Search results for: bone collagen stable isotope analysis

Authors: M. Tassi, E. Dotsika, P. Karalis, A. Trantalidou, A. Mazarakis Ainian

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The Kythnos Island in Greece is of particular archaeological interest, as it has been inhabited from the 12th BC until the 7th AD. From island excavations, numerous faunal and human skeletal remains have been recovered. This work is the first attempt at the paleoenvironmental reconstruction of the island via stable isotope analysis. Specifically, we perform 13C and 18O isotope analysis in faunal bone apatite in order to investigate the climate conditions that prevailed in the area. Additionally, we conduct 13C and 15N isotope analysis in faunal bone collagen, which will constitute the baseline for the subsequent diet reconstruction of the ancient Kythnos population.

Keywords: stable isotopes analysis, bone collagen stable isotope analysis, bone apatite stable isotope analysis, paleodiet, palaeoclimate

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Authors: Mojtaba Heydarizad, Hamid Ghalibaf Mohammadabadi

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Iran is a semi-arid and arid country in south-western Asia in the Middle East facing intense climatological drought from the early times. Therefore, studying the precipitation events and the moisture sources and air masses causing precipitation has great importance in this region. In this study, the moisture sources and stable isotope content of precipitation moisture in three main events in 2015 have been studied in North-Eastern Iran. HYSPLIT model backward trajectories showed that the Caspian Sea and the mixture of the Caspian and Mediterranean Seas are dominant moisture sources for the studied events. This showed the role of cP (Siberian) and Mediterranean (MedT) air masses. Stable isotope studies showed that precipitation events originated from the Caspian Sea with lower Sea Surface Temperature (SST) have more depleted isotope values. However, precipitation events sourced from the mixture of the Caspian and the Mediterranean Seas (with higher SST) showed more enriched isotope values.

Keywords: HYSPLIT, Iran, Northern Khorasan, stable isotopes

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Authors: Efthimios Kokkotos, Kostakis Marios, Beis Alexandros, Angelos Patakas, Antonios Avgeris, Vassilios Triantafyllidis

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H, C, and O stable isotope ratios were measured in different olive oils and table olives originating from different regions of Greece. In particular, the stable isotope ratios of different olive oils produced in the Lakonia region (Peloponesse – South Greece) from different varieties, i.e., cvs ‘Athinolia’ and ‘koroneiki’, were determined. Additionally, stable isotope ratios were also measured in different table olives (cvs ‘koroneiki’ and ‘kalamon’) produced in the same region (Messinia). The aim of this study was to provide sufficient isotope ratio data regarding each variety and region of origin that could be used in discriminative studies of oil olives and table olives produced by different varieties in other regions. In total, 97 samples of olive oil (cv ‘Athinolia’ and ‘koroneiki’) and 67 samples of table olives (cvs ‘kalmon’ and ‘koroneiki’) collected during two consecutive sampling periods (2021-2022 and 2022-2023) were measured. The C, H, and O isotope ratios were measured using Isotope Ratio Mass Spectrometry (IRMS), and the results obtained were analyzed using chemometric techniques. The measurements of the isotope ratio analyses were expressed in permille (‰) using the delta δ notation (δ=Rsample/Rstandard-1, where Rsample and Rstandardis represent the isotope ratio of sample and standard). Results indicate that stable isotope ratios of C, H, and O ranged between -28,5+0,45‰, -142,83+2,82‰, 25,86+0,56‰ and -29,78+0,71‰, -143,62+1,4‰, 26,32+0,55‰ in olive oils produced in Lakonia region from ‘Athinolia’ and ‘koroneiki ‘varieties, respectively. The C, H, and O values from table olives originated from Messinia region were -28,58+0,63‰, -138,09+3,27‰, 25,45+0,62‰ and -29,41+0,59‰,-137,67+1,15‰, 24,37+0,6‰ for ‘Kalamon’ and ‘koroneiki’ olives respectively. Acknowledgments: This research has been co-financed by the European Regional Development Fund of the European Union and Greek national funds through the Operational Program Competitiveness, Entrepreneurship and Innovation, under the call RESEARCH—CREATE—INNOVATE (Project code: T2EDK-02637; MIS 5075094, Title: ‘Innovative Methodological Tools for Traceability, Certification and Authenticity Assessment of Olive Oil and Olives’).

Keywords: olive oil, table olives, Isotope ratio, IRMS, geographical origin

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Authors: P. Vosynek, T. Návrat, M. Peč, J. Pořízka, P. Diviš

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Changes of mechanical properties and response behavior of bones is an important external sign of medical problems like osteoporosis, bone remodeling after fracture or surgery, osteointegration, or bone tissue loss of astronauts in space. Measuring of mechanical behavior of bones in physiological and osteoporotic states, quantified by different degrees of protein (collagen) and mineral loss, is thus an important topic in biomechanical research. This contribution deals with the relation between mechanical properties of the turkey long bone–tibia in physiological, demineralized, and deproteinized state. Three methods for comparison were used: densitometry, three point bending and harmonic response analysis. The results help to find correlations between the methods and estimate their possible application in medical practice.

Keywords: bone properties, long bone, osteoporosis, response behavior

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Authors: Lucille McWilliams

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As water temperatures drop in November, Kemp’s Ridley, Loggerhead, and Green sea turtles cold-stun in Cape Cod Bay. The foraging ecology of these sea turtles remains an understudied area of research. In this study, we aim to assess the diet of these turtles using a multi-tissue stable isotope analysis of cold-stunned kemp’s ridley, loggerhead, and green sea turtles stranded from 2015 to 2020. Stable isotope ratios of carbon and nitrogen were measured in blood, front and rear flipper, liver, muscle, skin, and scute tissue samples. We predict an elevated level of Nitrogen isotope ratios in kemp’s ridley and loggerhead turtles compared to green turtles due to the carnivorous loggerheads and kemp ridleys’ carnivorous diet and the greens herbivorous diet. We anticipate empty stomachs due to starvation while stranded, and a variety of foraging strategies, migration patterns, and trophic positions between these species. Data collected from this study will add to the knowledge of these turtles’ prey species and aid managers in the preservation of these species as a mitigation strategy for these turtles' extinction.

Keywords: sea turtles, kemp's ridleys, greens, loggerheads, cold-stunning, diet analysis, stable isotope analysis, environmental science, marine biology

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Authors: P. Hashim, M. S. Mohd Ridzwan, J. Bakar

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Collagen was isolated from chicken feet by using papain and pepsin enzymes in acetic acid solution at 4°C for 24h with a yield of 18.16% and 22.94% by dry weight, respectively. Chemical composition and characteristics of chicken feet collagen such as amino acid composition, SDS-PAGE patterns, FTIR spectra and thermal properties were evaluated. The chicken feet collagen is rich in the amino acids glycine, glutamic acid, proline and hydroxyproline. Electrophoresis pattern demonstrated two distinct α-chains (α1 and α2) and β chain, indicating that type I collagen is a major component of chicken feet collagen. The thermal stability of collagen isolated by papain and pepsin revealed stable denaturation temperatures of 48.40 and 53.35°C, respectively. The FTIR spectra of both collagens were similar with amide regions in A, B, I, II, and III. The study demonstrated that chicken feet collagen using papain isolation method is possible as commercial alternative ingredient.

Keywords: chicken feet, collagen, papain, pepsin

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Authors: A. M. Malagon Escandon, J. A. Arenas Alatorre, C. P. Chaires Rosas, N. A. Vazquez Torres, B. Hernandez Tellez, G. Pinon Zarate, M. Herrera Enriquez, A. E. Castell Rodriguez

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The current approach to the treatment of bone defects involves the use of scaffolds that provide a biological and mechanically stable niche to favor tissue repair. Despite the significant progress in the field of bone tissue engineering, several main problems associated are attributed to giving a low biodegradation degree, does not promote osseointegration and regeneration, if the bone is not healing as well as expected or fails to heal, will not be given a proper ossification or new bone formation. The actual approaches of bone tissue regeneration are directed to the use of decellularized native extracellular matrices, which are able of retain their own architecture, mechanic properties, biodegradability and promote new bone formation because they are capable of conserving proteins and other factors that are founded in physiological concentrations. Therefore, we propose an extracellular matrix-based bioscaffolds derived from bovine cancellous bone, which is processed by decellularization, demineralization, and hydrolysis of the collagen protein, these protocols have been successfully carried out in other organs and tissues; the effectiveness of its biosafety has also been previously evaluated in vivo and Food and Drug Administration (FDA) approved. In the specific case of bone, a more complex treatment is needed in comparison with other organs and tissues because is necessary demineralization and collagen denaturalization. The present work was made in order to obtain a temporal scaffold that succeed in degradation in an inversely proportional way to the synthesis of extracellular matrix and the maturation of the bone by the cells of the host.

Keywords: bioactive, biodegradable, bone, extracellular matrix-based bioscaffolds, stem cells, tissue engineering

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Authors: Hye Kyung Kim, Myung-Gyou Kim, Kang-Hyun Leem

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Postmenopausal osteoporosis is characterized by low bone density which leads to increased bone fragility and greater susceptibility to fracture. Current treatments for osteoporosis are dominated by drugs that inhibit bone resorption although they also suppress bone formation that may contribute to pathogenesis of osteonecrosis. To restore the extensive bone loss, there is a great need for anabolic treatments that induce osteoblasts to build new bone. Pre-osteoblastic cells produce proteins of the extra-cellular matrix, including type I collagen at first, and then to successively produce alkaline phosphatase (ALP) and osteocalcin during differentiation to osteoblasts. Finally, osteoblasts deposit calcium. Present study investigated the effects of egg yolk peptide (EYP) on osteogenic activities and bone matrix gene expressions in human osteoblastic MG-63 cells. The effects of EYP on cell proliferation, alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization were measured. The expression of osteogenic genes including COL1A1 (collagen, type I, alpha 1), ALP, BGLAP (osteocalcin), and SPP1 (secreted phosphoprotein 1, osteopontin) were measured by quantitative realtime PCR. EYP dose-dependently increased MG-63 cell proliferation, ALP activity, collagen synthesis, and calcium deposition. Furthermore, COL1A1, ALP, and SPP1 gene expressions were increased by EYP treatment. Present study suggested that EYP treatment enhanced osteogenic activities and increased bone matrix osteogenicgenes. These results could provide a mechanistic explanation for the bone-strengthening effects of EYP.

Keywords: egg yolk peptide, osteoblastic MG-63 cells, alkaline phosphatase, collagen synthesis, osteogenic genes, COL1A1, osteocalcin, osteopontin

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Authors: Hye Kyung Kim, Myung-Gyou Kim, Kang-Hyun Leem

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It has been reported that deer antler extract has bone-strengthening activity and effectively used in bone diseases therapy. However, little is known about the cellular and molecular mechanism of this effect. The upper section, mid section, and base of the antler has been known to exhibit different biological properties. Present study investigated the effects of these three parts of deer antler extracts on bone formation and resorption. The effects of deer antler extracts (DH) on bone formation were determined by cell proliferation, alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization in human osteoblastic MG-63 cells. The effect on bone resorption was determined by osteoclastogenesis from bone marrow-derived precursor cells driven by RANKL. Ethanol extracts of DH (50 ~ 100 µg/ml) dose-dependently increased cell proliferation, and upper part increased the cell proliferation by 118.4% while mid and base parts increased proliferation by 107.8% and 102.3%, respectively. ALP activity was significantly increased by upper part of the DH treatment. After enhancement of ALP activity, significant augmentation of collagen synthesis and calcification assessed by Sirus red and Alzarin red staining, respectively, was observed in upper part of the DH treatment. The effect of DH on bone resorption was not observed in all three parts of the DH. These results could provide a mechanistic explanation for the bone-strengthening effects of DH.

Keywords: alkaline phosphatase, collagen synthesis, deer antler, osteoblastic MG-63 cells

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Authors: Benedetta Isella, Aleksander Drinic, Alissa Heim, Phillip Czichowski, Lisa Lauts, Hans Leemhuis

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Introduction: Membranes for guided bone regeneration are essential to perform a barrier function between the soft and the regenerating bone tissue. Bioabsorbable membranes are desirable in this field as they do not require a secondary surgery for removal, decreasing patient surgical risk. Collagen was the first bioabsorbable alternative introduced on the market, but its degradation time may be too fast to guarantee bone regeneration, and optimisation is needed. Silk fibroin, being biocompatible, slowly bioabsorbable, and processable into different scaffold types, could be a promising alternative. Objectives: The objective is to compare the general performance of a silk fibroin membrane for guided bone regeneration to current collagen alternatives developing suitable standardized tests for the mechanical and morphological characterization. Methods: Silk fibroin and collagen-based membranes were compared from the morphological and chemical perspective, with techniques such as SEM imaging and from the mechanical point of view with techniques such as tensile and suture retention strength (SRS) tests. Results: Silk fibroin revealed a high degree of reproducibility in surface density. The SRS of silk fibroin (0.76 ± 0.04 N), although lower than collagen, was still comparable to native tissues such as the internal mammary artery (0.56 N), and the same can be extended to general mechanical behaviour in tensile tests. The SRS could be increased by an increase in thickness. Conclusion: Silk fibroin is a promising material in the field of guided bone regeneration, covering the interesting position of not being considered a product containing cells or tissues of animal origin from the regulatory perspective and having longer degradation times with respect to collagen.

Keywords: guided bone regeneration, mechanical characterization, membrane, silk fibroin

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Authors: Mojtaba Heydarizad

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Zagros mountain range is a very important precipitation zone in Iran as it receives high average annual precipitation compared to other parts of this country. Although this region is important precipitation zone in semi-arid an arid country like Iran, accurate method to study water resources in this region has not been applied yet. In this study, stable isotope δ¹⁸O content of precipitation and groundwater resources showed spatial variations across Zagros region as southern parts of Zagros region showed more enriched isotope values compared to the northern parts. This is normal as southern Zagros region is much drier with higher air temperature and evaporation compared to northern parts. In addition, the spatial variations of stable isotope δ¹⁸O in precipitation in Zagros region have been simulated by the models which consider the altitude and latitude variations as input to simulate δ¹⁸O in precipitation.

Keywords: groundwater, precipitation, simulation, stable isotopes, Zagros region

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Authors: Omid Gholipoor Bashiri, Ghafur Mosavi, Aliasghar Behnamghader, Seyed Mahmood Rabiee

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Objective: To determine the effect of selected bone graft on the compression properties of radial bone in rabbit. Design-Experimental in vivo study. Animals: A total of 45 adult male New Zealand white rabbits. Procedures: The rabbits were anesthetized and a one-cm-full thickness piece of radial bone was removed using oscillating saw in the all rabbit. The rabbits were divided into 5 groups on the basis of the material used to fill the bone defect: group 1: the paste of bone cement calcium phosphate; group II: the paste of calcium phosphate mixture with type I collagen; group III: tricalcium phosphate mixed with hydroxyapatite (TCP & HP) with 5% porosity; group IV: the same scaffold as group III with 10% porosity; and group V: the same scaffold as group III and IV with 20% porosity, with 9 rabbits in each group. Subsequently subdivided into 3 subgroups of 3 rabbits each. Results: There was a significant increase in compression properties of radial bone in the group II and V in 2nd and 3rd months as compared with groups I, III and IV. The mean endurable crack-strength in group II and V were slightly higher than that of normal radius (P<0.05). Conclusion and clinical relevance: Application of calcium phosphate paste with type I collagen and scaffold of tricalcium phosphate with hydroxyapatite having 20% porosity indicated to have positive effect in integral formation of qualitative callus at the site of fracture and early re-organization of callus to regain mechanical strength too.

Keywords: calcium phosphate, tricalcium phosphate, hydroxyapatite, radial bone, compressive properties, porosity, type i collagen, rabbit

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Authors: Solehuddin Shuib, Koay Boon Aik, Zainul Ahmad Rajion

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There are many types of mechanical failure on the dental implant. In this project, the failure that needs to take into consideration is the bone resorption on the dental implant. Human bone has its ability to remodel after the implantation. As the dental implant is installed into the bone, the bone will detect and change the bone structure to achieve new biomechanical environment. This phenomenon is known as bone remodeling. The objective of the project is to improve the performance of dental implant by using different types of design. These designs are used to analyze and predict the failure of the dental implant by using finite element analysis (FEA) namely ANSYS. The bone is assumed to be fully attached to the implant or cement. Hence, results are then compared with other researchers. The results were presented in the form of Von Mises stress, normal stress, shear stress analysis, and displacement. The selected design will be analyzed further based on a theoretical calculation of bone remodeling on the dental implant. The results have shown that the design constructed passed the failure analysis. Therefore, the selected design is proven to have a stable performance at the recovery stage.

Keywords: dental implant, FEA, bone remodeling, design

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Authors: Ahmed Mohamed El-Shamy, Akram Abbas El-Awady, Mahmoud Taha Eldestawy

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Background: The present study was to evaluate clinically and radiographically the combined effect of split crest technique by ultrasonic bone surgery and platelet concentrates in implant site development. Methods: Forty patients with narrow ridge were participated in this study. Patients were assigned randomly into one of the following four groups according to treatment: Group 1: Patients received split-crest technique by ultrasonic bone surgery with implant placement. Group 2: Patients received split-crest technique by ultrasonic bone surgery with implant placement and PRF. Group 3: Patients received split-crest technique by ultrasonic bone surgery with implant placement and PRP. Group 4: Patients received split-crest technique by ultrasonic bone surgery with implant placement and collagen membrane. Modified plaque index, modified sulcus bleeding index, and implant stability were recorded as a baseline and measured again at 3 and 6 months. CBCT scans were taken immediately after surgery completion and at 9 months to evaluate bone density at the bone-implant interface. Results after 6 months; collagen group showed statistically significantly lower mean modified bleeding index than the other groups. After 3 months, the PRF group showed statistically significantly higher mean implant stability with ostell ISQ units' than the other groups. After 6 months, the PRF group showed statistically significantly higher mean implant stability with ostell ISQ units' than the other groups. After 6 months, the PRF group showed statistically significantly higher mean bone density than the collagen group. Conclusion: Ultrasonic bone surgery in split-crest technique can be a successful option for increasing implant stability values throughout the healing period. The use of a combined technique of ultrasonic bone surgery with PRF and simultaneous implant placement potentially improves osseointegration (bone density). PRF membranes represent advanced technology for the stimulation and acceleration of bone regeneration.

Keywords: dental implants, split-crest, PRF, PRP

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Authors: Ling-Ling E., Hong-Chen Liu, Dong-Sheng Wang, Fang Su, Xia Wu, Zhan-Ping Shi, Yan Lv, Jia-Zhu Wang

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Objective: The objective of the present study is to evaluate the capacity of a tissue-engineered bone complex of recombinant human bone morphogenetic protein 2 (rhBMP-2) mediated dental pulp stem cells (DPSCs) and nano-hydroxyapatite/collagen/poly(L-lactide)(nHAC/PLA) to reconstruct critical-size alveolar bone defects in New Zealand rabbit. Methods: Autologous DPSCs were isolated from rabbit dental pulp tissue and expanded ex vivo to enrich DPSCs numbers, and then their attachment and differentiation capability were evaluated when cultured on the culture plate or nHAC/PLA. The alveolar bone defects were treated with nHAC/PLA, nHAC/PLA+rhBMP-2, nHAC/PLA+DPSCs, nHAC/PLA+DPSCs+rhBMP-2, and autogenous bone (AB) obtained from iliac bone or were left untreated as a control. X-ray and a polychrome sequential fluorescent labeling were performed post-operatively and the animals were sacrificed 12 weeks after operation for histological observation and histomorphometric analysis. Results: Our results showed that DPSCs expressed STRO-1 and vementin, and favoured osteogenesis and adipogenesis in conditioned media. DPSCs attached and spread well, and retained their osteogenic phenotypes on nHAC/PLA. The rhBMP-2 could significantly increase protein content, alkaline phosphatase (ALP) activity/protein, osteocalcin (OCN) content, and mineral formation of DPSCs cultured on nHAC/PLA. The X-ray graph, the fluorescent, histological observation and histomorphometric analysis showed that the nHAC/PLA+DPSCs+rhBMP-2 tissue-engineered bone complex had an earlier mineralization and more bone formation inside the scaffold than nHAC/PLA, nHAC/PLA+rhBMP-2 and nHAC/PLA+DPSCs, or even autologous bone. Implanted DPSCs contribution to new bone were detected through transfected eGFP genes. Conclutions: Our findings indicated that stem cells existed in adult rabbit dental pulp tissue. The rhBMP-2 promoted osteogenic capability of DPSCs as a potential cell source for periodontal bone regeneration. The nHAC/PLA could serve as a good scaffold for autologous DPSCs seeding, proliferation and differentiation. The tissue-engineered bone complex with nHAC/PLA, rhBMP-2, and autologous DPSCs might be a better alternative to autologous bone for the clinical reconstruction of periodontal bone defects.

Keywords: nano-hydroxyapatite/collagen/poly (L-lactide), dental pulp stem cell, recombinant human bone morphogenetic protein, bone tissue engineering, alveolar bone

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Authors: Monika Bansal

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Background: Placement of dental implant in narrow alveolar ridge is challenging to be treated. GBR procedure is currently most widely used to augment the deficient alveolar ridges and to treat the fenestration and dehiscence around dental implants. Thus, the objectives of the present study were to evaluate as well as compare the clinical performance of collagen membrane and titanium mesh for horizontal bone augmentation at dehisced implant sites. Methods and material: Total 12 single edentulous implant sites with buccal bone deficiency in 8 subjects were equally divided and treated simultaneously with either of the two membranes and DBBM(Bio-Oss) bone graft. Primary outcome measurements in terms of defect height and defect width were made using a calibrated plastic periodontal probe. Re-entry surgery was performed to remeasure the augmented site and to remove Ti-mesh at 6th month. Independent paired t-tests for the inter-group comparison and student-paired t-tests for the intra-group comparison were performed. The differences were considered to be significant at p ≤ 0.05. Results: Mean defect fill with respect to height and width was 3.50 ± 0.54 mm (87%) and 2.33 ± 0.51 mm (82%) for collagen membrane and 3.83 ± 0.75 mm (92%) and 2.50 ± 0.54 mm (88%) for Ti-mesh group respectively. Conclusions: Within the limitation of the study, it was concluded that mean defect height and width after 6 months were statistically significant within the group without significant difference between them, although defect resolution was better in Ti-mesh.

Keywords: collagen membrane, dehiscence, dental implant, horizontal bone, augmentation, ti-mesh

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Authors: Hye Kyung Kim, Myung-Gyou Kim, Kang-Hyun Leem

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The deer velvet antler is well known for its traditional medicinal value and is widely used in the clinic. It has been considered to possess bone-strengthening activity. The goal of this study was to investigate the anti-osteoporotic effect of deer antler velvet on ovariectomized rats (OVX), and their possible mechanism of the action. In the first step, the in vitro effects of DAE on bone loss were determined. The proliferation, collagen content and alkaline phosphatase (ALP) activity of human osteoblastic MG-63 cells and osteoclastogenesis from bone marrow-derived precursor cells were measured. The in vivo experiment confirmed the positive effect of DAE on bone tissue. 3-month old female Sparague-Dawley rats were either sham operated or OVX, and administered DAE (20 and 100 mg/kg) for 4 weeks. DAE increased MG-63 cell proliferation and ALP activity in a dose-dependent manner. Collagen content was also increased by DAE treatment. However, the effect of DAE on bone resorption was not observed. OVX rats supplemented with DAE showed osteoprotective effects as the bone ALP level was increased and c-terminal telopeptide level was decreased by 100 mg/kg DAE treatment compared with OVX controls. Moreover, the tartrate-resistant acid phosphatase-5b level was also decreased by DAE treatment. The present study suggests that DAE is effective in preventing bone loss in OVX rats, and may be potential therapeutic agents for the treatment of postmenopausal osteoporosis.

Keywords: bone ALP, c-terminal telopeptide, deer antler, osteoporosis, ovariectomy, tartrate-resistant acid phosphatase-5b

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Authors: A. Bajek, J. Skopinska-Wisniewska, A. Rynkiewicz, A. Jundzill, M. Bodnar, A. Marszalek, T. Drewa

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Traumatic injury and age-related degenerative diseases associated with cartilage are major health problems worldwide. The articular cartilage is comprised of a relatively small number of cells, which have a relatively slow rate of turnover. Therefore, damaged articular cartilage has a limited capacity for self-repair. New clinical methods have been designed to achieve better repair of injured cartilage. However, there is no treatment that enables full restoration of it. The aim of this study was to evaluate how collagen gel with bone marrow mesenchymal stem cells (MSCs) and collagen gel alone will influence on the hip cartilage repair after injury. Collagen type I was isolated from rats’ tails and cross-linked with N-hydroxysuccinimide in 24-hour process. MSCs were isolated from rats’ bone marrow. The experiments were conducted according to the guidelines for animal experiments of Ethics Committee. Fifteen 8-week-old Wistar rats were used in this study. All animals received hip joint surgery with a total of 30 created cartilage defects. Then, animals were randomly divided into three groups and filled, respectively, with collagen gel (group 1), collagen gel cultured with MSCs (group II) or left untreated as a control (control group). Immunohistochemy and radiological evaluation was carried out 11 weeks post implantation. It has been proved that the surface of the matrix is non-toxic, and its porosity promotes cell adhesion and growth. However, the in vivo regeneration process was poor. We observed the low integration rate of biomaterial. Immunohistochemical evaluation of cartilage after 11 weeks of treatment showed low II and high X collagen expression in two tested groups in comparison to the control one, in which we observed the high II collagen expression. What is more, after radiological analysis, we observed the best regeneration process in control group. The biomaterial construct and mesenchymal stem cells, as well as the use of the biomaterial itself was not sufficient to regenerate the hip cartilage surfaces. These results suggest that the collagen gel based biomaterials, even with MSCs, are not satisfactory in repar of hip cartilage defect. However, additional evaluation is needed to confirm these results.

Keywords: collafen gel, MSCs, cartilage repair, hip cartilage

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Authors: Anna-Akrivi Thomatou, Anastasios Zotos, Eleni C. Mazarakioti, Efthimios Kokkotos, Achilleas Kontogeorgos, Athanasios Ladavos, Angelos Patakas

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It is well known that accurate determination of geographic origin to confront mislabeling and adulteration of foods is considered as a critical issue worldwide not only for the consumers, but also for producers and industries. Among agricultural products, rice (Oryza sativa L.) is the world’s third largest crop, providing food for more than half of the world’s population. Consequently, the quality and safety of rice products play an important role in people’s life and health. Despite the fact that rice is predominantly produced in Asian countries, rice cultivation in Greece is of significant importance, contributing to national agricultural sector income. More than 25,000 acres are cultivated in Greece, while rice exports to other countries consist the 0,5% of the global rice trade. Although several techniques are available in order to provide information about the geographical origin of rice, little data exist regarding the ability of these methodologies to discriminate rice production from Greece. Thus, the aim of this study is the comparative evaluation of stable isotope ratio methodology regarding its discriminative ability for geographical origin determination of rice samples produced in Greece compared to those from three other Asian countries namely Korea, China and Philippines. In total eighty (80) samples were collected from selected fields of Central Macedonia (Greece), during October of 2021. The light element (C, N, S) isotope ratios were measured using Isotope Ratio Mass Spectrometry (IRMS) and the results obtained were analyzed using chemometric techniques, including principal components analysis (PCA). Results indicated that the 𝜹 15N and 𝜹 34S values of rice produced in Greece were more markedly influenced by geographical origin compared to the 𝜹 13C. In particular, 𝜹 34S values in rice originating from Greece was -1.98 ± 1.71 compared to 2.10 ± 1.87, 4.41 ± 0.88 and 9.02 ± 0.75 for Korea, China and Philippines respectively. Among stable isotope ratios studied, values of 𝜹 34S seem to be the more appropriate isotope marker to discriminate rice geographic origin between the studied areas. These results imply the significant capability of stable isotope ratio methodology for effective geographical origin discrimination of rice, providing a valuable insight into the control of improper or fraudulent labeling. Acknowledgement: This research has been financed by the Public Investment Programme/General Secretariat for Research and Innovation, under the call “YPOERGO 3, code 2018SE01300000: project title: ‘Elaboration and implementation of methodology for authenticity and geographical origin assessment of agricultural products.

Keywords: geographical origin, authenticity, rice, isotope ratio mass spectrometry

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Authors: Mehdi Ghanavati

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The Effects of salinity level and duration on carbon isotope discrimination (Δ) of Matricaria chamomilla and Matricaria aurea were evaluated. Four ecotypes of M. chamomilla and four ecotypes of M. aurea were grown at different NaCl concentrations (control, 6, 12 and 18 dS/m) in sand culture condition. Carbon isotope discrimination (Δ) varied significantly (p<0.001) among ecotypes. The amount of carbon isotope discrimination (Δ) increased in first salinity level (6 dS/m), but in other levels (12 and 18 dS/m) it did not increase. Stages of salinity treatments (two stages: first from seedling stage until the end of the experiment and second stage of stress exertion began at stem elongation and seedlings emergence from rosette stage to harvest) had not a significant difference. Study of two spices of chamomile showed the M. aurea had a higher amount of carbon isotope discrimination (Δ) (22.9%) than M. chamomilla (22.48%).

Keywords: salinity, carbon isotope discrimination, Matricaria chamomilla, Matricaria aurea

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Authors: Beata Kaczmarek-Szczepańska, Lidia Zasada

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Marine collagens such as fish skin, bone, cartilage, and scales, including both marine vertebrates and invertebrates sources, are more bioavailable compared to bovine or porcine collagen and have a higher absorption capability and more rapid bloodstream circulation due to their low molecular weight and small particle size. Fish skin may be used as a source of bioactive compounds. The advantage is that fish skin is a by-product of the food industry. The subject of the study is a lyophilizate consisting of a mixture of compounds from the group of glycosaminoglycans and collagen obtained as a result of processing fish skins. Bioactive compounds may find biomedical use in the production of dressing materials for wounds or in tissue engineering for the production of scaffolds for cells. Collagen and glycosaminoglycans were isolated from Salmo salar skin. The final mixture was obtained by the freeze-drying method. In the obtained lyophilizate, the content of amino acids was studied as well as the presence of polysaccharides. The studies showed the presence of glycine, proline, and hydroxyproline, which are the main amino acids of collagen. The HPLC analysis showed the presence of glucose which is a product of polysaccharides hydrolyzation and then reduction of glucuronic acid. It may be assumed that the lyophilizate contains both collagen and polysaccharide, which is probably hyaluronic acid. Acknowledgment: This work was carried out as a result of research project no. TANGO-V-A/0020/2021 financed by the National Centre for Research and Development.

Keywords: collagen, glycosaminoglycans, bioactive compounds, fish skin

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Authors: Temesgen Oljira, Olugbenga Akindeji Okunlola, Akinade Shadrach Olatunji, Dereje Ayalew, Bekele A. Bedada, Tasin Godlove Bafon

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The Megele area gold-bearing Neoproterozoic rocks in the Western Ethiopian Shield has been under exploration for the last few decades. The geochemical and ore petrological characterization of the gold-bearing granite gneiss and associated quartz vein is crucial in understanding the gold's genesis. The present study concerns the ore petrological, geochemical, and stable O2 and S characterization of the gold-bearing granite gneiss and associated quartz vein. This area is known for its long history of placer gold mining. The presence of quartz veins of different generations and orientations, visible sulfide mineralization, and oxidation suggests that the Megele area is geologically fertile for mineralization. The Au and base metals analysis also indicate that Megele area rocks are characterized by Cu (2-22 ppm av. 7.83 ppm), Zn (2-53 ppm av. 29.33 ppm), Co (1-27 ppm av. 13.33 ppm), Ni (2-16 ppm av. 10 ppm), Pb (5-10 ppm av. 8.33 ppm), Au (1-5 ppb av. 2.11 ppb), Ag (0.5 ppm), As (5-12 ppm av. 7.83 ppm), Cd (0.5ppm), Li (0.5 ppm), Mo (1-4 ppm av. 1.6 ppm), Sc (5-13 ppm av. 9.3 ppm), and Tl (10 ppm). The oxygen isotope (δ18O) values of gold-bearing granite gneiss and associated quartz veins range from +8.6 to +11.5 ‰, suggesting the mixing of metamorphic water with magmatic water within the ore-forming fluid. The Sulfur isotope (δ34S) values of gold-bearing granite gneiss range from -1.92 to -0.45 ‰ (mean value of -1.13 ‰) indicating the narrow range of value. This suggests that the sulfides have been precipitated from the fluid system originating from a single source of the magmatic component under sulfur isotopic fractionation equilibrium condition. The tectonic setting of the host rocks, the occurrence of ore bodies, mineral assemblages of the host rocks and proposed ore-forming fluids of the Megele area gold prospects have similarities with features of orogenic gold deposit. The δ18O and δ34S isotopic values also suggested a metamorphic origin with the magmatic components. Thus, the Megele gold prospect could be related to an orogenic gold deposit related to metamorphism and associated intrusions.

Keywords: fluid source, gold mineralization, oxygen isotope, stable isotope, sulfur isotope

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Authors: Moetazza M. Al-Shafei, Hala Abdel Karim, Eitedal M. Daoud, Hassan Zaki Hassuna

Abstract:

Epilepsy is a common neurological disorder affecting all age groups. It is one of the world's most prevalent non-communicable diseases. Increased evidence suggesting that long term usage of anti-epileptic drugs can have adverse effects on bone mineralization and bone molding .Aiming to study these effects and to give guide lines to support bone health through early intervention. From Neurology Out-Patient Clinic kaser Elaini University Hospital, 60 Patients were enrolled, 40 patients on antiepileptic drugs for at least two years and 20 controls matched with age and sex, epileptic but before starting treatment both chosen under specific criteria. Patients were divided into four groups, three groups with monotherapy treated with either Phynetoin, Valporic acid or Carbamazipine and fourth group treated with both Valporic acid and Carbamazipine. Estimation of serum Carboxy-Terminal Telopeptide of Type I- Collagen(ICTP) bone resorption marker, serum 25(OH )vit D3, calcium ,magnesium and phosphorus were done .Results showed that all patients on AED had significant low levels of 25(OH) vit D3 (p<0.001) ,with significant elevation of ICTP (P<0.05) versus controls. In group treated with Phynotoin highly significant elevation of (ICTP) marker and decrease of both serum 25(OH) vit D3 (P<0, 0001) and serum calcium(P<0.05)versus control. Double drug group showed significant decrease of serum 25(OH) vit D3 (P<0.0001) and decrease in Phosphorus (P<0.05) versus controls. Serum magnesium showed no significant differences between studied groups. We concluded that Anti- epileptic drugs appears to be an aggravating factor on bone mineralization ,so therapeutically it can be worth wile to supplement calcium and vitamin D even before initiation of antiepileptic therapy. ICTP marker can be used to evaluate change in bone resorption before and during AED therapy.

Keywords: antiepileptic drugs, bone minerals, carboxy teminal telopeptidetype-1-collagen bone resorption marker, vitamin D

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Authors: Ajit Kumar Behera, Saranya P., Sudhir Kumar, Krishnakumar A

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The stable isotope (δ¹⁸ O and δ²H) composition of groundwater of the coastal areas of Periyar and Mahanadi basins falling along East and West coast of India during North-East (NE) monsoon season have been studied. The east and west coast regions are surrounded by the Bay of Bengal and the Arabian Sea respectively, which are considered to be the primary sources for precipitation over India. The major difference between the Bay of Bengal and the Arabian Sea is that a number of large rivers feed the Bay of Bengal, whereas the Arabian Sea is fed by very few small rivers, resulting in enriched stable isotopic composition of the Arabian Sea than the Bay of Bengal. Previous studies have reported depleted ratios of stable isotopes during Northeast monsoon along East and West coasts due to the influence of the Bay of Bengal moisture source. The isotopic composition of groundwater of the Mahanadi delta in the east coast region varies from -6.87 ‰ to -3.40 ‰ for δ¹⁸ O and -45.42 ‰ to -22.43‰ for δ²H. However, the groundwater of the Periyar basin in the west coast has enriched stable isotope value varying from -4.3‰ to -2.5 ‰ for δ¹⁸ O and for δ²H from -23.7 to -6.4 ‰ which is a characteristic of South-West monsoon season. This suggests the groundwater system of the Mahanadi delta and the Periyar basins are influenced by dissimilar moisture sources. The δ¹⁸ O and δ² H relationship (δ²H= 6.513 δ¹⁸ O - 1.39) and d-excess value (< 10) in the east coast region indicates the influence of NE monsoon implying the quick groundwater recharge after precipitation with significant amount of evaporation. In contrast, the δ¹⁸ O and δ²H regression line (δ²H= 8.408 δ¹⁸ O + 11.71) with high d-excess value (>10) in the west coast region implies delayed recharge due to SW monsoon. The observed isotopic enrichment in west coast suggests that NE winter monsoon rainfall does not replenish groundwater quick enough to produce isotopic depletion during the season.

Keywords: Arabian sea, bay of Bengal, groundwater, monsoon, stable isotope

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Authors: Kira Rysakova, Vitaly Novikov

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An improved method of obtaining enzymatic collagen hydrolysate from the tissues of marine hydrobionts is proposed, which allows to obtain hydrolysate without pre-isolation of pure collagen. The method can be used to isolate enzymatic collagen hydrolysate from the waste of industrial processing of Red King crab and non-traditional objects - marine holothurias. Comparative analysis of collagen hydrolysates has shown the possibility of their use in a number of nutrient media, but this requires additional optimization of their composition and biological tests on wide sets of test strains of microorganisms.

Keywords: collagen hydrolysate, marine hydrobionts, red king crab, marine holothurias, enzymes, exclusive HPLC

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Authors: Jian Peng, Wenhui Xiong, Zheng Lu

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An optimal recipe of amendment (nutrients and electron acceptors) was developed and dominant indigenous benzene-degrading microorganisms were characterized in this study. Lessons were learnt from the development of the optimal amendment recipe: (1) salinity and substantial initial concentration of benzene were detrimental for benzene biodegradation; (2) large dose of amendments can shorten the lag time for benzene biodegradation occurrence; (3) toluene was an essential co-substance for promoting benzene degradation activity. The stable isotope probing study identified incorporation 13C from 13C-benzene into microorganisms, which can be considered as a direct evidence of the occurrence of benzene biodegradation. The dominant mechanism for benzene removal was identified by quantitative polymerase chain reaction analysis to be nitrate reduction. Microbial analyses (denaturing gradient gel electrophoresis and 16S ribosomal RNA) demonstrated that members of genus Dokdonella spp., Pusillimonas spp., and Advenella spp. were predominant within the microbial community and involved in the anaerobic benzene bioremediation.

Keywords: benzene, enhanced anaerobic bioremediation, stable isotope probing, biosep biotrap

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Authors: Ghodratollah Rostami Paydar

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The Baba-Ali and Galali iron deposits are located in northwest Hamedan and the Iranian Sanandaj-Sirjan geological structural zone. The host rocks of these deposits are metavolcanosedimentary successions of Songhor stratigraphic series with permo-trriassic age. Field investigation, ore geometry, textures and structures and paragenetic sequence of minerals, all indicate that the ore minerals are crystallized in four stages: primary volcanosedimentary stage, secondary regional metamorphism with formation of ductile shear zones, contact metamorphism and metasomatism stage and the finally late hydrothermal mineralization within uplift and exposure. Totally 29 samples of sulfide, oxide-silicate and carbonate minerals of iron orees and gangue has been purified for stable isotope analysis. The isotope ratio data assure that occurrence of dynamothermal metamorphism in these areas typically involves a lengthy period of time, which results in a tendency toward isotopic homogenization specifically in O and H stable isotopes and showing the role of metamorphic waters in mineralization process. Measurement of δ34S (CDT) in first generation of pyrite is higher than another ones, so it confirms the volcanogenic origin of primary iron mineralization. δ13C data measurements in Galali carbonate country rocks show a marine origin. δ18O in magnetite and skarn forming silicates, δ18O and δ13C in limestone and skarn calcite and δ34S in sulphides are all consistent with the interaction of a magmatic-equilibrated fluid with Galali limestone, and a dominantly magmatic source for S. All these data imply skarn formation and mineralisation in a magmatic-hydrothermal system that maintained high salinity to relatively late stages resulting in the formation of the regional Na metasomatic alteration halo. Late stage hydrothermal quartz-calcite veinlets are important for gold mineralization, but the economic evaluation is required to detailed geochemical studies.

Keywords: iron, polygenetic, stable isotope, BabaAli, Galali

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Authors: Kasun Gayashan Samarawickrama

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Bone diseases, defects, and fractions are commonly seen in modern life. Since bone is regenerating dynamic living tissue, it will undergo healing process naturally, it cannot recover from major bone injuries, diseases and defects. In order to overcome them, bone grafting technique was introduced. Gold standard was the best method for bone grafting for the past decades. Due to limitations of gold standard, alternative methods have been implemented. Apart from them artificial bone substitutes and bone tissue engineering have become the emerging methods with technology for bone grafting. Many bone diseases and defects will be healed permanently with these promising techniques in future.

Keywords: bone grafting, gold standard, bone substitutes, bone tissue engineering

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Authors: Kang-Hyun Leem, Myung-Gyou Kim, Hye Kyung Kim

Abstract:

Deer antler, traditionally used as a tonic and valuable drug in oriental medicine, has been considered to possess bone-strengthening activity. The upper section, mid section, and base of the antler has been known to exhibit different biological properties. Present study was performed to examine the effects of different parts of deer antler extract (DH) on osteogenic gene expressions in MG-63 cells and longitudinal bone growth in adolescent male rats. The expressions of osteogenic genes, collagen, alkaline phosphatase, osteocalcin, and osteopontin, were measured by quantitative real-time PCR. Longitudinal bone growth was measured in 3-week-old male Sprague-Dawley rats using fluorescence microscopy. To examine the effects on the growth plate metabolism, the total height of growth plate and bone morphogenetic protein-2 (BMP-2) were measured. Collagen and osteocalcin mRNA expressions were increased by all three parts of the DH treatment while osteopontin gene expression was not affected by any of the DH treatment. Alkaline phosphatase gene expression was increased by upper and mid part of DH while base part of DH fails to affect alkaline phosphatase gene expression. The upper and mid parts of the DH treatment enhanced longitudinal bone growth and total height of growth plate. The induction of BMP-2 protein expression in growth plate assessed by immunostaining was also promoted by upper and mid parts of the DH treatment. These results suggest that DH, especially upper and mid parts, stimulate osteogenic gene expressions and have the effect on bone growth in adolescent rats and might be used for the growth delayed adolescent and inherent growth failure patient.

Keywords: bone morphogenetic protein-2, deer antler, longitudinal bone growth, osteogenic genes

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Authors: Y. S. Darmanto, P. H. Riyadi, S. Susanti

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Recently, approximately 9.1 million people of 237.56 million of Indonesian population suffer diabetes. Such condition was caused by high rice consumption of most Indonesian people. It has been known that rice contains low amylose, high calorie, and possesses hyperglycemic properties. Through this study, we tried to solve that problem by creating a super food in order to provide an alternative healthy and balanced diet. We formulated Taro and Seaweed flour based analog rice that fortified by various fishes bone collagens. Corms of Taro contain easily digestible starch and seaweed is rich in fiber, vitamin, and mineral. That mixture was fortified with collagen-containing unique amino acids such as glysine, lysine, alanine, arginine, proline, and hydroxyprolin. Subsequently, super analog rice was characterized about its nutritional composition such are proximate analyses, water, dietary fiber and amylose content. Furthermore, its morphological structure was analyzed by using scanning electron microscopy while the level of consumer preferences was performed by hedonic test. Results demonstrated that fortification by using various fishes bone collagen into analog rice were significantly different in nutritional composition, morphological structure as well as its preferences. Thus, this study was expected as new avenue in functional food discovery especially in the treatment and prevention of diabetic diseases.

Keywords: analogue rice, taro, seaweed, collagen

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