osteoporotic bone. In this study, tocotrienol was combined with a polymer carrier (PLGA), and injected to the fracture site. The slow and constant release of tocotrienol particles would promote fracture healing of post-menopausal osteoporosis rat model. Fracture healing was assessed using micro-CT. Twenty-four Sprague-Dawley rats were ovariectomised or sham-operated and the left tibiae were fractured and fixed with plate and screws. The fractures were created at the upper third of the left tibiae. The rats were divided into 3 groups: sham-operated (SO), ovariectomised-control (OVxC) and PLGA-incorporated tocotrienol treatment (OVx + TT) groups. After 4 weeks, the OVx + TT group showed significantly better callus fracture healing than the OVxC group. In conclusion, tocotrienol-incorporated PLGA was able to promote fracture healing of osteoporotic bone.

Keywords: osteoporosis, micro-CT, tocotrienol, PLGA, fracture

Procedia PDF Downloads 647

Authors: Georgeta Voicu, Cristina Daniela Ghitulica, Andreia Cucuruz, Cristina Busuioc

Abstract:

In the field of tissue engineering, the compositional and microstructural features of the employed materials play an important role, with implications on the mechanical and biological behaviour of the medical devices. In this context, the development of apatite - natural biopolymer composites represents a choice of many scientific groups. Thus, hydroxyapatite powders were synthesized by a wet method, namely co-precipitation, starting from high purity reagents (CaO, MgO, and H3PO4). Moreover, the substitution of calcium with magnesium have been approached, in the 5 - 10 wt.% range. Afterward, the phosphate powders were integrated in two types of composites with chitosan, different from morphological point of view. First, 3D porous scaffolds were obtained by a freeze-drying procedure. Second, uniform, compact films were achieved by film casting. The influence of chitosan molecular weight (low, medium and high), as well as apatite powder to polymer ratio (1:1 and 1:2) on the morphological properties, were analysed in detail. In conclusion, the reported biocomposites, prepared by a straightforward route are suitable for bone substitution or repairing applications.

Keywords: bone reconstruction, chitosan, composite scaffolds, hydroxyapatite

Procedia PDF Downloads 299

Authors: Arnab Hazra

Abstract:

Breath analysis is a quick, noninvasive and inexpensive technique for disease diagnosis can be used on people of all ages without any risk. Only a limited number of volatile organic compounds (VOCs) can be associated with the occurrence of specific diseases. These VOCs can be considered as disease markers or breath markers. Selective detection with specific concentration of breath marker in exhaled human breath is required to detect a particular disease. For example, acetone (C₃H₆O), ethanol (C₂H₅OH), ethane (C₂H₆) etc. are the breath markers and abnormal concentrations of these VOCs in exhaled human breath indicates the diseases like diabetes mellitus, renal failure, breast cancer respectively. Nanomaterial-based vapor sensors are inexpensive, small and potential candidate for the detection of breath markers. In practical measurement, selectivity is the most crucial issue where trace detection of breath marker is needed to identify accurately in the presence of several interfering vapors and gases. Current article concerns a novel technique for selective and lower ppb level detection of breath markers at very low temperature based on TiO₂ nanotube array based vapor sensor devices. Highly ordered and oriented TiO₂ nanotube array was synthesized by electrochemical anodization of high purity tatinium (Ti) foil. 0.5 wt% NH₄F, ethylene glycol and 10 vol% H₂O was used as the electrolyte and anodization was carried out for 90 min with 40 V DC potential. Au/TiO₂ Nanotube/Ti, sandwich type sensor device was fabricated for the selective detection of VOCs in low concentration range. Initially, sensor was characterized where resistive and capacitive change of the sensor was recorded within the valid concentration range for individual breath markers (or organic vapors). Sensor resistance was decreased and sensor capacitance was increased with the increase of vapor concentration. Now, the ratio of resistive slope (mR) and capacitive slope (mC) provided a concentration independent constant term (M) for a particular vapor. For the detection of unknown vapor, ratio of resistive change and capacitive change at any concentration was same to the previously calculated constant term (M). After successful identification of the target vapor, concentration was calculated from the straight line behavior of resistance as a function of concentration. Current technique is suitable for the detection of particular vapor from a mixture of other interfering vapors.

Keywords: breath marker, vapor sensors, selective detection, TiO₂ nanotube array

Procedia PDF Downloads 137

Authors: H. M. Abdelmoneim, N. A. Babtain, A. S. Barhamain, A. Z. Kufiah, A. S. Malibari, S. F. Munassar, R. S. Rawa

Abstract:

Introduction: Prostate cancer is one of the most common causes of morbidity and mortality in men in developed countries. Cancer Stem Cells (CSCs) could be responsible for the progression and relapse of cancer. Therefore, CSCs markers could provide a prognostic strategy for human malignancies. Aldehyde dehydrogenase 1A1 (ALDH1A1) activity has been shown to be associated with tumorigenesis and proposed to represent a functional marker for tumor initiating cells in various tumor types including prostate cancer. Material & Methods: We analyzed the immunohistochemical expression of ALDH1A1 in benign prostatic hyperplasia (BPH), prostatic intraepithelial neoplasia (PIN) and prostatic adenocarcinoma and assessed their significant correlations in 50 TURP sections. They were microscopically interpreted and the results were correlated with histopathological types and tumor grade. Results: In different prostatic histopathological lesions we found that ALDH1A1 expression was low in BPH (13.3%) and PIN (6.7%) and then its expression increased with prostatic adenocarcinoma (40%), and this was statistically highly significant (P value = 0.02). However, in different grades of prostatic adenocarcinoma we found that the higher the Gleason grade the higher the expression for ALDH1A1 and this was statistically significant (P value = 0.02). We compared the expression of ALDH1A1 in PIN and prostatic adenocarcinoma. ALDH1A1 expression was decreased in PIN and highly expressed in prostatic adenocarcinoma and this was statistically significant (P value = 0.04). Conclusion: Increasing ALDH1A1 expression is correlated with aggressive behavior of the tumor. Immunohistochemical expression of ALDH1A1 might provide a potential approach to study tumorigenesis and progression of primary prostate carcinoma.

Keywords: ALDH1A1, BPH, PIN, prostatic adenocarcinoma

Procedia PDF Downloads 238

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

Abstract:

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

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

Procedia PDF Downloads 183

Authors: Masoud Sakhinia, Sajjad Ahmad

Abstract:

Introduction: Though knowledge of the compositional makeup and structure of the limbal niche has progressed exponentially during the past decade, much is yet to be understood. Identifying the precise profile and role of the stromal makeup which spans the ocular surface may inform researchers of the most optimum conditions needed to effectively expand LESCs in vitro, whilst preserving their differentiation status and phenotype. Limbal fibroblasts, as opposed to corneal fibroblasts are thought to form an important component of the microenvironment where LESCs reside. Methods: The corneal stroma was tissue engineered in vitro using both limbal and corneal fibroblasts embedded within a tissue engineered 3D collagen matrix. The effect of these two different fibroblasts on LESCs and hTCEpi corneal epithelial cell line were then subsequently determined using phase contrast microscopy, histolological analysis and PCR for specific stem cell markers. The study aimed to develop an in vitro model which could be used to determine whether limbal, as opposed to corneal fibroblasts, maintained the stem cell phenotype of LESCs and hTCEpi cell line. Results: Tissue culture analysis was inconclusive and required further quantitative analysis for remarks on cell proliferation within the varying stroma. Histological analysis of the tissue-engineered cornea showed a comparable structure to that of the human cornea, though with limited epithelial stratification. PCR results for epithelial cell markers of cells cultured on limbal fibroblasts showed reduced expression of CK3, a negative marker for LESC’s, whilst also exhibiting a relatively low expression level of P63, a marker for undifferentiated LESCs. Conclusion: We have shown the potential for the construction of a tissue engineered human cornea using a 3D collagen matrix and described some preliminary results in the analysis of the effects of varying stroma consisting of limbal and corneal fibroblasts, respectively, on the proliferation of stem cell phenotype of primary LESCs and hTCEpi corneal epithelial cells. Although no definitive marker exists to conclusively illustrate the presence of LESCs, the combination of positive and negative stem cell markers in our study were inconclusive. Though it is less traslational to the human corneal model, the use of conditioned medium from that of limbal and corneal fibroblasts may provide a more simple avenue. Moreover, combinations of extracellular matrices could be used as a surrogate in these culture models.

Keywords: cornea, Limbal Stem Cells, tissue engineering, PCR

Procedia PDF Downloads 259