RESUMEN
Chondrocytes are the only resident cell types that form the extracellular matrix of cartilage. Inflammation alters the anabolic and catabolic regulation of chondrocytes, resulting in the progression of osteoarthritis (OA). The potential of TMMG, a glucuronated flavone, was explored against the pathophysiology of OA in both in vitro and in vivo models. The effects of TMMG were evaluated on chondrocytes and the ATDC5 cell line treated with IL-1ß in an established in vitro inflammatory OA model. An anterior cruciate ligament transection (ACLT) model was used to simulate post-traumatic injury in vivo. Micro-CT and histological examination were employed to examine the micro-architectural status and cartilage alteration. Further, serum biomarkers were measured using ELISA to assess OA progression. In-vitro, TMMG reduced excessive ROS generation and inhibited pro-inflammatory IL-1ß secretion by mouse chondrocytes and macrophages, which contributes to OA progression. This expression pattern closely mirrored osteoclastogenesis prevention. In-vivo results show that TMMG prevented chondrocyte apoptosis and degradation of articular cartilage thickness, subchondral parameters, and elevated serum COMP, CTX-II, and IL-1ß which were significantly restored in 5 and 10 mg.kg-1day-1 treated animals and comparable to the positive control Indomethacin. In addition, TMMG also improved cartilage integrity and decreased the OARSI score by maintaining chondrocyte numbers and delaying ECM degradation. These findings suggest that TMMG may be a prospective disease-modifying agent that can mitigate OA progression.
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Cartílago Articular , Flavonas , Osteoartritis , Ratones , Animales , Condrocitos/metabolismo , Estudios Prospectivos , Osteoartritis/metabolismo , Cartílago Articular/metabolismo , Interleucina-1beta/metabolismo , Flavonas/farmacología , Flavonas/uso terapéutico , Células CultivadasRESUMEN
SARS coronavirus2 is the primary cause of new Coronavirus illness (COVID-19) (SARS- COV-2). Today, COVID-19 is a global epidemic. Coronavirus illness may be diagnosed using a variety of approaches. The gold standard is RT-PCR, which is used in all of these assays. Swabs from the nose, pharynx, or mouth are the most often used sampling methods for coronavirus detection. For COVID-19 testing, saliva may be utilized as an alternate sample. When compared to a nasopharyngeal swab, saliva samples have a number of advantages and disadvantages. Saliva has also been reviewed as a non-invasive diagnostic tool for the detection of COVID-19. The affordability of the salivary diagnostic process makes it an effective process for detecting the COVID-19 viruses. The researchers have found that salivary diagnostic processes have greater chances of success than other processes of Coronavirus detection. However, healthcare professionals need to make positive changes to their working processes to ensure the sustainability of the salivary diagnosis processes.
RESUMEN
In our previous study, a novel BMP2 secretagogue was synthesized belonging to a class of galloyl conjugates of flavanones, with remarkable osteogenic potential that promoted bone regeneration. We aimed to establish the protective effect of our compound against bone loss that co-exists with excess Glucocorticoid (GC) therapy. GC therapy induces osteoblast damage leading to apoptosis by increasing reactive oxygen species (ROS). Our results delineate that compound 5e (a BMP2 secretagogue) activates NRF2 signalling to counter the disturbed cellular redox homeostasis and escalate osteoblast survival as assessed by Western blot and immunocytochemistry. Depletion of NRF2 by siRNA blocked activation of the NRF2/HO-1 pathway, magnified oxidative stress, increased apoptosis and abrogated the protective effects of compound 5e. 5e, on the other hand, increased ALP, mineralization activity, and promoted osteoblast differentiation by activating WNT/ß-catenin signalling in BMP2 dependent manner, validated by Western blot of WNT3A, SOST, GSK3-ß and ß-catenin nuclear translocation. Treatment of 5e in presence of BMP inhibitor noggin attenuated the osteogenic efficacy and minimized Wnt//ß-catenin signalling in presence of dexamethasone. Our compound prevents GC challenged trabecular and cortical bone loss assessed by micro-CT and promotes bone formation and osteocyte survival determined by calcein labelling and TUNEL assay in GC treated animals. The osteogenic potential of the compound was authenticated by bone turnover markers. On a concluding note, compounds with BMP upregulation can be potential therapeutics for the prevention and treatment of glucocorticoid-induced osteoporosis.
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Osteogénesis , beta Catenina , Animales , Diferenciación Celular , Glucocorticoides/farmacología , Glucógeno Sintasa Quinasa 3/metabolismo , Glucógeno Sintasa Quinasa 3/farmacología , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , Osteoblastos , Osteogénesis/genética , Estrés Oxidativo , Secretagogos/metabolismo , Secretagogos/farmacología , Vía de Señalización Wnt , beta Catenina/genética , beta Catenina/metabolismoRESUMEN
AIMS: Postmenopausal osteoporosis is one of the world's biggest yet unnoticed health issues. After ovariectomy, declined estrogen level significantly contributes to the elevation of bone marrow adiposity and bone loss leading to osteoporosis. Therapeutics to prevent osteoporosis addressing various aspects are now in short supply. In this study we made an approach to synthesize nanoparticles of naturally occurring PPAR-γ inhibitor, betulinic acid (BA/NPs) and tested the same in altered bone metabolisms developed after ovariectomy. MAIN METHODS: The osteogenic efficacy of BA/NPs was established in human and rat primary osteoblast cells using qRT-PCR and immunoblot analysis. Furthermore, lineage allocations of multipotent bone marrow stromal cells were evaluated. Various aspects of altered bone metabolism after ovariectomy such as bone marrow adiposity and pathological bone loss were evaluated using µCT and histological assessments. KEY FINDINGS: BA/NPs exert potential osteogenic efficacy by modulating key osteogenic markers such as RUNX2 and BMP2. Mechanistically BA/NPs regulate osteoblastogenesis through Wnt/ß-catenin signaling. Further, BA/NPs showed the potential to inhibit the differentiation of multipotent BMSCs towards adipogenesis while favouring the osteogenic lineage via Wnt/ß-catenin pathway. In the in vivo study, increased bone marrow adiposity was reduced in ovariectomized rats after BA/NPs treatment as assessed by histology and µCT analysis. Loss of bone mineral density as a hallmark of pathological bone loss was also abrogated by BA/NPs. Furthermore, increased obesity after OVX was also prevented in BA/NPs treated animals. SIGNIFICANCE: Our findings imply that BA/NPs could be used further as a viable drug lead to counteract various pathophysiological challenges after menopause.
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Nanopartículas , Osteoporosis , Femenino , Ratas , Humanos , Animales , beta Catenina/metabolismo , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Médula Ósea/metabolismo , Adiposidad , PPAR gamma/metabolismo , Vía de Señalización Wnt , Osteogénesis , Diferenciación Celular , Osteoporosis/tratamiento farmacológico , Osteoporosis/etiología , Osteoporosis/metabolismo , Ovariectomía , Estrógenos/uso terapéutico , Obesidad , Ácido BetulínicoRESUMEN
In search of novel osteogenic entities, a series of twenty-seven quinazolinone-benzopyran-indole hybrids were designed and synthesized using molecular hybridization approach. All the compounds were scrutinized for their osteogenic potential, primarily based on alkaline phosphatase assay as one of the major anabolic markers. From the primary screening, four osteogenic compounds were sorted from the series and were found nontoxic to the osteoblasts. Further, increased osteoblast differentiation and osteogenic mRNA upregulations suggest compound 47 as the most potent osteoanabolic agent. Immunoblot and ELISA analysis demonstrated that compound 47 promotes osteogenesis via RUNX2 and BMP2 mediated non-canonical p38 pathway. In vivo studies in BALB/c mice inferred that compound 47 stimulates bone anabolism as evident from histological and gene expression studies at 5 mg. kg-1. day-1 dose. Furthermore, structural activity relationship (SAR) and pharmacokinetic studies suggest compound 47 as a BMP2 upregulator and a potential bone anabolic lead for combating future bone metabolic disorders.
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Benzopiranos , Osteogénesis , Ratones , Animales , Regulación hacia Arriba , Benzopiranos/metabolismo , Quinazolinonas/farmacología , Quinazolinonas/metabolismo , Proteína Morfogenética Ósea 2/metabolismo , Osteoblastos/metabolismo , Indoles/metabolismo , Diferenciación CelularRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Dalbergia sissoo DC. (Indian rosewood or Sheesham) is a traditional medicinal plant, reported since time immemorial for its analgesic, anti-nociceptive, anti-inflammatory, and immuno-modulatory properties. D. sissoo DC (DS). is being used traditionally to cure joint inflammation and joint pain. AIM: To study the potential of DS leaves and its derived novel compound CAFG to treat the clinical symptoms of osteoarthritis (OA) and its underlying mechanism. METHODS: The chemical profile of DS extract (DSE) with isoflavonoids and isoflvaonoid glycosides from the DS was established by UHPLC-PDA and UHPLC-MS/MS. Monosodium iodoacetate (MIA) was injected into the knee joint to develop the OA model in rats. DSE was given orally for 28 days daily at 250 and 500 mg.kg-1day-1. For in-vitro experiments, chondrocytes isolated from joint articular cartilage were negatively induced with interleukin-1ß (IL-1ß) and CAFG was given to the cells as a co-treatment. RESULTS: Chondrocytes undergo apoptosis following inflammation and proteoglycan synthesis affected in MIA injected knees. DSE administration prevented these effects as assessed by H&E and Toluidine blue staining. Micro-CT analysis showed that subchondral bone loss was restored. DSE decreased elevated serum levels of cartilage-bone degradation (CTX-I, CTX-II, and COMP), inflammation markers IL-1ß, and matrix-degrading MMP-3 and 13. The effects of IL-1ß on gene expression of chondrocytes were reversed by CAFG treatment at 1 µM. CONCLUSION: Data showed that DSE protected joint cartilage and deterioration in subchondral bone in vivo while in in-vitro, its active ingredient CAFG prevented interleukin-1ß induced effects and inhibited OA. This finding suggest that DSE and CAFG could be used as a possible therapeutic to treat osteoarthritis.
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Artralgia/tratamiento farmacológico , Dalbergia , Glicósidos/farmacología , Isoflavonas/farmacología , Osteoartritis/tratamiento farmacológico , Administración Oral , Analgésicos/farmacología , Animales , Antiinflamatorios/farmacología , Cartílago Articular/efectos de los fármacos , Células Cultivadas , Condrocitos/efectos de los fármacos , Modelos Animales de Enfermedad , Flavonoides/farmacología , Fitoterapia/métodos , Extractos Vegetales/farmacología , Ratas , Resultado del TratamientoRESUMEN
Obesity and ageing increases bone marrow fat which in turn is associated with lower bone mass. Marrow adipocytes by secreting cytokines, adipokines and free fatty acids change the bone marrow milieu and thus the number of osteoblasts. Palmitate is the common saturated fatty acid, an unavoidable ingredient we consume with food, which kindles cell apoptosis. Compound 4e is osteogenic in nature. We examine the effect of compound 4e in palmitate induced lipotoxicity in rat osteoblasts. Design of benzofuran Pyran hybrid compound (4e) was found to be effective in inhibiting palmitate induced cell apoptosis. In this study an in vitro model of palmitate was contrived. Anti-apoptotic effect of compound 4e was assessed by Annexin/PI and LDH (Lactate dehydrogenase) assay. Compound 4e also increased osteoblast differentiation and mineralization. It also increased expression of osteogenic markers (RUNX2 and BMP2), assessed by Real time PCR and immunofluorescence, which was impeded by palmitate. Acetyl Co-Carboxylase (ACC) and Fatty acid synthase (FAS), two prominent mediators of lipid biosynthesis were increased by palmitate exposure. Compound 4e modulated lipid biosynthesis by inhibiting ACC and FAS as reflected visually and after quantification of less lipid droplet formation suggesting that 4e is osteogenic and simultaneously anti-lipotoxic.
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Benzofuranos/farmacología , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Lipogénesis/efectos de los fármacos , Osteoblastos/efectos de los fármacos , Palmitatos/toxicidad , Piranos/farmacología , Fosfatasa Alcalina/metabolismo , Animales , Apoptosis/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Humanos , Osteoblastos/metabolismo , RatasRESUMEN
Osteoarthritis (OA) is an aging disorder characterized by degenerated cartilage and sub-chondral bone alteration in affected knee joints. Globally, millions of people suffer from this disease. However, there is a lack of safe and promising therapeutics, making the exploration and development of leads from natural sources urgent. Accordingly, food as medicine may be the most suitable approach for the treatment of this degenerative disease. Herein, we elucidated the protective role of Spinacia oleracea extract (SOE) in an anterior cruciate ligament transection (ACLT) model of osteoarthritis as a mimic of the human condition. ACL transection was done in the tibio-femoral joints of rats. SOE was orally administered at the dosage of 125 and 250 mg kg-1 day-1 for four weeks. It was shown that the animals with SOE treatment had better joint morphology than the ACLT animals, as evident by the shiny appearance of their cartilage. Hematoxylin and safranin-o staining showed that the number of chondrocytes was significantly reduced in the OA model, which was prevented with SOE treatment. The reduction in the cartilage thickness was well observed by toluidine blue staining. The reduced stain by safranin-o and toluidine blue, indicated proteoglycan loss in the ACLT-induced osteoarthritis model. The proteoglycan content and cartilage thickness were restored in the SOE group upon treatment at an SOE dosage of 125 and 250 mg kg-1 day-1. The micro-CT parameters of subchondral bone (SCB) and cartilage degradation markers in the serum corroborated our findings of the protective effects of SOE. In summary, our study suggests that SOE has therapeutic potential, which if taken regularly as a food supplement, can have beneficial effects.
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Ligamento Cruzado Anterior/cirugía , Osteoartritis/tratamiento farmacológico , Extractos Vegetales/administración & dosificación , Spinacia oleracea/química , Animales , Huesos/metabolismo , Huesos/fisiopatología , Cartílago Articular/crecimiento & desarrollo , Cartílago Articular/fisiopatología , Modelos Animales de Enfermedad , Femenino , Humanos , Articulación de la Rodilla/metabolismo , Articulación de la Rodilla/fisiopatología , Osteoartritis/metabolismo , Osteoartritis/fisiopatología , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND AND AIMS: Transversus abdominis plane (TAP) block is a popular technique for post-operative analgesia in abdominal surgeries. The aim of the study was to evaluate the relative efficacy of bupivacaine versus ropivacaine for post-operative analgesia using ultrasound-guided TAP block in laparoscopic cholecystectomies. METHODS: Sixty adults undergoing elective laparoscopic cholecystectomy were randomised to receive ultrasound-guided TAP block at the end of the surgical procedure with either 0.25% bupivacaine (Group I, n = 30) or 0.375% ropivacaine (Group II, n = 30). All patients were assessed for post-operative pain and rescue analgesic consumption at 10 min, 30 min, 1 h, 4 h, 8 h, 12 h and 24 h time points. Means for normally distributed data were compared using Student's t-test, and proportions were compared using Chi-square or Fisher's exact test whichever was applicable. RESULTS: Patients receiving ultrasound-guided TAP block with ropivacaine (Group II) had significantly lower pain scores when compared to patients who received the block with bupivacaine (Group I) at 10 min, 30 min and 1 h. However, both the drugs were equivalent for post-operative analgesia and 24 h cumulative rescue analgesic requirement (median [interquartile range]) (75.00 [75.00-75.00] in Group I vs. 75.00 [75.00-93.75] in Group II, P = 0.366). CONCLUSION: Ultrasound-guided TAP block with ropivacaine provides effective analgesia in the immediate post-operative period up to 1 h as compared to bupivacaine. However, both the drugs are similar in terms of 24 h cumulative rescue analgesic requirement.
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Tejido Adiposo/patología , Páncreas/patología , Enfermedades Pancreáticas/patología , Tejido Adiposo/diagnóstico por imagen , Femenino , Humanos , Persona de Mediana Edad , Páncreas/diagnóstico por imagen , Enfermedades Pancreáticas/diagnóstico por imagen , Tomografía Computarizada por Rayos XRESUMEN
Idiopathic spinal cord herniation (ISCH) is a rare cause of progressive myelopathy. This condition has recently seen an increased frequency of diagnosis, possibly due to increased awareness and the use of magnetic resonance (MR) imaging. ISCH is characterised by herniation of the thoracic spinal cord through an anterior or anterolateral dural defect. Patients usually present with a Brown-Séquard-like syndrome, which is gradually progressive and may evolve into severe paraparesis. This disease has a characteristic radiological appearance, and in most cases, excellent postsurgical outcome. We report ISCH and its imaging appearance in a 31-year-old woman with classical presentation, and discuss the current concepts regarding the aetiopathogenesis, radiological features and management of the disease.