Search results for: tissue engineering

Authors: Luiz Carlos Wrobel, Matjaz Hribersek, Jure Marn, Jurij Iljaz

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Dynamic thermography has been clinically proven to be a valuable diagnostic technique for skin tumor detection as well as for other medical applications such as breast cancer diagnostics, diagnostics of vascular diseases, fever screening, dermatological and other applications. Thermography for medical screening can be done in two different ways, observing the temperature response under steady-state conditions (passive or static thermography), and by inducing thermal stresses by cooling or heating the observed tissue and measuring the thermal response during the recovery phase (active or dynamic thermography). The numerical modelling of heat transfer phenomena in biological tissue during dynamic thermography can aid the technique by improving process parameters or by estimating unknown tissue parameters based on measured data. This paper presents a nonlinear numerical model of multilayer skin tissue containing a skin tumor, together with the thermoregulation response of the tissue during the cooling-rewarming processes of dynamic thermography. The model is based on the Pennes bioheat equation and solved numerically by using a subdomain boundary element method which treats the problem as axisymmetric. The paper includes computational tests and numerical results for Clark II and Clark IV tumors, comparing the models using constant and temperature-dependent thermophysical properties, which showed noticeable differences and highlighted the importance of using a local thermoregulation model.

Keywords: boundary element method, dynamic thermography, static thermography, skin tumor diagnostic

Procedia PDF Downloads 63

Authors: Vladimir Ryabov, Mikhail Baryshevs, Mikhail Voskresenskey, Boris Popov

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The human prostate gland (PG) samples were obtained from patients who had undergone radical prostatectomy for prostate cancer (PC) and used to extract total RNA and prepare the prostate stromal cell cultures (PSCC) and patients-derived organoids (PDO). Growth of the cell cultures was accessed under microscopic evaluation in transmitted light and the marker expression by reverse polymerase chain reaction (RT-PCR), immunofluorescence, and immunoblotting. Some PCR products from prostate tissue, PSCC, and PDO were cloned and sequenced. We found that the cells of early and late passages of PSCC and corresponding PDO expressed luminal (androgen receptor, AR; cytokeratin 18, CK18) and basal (CK5, p63) epithelial markers, the production of which decreased or disappeared in late PSCC and PDO. The PSCC and PDO of early passages from cancer tissue additionally produced cancer markers AMACR, TMPRSS2-ERG, and Ezh2. The expression of TMPRSS2-ERG fusion transcripts was verified by cloning and sequencing the PCR products. The results obtained suggest that early passages of PSCC might be used as a pre-clinical model for the evaluation of early markers of prostate cancer.

Keywords: localized prostate cancer, prostate epithelial markers, prostate cancer markers, AMACR, TMPRSS2-ERG, prostate stromal cell cultures, PDO

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Authors: E. Roshan Ara Begum, K. Bhavani, K. Subachitra, C. Kirthika, R. Shenbagarathai

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In recent years, there has been a growing concern for the production of chitosan blend nanofibrous scaffold for its favorable physicochemical properties which mimic the native extracellular matrix (ECM) both morphologically and chemically. Therefore, this study focused on production of β-chitosan(β-Cts) and Poly(vinyl alcohol)(PVA) blend nanofibrous scaffold by electrospinning. β-Cts was extracted from the squid pen waste of locally available squid variety Loligo duvauceli (Indian Squid). To the best of our knowledge, there are no reports on nanofibers preparation from the extracted β-Cts. Both the β-Cts and PVA polymers were mixed in two different proportions (30:70 and 40:60 respectively. The electrospun nanofibrous scaffolds were characterized by SEM, swelling property, in vitro enzymatic degradation, and hemo, biocompatibility properties. β-Cts/PVA nanofibers scaffolds had an average fiber diameter of 120 to 550nm.Among the two different β-Cts/PVA blends nanofibers the β-Cts/PVA (40:60) blend fibers demonstrated favourable tissue engineering properties. The β-Cts/PVA (40:60) blend nanofibers exhibited a swelling ratio of 36 ± 2.5% with mass loss percentage of 20 ± 2.71% after 4 weeks of degradation. It has exhibited good hemocompatible properties. HEK-293(Human Embryonic Kidney) cells lines were able to adhere and proliferate well in the β-Cts/PVA blends nanofibers. All these results indicated that electrospun β-Cts/PVA blends nanofibers are a suitable scaffold to be used for tissue engineering purposes.

Keywords: β-chitosan, electrospinning, nanofibers, poly(vinyl alcohol) (PVA)

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Authors: P. Shrivastava, S. Vijayalakshmi, A. K. Singh, S. Dalai, R. Teotia, P. Sharma, J. Bellare

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This paper focuses on the synthesis and application of nanobioactive glass for bone regeneration studies. Nanobioactive glass has been synthesized by sol gel method having a combination of silicon, calcium and phosphorous in the molar ratio of 75:21:4. The prepared particles were analyzed for surface morphology by FEG SEM and FEG TEM. Physiochemical properties were investigated using ICP AES, FTIR spectroscopy and X-ray diffraction (XRD) techniques. To ascertain their use for therapeutic use, biocompatibility evaluation of the particles was done by performing soaking studies in SBF and in vitro cell culture studies on MG63 cell lines. Cell morphology was observed by FE SEM and phase contrast microscopy. Nanobioactive glasses (NBG) thus prepared were of 30-200 nm in size, which makes them suitable for nano-biomedical applications. The spherical shape of the particles imparts high surface to volume ratio, promoting fast growth of hydroxyapatite (HA), which is the mineral component of bone. As evaluated by in vitro cell culture studies the NBG was found to enhance the surface activation which enhances osteoblast adhesion. This is an essential parameter to improve bone tissue integration, thereby making nanobioactive glass therapeutically suitable for correcting bone defects.

Keywords: biocompatibility, bone tissue engineering, hydroxyapatite, nanobioactive glass

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Authors: Haluk Özparlak, Murad Aydın Şanda, Gülşin Arslan

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Phoxinellus anatolicus, Carassius gibelio, Sander lucioperca, Vimba vimba tenella, Capoeta capoeta, Tinca tinca from Suğla Lake (Turkey) and Phoxinellus anatolicus, Scardinius erythrophthalmus, Tinca tinca from Beyşehir Lake (Turkey) are economically important fish species and these fish have been consumed as food by local people. P. anatolicus is also endangered and endemic species from Turkey. In this study, concentrations of Cd, Co, Cr, Fe, Mn, Ni, Pb and Zn were determined in muscle tissue of these fish by using atomic absorption spectrophotometer. Levels of metals in the muscle tissue of all the fish specimens were compared with results of previous studies, the tolerance levels of national and international guidelines and the levels of Provisional Tolerable Weekly Intake (PTWI) limits set by FAO/WHO. Concentrations of Cd, Cr, Ni and Pb in the muscle tissue of all the fish specimens from Suğla and Beyşehir Lakes exceeded the tolerance levels of national and international guidelines. However, concentrations of Cd, Fe, Pb and Zn were below PTWI limits. Therefore, in terms of these metal levels, consumption of fresh filet of examined seven fish species (weekly up to about 300 g/person) doesn’t seem to be objectionable for human health.

Keywords: Beyşehir Lake, fish, metal levels, Suğla Lake

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Authors: Soumya S., Manju Nidagodu Jayakumar, Vellore Kannan Gopinath

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Dental pulp inflammation resulting from dental caries often leads to a pathologic condition known as irreversible pulpitis and the currently managed by root canal treatment. Extirpation of the entire pulp tissue is done during this procedure, and the canal space is filled with synthetic materials. Recent studies in the stem cell biology state that some portion of the irreversibly inflamed pulp tissue could be viable with progenitor cells, having the properties similar to that of Mesenchymal stem cells. Hence, we aim to isolate Dental Pulp Stem Cells (DPSCs) from patients diagnosed with severe irreversible pulpitis and characterize the cells for the MSC specific markers. The pulp tissue was collected from the dental clinic and subjected to collagenase/dispase digestion. The isolated cells were expanded in culture, and the phenotypic characterization was done using flow cytometry. MSC specific markers such as CD-90, CD-73, and CD-105 were analysed along with negative markers such as CD-14 and CD-45. The isolated cells expressed positive expression for CD markers with CD90 and CD105 ( > 95%) and CD73 (19%). The cells did not express the negative markers CD-14 and CD-45. The commercially available DPSCs from vital extracted teeth, preferably molar/wisdom teeth with large pulp cavity or incomplete root growth in young patients (aged 15-30 years) showed more than 90% expression for all the CD markers such as CD-90, 73 and 105, whereas negative for CD-14 and CD-45. The DPSCs isolated from inflamed pulp tissue showed a less expression for CD-73 compared to the commercially available DPSCs whereas, as the other two markers were found to show similar percentage of positive expression. This could be attributed to the fact that the pulp population is very heterogeneous and we used the pooled tissue from different patients. Hence the phenotypic characterization and comparison with the commercially available DPSCs proved that the inflamed pulp tissue is a good source of MSC like cells which can be utilized further for regenerative application.

Keywords: collagenase/dispase, dental pulp stem cells, flow cytometry, irreversible pulpitis

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Authors: Shirin jalili, Hadi Shirzad, Mahasti Modarresi, Samaneh Nabavi, Somayeh Khanjani

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Identifying the source tissue of biological material found at crime scenes can be very informative in a number of cases. Despite their usefulness, current visual, catalytic, enzymatic, and immunologic tests for presumptive and confirmatory tissue identification are applicable only to a subset of samples, might suffer limitations such as low specificity, lack of sensitivity, and are substantially impacted by environmental insults. In addition their results are operator-dependent. Recently the possibility of discriminating body fluids using mRNA expression differences in tissues has been described but lack of long term stability of that Molecule and the need to normalize samples for each individual are limiting factors. The use of DNA should solve these issues because of its long term stability and specificity to each body fluid. Cells in the human body have a unique epigenome, which includes differences in DNA methylation in the promoter of genes. DNA methylation, which occurs at the 5′-position of the cytosine in CpG dinucleotides, has great potential for forensic identification of body fluids, because tissue-specific patterns of DNA methylation have been demonstrated, and DNA is less prone to degradation than proteins or RNA. Previous studies have reported several body fluid-specific DNA methylation markers.The presence or absence of a methyl group on the 5’ carbon of the cytosine pyridine ring in CpG dinucleotide regions called ‘CpG islands’ dictates whether the gene is expressed or silenced in the particular body fluid. Were described methylation patterns at tissue specific differentially methylated regions (tDMRs) to be stable and specific, making them excellent markers for tissue identification. The results demonstrate that methylation-based tissue identification is more than a proof-of-concept. The methodology holds promise as another viable forensic DNA analysis tool for characterization of biological materials.

Keywords: DNA methylation, forensic science, epigenome, tDMRs

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Authors: Ozan Ozkan, Hilal Turkoglu Sasmazel

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Natural polymers are widely used in tissue engineering applications, because of their biocompatibility, biodegradability and solubility in the physiological medium. On the other hand, synthetic polymers are also widely utilized in tissue engineering applications, because they carry no risk of infectious diseases and do not cause immune system reaction. However, the disadvantages of both polymer types block their individual usages as tissue scaffolds efficiently. Therefore, the idea of usage of natural and synthetic polymers together as a single 3D hybrid scaffold which has the advantages of both and the disadvantages of none has been entered to the literature. On the other hand, even though these hybrid structures support the cell adhesion and/or proliferation, various surface modification techniques applied to the surfaces of them to create topographical changes on the surfaces and to obtain reactive functional groups required for the immobilization of biomolecules, especially on the surfaces of synthetic polymers in order to improve cell adhesion and proliferation. In a study presented here, to improve the surface functionality and topography of the layer by layer electrospun 3D poly-epsilon-caprolactone/chitosan/poly-epsilon-caprolactone hybrid tissue scaffolds by using atmospheric pressure plasma method, thus to improve cell adhesion and proliferation of these tissue scaffolds were aimed. The formation/creation of the functional hydroxyl and amine groups and topographical changes on the surfaces of scaffolds were realized by using two different atmospheric pressure plasma systems (nozzle type and dielectric barrier discharge (DBD) type) carried out under different gas medium (air, Ar+O2, Ar+N2). The plasma modification time and distance for the nozzle type plasma system as well as the plasma modification time and the gas flow rate for DBD type plasma system were optimized with monitoring the changes in surface hydrophilicity by using contact angle measurements. The topographical and chemical characterizations of these modified biomaterials’ surfaces were carried out with SEM and ESCA, respectively. The results showed that the atmospheric pressure plasma modifications carried out with both nozzle type plasma and DBD plasma caused topographical and functionality changes on the surfaces of the layer by layer electrospun tissue scaffolds. However, the shelf life studies indicated that the hydrophilicity introduced to the surfaces was mainly because of the functionality changes. Therefore, according to the optimized results, samples treated with nozzle type air plasma modification applied for 9 minutes from a distance of 17 cm and Ar+O2 DBD plasma modification applied for 1 minute under 70 cm3/min O2 flow rate were found to have the highest hydrophilicity compared to pristine samples.

Keywords: biomaterial, chitosan, hybrid, plasma

Procedia PDF Downloads 244

Authors: Pegah Ebrahimi, Farshad Oveissi, Iman Manavi-Tehrani, Sina Naficy, David F. Fletcher, Fariba Dehghani, David S. Winlaw

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Despite advances in modern surgery, congenital heart disease remains a medical challenge and a major cause of infant mortality. Cardiovascular prostheses are routinely used in surgical procedures to address congenital malformations, for example establishing a pathway from the right ventricle to the pulmonary arteries in pulmonary valvar atresia. Current off-the-shelf options including human and adult products have limited biocompatibility and durability, and their fixed size necessitates multiple subsequent operations to upsize the conduit to match with patients’ growth over their lifetime. Non-physiological blood flow is another major problem, reducing the longevity of these prostheses. These limitations call for better designs that take into account the hemodynamical and anatomical characteristics of different patients. We have integrated tissue engineering techniques with modern medical imaging and image processing tools along with mathematical modeling to optimize the design of cardiovascular grafts in a patient-specific manner. Computational Fluid Dynamics (CFD) analysis is done according to models constructed from each individual patient’s data. This allows for improved geometrical design and achieving better hemodynamic performance. Tissue engineering strives to provide a material that grows with the patient and mimic the durability and elasticity of the native tissue. Simulations also give insight on the performance of the tissues produced in our lab and reduce the need for costly and time-consuming methods of evaluation of the grafts. We are also developing a methodology for the fabrication of the optimized designs.

Keywords: computational fluid dynamics, cardiovascular grafts, design optimization, tissue engineering

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Authors: F. M. El-Domyati, A. Atef, S. Edris, N. O. Gadalla, M. A. Al-Kordy, A. M. Ramadan, Y. M. Saad, H. S. Al-Zahrani, A. Bahieldin

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Transcriptome retrieved from SRA database of different tissues and treatments of C. roseus was assembled in order to detect tissue-specific transcription factors (TFs) and TFs possibly related to terpenoid indole alkaloids (TIA) pathway. A number of 290 TF-like transcripts along with 12 transcripts related to TIA biosynthetic pathway were divided in terms of co-expression in the different tissues, treatments and genotypes. Three transcripts encoding peroxidases 1 and 12 were downregulated in hairy root, while upregulated in mature leaf. Eight different transcripts of the TIA pathway co-expressed with TFs either functioning downstream tryptophan biosynthesis, e.g., tdc, str1 and sgd, or upstream vindoline biosynthesis, e.g., t16h, omt, nmt, d4h and dat. The results showed no differential expression of TF transcripts in hairy roots knocked down for tdc gene (TDCi) as compared to their wild type controls. There were several evidences of tissue-specific expression of TF transcripts in flower, mature leaf, root/hairy root, stem, seedling, hairy root and immature/mature leaves. Regulation included transcription factor families, e.g., bHLH, MYB and WRKY mostly induced by ABA and/or JA (or MeJA) and regulated during abiotic or biotic stress. The information of tissue-specific regulation and co-expression of TFs and genes in the TIA pathway can be utilized in manipulating alkaloid biosynthesis in C. roseus.

Keywords: SRA database, bHLH, MYB, WRKY, co-expression

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Authors: Xiao Wei, Dandan Sheng, Xiaoyan Jiang, Lili Gui, Huizhu Wang, Xi Yu, Hailiang Liu, Min Hong

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Objective: Mice Type2 allergic contact dermatitis was utilized in this study to explore the effect of AS-IV on Type 2 allergic inflammatory. Methods: The mice were topically sensitized on the shaved abdomens with 1.5% FITC solution on abdominal skin in the day 1 and day 2 and elicited on the right ear with 0.5% FITC solution at day 6. Mice were treated with either AS-IV or normal saline from day 1 to day 5 (induction phase). Auricle swelling was measured 24 h after the elicitation. Ear pathohistological examination was carried out by HE staining. IL-4\IL-13, and IL-9 levels of ear tissue were detected by ELISA. Mice were treated with AS-IV at the initial stage of induction phase, ear tissue was taked at day 3.TSLP level of ear tissue was detected by ELISA and TSLPmRNA\NF-kBmRNA\TLRs（TLR2\TLR3\TLR8\TLR9）mRNA were detected by PCR. Results: AS-IV induction phase evidently inhibited the auricle inflam-mation of the models; pathohistological results indicated that AS-IV induction phase alleviated local edema and angiectasis of mice models and reduced lymphocytic infiltration. AS-IV induction phase markedly decreased IL-4\IL-13, and IL-9 levels in ear tissue. Moreover, at the initial stage of induction pha-se, AS-IV significantly reduced TSLP\TSLPmRNA\NF-kBmRNA\TLR2mRNA\TLR8 mRNA levels in ear tissue. Conclusion: Administration with AS-IV in induction phase could inhibit Type 2 allergic contact dermatitis in mice significantly, and the mechanism may be related with regulating TSLP through TLRs-NF-kB pathway.

Keywords: Astragaioside IV, allergic contact dermatitis, TSLP, interleukin-4, interleukin-13, interleukin-9

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Authors: Leila Anoosheh

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Aim: The aim of this study was to assess The effects of aerobic training on microRNA let-7a expression and levels of tumor tissue IL-6 in mice with breast cancer. Method: Twenty BALB/c c mice (4-5 weeks,17 gr mass) were cancerous by injection of estrogen-dependent receptor breast cancer cells MC4-L2 and divided into two groups: tumor-training(TT) and tumor-control(TC) group. Then TT group completed aerobic training for 6 weeks, 5 days per week (14-18 m/min). After tumor emersion, tumor width and length were measured by digital caliper every week. 48 hours after the last exercise subjects were killed. Tissue sampling were collected and stored in -70ᵒ. Tumor tissue was homogenized and let-7a expression and IL-6 levels were accounted with Real time-PCR and ELISA Kit respectively. Statistical analysis of let-7a was conducted by the REST software. Repeated measures and independent tests were used to assess tumor size and IL-6, respectively. Results: Tumor size and IL-6 levels were significantly decreased in TT group compare with TC group (p<0.05). microRNA let-7a was increased significantly in TT against control group respectively (p=0/000). Conclusion: Reduction in tumor size, followed by aerobic exercise can be attributed to the loss of inflammatory factors such as IL-6; It seems that regarding to up regulation effects of aerobic exercise training on let-7a and down regulation effects of that on IL-6 in mice with breast cancer, This type of training can be used as adjuvant therapy in conjunction with other therapies for breast cancer.

Keywords: breast cancer, aerobic training, microRNA let-7a, IL-6

Procedia PDF Downloads 395

Authors: MyungGu Yeo, JongHan Ha, Gi-Hoon Yang, JaeYoon Lee, SeungHyun Ahn, Hyeongjin Lee, HoJun Jeon, YongBok Kim, Minseong Kim, GeunHyung Kim

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Tissue engineering is a rapidly growing interdisciplinary research area that may provide options for treating damaged tissues and organs. As a promising technique for regenerating various tissues, this technology requires biomedical scaffolds, which serve as an artificial extracellular matrix (ECM) to support neotissue growth. Electrospun micro/nanofibers have been used widely in tissue engineering because of their high surface-area-to-volume ratio and structural similarity to extracellular matrix. However, low mechanical sustainability, low 3D shape-ability, and low cell infiltration have been major limitations to their use. In this work, we propose new hybrid scaffolds interlayered with cell-laden electrospun micro/nano fibers and poly(caprolactone) microstructures. Also, we applied various concentrations of alginate and electric field strengths to determine optimal conditions for the cell-electrospinning process. The combination of cell-laden bioink (2 ⅹ 10^5 osteoblast-like MG63 cells/mL, 2 wt% alginate, 2 wt% poly(ethylene oxide), and 0.7 wt% lecithin) and a 0.16 kV/mm electric field showed the highest cell viability and fiber formation in this process. Using these conditions and PCL microstructures, we achieved mechanically stable hybrid scaffolds. In addition, the cells embedded in the fibrous structure were viable and proliferated. We suggest that the cell-embedded hybrid scaffolds fabricated using the cell-electrospinning process may be useful for various soft- and hard-tissue regeneration applications.

Keywords: bioink, cell-laden scaffold, micro/nanofibers, poly(caprolactone)

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Authors: Yimei Huang, Harvey Lui, Jianhua Zhao, Zhenguo Wu, Haishan Zeng

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Conventional laser treatment of skin diseases and cosmetic surgery is based on the principle of one-photon absorption selective photothermolysis which relies strongly on the difference in the light absorption between the therapeutic target and its surrounding tissue. However, when the difference in one-photon absorption is not sufficient, collateral damage would occur due to indiscriminate and nonspecific tissue heating. To overcome this problem, we developed a spatially selective photothermolysis method based on multiphoton absorption in which the heat generation is restricted to the focal point of a tightly focused near-infrared femtosecond laser beam aligned with the target of interest. A multimodal optical microscope with co-registered reflectance confocal imaging (RCM), two-photon fluorescence imaging (TPF), and second harmonic generation imaging (SHG) capabilities was used to perform and monitor the spatially selective photothermolysis. Skin samples excised from the shaved backs of euthanized NODSCID mice were used in this study. Treatments were performed by focusing and scaning the laser beam in the dermis with a 50µm×50µm target area. Treatment power levels of 200 mW to 400 mW and modulated pulse trains of different duration and period were experimented. Different treatment parameters achieved different degrees of spatial confinement of tissue alterations as visualized by 3-D RCM/TPF/SHG imaging. At 200 mW power level, 0.1 s pulse train duration, 4.1 s pulse train period, the tissue damage was found to be restricted precisely to the 50µm×50µm×10µm volume, where the laser focus spot had scanned through. The overlying epidermis/dermis tissue and the underneath dermis tissue were intact although there was light passing through these regions.

Keywords: multiphoton absorption photothermolysis, reflectance confocal microscopy, second harmonic generation microscopy, spatially selective photothermolysis, two-photon fluorescence microscopy

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Authors: Patcharakamon Nooeaid, Piyachat Chuysrinuan

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Electrospun natural polymers-based nanofibers are one of the most interesting materials used in tissue engineering and drug delivery applications. Bead-on-string nanofibers have gained considerable interest for sustained drug release. Vancomycin was used as the model drug and sodium alginate (SA)/soy protein isolated (SPI) as the polymer blend to fabricate the bead-on-string nanofibers by aqueous-based electrospinning. The bead-on-string SA/SPI nanofibers were successfully fabricated by the addition of poly(ethylene oxide) (PEO) as a co-blending polymer. SA-PEO with mass ratio of 70/30 showed the best spinnability with continuous nanofibers without the occurrence of beads. Bead structure formed with the addition of SPI and bead number increased with increasing SPI content. The electrospinning of 80/20 SA-PEO/SPI was obtained as a great promising bead-on-string nanofibers for drug loading, while the solution of 50/50 was not able to obtain continuous fibers. In vitro release tests showed that a more sustainable release profile up to 14 days with less initial burst release on day 1 could be obtained from the bead-on-string fibers than from smooth fibers with uniform diameter. In addition, vancomycin-loaded beaded fibers inhibited the growth of Staphylococcus aureus (S. aureus) bacteria. Therefore, the SA-PEO/SPI nanofibers showed the potential to be used as biomaterials for tissue engineering and drug delivery.

Keywords: bead-on-string fibers, electrospinning, drug delivery, tissue engineering

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Authors: Giriprasath Ramanathan, Sivakumar Singaravelu, M. D. Raja, Uma Tirichurapalli Sivagnanam

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Collagen is the abundant protein found in the skin of the animal body that has been designed to provide adequate structural support for the adhesion of cells. The dressing material widely used for tissue engineering and biomedical application has to posses good swelling and biological property for the absorption of exudates and cell proliferation. Acid solubilised collagen from the fish skin of the Arothron stellatus was extracted. The collagen with hydroxypropyl and carboxy methyl cellulose has the better biological property to enhance the healing efficiency. The inter property of collagen with interesting perspectives in the tissue engineering process leads to the development of biomaterial with natural polymer with biologically derived collagen. Keeping this as an objective, the composite biomaterial was fabricated to improve the wound healing and biological properties. In this study the collagen from Arothron stellatus fish skin (ACO) was uniformly blended separately with hydroxypropyl methyl cellulose (HPMC) and carboxyl methyl cellulose (CMC) as biosheets. The casted biosheets were impregnated with mupirocin to get rid of infection from the microbes. Further, the results obtained from differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), tensile studies and biocompatibility of the biosheets were assessed. The swelling, porosity and degradation of the casted biosheets were studied to make the biosheets as a suitable wound dressing material. ACO-HPMC and ACO-CMC biosheets both showed good results, but ACO-HPMC biosheet showed better results than ACO-CMC and hence it can be used as a potential dermal substitute in skin tissue engineering.

Keywords: arothron stellatus, biocompatibility, collagen, tensile strenght

Procedia PDF Downloads 279

Authors: Do Khanh Vy, Vuong Cat Khanh, Osamu Ohneda

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During atherosclerosis (AS) progression, perivascular adipose tissue-derived mesenchymal stem cells (PVAT-MSCs) are exposed to the hypoxic environment due to the oxygenic deprivation which might influence the adipose tissue-derived mesenchymal stem cells (AT-MSCs) function. Additionally, it has been reported that the angiogenic ability of subcutaneous AT-MSCs (SAT-MSCs) was impaired in the AS patients. However, up to now, the effects of AS on the characteristics and function of PVAT-MSCs have not been clarified yet. In the present study, we analyzed the AS microenvironment effects on the characteristics and function of AT-MSCs. We found that there was no significant difference in cellular morphology and differentiation ability between SAT-MSCs and PVAT-MSCs in AS patients. However, the proliferation of AS-derived PVAT-MSCs was less than those of AS-derived SAT-MSCs. Importantly, the migration of AS-derived PVAT-MSCs was faster than AS-derived SAT-MSCs. Of note, AS-derived PVAT-MSCs showed the upregulation of SDF1, which is related to the homing, and VEGF, which is related to the angiogenesis compared to those of AS-derived SAT-MSCs. Consistent with these results, AS-derived PVAT-MSCs showed the higher ability to recruit EPCs and ECs than AS-derived SAT-MSCs. In addition, EPCs and ECs which cultured in the presence of AS-derived PVAT-MSC conditioned medium showed the higher angiogenic function of the tube formation compared to those cultured in AS-derived SAT-MSC conditioned medium. This result suggests that the higher paracrine effects of AS-derived PVAT-MSCs support the angiogenic function of the target cells. Our data showed the different characteristics and functions of AT-MSCs derived from different sources of tissues. Under the AS microenvironment, it seems that the characteristics and functions of PVAT-MSCs might reflect the progression of AS. Further study will be necessary to clarify the mechanism in the future.

Keywords: atherosclerosis, mesenchymal stem cells, perivascular adipose tissue, subcutaneous adipose tissue

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Authors: Ratchada Sopakayang, Gerhard A. Holzapfel

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In this study, a new constitutive model is developed to describe the hyperelastic behavior of collagenous tissues with a parallel arrangement of collagen fibers such as ligaments and tendons. The model is formulated using a continuum approach incorporating the structural changes of the main tissue components: collagen fibers, proteoglycan-rich matrix and fiber-matrix interaction. The mechanical contribution of the interaction between the fibers and the matrix is simply expressed by a coupling term. The structural change of the collagen fibers is incorporated in the constitutive model to describe the activation of the fibers under tissue straining. Finally, the constitutive model can easily describe the stress-stretch nonlinearity which occurs when a ligament/tendon is axially stretched. This study shows that the interaction between the fibers and the matrix contributes to the mechanical tissue response. Therefore, the model may lead to a better understanding of the physiological mechanisms of ligaments and tendons under axial loading.

Keywords: constitutive model, fiber-matrix, hyperelasticity, interaction, ligament, tendon

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Authors: Naeim Akbari Shahkhosravi, Reza Kakavand, Helen M. S. Davies, Amin Komeili

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Equine locomotion and performance are significantly affected by hoof health. One of the most critical diseases of the hoof is laminitis, which can lead to horse lameness in a severe condition. This disease exhibits the mechanical properties degradation of the laminar junction tissue within the hoof. Therefore, it is essential to investigate the biomechanics of the hoof, focusing specifically on excessive and cumulatively accumulated stresses within the laminar junction tissue. For this aim, the current study generated a novel equine hoof Finite Element (FE) model under dynamic physiological loading conditions and employing a hyperelastic material model. Associated tissues of the equine hoof were segmented from computed tomography scans of an equine forelimb, including the navicular bone, third phalanx, sole, frog, laminar junction, digital cushion, and medial- dorsal- lateral wall areas. The inner tissues were connected based on the hoof anatomy, and the hoof was under a dynamic loading over cyclic strides at the trot. The strain distribution on the hoof wall of the model was compared with the published in vivo strain measurements to validate the model. Then the validated model was used to study the development of laminitis. The ultimate stress tolerated by the laminar junction before rupture was considered as a stress threshold. The tissue damage was simulated through iterative reduction of the tissue’s mechanical properties in the presence of excessive maximum principal stresses. The findings of this investigation revealed how damage initiates from the medial and lateral sides of the tissue and propagates through the hoof dorsal area.

Keywords: horse hoof, laminitis, finite element model, continuous damage

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Authors: Martyna Ekiert, Andrzej Mlyniec

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Tendons are the biological structures, which primary function is to transfer force generated by muscles to the bones. Unfortunately, damages of tendons are also one of the most common injuries of the human musculoskeletal system. For the most severe cases of tendon rupture, such as the tear of calcaneus tendon or anterior cruciate ligament of the knee, a surgical procedure is the only possible way of full recovery. Tendons used as biological grafts are usually subjected to the process of deep freezing and subsequent thawing. This, in particular for multiple freezing/thawing cycles, may result in changes of tendon internal structure causing deterioration of mechanical properties of the tissue. Therefore, studies on the influence of freezing on tendons biomechanics, including internal water content in soft tissue, seems to be greatly needed. An experimental study of the influence of freezing on mechanical properties of the tendon was performed on fascicles samples dissected form bovine flexor tendons. The preparation procedure was performed with the presence of 0.9% saline solution in order to prevent an excessive tissue drying. All prepared samples were subjected to the different number of freezing/thawing cycles. For freezing part of the protocol we used -80°C temperature while for slow thawing we used fridge temperature (4°C) combined with equalizing temperatures in the standard state (25°C). After final thawing, the mechanical properties of each sample was examined using cyclic loading test. Our results may contribute for better understanding of negative effects of soft tissues freezing, resulting from abnormal thermal expansion of water. This also may help to determine the limit of freezing/thawing cycles disqualifying tissue for surgical purposes and thus help optimize tissues storage conditions.

Keywords: freezing, soft tissue, tendon, bovine fascicles

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Authors: Masoud Sakhinia, Sajjad Ahmad

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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

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Authors: Musarat Ishaq, Tara Karnezis, Ramin Shayan

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Lipedema, a poorly understood chronic disease of adipose hyper-deposition, is often mistaken for obesity and causes significant impairment to mobility and quality-of-life. To identify molecular mechanisms underpinning lipedema, we employed comprehensive omics-based comparative analyses of whole tissue, adipocyte precursors (adipose-derived stem cells (ADSCs)), and adipocytes from patients with or without lipedema. Transcriptional profiling revealed significant differences in lipedema tissue, adipocytes, and ADSCs, with altered levels of mRNAs involved inproliferation and cell adhesion. One highly up-regulated gene in lipedema adipose tissue, adipocytes and ADSCs, ZIC4, encodes Zinc Finger Protein ZIC 4, a class of transcription factor which may be involved in regulating metabolism and adipogenesis. ZIC4 inhibition impaired the adipogenesis of ADSCs into mature adipocytes. Epigenetic regulation study revealed overexpression of ZIC4 is involved in decreased promoter DNA methylation and subsequent decrease in adipogenesis. These epigenetic modifications can alter adipocytes microenvironment and adipocytes differentiation. Our study show that epigenetic events regulate the ability of ADSCs to commit and differentiate into mature adipocytes by modulating ZIC4.

Keywords: lipedema, adipose-derived stem cells, adipose tisue, adipocytes, zinc finger protein, epigenetic

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Authors: Rassoli Aisa, Abrishami Movahhed Arezu, Faturaee Nasser, Seddighi Amir Saeed, Shafigh Mohammad

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Cardiovascular diseases are one of the most common causes of mortality in developed countries. Coronary artery abnormalities and carotid artery stenosis, also known as silent death, are among these diseases. One of the treatment methods for these diseases is to create a deviatory pathway to conduct blood into the heart through a bypass surgery. The saphenous vein is usually used in this surgery to create the deviatory pathway. Unfortunately, a re-surgery will be necessary after some years due to ignoring the disagreement of mechanical properties of graft tissue and/or applied prostheses with those of host tissue. The objective of the present study is to clarify the viscoelastic behavior of human saphenous tissue. The stress relaxation tests in circumferential and longitudinal direction were done in this vein by exerting 20% and 50% strains. Considering the stress relaxation curves obtained from stress relaxation tests and the coefficients of the standard solid model, it was demonstrated that the saphenous vein has a non-linear viscoelastic behavior. Thereafter, the fitting with Fung’s quasilinear viscoelastic (QLV) model was performed based on stress relaxation time curves. Finally, the coefficients of Fung’s QLV model, which models the behavior of saphenous tissue very well, were presented.

Keywords: Viscoelastic behavior, stress relaxation test, uniaxial tensile test, Fung’s quasilinear viscoelastic (QLV) model, strain rate

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Authors: Amin Jabbari

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The state of the art in major 3D printing technologies, such as powder-based and slurry based, has led researchers to investigate the ability to fabricate bone scaffolds for bone tissue engineering using biomaterials. In addition, 3D printing technology can simulate mechanical and biological surface properties and print with high precision complex internal and external structures that match their functional properties. Polymer matrix composites reinforced with particulate bioceramics, hydrogels reinforced with particulate bioceramics, polymers coated with bioceramics, and non-porous bioceramics are among the materials that can be investigated for bone scaffold printing. Furthermore, it was shown that the introduction of high-density micropores into the sparingly dissolvable CSiMg10 and dissolvable CSiMg4 shell layer inevitably leads to a nearly 30% reduction in compressive strength, but such micropores can easily influence the ion release behavior of the scaffolds. Also, biocompatibility tests such as cytotoxicity, hemocompatibility and genotoxicity were tested on printed parts. The printed part was tested in vitro, and after 24-26 h for cytotoxicity, and 4h for hemocompatibility test, the CSiMg4@CSiMg10-p scaffolds were found to have significantly higher osteogenic capability than the other scaffolds of implantation. Overall, these experimental studies demonstrate that 3D printed, additively-manufactured bioceramic calcium (Ca)-silicate scaffolds with appropriate pore dimensions are promising to guide new bone ingrowth.

Keywords: AM, 3D printed implants, bioceramic, tissue engineering

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Authors: Anna Lierova, Jitka Kasparova, Marcela Jelicova, Lucie Korecka, Zuzana Bilkova, Zuzana Sinkorova

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Hyaluronic acid (HA) is a simple linear, unbranched polysaccharide with a lot of exceptional physiological and chemical properties such as high biocompatibility and biodegradability, strong hydration and viscoelasticity that depend on the size of the molecule. It plays the important role in a variety of molecular events as tissue hydration, mechanical protection of tissues and as well as during inflammation, leukocyte migration, and extracellular matrix remodeling. Also, HA-based biomaterials, including HA scaffolds, hydrogels, thin membranes, matrix grafts or nanoparticles are widely use in various biomedical applications. Our goal is to determine the radioprotective effect of hyaluronic acid nanoparticles (HA NPs). We are investigating effect of ionizing radiation on stability of HA NPs, in vitro relative toxicity of nanoscale as well as effect on cell lines and specific surface receptors and their response to ionizing radiation. An exposure to ionizing radiation (IR) can irreversibly damage various cell types and may thus have implications for the level of the whole tissue. Characteristic manifestations are formation of over-granulated tissue, remodeling of extracellular matrix (ECM) and abortive wound healing. Damages are caused by either direct interaction with DNA and IR proteins or indirectly by radicals formed during radiolysis of water Accumulation and turnover of ECM are a hallmark of radiation induces lung injury, characterized by inflammation, repair or remodeling health pulmonary tissue. HA is a major component of ECM in lung and plays an important role in regulating tissue injury, accelerating tissue repair, and controlling disease outcomes. Due to that, HA NPs were applied to in vivo model (C57Bl/6J mice) before total body or partial thorax irradiation. This part of our research is targeting on effect of exogenous HA on the development and/or mitigating acute radiation syndrome and radiation induced lung injuries.

Keywords: hyaluronic acid, ionizing radiation, nanoparticles, radiation induces lung damages

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Authors: Djamel Remache, Serge Dos Santos, Michael Cliez, Michel Gratton, Patrick Chabrand, Jean-Marie Rossi, Jean-Louis Milan

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Skin tissue is an inhomogeneous and anisotropic material. Uniaxial tensile testing is one of the primary testing techniques for the mechanical characterization of skin at large scales. In order to predict the mechanical behavior of materials, the direct or inverse analytical approaches are often used. However, in case of an inhomogeneous and anisotropic material as skin tissue, analytical approaches are not able to provide solutions. The numerical simulation is thus necessary. In this work, the uniaxial tensile test and the FEM (finite element method) based inverse method were used to identify the anisotropic mechanical properties of porcine skin tissue. The uniaxial tensile experiments were performed using Instron 8800 tensile machine®. The uniaxial tensile test was simulated with FEM, and then the inverse optimization approach (or the inverse calibration) was used for the identification of mechanical properties of the samples. Experimentally results were compared to finite element solutions. The results showed that the finite element model predictions of the mechanical behavior of the tested skin samples were well correlated with experimental results.

Keywords: mechanical skin tissue behavior, uniaxial tensile test, finite element analysis, inverse optimization approach

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Authors: Hande Yesil, Seda Aycan Altan, M. Vehbi Bal, Alper Uyar, O. Cumhur Sipahi

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Purpose: Fixed partial dentures (FPDs) must fulfill functional requirements such as phonetics, chewing efficiency and esthetics especially in the anterior region. A convex type tissue surface is usually recommended for pontics of FPDs. That pontic design also provides suitable oral hygiene and ease of cleaning. However, high esthetic requirements and correct emergence profile are not always achievable because of the convex shape of adjacent soft tissues. Therefore, the ovate type pontic which fulfills the high esthetic demands of the patients may be a good alternative to the modified ridge lap pontic design. Clinical Report: A female patient referred with the complaint of anterior upper edentulousness. In the oral examination it was determined that teeth 11, 12, 21, 22 were deficient. A thick and convex gingival tissue that may cause aesthetic problems was also observed.. Periodontal augmentation surgery was performed to ensure proper papillary configuration and gingival contour. An interim removable partial denture (IRPD) which applied pressure to operated gingival tissues was fabricated postoperatively. The IRPD was used for 4 weeks and after completion of tissue sculpting, the permanent FPD with an ovate pontic was fabricated and cemented. After a follow-up period of 6 months, not any esthetical and hygienic problem was detected and the patient was satisfied with her prosthesis. Conclusion: It was concluded that shaping of gingival contours with IRPD and use of a FPD with ovate pontic fulfills all esthetic and hygienic requirements.

Keywords: interim appliance, ovate pontic, tissue sculpturing, fixed partial denture

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Authors: Vladyslav Povoroznyuk, Oksana Ivanyk, Nataliia Dzerovych

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In the diagnostics of osteoporosis, the gold standard is considered to be bone mineral density; however, X-ray densitometry is not an accurate indicator of osteoporotic fracture risk under all circumstances. In this regard, the search for new methods that could determine the indicators not only of the mineral density, but of the bone tissue quality, is a logical step for diagnostic optimization. One of these methods is the evaluation of trabecular bone quality. The aim of this study was to examine the quality and mineral density of spine bone tissue, femoral neck, and body composition of women depending on the duration of the postmenopausal period, to determine the correlation of body fat with indicators of bone mineral density and quality. The study examined 179 women in premenopausal and postmenopausal periods. The patients were divided into the following groups: Women in the premenopausal period and women in the postmenopausal period at various stages (early, middle, late postmenopause). A general examination and study of the above parameters were conducted with General Electric X-ray densitometer. The results show that bone quality and mineral density probably deteriorate with advancing of postmenopausal period. Total fat and lean mass ratio is not likely to change with age. In the middle and late postmenopausal periods, the bone tissue mineral density of the spine and femoral neck increases along with total fat mass.

Keywords: osteoporosis, bone tissue mineral density, bone quality, fat mass, lean mass, postmenopausal osteoporosis

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Authors: C. Busuioc, I. Stancu, A. Nicoara, A. Zamfirescu, A. Evanghelidis

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The field of tissue engineering has expanded its potential due to the use of composite biomaterials belonging to increasingly complex systems, leading to bone substitutes with properties that are continuously improving to meet the patient's specific needs. Furthermore, the development of biomaterials based on ceramic and polymeric phases is an unlimited resource for future scientific research, with the final aim of restoring the original tissue functionality. Thus, in the first stage, composite scaffolds based on polycaprolactone (PCL) or polylactic acid (PLA) and inorganic powders were prepared by employing the electrospinning technique. The targeted powders were: commercial and laboratory synthesized hydroxyapatite (HAp), as well as barium titanate (BT). By controlling the concentration of the powder within the precursor solution, together with the processing parameters, different types of three-dimensional architectures were achieved. In the second stage, both the mineral powders and hybrid composites were investigated in terms of composition, crystalline structure, and microstructure so that to demonstrate their suitability for tissue engineering applications. Regarding the scaffolds, these were proven to be homogeneous on large areas and loaded with mineral particles in different proportions. The biological assays demonstrated that the addition of inorganic powders leads to modified responses in the presence of simulated body fluid (SBF) or cell cultures. Through SBF immersion, the biodegradability coupled with bioactivity were highlighted, with fiber fragmentation and surface degradation, as well as apatite layer formation within the testing period. Moreover, the final composites represent supports accepted by the cells, favoring implant integration. Concluding, the purposed fibrous materials based on bioresorbable polymers and mineral powders, produced by the electrospinning technique, represent candidates with considerable potential in the field of tissue engineering. Future improvements can be attained by optimizing the synthesis process or by simultaneous incorporation of multiple inorganic phases with well-defined biological action in order to fabricate multifunctional composites.

Keywords: barium titanate, electrospinning, fibre networks, hydroxyapatite, smart scaffolds

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Authors: Lizeth Fuentes-Mera, Vanessa Perez-Silos, Nidia K. Moncada-Saucedo, Alejandro Garcia-Ruiz, Alberto Camacho, Jorge Lara-Arias, Ivan Marino-Martinez, Victor Romero-Diaz, Adolfo Soto-Dominguez, Humberto Rodriguez-Rocha, Hang Lin, Victor Pena-Martinez

Abstract:

Biphasic scaffolds in cartilage tissue engineering have been designed to influence not only the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone to promote the implant integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a biphasic scaffold based on the assembly of a cartilage phase constituted by fibroin biofunctionalized with bovine cartilage matrix; cellularized with differentiated pre-chondrocytes from adipose tissue stem cells (autologous) and well attached to a bone phase (bone bovine decellularized) to mimic the structure of the nature of native tissue and to promote the cartilage regeneration in a model of joint damage in pigs. Biphasic scaffolds were assembled by fibroin crystallization with methanol. The histological and ultrastructural architectures were evaluated by optical and scanning electron microscopy respectively. Mechanical tests were conducted to evaluate Young's modulus of the implant. For the biological evaluation, pre-chondrocytes were loaded onto the scaffolds and cellular adhesion, proliferation, and gene expression analysis of cartilage extracellular matrix components was performed. The scaffolds that were cellularized and matured for 10 days were implanted into critical 3 mm in diameter and 9-mm in depth osteochondral defects in a porcine model (n=4). Three treatments were applied per knee: Group 1: monophasic cellular scaffold (MS) (single chondral phase), group 2: biphasic scaffold, cellularized only in the chondral phase (BS1), group 3: BS cellularized in both bone and chondral phases (BS2). Simultaneously, a control without treatment was evaluated. After 4 weeks of surgery, integration and regeneration tissues were analyzed by x-rays, histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular biphasic composites exhibited Young's modulus of 805.01 kPa similar to native cartilage (400-800 kPa). In vitro biological studies revealed the chondroinductive ability of the biphasic implant, evidenced by an increase in sulfated glycosaminoglycan (GAGs) and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, In group 1, the defects were not reconstructed. In group 2 and 3 a good integration of the implant with the surrounding tissue was observed. Defects in group 2 were fulfilled by hyaline cartilage and normal bone. Group 3 defects showed fibrous repair tissue. In conclusion; our findings demonstrated the efficacy of biphasic and bioactive scaffold based on silk fibroin, which entwined chondroinductive features and biomechanical capability with appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.

Keywords: biphasic scaffold, extracellular cartilage matrix, silk fibroin, osteochondral tissue engineering

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