Effect of nitrogen-rich cell culture surfaces on type X collagen expression by bovine growth plate chondrocytes

Background: Recent evidence indicates that osteoarthritis (OA) may be a systemic disease since mesenchymal stem cells (MSCs) from OA patients express type X collagen, a marker of late stage chondrocyte hypertrophy (associated with endochondral ossification). We recently showed that the expression of type X collagen was suppressed when MSCs from OA patients were cultured on nitrogen (N)-rich plasma polymer layers, which we call "PPE:N" (N-doped plasma-polymerized ethylene, containing up to 36 atomic percentage (at.%) of N.Methods: In the present study, we examined the expression of type X collagen in fetal bovine growth plate chondrocytes (containing hypertrophic chondrocytes) cultured on PPE:N. We also studied the effect of PPE: N on the expression of matrix molecules such as type II collagen and aggrecan, as well as on proteases (matrix metalloproteinase-13 (MMP-13) and molecules implicated in cell division (cyclin B2). Two other culture surfaces, "hydrophilic" polystyrene (PS, regular culture dishes) and nitrogen-containing cation polystyrene (Primaria (R)), were also investigated for comparison.Results: Results showed that type X collagen mRNA levels were suppressed when cultured for 4 days on PPE: N, suggesting that type X collagen is regulated similarly in hypertrophic chondrocytes and in human MSCs from OA patients. However, the levels of type X collagen mRNA almost returned to control value after 20 days in culture on these surfaces. Culture on the various surfaces had no significant effects on type II collagen, aggrecan, MMP-13, and cyclin B2 mRNA levels.Conclusion: Hypertrophy is diminished by culturing growth plate chondrocytes on nitrogen-rich surfaces, a mechanism that is beneficial for MSC chondrogenesis. Furthermore, one major advantage of such "intelligent surfaces" over recombinant growth factors for tissue engineering and cartilage repair is potentially large cost-saving

The impacts of discriminatory experiences on lesbian, gay and bisexual people in sport

This study examines the nature and impact of sexist and homophobic discrimination experienced by lesbians, gays and bisexuals (LGB) in Australian sporting settings. A mixed methods online survey was utilized to collate participant experiences. The findings suggest that, in sport, participants experienced sexism directly and systemically, and homophobia explicitly and implicitly. Women experienced sexism and homophobia, whilst men reported more homophobic events. The most mentioned impacts of discrimination were negative emotions such as sadness, anger, distress and shame, followed by negative engagement with sport such as disliking sport, or avoiding or leaving sport. The well-recognized benefits of sport such as physical and mental well-being, social connections, enjoyment, positive identity and achievement may be more difficult to realize within this context of significant social stress

Quantitative magnetic resonance imaging of enzymatically induced degradation of the nucleus pulposus of intervertebral discs

STUDY DESIGN: The structural integrity of the nucleus pulposus (NP) of intervertebral discs was targeted by enzyme-specific degradations to correlate their effects to the magnetic resonance (MR) signal. OBJECTIVE: To develop quantitative MR imaging as an accurate and noninvasive diagnostic tool to better understand and treat disc degeneration. SUMMARY OF BACKGROUND DATA: Quantitative MR analysis has been previously shown to reflect not only the disc matrix composition, but also the structural integrity of the disc matrix. Further work is required to identify the contribution of the structural integrity versus the matrix composition to the MR signal. METHODS: The bovine coccygeal NPs were injected with either enzyme or buffer, incubated at 37 degrees C as static, unloaded and closed 3-disc segments, and analyzed by a 1.5-Tesla MR scanner to measure MR parameters. RESULTS: Collagenase degradation of the NP significantly decreased the relaxation times, slightly decreased the magnetization transfer ratio, and slightly increased the apparent diffusion coefficient. Targeting the proteoglycan and/or hyaluronan integrity by trypsin and hyaluronidase did not significantly affect the MR parameters, except for an increase in the apparent diffusion coefficient of the disc after trypsin treatment. CONCLUSIONS: Our results demonstrate that changes in the structural integrity of matrix proteins can be assessed by quantitative MR

Effect of nitrogen-rich cell culture surfaces on type X collagen expression by bovine growth plate chondrocytes

Abstract Background Recent evidence indicates that osteoarthritis (OA) may be a systemic disease since mesenchymal stem cells (MSCs) from OA patients express type X collagen, a marker of late stage chondrocyte hypertrophy (associated with endochondral ossification). We recently showed that the expression of type X collagen was suppressed when MSCs from OA patients were cultured on nitrogen (N)-rich plasma polymer layers, which we call "PPE:N" (N-doped plasma-polymerized ethylene, containing up to 36 atomic percentage (at.% ) of N. Methods In the present study, we examined the expression of type X collagen in fetal bovine growth plate chondrocytes (containing hypertrophic chondrocytes) cultured on PPE:N. We also studied the effect of PPE:N on the expression of matrix molecules such as type II collagen and aggrecan, as well as on proteases (matrix metalloproteinase-13 (MMP-13) and molecules implicated in cell division (cyclin B2). Two other culture surfaces, "hydrophilic" polystyrene (PS, regular culture dishes) and nitrogen-containing cation polystyrene (Primaria®), were also investigated for comparison. Results Results showed that type X collagen mRNA levels were suppressed when cultured for 4 days on PPE:N, suggesting that type X collagen is regulated similarly in hypertrophic chondrocytes and in human MSCs from OA patients. However, the levels of type X collagen mRNA almost returned to control value after 20 days in culture on these surfaces. Culture on the various surfaces had no significant effects on type II collagen, aggrecan, MMP-13, and cyclin B2 mRNA levels. Conclusion Hypertrophy is diminished by culturing growth plate chondrocytes on nitrogen-rich surfaces, a mechanism that is beneficial for MSC chondrogenesis. Furthermore, one major advantage of such "intelligent surfaces" over recombinant growth factors for tissue engineering and cartilage repair is potentially large cost-saving.</p

Possíveis efeitos adversos dos campos eletromagnéticos (50/60 Hz) em humanos e em animais Potential adverse effects of electromagnetic fields (50/60 Hz) on humans and animals

Os avanços tecnológicos têm aumentado o número de equipamentos elétricos e eletrônicos, seja nas residências ou mesmo no ambiente de trabalho, fazendo com que a população conviva com grande número de fontes de irradiação eletromagnética, com os mais diversos níveis de potência e freqüência. Por muitos anos, alguns cientistas e engenheiros acreditaram que o campo eletromagnético (CEM) com freqüência extremamente baixa não pudesse causar efeitos e alterações significantes no material biológico. O objetivo deste trabalho é verificar os possíveis efeitos adversos dos CEMs em humanos e animais, que foram publicados nos últimos anos, através de uma revisão da literatura disponível em Medline, revistas nacionais e internacionais e catálogos de obras de referência na área dos CEM (50/60 Hz). Como resultado foi observado que o CEM (50/60 Hz) é capaz de produzir diversos efeitos adversos em humanos e animais, como por exemplo: distúrbios na reprodução, doenças degenerativas, efeitos psiquiátricos e psicológicos, alterações citogenéticas, alterações no sistema cardiovascular, nervoso e neuroendócrino, bem como nos parâmetros biológicos e bioquímicos. Apesar de todas estas constatações e devido a muitas controvérsias entre vários autores, faz-se necessário um estudo mais específico e aprofundado sobre o assunto.<br>The technologic development has increased the number of electric and electronic devices for household and work environment applications. In this way, we have to cope with a diverse quantity of electromagnetic irradiation sources, with different power and frequency ranges. For many years, some scientists and engineers believed that low-frequencies electromagnetic field (EMF) could not cause any bad effect or substantial alterations on the biologic livings. This work has the objective to perform a literature review of the possible effects of EMF in human beings and animals, that was published in the past years on Medline, international, and national journals about the EMF (50/60Hz). The results showed that extremely low EMF might produce adverse effects, i.e. cancer, reproduction disruption, degenerative illnesses, citogenetic alterations, and cardiovascular, neurologic and neuroendocrine system alterations in humans and animals. The biologic and biochemical parameters suffered interference as well. Despite all these findings, we can find some disagreements among the authors. Hence it is necessary to extend the research about this issue

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field