Evaluation and Validation of Two Chromatographic Methods (HPLC-Fluorescence and LC–MS/MS) for the Determination and Confirmation of Ochratoxin A in Pig Tissues

Milićević, Dragan; Jurić, Verica; Stefanović, Sofija; Baltić, Tatjana; Janković, Saša

doi:10.1007/s00244-009-9436-2

Cited by 15 publications

(7 citation statements)

References 21 publications

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“…It is worth noting that these prior studies used an HPLC fluorescence detection assay for pyridinoline quantification as opposed to the LC-MS technique used in this paper. Because LC-MS methods have repeatedly been shown to be more precise and accurate than HPLC fluorescence detection methods [54][55][56] , future characterization studies on native fibrocartilage crosslinks using LC-MS would yield more comparable data to those collected by our study. Overall, TCL + LPA treated neomenisci exhibited crosslink content that appears to meet or exceed values reported in the literature, though characterization of native tissue via LC-MS methods is warranted.…”

Section: Discussionmentioning

confidence: 70%

Engineering self-assembled neomenisci through combination of matrix augmentation and directional remodeling

Gonzalez-Leon

Bielajew

et al. 2020

Acta Biomaterialia

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Knee meniscus injury is frequent, resulting in over 1 million surgeries annually in the United States and Europe. Because of the near-avascularity of this fibrocartilaginous tissue and its intrinsic lack of healing, tissue engineering has been proposed as a solution for meniscus repair and replacement. This study describes an approach employing bioactive stimuli to enhance both extracellular matrix content and organization of neomenisci toward augmenting their mechanical properties. Self-assembled fibrocartilages were treated with TGF-β1, chondroitinase ABC, and lysyl oxidase-like 2 (collectively termed TCL) in addition to lysophosphatidic acid (LPA). TCL + LPA treatment synergistically improved circumferential tensile stiffness and strength, significantly enhanced collagen and pyridinoline crosslink content per dry weight, and achieved tensile anisotropy (circumferential/radial) values of neomenisci close to 4. This study utilizes a combination of bioactive stimuli for use in tissue engineering studies, providing a promising path toward deploying these neomenisci as functional repair and replacement tissues. Statement of SignificanceThis study utilizes a scaffold-free approach, which strays from the tissue engineering paradigm of using scaffolds with cells and bioactive factors to engineer neotissue. While self-assembled neomenisci have attained compressive properties akin to native tissue, tensile properties still require improvement before being able to deploy engineered neomenisci as functional tissue repair or replacement options. In order to augment tensile properties, this study utilized bioactive factors known to augment matrix content in combination with a soluble factor that enhances matrix organization and anisotropy via cell traction forces. Using a bioactive factor to enhance matrix organization mitigates the need for bioreactors used to apply mechanical stimuli or scaffolds to induce proper fiber alignment.

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Section: Discussionmentioning

confidence: 70%

Engineering self-assembled neomenisci through combination of matrix augmentation and directional remodeling

Gonzalez-Leon

Bielajew

et al. 2020

Acta Biomaterialia

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“…Pyridinoline crosslink content normalized to dry weight trended highest in the central region of the medial minipig meniscus and was measured at approximately 0.16%, which is higher than levels obtained in human menisci using an HPLC fluorescence detection assay at 0.12% ( Takahashi et al, 1998 ). It should be noted that values in the present study were obtained using an LC-MS method, which has been shown to be more precise and accurate than HPLC fluorescence detection methods ( Milićević et al, 2010 ; Wang et al, 2012 ; Bandeira et al, 2013 ; Bielajew et al, 2020 , 2021 ). Pyridinoline crosslink content, for example, has been quantified using LC-MS techniques in bovine articular cartilage, showing crosslink levels of 0.12% of total dry weight, which were on par with values recorded in this study ( Espinosa et al, 2021a ).…”

Section: Discussionmentioning

confidence: 80%

Yucatan Minipig Knee Meniscus Regional Biomechanics and Biochemical Structure Support its Suitability as a Large Animal Model for Translational Research

Gonzalez-Leon

Athanasiou

2022

Front. Bioeng. Biotechnol.

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Knee meniscus injuries are the most frequent causes of orthopedic surgical procedures in the U.S., motivating tissue engineering attempts and the need for suitable animal models. Despite extensive use in cardiovascular research and the existence of characterization data for the menisci of farm pigs, the farm pig may not be a desirable preclinical model for the meniscus due to rapid weight gain. Minipigs are conducive to in vivo experiments due to their slower growth rate than farm pigs and similarity in weight to humans. However, characterization of minipig knee menisci is lacking. The objective of this study was to extensively characterize structural and functional properties within different regions of both medial and lateral Yucatan minipig knee menisci to inform this model’s suitability as a preclinical model for meniscal therapies. Menisci measured 23.2–24.8 mm in anteroposterior length (33–40 mm for human), 7.7–11.4 mm in width (8.3–14.8 mm for human), and 6.4–8.4 mm in peripheral height (5–7 mm for human). Per wet weight, biochemical evaluation revealed 23.9–31.3% collagen (COL; 22% for human) and 1.20–2.57% glycosaminoglycans (GAG; 0.8% for human). Also, per dry weight, pyridinoline crosslinks (PYR) were 0.12–0.16% (0.12% for human) and, when normalized to collagen content, reached as high as 1.45–1.96 ng/µg. Biomechanical testing revealed circumferential Young’s modulus of 78.4–116.2 MPa (100–300 MPa for human), circumferential ultimate tensile strength (UTS) of 18.2–25.9 MPa (12–18 MPa for human), radial Young’s modulus of 2.5–10.9 MPa (10–30 MPa for human), radial UTS of 2.5–4.2 MPa (1–4 MPa for human), aggregate modulus of 157–287 kPa (100–150 kPa for human), and shear modulus of 91–147 kPa (120 kPa for human). Anisotropy indices ranged from 11.2–49.4 and 6.3–11.2 for tensile stiffness and strength (approximately 10 for human), respectively. Regional differences in mechanical and biochemical properties within the minipig medial meniscus were observed; specifically, GAG, PYR, PYR/COL, radial stiffness, and Young’s modulus anisotropy varied by region. The posterior region of the medial meniscus exhibited the lowest radial stiffness, which is also seen in humans and corresponds to the most prevalent location for meniscal lesions. Overall, similarities between minipig and human menisci support the use of minipigs for meniscus translational research.

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“…Other solvents have been used in recent years for mycotoxin LLE from organs and tissues. For instance, OTA was extracted using CH 2 Cl 2 [ 120 ] and by a solvent mixture of ice-cold absolute ethanol/trichloroacetic acid [ 121 ] from pig and rat samples; plasma, liver, and kidney, respectively.…”

Section: Resultsmentioning

confidence: 99%

Studies on the Presence of Mycotoxins in Biological Samples: An Overview

Escrivá¹,

Font²,

Manyes³

et al. 2017

Toxins

109

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Mycotoxins are fungal secondary metabolites with bioaccumulation levels leading to their carry-over into animal fluids, organs, and tissues. As a consequence, mycotoxin determination in biological samples from humans and animals has been reported worldwide. Since most mycotoxins show toxic effects at low concentrations and considering the extremely low levels present in biological samples, the application of reliable detection methods is required. This review summarizes the information regarding the studies involving mycotoxin determination in biological samples over the last 10 years. Relevant data on extraction methodology, detection techniques, sample size, limits of detection, and quantitation are presented herein. Briefly, liquid-liquid extraction followed by LC-MS/MS determination was the most common technique. The most analyzed mycotoxin was ochratoxin A, followed by zearalenone and deoxynivalenol—including their metabolites, enniatins, fumonisins, aflatoxins, T-2 and HT-2 toxins. Moreover, the studies were classified by their purpose, mainly focused on the development of analytical methodologies, mycotoxin biomonitoring, and exposure assessment. The study of tissue distribution, bioaccumulation, carry-over, persistence and transference of mycotoxins, as well as, toxicokinetics and ADME (absorption, distribution, metabolism and excretion) were other proposed goals for biological sample analysis. Finally, an overview of risk assessment was discussed.

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Evaluation and Validation of Two Chromatographic Methods (HPLC-Fluorescence and LC–MS/MS) for the Determination and Confirmation of Ochratoxin A in Pig Tissues

Cited by 15 publications

References 21 publications

Engineering self-assembled neomenisci through combination of matrix augmentation and directional remodeling

Engineering self-assembled neomenisci through combination of matrix augmentation and directional remodeling

Yucatan Minipig Knee Meniscus Regional Biomechanics and Biochemical Structure Support its Suitability as a Large Animal Model for Translational Research

Studies on the Presence of Mycotoxins in Biological Samples: An Overview

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