249 research outputs found
TMJ Disorders: Future Innovations in Diagnostics and Therapeutics
Because their etiologies and pathogenesis are poorly understood, temporomandibular joint (TMJ) diseases are difficult to diagnose and manage. All current approaches to treatments of TMJ diseases are largely palliative. Definitive and rational diagnoses or treatments can only be achieved through a comprehensive understanding of the etiologies, predisposing factors, and pathogenesis of TMJ diseases. While much work remains to be done in this field, novel findings in biomedicine and developments in imaging and computer technologies are beginning to provide us with a vision of future innovations in the diagnostics and therapeutics of TMJ disorders. These advances include the identification and use of local or systemic biomarkers to diagnose disease or monitor improvements in therapy; the use of imaging technologies for earlier and more sensitive diagnostics; and the use of biomedicine, biomimetics, and imaging to design and manufacture bioengineered joints. Such advances are likely to help to customize and enhance the quality of care we provide to patients with TMJ disorders.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/153727/1/jddj002203372008728tb04569x.pd
Identification of Tectorigenin as a natural pro-hypoxia compound: implications in modulation of cellular differentiation and senescence
Background: Hypoxia, a suboptimal level of oxygen, evokes stress response in cells and activated hypoxia signaling has been largely established as a pro-metastasis and pro-angiogenic factor for tumor cells. On the other hand, age-related neurodegenerative disorders are characterized by hypoxic environment, accumulation of molecular garbage and induction of premature senescence. Several recent studies have reported anti-stress impact of the intermittent induction of hypoxia signaling in these cells.
Methods: Screening of a phytochemical library using Hypoxia Responsive Element (HRE) driven luciferase as a reporter was carried out to identify hypoxia-modulating phytochemicals. Activation of HIF-1a (master regulator of hypoxia signaling) was validated by Western Blotting and immunostaining using specific antibodies. Short-term and long-term effect of the selected compounds on cell viability were determined by cell viability and colony forming assays, respectively. Furthermore, in vitro wound-scratch assays, protein aggregation models, and replicative senescence models were recruited to determine the effect of the selected compound on these phenotypes.
Results: Tectorigenin (TEC) (iso-flavone obtained from leopard lily or Iris domestica) was selected as a pro-hypoxia factor. TEC treated cells showed significant activation HRE-driven luciferase reporter and upregulation of endogenous HIF-1a. On these lines, it was found that TEC resulted in de-aggregation of induced aggregation of protein reporters. cDNA microarray data revealed that TEC modulated the expression of genes involved in cell migration and differentiation. We used cellular senescence and astrocytic differentiation models and found lifespan extension of normal human fibroblasts and differentiation of rat-glioma cells, respectively.
Conclusions: TEC could be defined as an anti-stress and anti-aging phytochemical that could be useful to manage hypoxia-driven ailments, involving protein aggregation and neurodegeneration. Further studies are warranted to support these claims and to dissect their molecular mechanism(s) of action
Cell-based screening of antistress activity of some phytochemicals: Identification, validation, and relevance to old-age related pathologies
Background: A variety of environmental stresses have been shown to contribute to poor quality of life, tissue dysfunctions and ailments including metabolic disorders, cognitive impairment, and accelerated aging. Oxidative stress (an imbalance between the production and processing of highly reactive oxygen species) is largely associated with these phenotypes. Whereas drug development and disease therapeutics have advanced remarkably in last three decades, there are still limited options for stress management. Since the later can effectively decrease the disease burden, we aimed to screen phytochemicals with anti-oxidative stress activity using cell-based assays.
Methods: Brain-derived cells were subjected to chemical models of oxidative (paraquat), metal (cadmium nitrate) or hypoxia (cobalt chloride) stresses. Stressed cells were allowed to recover either in the control or phytochemical supplemented culture medium. Cell survival and protein expression/signaling were analyzed to select the useful compounds and/or plant extracts.
Results: Cells subjected to paraquat stress showed decrease in their viability. Three rounds of blind screening of the 24 phytochemicals resulted in identification of 5 compounds that caused better recovery of cells. The selected compounds were examined for their ability for protection against metal and hypoxia stresses induced by cadmium nitrate and cobalt chloride, respectively. Based on these 3D-anti stress protection ability, Withanone (Wi-N) and triethylene glycol (TEG) were selected for molecular validation. We found that whereas stress caused increase in (i) apoptosis (ii) ROS accumulation coupled with mitochondrial depolarization (iii) DNA double-strand break (iiii) protein aggregation, the selected compounds and the Ashwagandha extracts (known to possess these compounds) caused remarkable protection. Furthermore, both Wi-N and TEG caused differentiation of C6 glioblastoma and IMR-32 neuroblastoma as evidenced by the respective differentiated cell morphology and increased expression of biomarkers.
Conclusion: The results suggested that Wi-N, TEG, their mixture, and the natural resource (Ashwagandha) possess potent antistress activity that may be useful for management of old-age-related ailments
Folate receptor mediated targeting enhances selective cytotoxicity of Ashwagandha derived drugs to cancer cells
Background: Folate receptors (FRs) have been shown to be overexpressed on the surface of a variety of cancer cells and their expression are limited in normal cells and tissues. Since FR strongly binds to folic acid (FA), FA-functionalized nanocarriers have been proposed as a reliable strategy for delivery of anticancer drugs. We have earlier reported that the alcoholic extract of Ashwagandha leaves (i-Extract) and its major cytotoxic component, Withaferin A (Wi-A), have cancer cell killing activity. In the present study, we synthesized a FR-targeting i-Extract nanocomplex (FRi-ExNC) and a FR-targeting Wi-A nanocomposite (FRWi-ANC), by conjugating FA to polyethylene glycol and amphiphilic nanoframeworks, respectively. We investigated their anticancer potentials in in vitro and in vivo assays.
Methods: Selective cellular uptake of FRi-ExNC and FRWi-ANC were evaluated by immunofluorescent microscopy. Cytotoxic effect of FRi-ExNC and FRWi-ANC in cancer cells were detected by assays including cell viability, apoptosis and biochemical determination of proteins involved in these phenotypes. The antitumor efficacy of FRi-ExNC and FRWi-ANC were investigated by in vivo tumor formation assays in nude mice.
Results: We found that FRi-ExNC and FRWi-ANC caused stronger cytotoxicity as seen by induction of apoptosis. It was confirmed by cell cycle and protein expression analyses. In vivo tumor growth assays for subcutaneous xenografts in nude mice also revealed significantly enhanced suppression of tumor growth in the treated groups.
Conclusions: Our results suggested that these two kinds of nanoparticles serve as useful nanomedical tools for selective targeting of drugs to the cancer cells and enhanced anticancer activity
Merger of Ayurveda and Tissue Culture-Based Functional Genomics: Inspirations from Systems Biology
Ayurveda is one of the ancient systems of health care of Indian origin. Roughly translated into "Knowledge of life", it is based on the use of natural herbs and herb products for therapeutic measures to boost physical, mental, social and spiritual harmony and improve quality of life. Although sheltered with long history and high trust, ayurveda principles have not entered laboratories and only a handful of studies have identified pure components and molecular pathways for its life-enhancing effects. In the post-genomic era, genome-wide functional screenings for targets for diseases is the most recent and practical approach. We illustrate here the merger of ayurveda and functional genomics in a systems biology scenario that reveals the pathway analysis of crude and active components and inspire ayurveda practice for health benefits, disease prevention and therapeutics
Cell-based experimental evidence to the anti-COVID-19 potential of Ashwagandha and honeybee propolis ingredients
Background: The COVID-19 pandemic emerged in December 2019 by a novel strain of SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) has led to new endeavours in repurposing of existing drugs, anti-COVID-19 vaccine and drug development. Natural products, due to their general safety and wider availability, have attracted research and public attention. In this study, we report anti-COVID potential of compounds from honeybee propolis and Ayurvedic herb, Ashwagandha. Effect of active ingredients was studied on human cell surface receptors (ACE-2:Angiotensin Converting Enzyme-2/Spike protein and TMPRSS2:Transmembrane Protease Serine 2), critical for virus infection and virus main protease (Mpro, essential for virus replication), through molecular simulations and in vitro experiments.
Methods: Structure-based computational analyses were performed to predict the effect of honeybee propolis (CAPE: Caffeic Acid Phenethyl Ester and ARC: Artepillin C), and Ashwagandha (Withanolides) ingredients on virus-host cell surface receptors. Cell-based assays were used to investigate the effect of these compounds on the expression level of the target proteins and virus replication.
Results: Ashwagandha-derived nine withanolides were tested in silico for their potential to target and inhibit (i) ACE-2 and TMPRSS2 receptors (ii) viral main protease Mpro. We found that most withanolides possess capacity to bind to ACE-2, TMPRSS2 and Mpro . On the other hand, CAPE and ARC showed stable interactions at the active site of ACE2 and Mpro . ARC, but not CAPE, showed stable interaction with TMPRSS2. Human cells treated with withanolides, CAPE or ARC showed downregulation of both the receptors. Furthermore, celland PCR-based SARS-CoV-2 replication assays endorsed their antiviral activity.
Conclusion: The findings suggest that the Ashwagandha-withanolides and honeybee propolis-derived compounds, CAPE, and ARC, could be helpful in the reduction of viral replication/infection, and hence warrant further experimental and clinical attention
Identification and characterization of anticancer potential of a novel small molecule, Mortaparibmild
Background: The development of new anticancer drugs and treatment modalities form a priority research field. The tumor suppressor protein p53 is frequently mutated or functionally inactivated in a large variety of cancers. Its inactivation by mortalin, a member of the heat shock 70 protein family, has been shown to contribute to carcinogenesis. The small molecule inhibitors of mortalin-p53 interactions have been shown to reactivate p53 yielding apoptosis/growth arrest in cancer cells. Therefore, abrogators of mortalin-p53 interaction have emerged as possible new therapeutic anticancer reagents.
Methods: We performed chemical library screening based on the imaging of mortalin-p53 interaction, leading to the identification of a novel triazole derivative 4-[(4-amino-5-thiophen-2-yl-1,2,4-triazol-3-yl)sulfanylmethyl]-N-(4-methoxyphenyl)-1,3-thiazol-2-amine. Bioinformatics and experimental analyses were conducted to assess the anti-cancer potency of this molecule, named Mortaparibmild.
Results: Mortaparibmild could bind to mortalin and p53 on their interaction sites. It caused downregulation of mortalin and PARP1 expression. However, a higher dose of Mortaparibmild was required for inducing apoptosis/growth arrest in cancer cells as compared to Mortaparib and MortaparibPlus, the previously reported molecules with similar properties [Elwakeel et. al. (2021) Cancers 13:3043; Sari et.al. (2021) Cancers 13:835 and Putri, et.al. (2019) J Exp Clin Cancer Res 38:1]. It was also effective for triggering apoptosis/growth arrest in p53null cancer cells suggesting its p53-independent activities. Molecular characterization of p53-dependent and independent Mortaparibmild activity and their relevance to cancer therapy will be discussed.
Conclusion: Mortaparibmild is a new small molecule capable of inhibiting mortalin and PARP1 and inducing apoptosis in cancer cells
Inhibition of lipid accumulation in hepatocytes by unique Ashwagandha extracts
Introduction: Ashwaganda (Withania Somnifera) is a popular ayurvedic herb, trusted for a variety of health benefits in Indian traditional home medicine system. Steroidal lactones, Withaferin A (Wi-A) and Withanone (Wi-N), have been characterized as its major bioactives with a variety of bioactivities. We investigated the effect of Ashwagandha extracts on steatosis, abnormal retention of fat within a cell or organ that often affects liver as non-alcoholic fatty liver disease (NAFLD).
Methods: We prepared extracts from Ashwagandha that varied in their Wi-A and Wi-N content. Cytotoxicity of these extracts on human hepatocytes (Huh-7 and Suit-2) was evaluated by cell viability assays. Nontoxic doses were used to treat the cells subjected to activated lipid accumulation by palmitic acid (PA). The lipolygenesis was evaluated by Oil Red O and triglyceride (TG) assays, and the expression of molecules involved in this process.
Results and Discussions: The four kinds of extracts with different amounts of total withanolides and Wi-A:Wi-N ratio were generated. Cells were treated with PA to induce lipid accumulation. We found that in cells pre-treated with specific Ashwahandha extracts, TG accumulation was decreased. Of note, Sterol regulatory element-binding protein-1c (SREBP-1c), and its downstream effector-Fas, the key regulators of lipogenesis showed downregulation in specific extract-treated cells. Furthermore, the expression of PPARγ, a key factor involved in hepatic lipogenesis, showed decrease in cells treated with some of these extracts.
Conclusion: Ashwagandha extracts may provide a useful natural resource with anti-steatosis activity, maintaining liver health and NAFLD prevention
Structure-Function Relationships of the Temporomandibular Joint in Response to Altered Loading
Aims:
To elucidate the effects of decreased occlusal loading (DOL), with or without reloading (RL), on the structure and bite force function of the mandibular condylar fibrocartilage in skeletally mature male mice.
Methods:
At 13 weeks old, 30 wild type (WT) male mice were subjected to: (1) 6 weeks normal loading (NL); (2) 6 weeks DOL; or (3) 4 weeks DOL + 2 weeks RL. Histomorphometry, cell metabolic activity, gene expression of chondrogenic markers, and bite force tests were performed.
Results:
DOL resulted in a significant increase in apoptosis (P .10), apoptosis (P > .999), and bite force (P > .90), but not in mandibular condylar fibrocartilage thickness (P > .05).
Conclusions:
DOL in skeletally mature mice induces mandibular condylar fibrocartilage atrophy at the hypertrophic cell layer with a corresponding decrease in bite force.R56 DE020097F32 DE026366French Federation of Orthodontic
Molecular insights to the dose-dependent activities of Ashwagandha extracts
Background: Stress is an inevitable component of life. Several herbs are known for their health supporting effects that range from treatment of stress, common cold to cancer. We investigated the dose-dependent effect of Ashwagandha (Withania somnifera) extracts on human normal and cancer cells, and have attempted to resolve the molecular mechanisms of their antistress activities.
Methods: Ashwagandha extracts were chemically profiled by HPLC. Cytotoxicity was determined by viability assays. Biochemical and immunoimaging assays were performed using specific antibodies.
Results: Human normal cells treated with low doses of the leaf extract or purified withanolides (Withaferin A or Withanone) showed no toxicity. Such non-toxic doses were selected for antistress, neurodifferentiation and neuroregenerative assays. We found that whereas normal cells exposed to oxidative and UV stresses showed poor viability/growth arrest/apoptosis, cells treated with low doses of Ashwagandha extracts were protected. Brain-derived cells exposed to glutamate and scopolamine stresses showed protection and strong differentiation as marked by expression of neurodifferentiation markers. Muscle-derived cells cultured in low doses of extract showed muscle differentiation as marked by expression of muscle differentiation markers. Most recently, using computational tools, we examined potential of Ashwagandha for anti-SARS-CoV2 virus activity, and found that most of the Ashwagandha Withanolides have potential to block cell surface receptors (ACE2 and TMPRSS2) that are involved in entry of virus to human cells. Furthermore, Ashwagandha treated cells showed decrease in ACE2 and TMPRSS2 expression suggesting its potential in blocking virus infection.
Conclusion: Ashwagandha extracts and withanolides possess useful bioactivities
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