Search results for: tissue invasion

Authors: Yuri V. Plugatar, Vladimir P. Koba, Vladimir V. Papelbu, Vladimir N. Gerasimchuk, Tatjana M. Sakhno

Abstract:

Using a supersonic shaft–wood tomography, the evaluation of a shaft-wood phyto-pathogenic damage level of Sequoiadendron giganteum (Lindl.) Buccholz was prosecuted. The digital bivariate reflections of the shaft tissue damage were obtained, the characteristics of comparative parameters of the wood-decay degree were given. The investigation results allowed to show up the role of some edaphic factors in their affection on a vital condition and the level of destructive processes while shaft tissue damaging of S.giganteum. It was pinned up that soil consolidation, and hydro-morphication equally make for a phyto-pathogenic damage of plants. While soil consolidation negative acting the shaft-wood damage is located in an underneath of a shaft. In the conditions of an enlarged hydro-morphication a tissue degradation runs less intensively, the destructive processes more active spread in a vertical section of a shaft. The use of a supersonic tomography method gives wide possibilities to diagnose a shaft-wood phyto-pathogenic damage.

Keywords: Sequoiadendron giganteum (Lindl.) Buccholz, supersonic tomography, diagnosis, phyto-pathogenic damage, a vital condition

Procedia PDF Downloads 213

Authors: Er-Yuan Chuang

Abstract:

Bio-mimetic matters have biological functionalities. This might be valuable in the development of versatile biomaterials. At biological fields, protein-based materials might be components to form a 3D network of extracellular biomolecules, containing growth factors. Also, the protein-based biomaterial provides biochemical and structural assistance of adjacent cells. In this study, we try to prepare protein based biomaterial, which was harvested from living animal. We analyzed it’s chemical, physical and biological property in vitro. Besides, in vivo bio-interaction of the prepared biomimetic matrix was tested in an animal model. The protein-based biomaterial has degradability and biocompatibility. This development could be used for tissue regenerations and be served as platform technologies.

Keywords: protein based, in vitro study, in vivo study, biomaterials

Procedia PDF Downloads 189

Authors: Mohammad Hamed Asosheh

Abstract:

The development of nanobiomaterials has opened new avenues in the field of biomedical engineering, offering unparalleled possibilities for advanced therapeutic applications. This study explores the synthesis and characterization of a distinct class of nanobiomaterials designed to enhance drug delivery systems and support tissue engineering. By integrating biodegradable polymers with bioactive nanoparticles, we have engineered a multifunctional platform that ensures controlled drug release, targeted delivery, and improved biocompatibility. Our findings demonstrate that these nanobiomaterials not only exhibit excellent mechanical properties but also promote cell proliferation and differentiation, making them ideal candidates for regenerative medicine. Furthermore, in vitro and in vivo assessments reveal that the engineered materials significantly reduce cytotoxicity while enhancing the therapeutic efficacy of encapsulated drugs. This research presents a promising approach to addressing current challenges in drug delivery and tissue regeneration, with the potential to revolutionize the treatment of chronic diseases and injury repair. Future work will focus on optimizing the material composition for specific clinical applications and conducting large-scale studies to evaluate long-term safety and effectiveness.

Keywords: nanobiomaterials, drug delivery systems, therapeutic efficacy, bioactive nanoparticles

Procedia PDF Downloads 28

Authors: Mahsa Ahmadi, Mehdi Mehdikhani-Nahrkhalaji, Jaleh Varshosaz, Shadi Farsaei

Abstract:

Gelatin and hyaluronic acid are commonly used in skin tissue engineering scaffolds, but because of their low mechanical properties and high biodegradation rate, adding a synthetic polymer such as polycaprolactone could improve the scaffold properties. Therefore, we developed a gelatin-hyaluronic acid-polycaprolactone scaffold, containing 0.5 % atorvastatin loaded nanostructured lipid carriers (NLCs) for skin tissue engineering. The atorvastatin loaded NLCs solution was prepared by solvent evaporation method and freeze drying process. Synthesized atorvastatin loaded NLCs was added to the gelatin and hyaluronic acid solution, and a membrane was fabricated with solvent evaporation method. Thereafter it was coated by a thin layer of polycaprolactone via spine coating set. The resulting scaffolds were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analyses. Moreover, mechanical properties, in vitro degradation in 7 days period, and in vitro drug release of scaffolds were also evaluated. SEM images showed the uniform distributed NLCs with an average size of 100 nm in the scaffold structure. Mechanical test indicated that the scaffold had a 70.08 Mpa tensile modulus which was twofold of tensile modulus of normal human skin. A Franz-cell diffusion test was performed to investigate the scaffold drug release in phosphate buffered saline (pH=7.4) medium. Results showed that 72% of atorvastatin was released during 5 days. In vitro degradation test demonstrated that the membrane was degradated approximately 97%. In conclusion, suitable physicochemical and biological properties of membrane indicated that the developed gelatin-hyaluronic acid-polycaprolactone nanocomposite scaffold containing 0.5 % atorvastatin loaded NLCs could be used as a good candidate for skin tissue engineering applications.

Keywords: atorvastatin, gelatin, hyaluronic acid, nano lipid carriers (NLCs), polycaprolactone, skin tissue engineering, solvent casting, solvent evaporation

Procedia PDF Downloads 252

Authors: Tuhina-khatun, Mohamed Hanafi Musa, Mohd Rafii Yosup, Wong Mui Yun, Aktar-uz-Zaman, Mahbod Sahebi

Abstract:

Suppression subtractive hybridization (SSH) method is valuable tool for identifying differentially regulated genes in disease specific or tissue specific genes important for cellular growth and differentiation. It is a widely used method for separating DNA molecules that distinguish two closely related DNA samples. SSH is one of the most powerful and popular methods for generating subtracted cDNA or genomic DNA libraries. It is based primarily on a suppression polymerase chain reaction (PCR) technique and combines normalization and subtraction in a solitary procedure. The normalization step equalizes the abundance of DNA fragments within the target population, and the subtraction step excludes sequences that are common to the populations being compared. This dramatically increases the probability of obtaining low-abundance differentially expressed cDNAs or genomic DNA fragments and simplifies analysis of the subtracted library. SSH technique is applicable to many comparative and functional genetic studies for the identification of disease, developmental, tissue specific, or other differentially expressed genes, as well as for the recovery of genomic DNA fragments distinguishing the samples under comparison.

Keywords: suppression subtractive hybridization, differentially expressed genes, disease specific genes, tissue specific genes

Procedia PDF Downloads 433

Authors: Anita Conti, Riccardo Ossanna, Lindsey A. Quintero, Giamaica Conti, Andrea Sbarbati

Abstract:

Ischemia/reperfusion (IR) injury induces muscle fiber atrophy and skeletal muscle fiber death with subsequently functionality loss. The heterogeneous pool of cells, especially mesenchymal stem cells, contained in the stromal vascular fraction (SVF) of adipose tissue could promote muscle fiber regeneration. To prevent SVF dispersion, it has been proposed the use of injectable biopolymers that work as cells carrier. A significant element of the extracellular matrix is hyaluronic acid (HA), which has been widely used in regenerative medicine as a cell scaffold given its biocompatibility, degradability, and the possibility of chemical functionalization. Connective tissue micro-fragments enriched with SVF obtained from mechanical disaggregation of adipose tissue were evaluated for IR muscle injury regeneration using low molecular weight HA as a scaffold. IR induction. Hindlimb ischemia was induced in 9 athymic nude mice through the clamping of the right quadriceps using a plastic band. Reperfusion was induced by cutting the plastic band after 3 hours of ischemic period. Contralateral (left) muscular tissue was used as healthy control. Treatment. Twenty-four hours after the IR induction, animals (n=3) were intramuscularly injected with 100 µl of SVF mixed with HA (SVF-HA). Animals treated with 100 µl of HA (n=3) and 100 µl saline solution (n=3) were used as control. Treatment monitoring. All animals were in vivo monitored by magnetic resonance imaging (MRI) at 5, 7, 14 and 18 days post-injury (dpi). High-resolution morphological T2 weighed, quantitative T2 map and Dynamic Contrast-Enhanced (DCE) images were acquired in order to assess the regenerative potential of SVF-HA treatment. Ex vivo evaluation. After 18 days from IR induction, animals were sacrificed, and the muscles were harvested for histological examination. At 5 dpi T2 high-resolution MR images clearly reveal the presence of an extensive edematous area due to IR damage for all groups identifiable as an increase of signal intensity (SI) of muscular and surrounding tissue. At 7 dpi, animals of the SVF-HA group showed a reduction of SI, and the T2relaxation time of muscle tissue of the HA-SVF group was 29±0.5ms, comparable with the T2relaxation time of contralateral muscular tissue (30±0.7ms). These suggest a reduction of edematous overflow and swelling. The T2relaxation time at 7dpi of HA and saline groups were 84±2ms and 90±5ms, respectively, which remained elevated during the rest of the study. The evaluation of vascular regeneration showed similar results. Indeed, DCE-MRI analysis revealed a complete recovery of muscular tissue perfusion after 14 dpi for the SVF-HA group, while for the saline and HA group, controls remained in a damaged state. Finally, the histological examination of SVF-HA treated animals exhibited well-defined and organized fibers morphology with a lateralized nucleus, similar to contralateral healthy muscular tissue. On the contrary, HA and saline-treated animals presented inflammatory infiltrates, with HA slightly improving the diameter of the fibers and less degenerated tissue. Our findings show that connective tissue micro-fragments enriched with SVF induce higher muscle homeostasis and perfusion restoration in contrast to control groups.

Keywords: ischemia/reperfusion injury, regenerative medicine, resonance imaging, stromal vascular fraction

Procedia PDF Downloads 127

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

Procedia PDF Downloads 153

Authors: Alireza Barari, Maryam Firozi-Niyaki, Maryam Kamarlouei

Abstract:

Background: Herbs have a strong anti-cancer effect. Also, exercise is one of several lifestyle factors known to lower the risk of developing cancer. The aim of this study was to investigate the effect of endurance training and taxol on cyclooxygenase-2 and prostaglandin E2 in the liver tissue of mice with cervical cancer. Materials and Methods: In this experimental study, 35 female C57 mice were randomly divided into 5 groups (n=7 in each group): control (healthy), control (cancer), complement (cancer), training-supplementary (cancer) and training (cancer). The implantation of cancerous tumors was performed under the skin of the upper pelvis. The training group completed the endurance training protocol, which included 3 sessions per week, 50 minutes per session, at a speed of 14-18 m/s for six weeks. A dose of 60 mg/kg/day of pure taxol was injected intra peritoneally. The dependent variables of this study were measured 24 hours after the last training session by ELISA. Results: The results showed that the use of taxol and endurance training reduced the levels of cyclooxygenase-2 and prostaglandin E2 in the liver tissues of C57 mice with cervical cancer. Conclusion: Induction of the cancerous tissue in mice with cervical cancer increases the levels of cyclooxygenase-2 and prostaglandin E2 and endurance training along with taxol may reduce these levels.

Keywords: cervical cancer, taxol, endurance training, cyclooxygenase-2, prostaglandin E2

Procedia PDF Downloads 238

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

Abstract:

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

Procedia PDF Downloads 242

Authors: Matej Patzelt, Jan Dudak, Frantisek Krejci, Jan Zemlicka, Vladimir Musil, Jitka Riedlova, Viktor Sykora, Jana Mrzilkova, Petr Zach

Abstract:

Introduction: Micro-CT is well used for examination of bone structures and teeth. On the other hand visualization of the soft tissues is still limited. The goal of our study was to create a new fixation method for soft tissue imaging in micro-CT. Methodology: We used organs of 18 mice - heart, lungs, kidneys, liver and brain, which we fixated in ethanol after meticulous preparation. We fixated organs in different concentrations of ethanol and for different period of time. We used three types of ethanol concentration - 97%, 50% and ascending ethanol concentration (25%, 50%, 75%, 97% each for 12 hours). Fixated organs were scanned after 72 hours, 168 hours and 336 hours period of fixation. We scanned all specimens in micro-CT MARS (Medipix All Resolution System). Results: Ethanol method provided contrast enhancement in all studied organs in all used types of fixation. Fixation in 97% ethanol provided very fast fixation and the contrast among the tissues was visible already after 72 hours of fixation. Fixation for the period of 168 and 336 hours gave better details, especially in lung tissue, where alveoli were visualized. On the other hand, this type of fixation caused organs to petrify. Fixation in 50% ethanol provided best results in 336 hours fixation, details were visualized better than in 97% ethanol and samples were not as hard as in fixation in 97% ethanol. Best results were obtained in fixation in ascending ethanol concentration. All organs were visualized in great details, best-visualized organ was heart, where trabeculae and valves were visible. In this type of fixation, organs stayed soft for whole time. Conclusion: New ethanol method is a great option for soft tissue fixation as well as the method for enhancing contrast among tissues in organs. The best results were obtained with fixation of the organs in ascending ethanol concentration, the best visualized organ was the heart.

Keywords: x-ray imaging, small animals, ethanol, ex-vivo

Procedia PDF Downloads 321

Authors: Sakinah Abdullah, Rosna Mat Taha

Abstract:

Antioxidants are compounds that protect cells against the damaging effects of reactive oxygen species such as singlet oxygen, superoxide, peroxyl radicals, and peroxynitrite which result in oxidative stress leading to cellular damage. Natural antioxidant are in high demand because of their potential in health promotion and disease prevention and their improved safety and consumer acceptability. Plants are rich sources of natural antioxidant. Dioscorea alata L. known as 'ubi badak' in Malaysia were well known for their antioxidant content, but this plant was seasonal. Thus, tissue culture technique was used to mass propagate this plant. In the present work, a comparative study between in vitro (from tissue culture) and in vivo (from intact plant) samples of Dioscorea alata L. for their antioxidant potential by 2,2-diphenil -1- picrylhydrazyl (DPPH) radical scavenging activity method and their total phenolic and flavonoid contents were carried out. All samples had better radical scavenging activity but in vivo samples had the strongest radical scavenging activity compared to in vitro samples. Furthermore, tubers from in vivo samples showed the greatest free radical scavenging effect and comparatively greater phenolic content than in vitro samples.

Keywords: Dioscorea alata, tissue culture, antioxidant, in vivo, in vitro, DPPH

Procedia PDF Downloads 469

Authors: Pennisi Stefania, Giuffrida Graziella, Coppa Federica, Iannello Giulia, Cartelli Simone, Lo Faro Riccardo, Ferruggia Greta, Brundo Maria Violetta

Abstract:

Tissue engineering aims to develop advanced medical devices to restore normal functions of damaged tissue. These devices, even more effective than conventional methods, are called skin substitutes and are configured as drugs to be applied to the damaged area, to heal extensive and deep wounds which could otherwise lead to chronic wounds lasting over time. Among the variety of commercially available skin substitutes, those that have proven to be most effective are those consisting of a bilayer scaffold. The aim of our research was to design a skin substitute which can promote cell proliferation, cell migration and angiogenesis, and which can guarantee timely closure of the wound with satisfactory aesthetic results, in order to avoid the patient excessive pain, risk of contracting infections and long-term hospitalization. The product was tested in vitro using the Scratch Assay. The assay was carried out both on the matrix modified with hyaluronic acid and on the matrix based only on collagen. In both cases, after 48 hours of exposure the wound scratch was almost completely closed in treated cells compared to untreated control.

Keywords: collagen, hyaluronic acid, scratch- wound-healing assay, tissue regeneration

Procedia PDF Downloads 25

Authors: Nam-Seok Joo, Da-Eun Jung, Beom-Hee Choi

Abstract:

Background and Objectives: Recently, the triglyceride/high-density lipoprotine cholesterol (TG/HDL) ratio and pericardial adipose tissue (PAT) has gained attention as an indicator related to metabolic syndrome (MS). To date, there has been no research on the relationship between TG/HDL and PAT, we aimed to investigate the association between the TG/HDL and PAT. Methods: In this cross-sectional study, we investigated 627 patients who underwent coronary multidetector computed tomography and metabolic parameters. We divided subjects into two groups according to the cut-off PAT volume associated with MS, which is 142.2 cm³, and we compared metabolic parameters between those groups. We divided the TG/HDL ratio into tertiles according to Log(TG/HDL) and compared PAT-related parameters by analysis of variance. Finally, we applied logistic regression analysis to obtain the odds ratio of high PAT (PAT volume≥142.2 cm³) in each tertile, and we performed receiver operating characteristic (ROC) analysis to get the cut-off of TG/HDL ratio according to high PAT. Results: The mean TG/ HDL ratio of the high PAT volume group was 3.6, and TG/ HDL ratio had a strong positive correlation with various metabolic parameters. In addition, in the Log (TG/HDL) tertile groups, the higher tertile had more metabolic derangements, including PAT, and showed higher odds ratios of having high PAT (OR=4.10 in the second tertile group and OR=5.06 in their third tertile group, respectively) after age, sex, smoking adjustments. TG/HDL ratio according to the having increased PAT by ROC curve showed 1.918 (p < 0.001). Conclusion: TG/HDL ratio and high PAT volume have a significant positive correlation, and higher TG/HDL ratio showed high PAT. The cut-off value of the TG/HDL ratio was 1.918 to have a high PAT.

Keywords: triglyceride, high-density lipoprotein, pericardial adipose tissue, cut-off value

Procedia PDF Downloads 15

Authors: Andrew Kanzunguze, Lusayo Mwabumba, Jason K. Gilbertson, Dominic B. Gondwe, George Z. Nxumayo

Abstract:

Knowledge about the spatio-temporal distribution of invasive plants in protected areas provides a base from which hypotheses explaining proliferation of plant invasions can be made alongside development of relevant invasive plant monitoring programs. The aim of this study was to investigate the spatio-temporal distribution of bracken fern on the grassland plateau of Nyika National Park over the past 30 years (1986-2016) as well as to determine the current extent of the invasion. Remote sensing, machine learning, and statistical modelling techniques (object-based image analysis, image classification and linear regression analysis) in geographical information systems were used to determine both the spatial and temporal distribution of bracken fern in the study area. Results have revealed that bracken fern has been increasing coverage on the Nyika plateau at an estimated annual rate of 87.3 hectares since 1986. This translates to an estimated net increase of 2,573.1 hectares, which was recorded from 1,788.1 hectares (1986) to 4,361.9 hectares (2016). As of 2017 bracken fern covered 20,940.7 hectares, approximately 14.3% of the entire grassland plateau. Additionally, it was observed that the fern was distributed most densely around Chelinda camp (on the central plateau) as well as in forest verges and roadsides across the plateau. Based on these results it is recommended that Ecological Niche Modelling approaches be employed to (i) isolate the most important factors influencing bracken fern proliferation as well as (ii) identify and prioritize areas requiring immediate control interventions so as to minimize bracken fern proliferation in Nyika National Park.

Keywords: bracken fern, image classification, Landsat-8, Nyika National Park, spatio-temporal distribution

Procedia PDF Downloads 179

Authors: Dins Sumerags, Mara Pilmane, Vita Konopecka, Gunta Sumeraga

Abstract:

Reinke’s oedema is a specific type of chronic laryngitis evolving only in smokers. Our study aimed to identify the presence and interaction of the immunohistochemical markers for inflammation [IL-1α] and [IL-10], proliferation [Ki-67] and immunoreactive innervation [PGP 9.5] in the laryngeal mucosa using biotin-streptavidin immunochemical staining method. The laryngeal tissue samples were taken from the vocal cord during the surgery of the Reinke’s oedema and compared to the control group from the tissue samples of the cadavers without any visual laryngeal disease. The study results confirm increased cellular proliferation and elevation of the inflammation markers in the laryngeal mucosa in the case of Reinke’s oedema by comparing with the control.

Keywords: reinke`s oedema, immunohistochemical markers, laryngeal mucosa, biotin-streptavidin

Procedia PDF Downloads 134

Authors: P. Afsharian, S. Mousazadeh, M. Shahhoseini, R. Aflatoonian

Abstract:

Introduction: Matrix MetalloProteinases (MMPs) degrade extracellular matrix components to provide normal remodeling and contribute to pathological tissue destruction and cell migration in endometriosis. It is accepted that MMPs are resistant to suppression by progesterone in endometriotic tissues. The physiological effects of progesterone are mediated by its two progesterone receptor (PGR) isoforms, namely PGR-A and PGR-B. The capacity of progesterone affect to gene expression is dependent on the PGR-A/PGR-B ratio. The imbalance ratio in endometriotic tissue may be an important mechanism to be resulted in Progesterone resistance and modify progesterone action via differential regulation of specific progesterone response genes and improve endometriosis disease. Material and methods: RNA was extracted from twenty ectopic (endometriotic) and eutopic (endometrial) tissue samples of women undergoing laparoscopy for endometriosis and 20 healthy fertile women at Royan Institute, Tehran, Iran. Analysis of PGR-A, PGR-B, MMP-2 and MMP-9 mRNA expression was performed using Real-time PCR in ectopic and eutopic tissues. Then, Statistical analysis was calculated according to the 2-ΔΔCT equation for all samples. Results: Quantitative RT–PCR analyses of PGR-A and PGR-B mRNA revealed that there were differences in both isoformes of PGRs mRNA expressions between ectopic and control eutopic tissues. We were able to demonstrate low expression levels of PGR-B isoforms in ectopic tissues. Although, PGR-A expression was significantly higher in the same ectopic samples compare to controls.This method permitted us to demonstrate significant overexpression of MMP-2 and MMP-9 in ectopic samples compared to control endometrial tissues, as well. Conclusions: Our data suggest that low expression levels of PGR-B and overexpression of PGR-A can alter PGR-A/PGR-B ratio in endometriotic ectopic tissues. Imbalance ratio of PGRs in endometriotic tissue may be able to consequence MMP-2 and MMP-9 overexpression which can be important in pathogenesis and treatment of disease.

Keywords: endometriosis, matrix metalloproteinases, progesterone receptor -A and -B, PGR-A/PGR-B ratio

Procedia PDF Downloads 318

Authors: J. H. Bang, H. J. Baek, K. I. Kim, B. J. Lee, H. J. Jung, H. J. Jang, S. K. Jung

Abstract:

Asthma is a chronic inflammatory lung disease characterized by airway hyper responsiveness (AHR), airway obstruction and airway wall remodeling responsible for significant morbidity and mortality worldwide. Genetic and environment factors may result in asthma, but there are no the exact causes of asthma. Chungpye-tang (CPT) has been prescribed as a representative aerosol agent for patients with dyspnea, cough and phlegm in the respiratory clinic at Kyung Hee Korean Medicine Hospital. This Korean herbal medicines have the effect of dispelling external pathogen and dampness pattern. CPT is composed of 4 species of herbal medicines. The 4 species of herbal medicines are Ephedrae herba, Pogostemonis(Agatachis) herba, Caryophylli flos and Zingiberis rhizoma crudus. CPT suppresses neutrophil infiltration and the production of pro-inflammatory cytokines in lipopolysaccharide (LPS)-induced acute lung injury (ALI) mouse model. Moreover, the anti-inflammatory effects of CPT on a mouse model of Chronic Obstructive Pulmonary Disease (COPD) was proved. Activation of the NF-κB has been proven that it plays an important role in inflammation via inducing transcription of pro-inflammatory genes. Over-expression of NF-κB has been believed be related to many inflammatory diseases such as arthritis, gastritis, asthma and COPD. So we firstly hypothesize whether CPT has an anti-inflammatory effect on asthmatic human airway epithelial tissue via inhibiting NF-κB pathway. In this study, CPT was extracted with distilled water for 3 hours at 100°C. After process of filtration and evaporation, it was freeze dried. And asthmatic human lung tissues were provided by MatTek Corp. We investigated the precise mechanism of the anti-inflammatory effect of CPT by western blotting analysis. We observed whether the decoction extracts could reduce NF-κB activation, COX-2 protein expression and NF-κB-mediated pro-inflammatory cytokines such as TNF-α, eotaxin, IL-4, IL-9 and IL-13 in asthmatic human lung tissue. As results of this study, there was a trend toward decreased NF-κB expression in asthmatic human airway epithelial tissue. We found that the inhibition effects of CPT on COX-2 expression was not determined. IL-9 and IL-13 secretion was significantly reduced in the asthmatic human lung tissue treated with CPT. Overall, our results indicate that CPT has an anti-inflammatory effect through blocking the signaling pathway of NF-κB, thereby CPT may be a potential remedial agent for allergic asthma.

Keywords: Chungpye-tang, allergic asthma, asthmatic human airway epithelial tissue, nuclear factor kappa B (NF-κB) pathway, COX-2

Procedia PDF Downloads 335

Authors: Nidhi Jindal, Ashok Chaudhury, Manisha Mangal

Abstract:

Andrographis paniculata, (Burm f.) Wallich ex. Nees (Family Acanthaceae) popularly known as King of Bitters, is an important medicinal herb. It has an astonishingly wide range of medicinal properties such as anti-inflammatory,antidiarrhoeal, antiviral, antimalarial, hepatoprotective, cardiovascular, anticancer, and immunostimulatory activities. It is widely cultivated in southern Asia. Though propagation of this herb generally occurs through seeds, it has many germination problems which intrigued scientists to work out on the alternative techniques for its mass production. The potential of tissue culture techniques as an alternative tool for AP multiplication was found to be promising. However, the high mortality rate of explants caused by phenolic browning of explants is one of the difficulties reported. Low multiplication rates were reported in the proliferation phase, as well as cultures decline characterized by leaf fall and loss of overall vigor. In view of above problems, a study was undertaken to overcome seed dormancy to improve germination potential and to investigate further on the possible means for successful proliferation of cultures via preventive approaches to overcome failures caused by phenolic browning. Experiments were conducted to improve germination potential and among all the chemical and mechanical trials, scarification of seeds with sand paper proved to be the best method to enhance the germination potential (82.44%) within 7 days. Similarly, several pretreatments and media combinations were tried to overcome browning of explants leading to the conclusion that addition of 0.1% citric acid and 0.2% of ascorbic acid in the media followed by rapid sub culturing of explants controlled browning and decline of explants by 67.45%.

Keywords: plant tissue culture, empirical measure, germination, tissue culture

Procedia PDF Downloads 414

Authors: Mesih Kocamuftuoglu, Gonca Ozan, Enver Ozan, Nalan Kaya, Sema Temizer Ozan

Abstract:

Metabolic Syndrome, one of the common metabolic disorder, occurs with co-development of insulin resistance, obesity, dislipidemia and hypertension problems. Insulin resistance appears to play a pathogenic role in the metabolic syndrome. Also, there is a relationship between insulin resistance and infertility as known. Turmeric (Curcuma longa L.) a polyphenolic chemical is widely used for its coloring, flavoring, and medicinal properties, and exhibits a strong antioxidant activity. In this study, we assess the effects of turmeric on rat uterine tissue in metabolic syndrome model induced by high fructose diet. Thirty-two adult female Wistar rats weighing 220±20 g were randomly divided into four groups (n=8) as follows; control, fructose, turmeric, and fructose plus turmeric. Metabolic syndrome was induced by fructose solution 20% (w/v) in tap water, and turmeric (C.Longa) administered at the dose of 80 mg/kg body weight every other day by oral gavage. After the experimental period of 8 weeks, rats were decapitated, serum and uterine tissues were removed. Serum lipid profile, glucose, insülin levels were measured. Uterine tissues were fixed for histological analyzes. The uterine tissue sections were stained with hematoxylin-eosin (H & E) stain, then examined and photographed on a light microscope (Novel N-800Mx20). As a result, fructose consumption effected serum lipids, insulin levels, and insulin resistance significantly. Endometrium and myometrium layers were observed in normal structure in control group of uterine tissues. Perivascular edema, peri glandular fibrosis, and inflammatory cell increase were detected in fructose group. Sections of the fructose plus turmeric group showed a significant improvement in findings when compared to the fructose group. Turmeric group cell structures were observed similar with the control group. These results demonstrated that high-fructose consumption could change the structure of the uterine tissue. On the other hand, turmeric administration has beneficial effects on uterine tissue at that dose and duration when administered with fructose.

Keywords: metabolic syndrome, rat, turmeric, uterus

Procedia PDF Downloads 177

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

Abstract:

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)

Procedia PDF Downloads 235

Authors: Krasimir I. Ivanov, Penka S. Zapryanova, Stefan V. Krustev, Violina R. Angelova

Abstract:

Three commonly used digestion methods (dry ashing, acid digestion, and microwave digestion) in different variants were compared for digestion of tobacco leaves. Three main macroelements (K, Ca and Mg) were analysed using AAS Spectrometer Spectra АА 220, Varian, Australia. The accuracy and precision of the measurements were evaluated by using Polish reference material CTR-VTL-2 (Virginia tobacco leaves). To elucidate the problems with elemental recovery X-Ray and SEM–EDS analysis of all residues after digestion were performed. The X-ray investigation showed a formation of KClO₄ when HClO4 was used as a part of the acids mixture. The use of HF at Ca and Mg determination led to the formation of CaF₂ and MgF₂. The results were confirmed by energy dispersive X-ray microanalysis. SPSS program for Windows was used for statistical data processing.

Keywords: digestion methods, plant tissue, determination of macroelements, K, Ca, Mg

Procedia PDF Downloads 317

Authors: Rajeswari Raguraman, Sowmya Parameswaran, Krishnakumar Subramanian, Jagat Kanwar, Rupinder Kanwar

Abstract:

Background: Tumor microenvironment has been implicated in several cancers to regulate cell growth, invasion and metastasis culminating in outcome of therapy. Tumor stroma consists of multiple cell types that are in constant cross-talk with the tumor cells to favour a pro-tumorigenic environment. Not much is known about the existence of tumor microenvironment in the pediatric intraocular malignancy, Retinoblastoma (RB). In the present study, we aim to understand the multiple stromal cellular subtypes and tumor stromal interactions expressed in RB tumors. Materials and Methods: Immunohistochemistry for stromal cell markers CD31, CD68, alpha-smooth muscle (α-SMA), vimentin and glial fibrillary acidic protein (GFAP) was performed on formalin fixed paraffin embedded tissues sections of RB (n=12). The differential expression of stromal target molecules; fibroblast activation protein (FAP), tenascin-C (TNC), osteopontin (SPP1), bone marrow stromal antigen 2 (BST2), stromal derived factor 2 and 4 (SDF2 and SDF4) in primary RB tumors (n=20) and normal retina (n=5) was studied by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. The differential expression was correlated with the histopathological features of RB. The interaction between RB cell lines (Weri-Rb-1, NCC-RbC-51) and Bone marrow stromal cells (BMSC) was also studied using direct co-culture and indirect co-culture methods. The functional effect of the co-culture methods on the RB cells was evaluated by invasion and proliferation assays. Global gene expression was studied by using Affymetrix 3’ IVT microarray. Pathway prediction was performed using KEGG and the key molecules were validated using qRT-PCR. Results: The immunohistochemistry revealed the presence of several stromal cell types such as endothelial cells (CD31+;Vim+/-); macrophages (CD68+;Vim+/-); Fibroblasts (Vim+; CD31-;CD68- );myofibroblasts (α-SMA+/ Vim+) and invading retinal astrocytes/ differentiated retinal glia (GFAP+; Vim+). A characteristic distribution of these stromal cell types was observed in the tumor microenvironment, with endothelial cells predominantly seen in blood vessels and macrophages near actively proliferating tumor or necrotic areas. Retinal astrocytes and glia were predominant near the optic nerve regions in invasive tumors with sparse distribution in tumor foci. Fibroblasts were widely distributed with rare evidence of myofibroblasts in the tumor. Both gene and protein expression revealed statistically significant (P<0.05) up-regulation of FAP, TNC and BST2 in primary RB tumors compared to the normal retina. Co-culture of BMSC with RB cells promoted invasion and proliferation of RB cells in direct and indirect contact methods respectively. Direct co-culture of RB cell lines with BMSC resulted in gene expression changes in ECM-receptor interaction, focal adhesion, IL-8 and TGF-β signaling pathways associated with cancer. In contrast, various metabolic pathways such a glucose, fructose and amino acid metabolism were significantly altered under the indirect co-culture condition. Conclusion: The study suggests that the close interaction between RB cells and the stroma might be involved in RB tumor invasion and progression which is likely to be mediated by ECM-receptor interactions and secretory factors. Targeting the tumor stroma would be an attractive option for redesigning treatment strategies for RB.

Keywords: gene expression profiles, retinoblastoma, stromal cells, tumor microenvironment

Procedia PDF Downloads 384

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

Abstract:

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 321

Authors: Husain S. Yawer, Vasim Raja Panwar, Nidhi Priya

Abstract:

The objective of this study is to evaluate and compare the role of nano-gelatin and Bioengineered Scaffolds on the attachment, proliferation, and osteogenic differentiation of human dental pulp stem cells (DPSCs). Tooth decay and early fall have each been one of the most prevailing dental disorders which cause physical and emotional suffering and compromise the patient's quality of life. The design of novel scaffolding materials will be based on mimicking the architecture of natural dental extracellular matrix which may provide as in vivo environments for proper cell growth. This methodology will involve the combination of nano-fibred gelatin as well as biodegradable hydrogel based tooth scaffold. We have measured and optimized the Dental Pulp Stem Cells growth profile in cultures carried out on collagen-coated plastic surface, however, for tissue regeneration study, we aim to develop an enhanced microenvironment for stem cell growth and dental tissue regeneration. We believe biomimetic cell adhesion and scaffolds might provide a near in vivo growth environment for proper growth and differentiation of human DPSCs, which further help in dentin/pulp tissue regeneration.

Keywords: nano-gelatin, stem cells, dental pulp, scaffold

Procedia PDF Downloads 330

Authors: Shalu Jain, Anju Bansal, Anup Kumar, Sunita Saxena

Abstract:

Objectives: The novel TMPRSS2:ERG gene fusion is a common somatic event in prostate cancer that in some studies is linked with a more aggressive disease phenotype. Thus, this study aims to determine whether clinical variables are associated with the presence of TMPRSS2:ERG-fusion gene transcript in Indian patients of prostate cancer. Methods: We evaluated the clinical variables with presence and absence of TMPRSS2:ERG gene fusion in prostate cancer and BPH association of clinical patients. Patients referred for prostate biopsy because of abnormal DRE or/and elevated sPSA were enrolled for this prospective clinical study. TMPRSS2:ERG mRNA copies in samples were quantified using a Taqman chemistry by real time PCR assay in prostate biopsy samples (N=42). The T2:ERG assay detects the gene fusion mRNA isoform TMPRSS2 exon1 to ERG exon4. Results: Histopathology report has confirmed 25 cases as prostate cancer adenocarcinoma (PCa) and 17 patients as benign prostate hyperplasia (BPH). Out of 25 PCa cases, 16 (64%) were T2: ERG fusion positive. All 17 BPH controls were fusion negative. The T2:ERG fusion transcript was exclusively specific for prostate cancer as no case of BPH was detected having T2:ERG fusion, showing 100% specificity. The positive predictive value of fusion marker for prostate cancer is thus 100% and the negative predictive value is 65.3%. The T2:ERG fusion marker is significantly associated with clinical variables like no. of positive cores in prostate biopsy, Gleason score, serum PSA, perineural invasion, perivascular invasion and periprostatic fat involvement. Conclusions: Prostate cancer is a heterogeneous disease that may be defined by molecular subtypes such as the TMPRSS2:ERG fusion. In the present prospective study, the T2:ERG quantitative assay demonstrated high specificity for predicting biopsy outcome; sensitivity was similar to the prevalence of T2:ERG gene fusions in prostate tumors. These data suggest that further improvement in diagnostic accuracy could be achieved using a nomogram that combines T2:ERG with other markers and risk factors for prostate cancer.

Keywords: prostate cancer, genetic rearrangement, TMPRSS2:ERG fusion, clinical variables

Procedia PDF Downloads 444

Authors: Choki, Supatchai Boonpratham, Suwannee Luppanapornlarp

Abstract:

The objective of this study was to find a correlation between cephalometric measurements and visual perception of facial profile in skeletal type II patients. In this study, 250 lateral cephalograms of female patients from age, 20 to 22 years were analyzed. The profile outlines of all the samples were hand traced and transformed into silhouettes by the principal investigator. Profile ratings were done by 9 orthodontists on Visual Analogue Scale from score one to ten (increasing level of convexity). 37 hard issue and soft tissue cephalometric measurements were analyzed by the principal investigator. All the measurements were repeated after 2 weeks interval for error assessment. At last, the rankings of visual perceptions were correlated with cephalometric measurements using Spearman correlation coefficient (P < 0.05). The results show that the increase in facial convexity was correlated with higher values of ANB (A point, nasion and B point), AF-BF (distance from A point to B point in mm), L1-NB (distance from lower incisor to NB line in mm), anterior maxillary alveolar height, posterior maxillary alveolar height, overjet, H angle hard tissue, H angle soft tissue and lower lip to E plane (absolute correlation values from 0.277 to 0.711). In contrast, the increase in facial convexity was correlated with lower values of Pg. to N perpendicular and Pg. to NB (mm) (absolute correlation value -0.302 and -0.294 respectively). From the soft tissue measurements, H angles had a higher correlation with visual perception than facial contour angle, nasolabial angle, and lower lip to E plane. In conclusion, the findings of this study indicated that the correlation of cephalometric measurements with visual perception was less than expected. Only 29% of cephalometric measurements had a significant correlation with visual perception. Therefore, diagnosis based solely on cephalometric analysis can result in failure to meet the patient’s esthetic expectation.

Keywords: cephalometric measurements, facial profile, skeletal type II, visual perception

Procedia PDF Downloads 138

Authors: Kara J. Coffman-Rea, Karen E. Samonds

Abstract:

Microplastic pollution is an urgent global threat to our planet and human health. Microplastic particles have been detected within our food, water, and atmosphere, and found within the human stool, placenta, and lung tissue. However, most spectrometric microplastic detection methods require chemical digestion which can alter or destroy microplastic particles and makes it impossible to acquire information about their in-situ distribution. MALDI TOF MS (Matrix-assisted laser desorption ionization-time of flight mass spectrometry) is an analytical method using a soft ionization technique that can be used for polymer analysis. This method provides a valuable opportunity to both acquire information regarding the in-situ distribution of microplastics and also minimizes the destructive element of chemical digestion. In addition, MALDI TOF MS allows for expanded analysis of the microplastics including detection of specific additives that may be present within them. MALDI TOF MS is particularly sensitive to sample preparation and has not yet been used to analyze environmental microplastics within their specific location (e.g., biological tissues, sediment, water). In this study, microplastics were created using polyethylene gloves, polystyrene micro-foam, and polyethylene terephthalate cable sleeving. Plastics were frozen using liquid nitrogen and ground to obtain small fragments. An artificial tissue was created using a cellulose sponge as scaffolding coated with a MaxGel Extracellular Matrix to simulate human lung tissue. Optimal preparation techniques (e.g., matrix, cationization reagent, solvent, mixing ratio, laser intensity) were first established for each specific polymer type. The artificial tissue sample was subsequently spiked with microplastics, and specific polymers were detected using MALDI-TOF-MS. This study presents a novel method for the detection of environmental polyethylene, polyethylene terephthalate, and polystyrene microplastics within a complex sample. Results of this study provide an effective method that can be used in future microplastics research and can aid in determining the potential threats to environmental and human health that they pose.

Keywords: environmental plastic pollution, MALDI-TOF MS, microplastics, polymer identification

Procedia PDF Downloads 256

Authors: Saeedeh Shahbazkhany

Abstract:

Cancer is a terrible disease that, if not diagnosed early, therapy can be difficult while it is easily medicable if it is diagnosed in early stages. So it is very important for cancer diagnosis that medical procedures are performed. In this paper we introduce a new method. In this method, we only need pictures of healthy cells and cancer cells. In fact, where we suspect cancer, we take a picture of cells or tissue in that area, and then take some pictures of the surrounding tissues. Then, fractal dimension of images are calculated and compared. Cancer can be easily detected by comparing the fractal dimension of images. In this method, we use Matlab software.

Keywords: Matlab software, fractal dimension, cancer, surrounding tissues, cells or tissue, new method

Procedia PDF Downloads 354

Authors: M. Dimitri, M. Ricci, F. Bigi, M. Romiti, A. Corvi

Abstract:

Tissue engineering has reached a significant milestone with the integration of 3D printing for the creation of complex bioconstructs equipped with vascular networks, crucial for cell maintenance and growth. This study aims to demonstrate the effectiveness of a portable microperfusion system designed to adapt dynamically to the evolving conditions of cell growth within 3D-printed bioconstructs. The microperfusion system was developed to provide a constant and controlled flow of nutrients and oxygen through the integrated vessels in the bioconstruct, replicating in vivo physiological conditions. Through a series of preliminary experiments, we evaluated the system's ability to maintain a favorable environment for cell proliferation and differentiation. Measurements of cell density and viability were performed to monitor the health and functionality of the tissue over time. Preliminary results indicate that the portable microperfusion system not only supports but optimizes cell growth, effectively adapting to changes in metabolic needs during the bioconstruct maturation process. This research opens perspectives in tissue engineering, demonstrating that a portable microperfusion system can be successfully integrated into 3D-printed bioconstructs, promoting sustainable and uniform cell growth. The implications of this study are far-reaching, with potential applications in regenerative medicine and pharmacological research, providing a platform for the development of functional and complex tissues.

Keywords: biofabrication, microfluidic perfusion system, 4D bioprinting

Procedia PDF Downloads 30

Authors: Holm Zerbe, Laura Macias, Hans-Joachim Schuberth, Wolfram Petzl

Abstract:

Mastitis is among the most important production diseases in cows. It accounts for large parts of antimicrobial drug use in the dairy industry worldwide. Due to the imminent normative to reduce the use of antimicrobial drugs in livestock, new ways for therapy and prophylaxis of mastitis are needed. Recently epigenetic regulation of inflammation by chromatin modifications has increasingly drawn attention. Currently, some epigenetic modifiers have already been approved for the use in humans, however little is known about their actions in the bovine system. The aim of our study was to investigate whether three selected epigenetic modifiers (Vitamin D3, SAHA and S2101) influence the initial immune response towards mastitis pathogens in bovine udder tissue in vitro. Tissue explants of the teat cistern and udder parenchyma were collected from 21 cows and were incubated for 36 hours in the absence and presence of epigenetic modifiers. Additionally, the tissue was stimulated with heat-inactivated particles of Escherichia coli and Staphylococcus aureus, which are regarded as two of the most important mastitis pathogens. After incubation, the explants were tested by RT-qPCR for transcript abundances of immune-related candidate genes. Gene expression was validated in culture supernatants by an AlphaLISA assay. Furthermore, the culture supernatants were analyzed for their chemotactic capacity through a chemotaxis assay. Statistical analysis of data was performed with the program ‘R’ version 3.2.3. Vitamin D3 had no effect on the immune response of udder tissue in vitro after stimulation with mastitis pathogens. The epigenetic modifiers SAHA and S2101 however significantly blocked the pathogen-induced upregulation of CXCL8, TNFα, S100A9 and LAP (P < 0.05). The regulation of IL10 was not affected by treatment with SAHA and S2101. Transcript abundances for CXCL8 were reflected by IL8 contents and chemotactic activity in culture supernatants. In conclusion, these data show the potential of epigenetic modifiers (SAHA and S2101) to block overshooting inflammation in the udder. Thus epigenetic modifiers may serve in future as immune modulators for the treatment and/or prophylaxis of clinical mastitis. (Funded by Deutsche Forschungsgemeinschaft PE 1495/2-1).

Keywords: mastitis, cattle, epigenetics, immunomodulation

Procedia PDF Downloads 235