Search results for: bovine milk

Authors: Alena Semeradtova, Marcel Stofik, Lucie Mareckova, Petr Maly, Ondrej Stanek, Jan Maly

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Recently, novel strategies based on so-called molecular evolution were shown to be effective for the production of various peptide ligand libraries with high affinities to molecular targets of interest comparable or even better than monoclonal antibodies. The major advantage of these peptide scaffolds is mainly their prevailing low molecular weight and simple structure. This study describes a new high-affinity binding molecules based immunesensor using a simple optical system for human serum albumin (HSA) detection as a model molecule. We present a comparison of two variants of recombinant binders based on albumin binding domain of the protein G (ABD) performed on micropatterned glass chip. Binding domains may be tailored to any specific target of interest by molecular evolution. Micropatterened glass chips were prepared using UV-photolithography on chromium sputtered glasses. Glass surface was modified by (3-aminopropyl)trietoxysilane and biotin-PEG-acid using EDC/NHS chemistry. Two variants of high-affinity binding molecules were used to detect target molecule. Firstly, a variant is based on ABD domain fused with TolA chain. This molecule is in vivo biotinylated and each molecule contains one molecule of biotin and one ABD domain. Secondly, the variant is ABD domain based on streptavidin molecule and contains four gaps for biotin and four ABD domains. These high-affinity molecules were immobilized to the chip surface via biotin-streptavidin chemistry. To eliminate nonspecific binding 1% bovine serum albumin (BSA) or 6% fetal bovine serum (FBS) were used in every step. For both variants range of measured concentrations of fluorescently labelled HSA was 0 – 30 µg/ml. As a control, we performed a simultaneous assay without high-affinity binding molecules. Fluorescent signal was measured using inverse fluorescent microscope Olympus IX 70 with COOL LED pE 4000 as a light source, related filters, and camera Retiga 2000R as a detector. The fluorescent signal from non-modified areas was substracted from the signal of the fluorescent areas. Results were presented in graphs showing the dependence of measured grayscale value on the log-scale of HSA concentration. For the TolA variant the limit of detection (LOD) of the optical immunosensor proposed in this study is calculated to be 0,20 µg/ml for HSA detection in 1% BSA and 0,24 µg/ml in 6% FBS. In the case of streptavidin-based molecule, it was 0,04 µg/ml and 0,07 µg/ml respectively. The dynamical range of the immunosensor was possible to estimate just in the case of TolA variant and it was calculated to be 0,49 – 3,75 µg/ml and 0,73-1,88 µg/ml respectively. In the case of the streptavidin-based the variant we didn´t reach the surface saturation even with the 480 ug/ml concentration and the upper value of dynamical range was not estimated. Lower value was calculated to be 0,14 µg/ml and 0,17 µg/ml respectively. Based on the obtained results, it´s clear that both variants are useful for creating the bio-recognizing layer on immunosensors. For this particular system, it is obvious that the variant based on streptavidin molecule is more useful for biosensing on glass planar surfaces. Immunosensors based on this variant would exhibit better limit of detection and wide dynamical range.

Keywords: high affinity binding molecules, human serum albumin, optical immunosensor, protein G, UV-photolitography

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Authors: Shahram Naghizadeh Raeisi, Ali Alghooneh

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The loss of curd cohesiveness and syneresis are two common problems in the ultrafiltered cheese industry. In this study, by using membrane technology and protein modification, a modified cheese was developed and its properties were compared with a control sample. In order to decrease the lactose content and adjust the protein, acidity, dry matter and milk minerals, a combination of ultrafiltration, nanofiltration and reverse osmosis technologies was employed. For protein modification, a two-stage chemical and enzymatic reaction was employed before and after ultrafiltration. The physicochemical and microstructural properties of the modified ultrafiltered cheese were compared with the control one. Results showed that the modified protein enhanced the functional properties of the final cheese significantly (pvalue< 0.05), even if the protein content was 50% lower than the control one. The modified cheese showed 21 ± 0.70, 18 ± 1.10 & 25±1.65% higher hardness, cohesiveness and water-holding capacity values, respectively, than the control sample. This behavior could be explained by the developed microstructure of the gel network. Furthermore, chemical-enzymatic modification of milk protein induced a significant change in the network parameter of the final cheese. In this way, the indices of network linkage strength, network linkage density, and time scale of junctions were 10.34 ± 0.52, 68.50 ± 2.10 & 82.21 ± 3.85% higher than the control sample, whereas the distance between adjacent linkages was 16.77 ± 1.10% lower than the control sample. These results were supported by the results of the textural analysis. A non-linear viscoelastic study showed a triangle waveform stress of the modified protein contained cheese, while the control sample showed rectangular waveform stress, which suggested a better sliceability of the modified cheese. Moreover, to study the shelf life of the products, the acidity, as well as molds and yeast population, were determined in 120 days. It’s worth mentioning that the lactose content of modified cheese was adjusted at 2.5% before fermentation, while the lactose of the control one was at 4.5%. The control sample showed 8 weeks shelf life, while the shelf life of the modified cheese was 18 weeks in the refrigerator. During 18 weeks, the acidity of modified and control samples increased from 82 ± 1.50 to 94 ± 2.20 °D and 88 ± 1.64 to 194 ± 5.10 °D, respectively. The mold and yeast populations, with time, followed the semicircular shape model (R2 = 0.92, R2adj = 0.89, RMSE = 1.25). Furthermore, the mold and yeast counts and their growth rate in the modified cheese were lower than those for control one; Aforementioned result could be explained by the shortage of the source of energy for the microorganism in the modified cheese. The lactose content of the modified sample was less than 0.2 ± 0.05% at the end of fermentation, while this was 3.7 ± 0.68% in the control sample.

Keywords: non-linear viscoelastic, protein modification, semicircular shape model, ultrafiltered cheese

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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

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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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Authors: C. I. Boshoff, S. Steenkamp-Jonker

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Cystic echinococcosis (CE), caused by Echinococcus granulosus is an important parasitic infection in livestock worldwide, with severe zoonotic potential. It is important to understand the variability of Echinococcus granulosus, as genotype variations may influence lifecycle patterns, development rate, and transmission. Cystic Echinococcus samples were collected from domestic animals in Eastern Cape Province, South Africa. A molecular study was performed on 14 hydatid cysts obtained from caprine, ovine and bovine livers in order to determine the Echinococcus granulosus strain present in these hosts. The sequencing of the mitochondrial cytochrome C oxidase subunit I (coxI) gene of the hydatid cysts produced sequences of 400 bp for each sample analysed. These sequences were aligned with those present in GenBank and a phylogenetic tree was constructed. Based on coxI genotype the isolates could be grouped into E. granulosus sensu stricto. The findings of the study represent a pilot molecular study on Echinococcus from domestic animals undertaken in South Africa.

Keywords: Echinococcus granulosus, genotypes, livestock, South Africa

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Authors: Hamid Abdulroof Saleh

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Background: Dioxins are one of the most widely distributed environmental pollutants. Dioxins consist of feedstock during the preparation of some industries, such as the paper industry as they can be produced in the atmosphere during the process of burning garbage and waste, especially medical waste. Dioxins can be found in the adipose tissues of animals in the food chain as well as in human breast milk. 2,3,7,8-Tetrachlorodibenzo-pdioxin (TCDD) is the most toxic component of a large group of dioxins. Humans are exposed to TCDD through contaminated food items like meat, fish, milk products, eggs etc. Recently, natural formulations relating to reducing or eliminating TCDD toxicity have been in focus. Ginger rhizome (Zingiber officinale R., family: Zingiberaceae), is used worldwide as a spice. Both antioxidative and androgenic activity of Z. officinale was reported in animal models. Researchers showed that ginger oil has dominative protective effect on DNA damage and might act as a scavenger of oxygen radical and might be used as an antioxidant. Aim of the work: The present study was undertaken to evaluate the toxic effect of TCDD on the structure and histoarchitecture of the testis and the protective role of co-administration of ginger root extract to prevent this toxicity. Materials & Methods: Male adult rats of Sprague-Dawley strain were assigned to four groups, eight rats in each; control group, dioxin treated group (given TCDD at the dose of 100 ng/kg Bwt/day by gavage), ginger treated group (given 50 mg/kg Bwt/day of ginger root extract by gavage), dioxin and ginger treated group (given TCDD at the dose of 100 ng/kg Bwt/day and 50 mg/kg Bwt/day of ginger root extract by gavages). After three weeks, rats were weighed and sacrificed where testis were removed and weighted. The testes were processed for routine paraffin embedding and staining. Tissue sections were examined for different morphometric and histopathological changes. Results: Dioxin administration showed a harmful effects in the body, testis weight and other morphometric parameters of the testis. In addition, it produced varying degrees of damage to the seminiferous tubules, which were shrunken and devoid of mature spermatids. The basement membrane was disorganized with vacuolization and loss of germinal cells. The co-administration of ginger root extract showed obvious improvement in the above changes and showed reversible morphometric and histopathological changes of the seminiferous tubules. Conclusion: Ginger root extract treatment in this study was successful in reversing all morphometric and histological changes of dioxin testicular damage. Therefore, it showed a protective effect on testis against dioxin toxicity.

Keywords: dioxin, ginger, rat, testis

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Authors: Binnur Sagbas

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Artificial joint replacements such as total knee and total hip prosthesis have been applied to the patients who affected by osteoarthritis. Although different material combinations are used for these joints, biopolymers are most commonly preferred materials especially for acetabular cup and tibial component of hip and knee joints respectively. The main limitation that shortens the service life of these prostheses is wear. Wear is complicated phenomena and it must be considered with friction and lubrication. In this study, micro wave (MW) induced argon+oxygen plasma surface modification were applied on ultra-high molecular weight polyethylene (UHMWPE) and vitamin E blended UHMWPE (VE-UHMWPE) biopolymer surfaces to improve surface wettability and wear resistance of the surfaces. Contact angel measurement method was used for determination of wettability. Ball-on-disc wear test was applied under 25% bovine serum lubrication conditions. The results show that surface wettability and wear resistance of both material samples were increased by plasma surface modification.

Keywords: artificial joints, plasma surface modification, UHMWPE, vitamin E, wear

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Authors: Meli̇sa Aşci, Berk Kiliç, Emine Ulusoy

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Ingestion of microplastics in humans has been increasing rapidly, as such hazardous materials are abundant in multiple food products, specifically milks. With increasing consumption rates, humans have been ingesting microplastics on a daily basis, making them prone to be intoxicated and even cause the disruption of intracellular pathways and liver cell disruption, and eventually tissue and organ damage. In this experiment, different milk types(animal-based and plant-based) were tested for microplastics. Results showed that animal-based milks contained a higher concentration of microplastics compared to plant-based milks. Research has shown that in addition to causing health issues in humans, microplastics can also affect livestock animals and plants.

Keywords: microplastics, plant-based milks, animal-based milks, preventive nutrition

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Authors: Mosaab A. Omar, Mohammad Saleh Al-Aboody, Mohmed A. Rizk, Shimaa M. Elsayed, Ahmed ElSify, Naoaki Yokoyama, Ikuo Igarashi

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Enoxacin is a broad-spectrum 6-fluoronaphthyridinone antibacterial agent (fluoroquinolones) structurally related to nalidixic acid used mainly in the treatment of urinary tract infections and gonorrhea. Also, it has been shown recently that it may have cancer inhibiting effect. The primary antibabesial effect of Enoxacin is due to inhibition of DNA gyrase subunit A, and DNA topoisomerase. In the present study, enoxacin was tested as a potent inhibitor against the in vitro growth of bovine and equine Piroplasms. The in vitro growth of five Babesia species that were tested was significantly inhibited (P<0.05) by micromolar concentrations of enoxacin (IC50 values= 13.5, 7.2, 7.5, and 24.2 µM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Enoxacin IC50 values for Babesia and Theileria parasites were satisfactory as the drug is a potent antibacterial drug with minimum side effects. Therefore, enoxacin might be used for the treatment of Babesiosis and Theileriosis especially in case of mixed infections with bacterial diseases or in the case of animal sensitivity against diminazin toxicity.

Keywords: enoxacin, Babesia, Theileria, IC50 and dimenazin

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Authors: Lubna Naz, Munir Ahmad, G. M. Arif

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This study analyzes static and dynamic welfare impacts of food price changes for various landholding groups in Pakistan. The study uses three classifications of land ownership, landless, small landowners and large landowners, for analysis. The study uses Panel Survey, Pakistan Rural Household Survey (PRHS) of Pakistan Institute of Development Economics Islamabad, of rural households from two largest provinces (Sindh and Punjab) of Pakistan. The study uses all three waves (2001, 2004 and 2010) of PRHS. This research work makes three important contributions in literature. First, this study uses Quadratic Almost Ideal Demand System (QUAIDS) to estimate demand functions for eight food groups-cereals, meat, milk and milk products, vegetables, cooking oil, pulses and other food. The study estimates food demand functions with Nonlinear Seemingly Unrelated (NLSUR), and employs Lagrange Multiplier and test on the coefficient of squared expenditure term to determine inclusion of squared expenditure term. Test results support the inclusion of squared expenditure term in the food demand model for each of landholding groups (landless, small landowners and large landowners). This study tests for endogeneity and uses control function for its correction. The problem of observed zero expenditure is dealt with a two-step procedure. Second, it creates low price and high price periods, based on literature review. It uses elasticity coefficients from QUAIDS to analyze static and dynamic welfare effects (first and second order Tylor approximation of expenditure function is used) of food price changes across periods. The study estimates compensation variation (CV), money metric loss from food price changes, for landless, small and large landowners. Third, this study compares the findings on welfare implications of food price changes based on QUAIDS with the earlier research in Pakistan, which used other specification of the demand system. The findings indicate that dynamic welfare impacts of food price changes are lower as compared to static welfare impacts for all landholding groups. The static and dynamic welfare impacts of food price changes are highest for landless. The study suggests that government should extend social security nets to landless poor and categorically to vulnerable landless (without livestock) to redress the short-term impact of food price increase. In addition, the government should stabilize food prices and particularly cereal prices in the long- run.

Keywords: QUAIDS, Lagrange multiplier, NLSUR, and Tylor approximation

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Authors: Rima Zakzouk, Noriko Hiraishi, Mohamed Mahdi Alshahni, Koichi Makimura, Junji Tagami

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Background and Purpose: Silver diamine fluoride (SDF) is used to prevent and arrest dental caries. The aim of this study is to evaluate the effect of SDF on reducing Candida albicans adhesion on dentin. Materials and Methods: Bovine dentin disks (6×6 mm) were cut by Isomet and polished using grit silicon carbide papers down to 2000 in order to obtain flat dentin surfaces. Samples were divided into two groups. The first group (SDF group) was treated with 38% SDF for 3 min, while the other group (control group) did not undergo SDF treatment. All samples were exposed to C. albicans suspension, washed after 6 hours incubation at 30 °C before to be tested using XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) and real time PCR approaches. Statistical analyses of the results were performed at the significance level α = 0.05. Results: SDF inhibited C. albicans adhesion onto dentin. A significant difference was found between the SDF and control groups in both XTT and real time PCR tests. Conclusion: Using SDF to arrest the caries, could inhibit the Candida growth on dentin.

Keywords: silver diamine fluoride, dentin, real time PCR, XTT

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Authors: Vidya Pratap, Seena Biju, Keshavdev A.

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In the educational town of Manipal (located in southern India) households dispose their wastes without segregation. Mixed wastes (organic, inorganic and hazardous items) are collected either by private collectors or by the local municipal body in trucks and taken to dump yards. These collectors select certain recyclables from the collected trash and sell them to scrap merchants to earn some extra money. Rag pickers play a major role in picking up card board boxes, glass bottles and milk sachets from dump yards and public areas and scrap iron from construction sites for recycling. In keeping with the Indian Prime Minister’s mission of Swachh Bharat (A Clean India), the local municipal administration is taking efforts to ensure segregation of domestic waste at source. With this in mind, each household in a residential area in Manipal was given two buckets – for wet and dry wastes (wet waste referred to organic waste while dry waste included recyclable and hazardous items). A study was conducted in this locality covering a cluster of 145 households to assess the residents’ knowledge of recyclable, organic and hazardous items commonly disposed by households. Another objective of this research was to evaluate the extent to which the residents actually dispose their wastes appropriately. Questionnaires were self-administered to a member of each household with the assistance of individuals speaking the local language whenever needed. Respondents’ knowledge of whether an item was organic, inorganic or hazardous was captured through a questionnaire containing a list of 50 common items. Their behaviour was captured by asking how they disposed these items. Results show that more than 70% of respondents are aware that banana and orange peels, potato skin, egg shells and dried leaves are organic; similarly, more than 70% of them consider newspapers, notebook and printed paper are recyclable. Less than 65% of respondents are aware that plastic bags and covers and plastic bottles are recyclable. However, the results of the respondents’ recycling behaviour is less impressive. Fewer than 35% of respondents recycle card board boxes, milk sachets and glass bottles. Unfortunately, since plastic items like plastic bags and covers and plastic bottles are not accepted by scrap merchants, they are not recycled. This study shows that the local municipal authorities must find ways to recycle plastic into products, alternate fuel etc.

Keywords: behaviour, knowledge, plastic waste management, recyclables

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Authors: Mohammad Asadpour, Gholamreza Razmi, Gholamreza Mohammadi, Abolghasem Naghibi

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Cryptosporidium Spp., protozoan parasites of the phylum Apicomplexa, have a wide spectrum of hosts including humans, domestic animals and wild mammals, birds, reptiles, amphibians and fish. Dairy cattle have been identified in numerous reports as a major source of environmental contamination with this pathogen. In this study, a Polymerase Chain Reaction (PCR), Restriction Fragment Length Polymorphism (RFLP) analysis of the Small-Subunit (SSU) rRNA gene was used to detect and identify Cryptosporidium Spp. in 300 fecal specimens from 1 to 30 days pre-wean calves in 10 farms in Mashhad, Iran. Eighty five (28.3%) and forty five (15%) of the specimens were positive for Cryptosporidium by microscopic and PCR examination respectively. Restriction digestion of the PCR products by VSPI and Ssp1 restriction enzymes and analysis of sequence data revealed the presence of C. parvum, bovine genotype in all isolates. Our findings suggest that cattle can be a source of Cryptosporidial infections for humans and animals in Mashhad area. This is the first published description of Cryptosporidium sub genotyping in Mashhad.

Keywords: cryptosporidium, genotype, dairy calves, 18S rRNA, Mashhad

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

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In recent years, a new field of basic research has emerged: the biology and physiology of extracellular vesicles and their application in diagnostics and therapy. In particular, exosomes attract the scientific community as nanovesicles of endosomal origin, which can be secreted by a variety of cells and are found in all biological fluids. Exosomes have recently gained attention also in the cosmetic field: in fact they are used in creams, serums and masks for topical use, proving to have a series of therapeutic and anti-aging benefits. To date, the orally administration of exosomes is the subject of attention because it represents a non-invasive and efficient method for delivering bioactive molecules into the intestine. We decided to focus our research on the creation of a food supplement that contains various bioactive factors, vitamins and a new technology called AMPLEX PLUS, containing a mixture of 20 different biologically active factors (GF20) and exosomes isolated and purified from bovine colostrum. We have demonstrated in vitro that this new supplement acts on telomerase, slowing down cell aging.

Keywords: AMPLEX PLUS, colostrum, exosomes, telomerase

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Authors: S. Md. Shaarani, J. Md. Jahim, R. A. Rahman, R. Md. Illias

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Modern developments in biotechnology have paved the way for extensive use of biocatalysis in industries. Although it offers immense potential, industrial application is usually hampered by lack of operational stability, difficulty in recovery as well as limited re-use of the enzyme. These drawbacks, however, can be overcome by immobilization. Cross-linked enzyme aggregates (CLEAs), a versatile carrier-free immobilization technique is one that is currently capturing global interest. This approach involves precipitating soluble enzyme with an appropriate precipitant and subsequent crosslinking by a crosslinking reagent. Without ineffective carriers, CLEAs offer high enzymatic activity, stability and reduced production cost. This study demonstrated successful CLEA synthesis of recombinant xylanase from Trichoderma reesei using ethanol as aggregating agent and glutaraldehyde (2% (v/v); 100 mM) as crosslinker. Effects of additives including proteic feeder such as bovine serum albumin (BSA) and poly-L-Lysine were investigated to reveal its significance in enhancing the performance of enzyme. Addition of 0.1 mg BSA/U xylanase showed considerable increment in CLEA development with approximately 50% retained activity.

Keywords: cross-linked, immobilization, recombinant, xylanase

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Authors: Jessica P. Fiorotti, Maria C. Freitas, Caio J. B. Coutinho-Rodrigues, Mariana G. Camargo, Emily S. Mesquita, Amanda R. C. Corval, Ricardo O. B. Bitencourt, Allan F. Marciano, Diva D. Spadacci-Morena, Patricia S. Golo, Isabele C. Angelo, Vania R. E. P. Bittencourt

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The bovine tick, Rhipicephalus microplus, is an arthropod of great importance in veterinary medicine leading to anemia, weight loss, animals' leather depreciation and also acting as a vector of many pathogens. In this way, the parasitism causes a loss of 3.24 billion dollars per year in Brazil. Knowingly, entomopathogenic fungi act as natural controller of some arthropods, acting mainly by active penetration through the cuticle. However, it can also act on the hemolymph and through the production of mycotoxins. Hemocytes are responsible for the cellular immune response and participate in the processes of phagocytosis, nodulation and encapsulation and may undergo changes when challenged by pathogens. The aim of the present study was to evaluate changes in R. microplus hemocytes after inoculation of Metarhizium robertsii using transmission electron microscopy. The isolate ARSEF 2575 and 200 engorged R. microplus females were used. The groups were divided into control, in which the females were inoculated with 5 μL of sterile distilled water solution and 0.1% Tween 80, and a group inoculated with 5 μL of fungal suspension at the concentration of 10⁷ conidia mL⁻¹. The experiment was performed in duplicate and each group contained 50 females. Twenty-four hours after fungal inoculation, hemolymph was collected through the cuticle dorsal surface perforation of the tick females. After collection, the hemolymph samples were centrifuged at 500 x g for 3 minutes at 4 °C, the plasma was discarded and the hemocyte pellet was resuspended in 50 μl PBS. The suspension material was fixed in 2% glutaraldehyde in Millonig buffer for three hours. After fixation, the material was centrifuged at 500 x g for 3 minutes, the supernatant was discarded and the cells were resuspended in a wash solution. Subsequently, the cells were post-fixed with 1% osmium tetroxide in phosphate buffer for one hour at room temperature and dehydrated in increasing concentrations of ethanol, and then embedded in Epon resin. The ultrathin sections were examined under the LEO EM 906E transmission electron microscopy at 80kV. The ultrastructural results revealed that.in control group, the cells were considered intact, in which the granulocytes were observed with granules of different electrodensities, intact mitochondria and cytoplasm without vacuolization. In addition, granulocytes showed plasma membrane projections similar to pseudopodia. Plasmatocytes presented as irregularly shaped cells, with the eccentric nucleus, agranular cytoplasm and some cells presented pseudopodia. Nevertheless, in the group exposed to the fungus, most of the cells presented in degeneration. The granulocytes found had fewer granules in the cytoplasm and more vacuoles. Plasmatocytes, after treatment, presented many vacuoles also in the cytoplasm and the lysosomes presented great amount of electrodense material in their interior. Thus, the results suggest that the fungus has a depressant action in the immune system of the tick, not only by the cell degranulation, but also suggesting that this leads to morphological changes in the hemocytes and may even trigger processes such as phagocytosis.

Keywords: bovine tick, cellular defense, entomopathogenic fungi, immune response

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Authors: J. Suwanprateeb, F. Thammarakcharoen

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Calcium phosphate coating (CaP) has been employed for protein delivery, but the typical direct protein adsorption on the coating led to low incorporation content and fast release of the protein from the coating. By using bovine serum albumin (BSA) as a model protein, rapid biomimetic co-precipitation between calcium phosphate and BSA was employed to control the distribution of BSA within calcium phosphate coating during biomimetic formation on titanium surface for only 6 h at 50 oC in an accelerated calcium phosphate solution. As a result, the amount of BSA incorporation and release duration could be increased by using a rapid biomimetic co-precipitation technique. Up to 43 fold increases in the BSA incorporation content and the increase from 6 h to more than 360 h in release duration compared to typical direct adsorption technique were observed depending on the initial BSA concentration used during co-precipitation (1, 10, and 100 microgram/ml). From X-ray diffraction and Fourier transform infrared spectroscopy studies, the coating composition was not altered with the incorporation of BSA by this rapid biomimetic co-precipitation and mainly comprised octacalcium phosphate and hydroxyapatite. However, the microstructure of calcium phosphate crystals changed from straight, plate-like units to curved, plate-like units with increasing BSA content.

Keywords: biomimetic, Calcium Phosphate Coating, protein, titanium

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Authors: Jong Kook Lee, Sangcheol Eum, Jaehong Kim

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Wollastonite/hydroxyapatite composite coatings on zirconia were obtained by room temperature spray process. Wollastonite powder was synthesized by solid-state reaction between calcite and silica powder. Hydroxyapatite powder was prepared from bovine bone by the calcination at 1200oC 1h. From two starting raw powders, three kinds of powder mixture were obtained by the ball milling for 24h. By using these powders, wollastonite/hydroxyapatite coatings were fabricated on zirconia substrates by a room temperature spray process, and their microstructure and biological behavior were investigated and compared with pure wollastonite and hydroxyapatite coatings. Wollastonite/hydroxyapatite coatings on zirconia substrates were homogeneously formed in microstructure and had a nanoscaled grain size. The phase composition of the resultant wollastonite/hydroxyapatite coatings was similar to that of the starting powders, however, the grain size of the wollastonite or hydroxyapatite particles was reduced to about 100 nm due to their formation by particle impaction and fracture. The wollastonite/hydroxyapatite coating layer exhibited bioactivity in a stimulated body fluid and forming ability of new hydroxyapatite precipitates of 25 nm during in vitro test in SBF solution, which was enhanced by the increasing wollastonite content.

Keywords: wollastonite, hydroxyapatite composite coatings, room temperature spay process, zirconia

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Authors: Roihatul Mutiah, Sukardiman, Aty Widyawaruyanti

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Calotropis gigantiea (L.) R. Br (Apocynaceae) commonly called as “Biduri” or “giant milk weed” is a well-known weed to many cultures for treating various disorders. Several studies reported that C.gigantea roots has anticancer activity. The main aim of this research was to isolate and identify an active marker compound of C.gigantea roots for quality control purpose of its extract in the development as anticancer natural product. The isolation methods was bioactivity guided column chromatography, TLC, and HPLC. Evaluated anticancer activity of there substances using MTT assay methods. Identification structure active compound by UV, 1HNMR, 13CNMR, HMBC, HMQC spectral and other references. The result showed that the marker active compound was identical as Calotropin.

Keywords: calotropin, Calotropis gigantea, anticancer, marker active

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Authors: Jesus Hernandez, Zead Elzoeiry, Md. S. Islam, Abel E. Navarro

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The adsorption of bovine serum albumin (BSA) and human hemoglobin (Hb) on naturally-occurring adsorbents was studied to evaluate the potential recovery of proteins from meat industry residues. Spent peppermint tea (PM), powdered purple corn cob (PC), natural clay (NC) and chemically-modified clay (MC) were investigated to elucidate the effects of pH, adsorbent dose, initial protein concentration, presence of salts and heavy metals. Equilibrium data were fitted according to isotherm models, reporting a maximum adsorption capacity at pH 8 of 318 and 344 mg BSA/g of PM and NC, respectively. Moreover, Hb displayed maximum adsorption capacity at pH 5 of 125 and 143 mg/g of PM and PC, respectively. Hofmeister salt effect was only observed for PM/Hb system. Salts tend to decrease protein adsorption, and the presence of Cu(II) ions had negligible impacts on the adsorption onto NC and PC. Desorption experiments confirmed that more than 85% of both proteins can be recovered with diluted acids and bases. SEM, EDX, and TGA analyses demonstrated that the adsorbents have favorable morphological and mechanical properties. The long-term goal of this study aims to recover soluble proteins from industrial wastewaters to produce animal food or any protein-based product.

Keywords: adsorption, albumin, clay, hemoglobin, spent peppermint leaf

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Authors: J. D. Cabral, E. Murray, P. Turner, E. Hewitt, A. Ali, M. McConnell

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Worldwide demand for organ replacement and tissue regeneration is progressively increasing. Three-dimensional (3D) bioprinting, where a physical construct is produced using computer-aided design, is a promising tool to advance the tissue engineering and regenerative medicine fields. In this paper we describe two different approaches to developing 3D bioprinted constructs for use in tissue regeneration. Bioink development is critical in achieving the 3D biofabrication of functional, regenerative tissues. Hydrogels, cross-linked macromolecules that absorb large amounts of water, have received widespread interest as bioinks due to their relevant soft tissue mechanics, biocompatibility, and tunability. In turn, not only is bioink optimisation crucial, but the creation of vascularized tissues remains a key challenge for the successful fabrication of thicker, more clinically relevant bioengineered tissues. Among the various methodologies, cell-laden hydrogels are regarded as a favorable approach; and when combined with novel core/shell 3D bioprinting technology, an innovative strategy towards creating new vessel-like structures. In this work, we investigate this cell-based approach by using human umbilical endothelial cells (HUVECs) entrapped in a viscoelastic chitosan/dextran (CD)-based core hydrogel, printed simulataneously along with a gelatin methacrylate (GelMA) shell. We have expanded beyond our previously reported FDA approved, commercialised, post-surgical CD hydrogel, Chitogel®, by functionalizing it with cell adhesion and proteolytic peptides in order to promote bone marrow-derived mesenchymal stem cell (immortalized BMSC cell line, hTERT) and HUVECs growth. The biocompatibility and biodegradability of these cell lines in a 3D bioprinted construct is demonstrated. Our studies show that particular peptide combinations crosslinked within the CD hydrogel was found to increase in vitro growth of BMSCs and HUVECs by more than two-fold. These gels were then used as a core bioink combined with the more mechanically robust, UV irradiated GelMA shell bioink, to create 3D regenerative, vessel-like scaffolds with high print fidelity. As well, microporous MEW scaffolds made from milk proteins blended with PCL were found to show promising bioactivity, exhibiting a significant increase in keratinocyte (HaCaTs) and fibroblast (normal human dermal fibroblasts, NhDFs) cell migration and proliferation when compared to PCL only scaffolds. In conclusion, our studies indicate that a peptide functionalized CD hydrogel bioink reinforced with a GelMA shell is biocompatible, biodegradable, and an appropriate cell delivery vehicle in the creation of regenerative 3D constructs. In addition, a novel 3D printing technique, melt electrowriting (MEW), which allows fabrication of micrometer fibre meshes, was used to 3D print polycaprolactone (PCL) and bioactive milk protein, lactorferrin (LF) and whey protein (WP), blended scaffolds for potential skin regeneration applications. MEW milk protein/PCL scaffolds exhibited high porosity characteristics, low overall biodegradation, and rapid protein release. Human fibroblasts and keratinocyte cells were seeded on to the scaffolds. Scaffolds containing high concentrations of LF and combined proteins (LF+WP) showed improved cell viability over time as compared to PCL only scaffolds. This research highlights two scaffolds made using two different 3D printing techniques using a combination of both natural and synthetic biomaterial components in order to create regenerative constructs as potential chronic wound treatments.

Keywords: biomaterials, hydrogels, regenerative medicine, 3D bioprinting

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Authors: V. Verma, Syed Riyaz-ul-Hassan

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Staphylococcus aureus is one of the most commonly found pathogenic bacteria and is hard to eliminate from the human environment. It is responsible for many nosocomial infections, besides being the main causative agent of food intoxication by virtue of its variety of enterotoxins. Routine detection of S. aureus in food is usually carried out by traditional methods based on morphological and biochemical characterization. These methods are time-consuming and tedious. In addition, misclassifications with automated susceptibility testing systems or commercially available latex agglutination kits have been reported by several workers. Consequently, there is a need for methods to specifically discriminate S. aureus from other staphylococci as quickly as possible. Data on protocols developed using molecular means like PCR technology will be presented for rapid and specific detection of this pathogen in food, clinical and environmental samples, especially milk.

Keywords: food Pathogens, PCR technology, rapid and specific detection, staphylococcus aureus

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Authors: Haiyang Su, Kun Qian

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We developed a novel plasmonic matrix assisted laser desorption/ionization mass spectrometry (MALDI MS) approach to detect small nutrients and toxin in complex biological emulsion samples. We used silver nanoshells (SiO₂@Ag) with optimized structures as matrices and achieved direct analysis of ~6 nL of human breast milk without any enrichment or separation. We performed identification and quantitation of small nutrients and toxins with limit-of-detection down to 0.4 pmol (for melamine) and reaction time shortened to minutes, superior to the conventional biochemical methods currently in use. Our approach contributed to the near-future application of MALDI MS in a broad field and personalized design of plasmonic materials for real case bio-analysis.

Keywords: plasmonic materials, laser desorption/ionization, mass spectrometry, small nutrients, toxins

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Authors: Nasiruddin Khan

Abstract:

The action of (-)-epicatechin, a cocoa (Theobroma cacao) flavanol that modulates redox/oxidative stress are contributed mainly to their antioxidant properties. The present study investigates the concentration and time dependent effect of (-)-epicatechin metabolites 3MeEc, 4MeEc, and 4SulEc on the production of ROS on BAEC using L-012, Lucigenin as chemiluminescence dye and XO/HX system. Our result demonstrates that 3MeEc shows significant (P <0.05) lowering effect of ROS production in BAEC with increasing concentration of metabolite while L-012 was used as chemiluminescence dye but not in the case of Lucigenin. In XO/HX system, using L-012 as chemiluminescence dye, 3MeEc and 4MeEc showed significant lowering effect on ROS production with increasing concentration from 100-500nM as compared to the positive control (SOD). When Lucigenin was used as chemiluminescence dye, 3MeEc exerted significant lowering effect with increasing concentration when compared to the positive control (SOD) whereas 4MeEc showed significant lowering effect in ROS production from 250 nM on as compared to positive control. For 4SulEc, a significant lowering effect of ROS production was only observed at 100 and 250 nM. Overall, although each metabolite shows considerable effect, 3MeEc exhibited more pronounced effect on decreasing the production of ROS as compared to other two metabolites.

Keywords: epicatechin metabolites, HO-1, Nrf2, ROS

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Authors: Noor-ul-Ain, Muhammad Thair Khan, Adeela Ajmal, Hamid Mustafa

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Pakistan is an agricultural country and livestock only share 11.8 percent to national GDP during 2015-16. Pakistan is a 3rd largest milk producing country having 41.2, 35.6, 29.4, 68.4 and 1.0 million head cattle, buffalo, sheep, goat and camel, respectively. Modern urbanization and shortage of feed resources for livestock species in a country is an alarming threat. The introduction of new technology and advanced techniques solve this issue. This includes drought feeding, increase production, aid to crop management, balance nutrition and easily storaged of wet feed products. It is therefore clear that silage has important role in animal feed and feeding. Financial model of this study clear the effectiveness of silage. Therefore, it is revealed from this study that silage is a cost-effective option for a profitable dairy farming in Pakistan.

Keywords: feed, silage, dairy, production, Pakistan

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Authors: M. J. Reis Lima, J. Oliveira, A. C. Sousa Pereira, M. C. Castilho, E. Teixeira-Lemos

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The increasing interest in plant sterol enriched foods is due to the fact that they reduce blood cholesterol concentrations without adverse side effects. In this context, enriched foods with phytosterols may be helpful in protecting population against atherosclerosis and cardiovascular diseases. The aim of the present work was to evaluate in a population of Viseu, Portugal, the consumption habits low-fat, plant sterol-enriched yoghurt. For this study, 577 inquiries were made and the sample was randomly selected for people shopping in various supermarkets. The preliminary results showed that the biggest consumers of these products were women aged 45 to 65 years old. Most of the people who claimed to buy these products consumed them once a day. Also, most of the consumers under antidyslipidemic therapeutics noticed positive effects on hypercholesterolemia.

Keywords: consumption habits, fermented milk, functional foods, low fat, phytosterols

Procedia PDF Downloads 447

Authors: Al-Braa Kashegari, Ali M. El-Halawany, Akram A. Shalabi, Sabrin R. M. Ibrahim, Hossam M. Abdallah

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Chemical investigation of Chrozophora oblongifolia resulted in the isolation of five major compounds that were identified as apeginin-7-O-glucoside (1), quercetin-3-O-glucuronic acid (2), quercetin-3-O-glacturonic acid (3), rutin (4), and 1,3,6-trigalloyl glucose (5). The identity of isolated compounds was assessed by different spectroscopic methods, including one- and two-dimensional NMR. The isolated compounds were tested for their antioxidant activity using different assays viz., DPPH, FRAP, ABTS, ORAC, and metal chelation effects. In addition, the inhibition of target enzymes involved in the metabolic syndrome, such as alpha-glucosidase and pancreatic lipase, were carried out. Moreover, the effect of the compounds on the advanced glycation end-products (AGEs) as one of the major complications of oxidative stress and hyperglycemia in metabolic syndromes were carried out using BSA‐fructose (bovine serum albumin), BSA-methylglyoxal, and arginine methylglyoxal models. The pure isolates showed a protective effect in metabolic syndromes as well as promising antioxidant activity. The results showed potent activity of compound 5 in all measured parameters meanwhile, none of the tested compounds showed activity against pancreatic lipase.

Keywords: Chrozophora oblongifolia, antioxidant, pancreatic lipase, metabolic syndromes

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Authors: Selda Uzal Seyfi

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Goat and sheep milk is quite significant in human nutrition. It is considered as more important day by day. This study was carried out in order to determine applied manure management system and their possibilities of improvement in goat and sheep farm in between 2012 and 2013 years. In the study, it was investigated manure management systems of 25 pieces of sheep and goat farms. It was analyzed the manure collecting, storage and treatment features of farms and whether or not they are suitable for animal breeding. As a result of the study, it was determined that the applied manure management systems in the farm were insufficient. Planning the manure management systems in goat and sheep breeding is appropriate technical criteria is useful in respect of the animal welfare, animal health, the health of workers in the barn and environmental pollution.

Keywords: goat farm, sheep farm, manure storage, manure management

Procedia PDF Downloads 394

Authors: Nikoo Soleimani

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Background: Reducing torque during bone drilling is one of the effective factors in reaching to an optimal drilling process. Methods: 15 bovine femurs were drilled in vitro with a drill bit with a diameter of 4 mm using two methods of torsional ultrasonic assisted drilling (T-UAD) and convent conventional drilling (CD) and the effects of changing the feed rate and rotational speed on the torque were compared in both methods. Results: There was no significant difference in the thrust force measured in both methods due to the direction of vibrations. Results showed that using T-UAD method for bone drilling at feed rates of 0.16, 0.24 and 0.32 mm/rev led for all rotational speeds to a decrease of at least 16.3% in torque compared to the CD method. Further, using T-UAD at rotational speeds of 355~1000 rpm with various feed rates resulted in a torque reduction of 16.3~50.5% compared to CD method. Conclusions: Reducing the feed rate and increasing the rotational speed, except for the rotational speed of 500 rpm and a feed rate of 0.32 mm/rev, resulted generally in torque reduction in both methods. However, T-UAD is a more effective and desirable option for bone drilling considering its significant torque reduction.

Keywords: torsional ultrasonic assisted drilling, torque, bone drilling, rotational speed, feed rate

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Authors: Muhammad Waqar Ashraf, Atiq A. Mian

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In the present study, accumulation of eight heavy metals, lead (Pb), cadmium (Cd), manganese (Mn), copper (Cu), zinc (Zn), iron (Fe), nickel (Ni) and chromium (Cr)was determined in kidney, heart, liver and muscle tissues of Lethrinus miniatus fish caught from Arabian Gulf. Metal concentrations in all the samples were measured using Atomic Absorption Spectroscopy. Analytical validation of data was carried out by applying the same digestion procedure to standard reference material (NIST-SRM 1577b bovine liver). Levels of lead (Pb) in the liver tissue (0.60µg/g) exceeded the limit set by European Commission (2005) at 0.30 µg/g. Zinc concentration in all tissue samples were below the maximum permissible limit (50 µg/g) as set by FAO. Maximum mean cadmium concentration was found 0.15 µg/g in the kidney tissues. Highest content of Mn in the studied tissues was seen in the kidney tissue (2.13 µg/g), whereas minimum was found in muscle tissue (0.87 µg/g). The present study led to the conclusion that muscle tissue is the least contaminated tissue in Lethrinus miniatus and consumption of organs should be avoided as much as possible.

Keywords: lethrinus miniatus, arabian gulf, heavy metals, atomic absorption spectroscopy

Procedia PDF Downloads 352

Authors: John O. Akindoyo, M. D. H. Beg, Suriati Binti Ghazali, Nitthiyah Jeyaratnam

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Regeneration of bone due to the many health challenges arising from traumatic effects of bone loss, bone tumours and other bone infections is fast becoming indispensable. Over the period of time, some approaches have been undertaken to mitigate this challenge. This includes but not limited to xenografts, allografts, autografts as well as artificial substitutions like bioceramics, synthetic cements and metals. However, most of these techniques often come along with peculiar limitation and problems such as morbidity, availability, disease transmission, collateral site damage or absolute rejection by the body as the case may be. Hydroxyapatite (HA) is very compatible and suitable for this application. However, most of the common methods for HA synthesis are expensive and environmentally unfriendly. Extraction of HA from bio-wastes have been perceived not only to be cost effective, but also environment-friendly. In this research, HA was produced from bio-waste: namely bovine bones through a combination of hydrothermal chemical processes and ordinary calcination techniques. Structure and property of the HA was carried out through different characterization techniques (such as TGA, FTIR, DSC, XRD and BET). The synthesized HA was found to possess similar properties to stoichiometric HA with highly desirable thermal, degradation, structural and porous properties. This material is unique for its potential minimal cost, environmental friendliness and property controllability. It is also perceived to be suitable for tissue and bone engineering applications.

Keywords: biomaterial, biopolymer, bone, hydroxyapatite

Procedia PDF Downloads 314