Traditional Therapeutic Agents Research Articles

Development of hyaluronate tethered magnetic nanoparticlesfor targeted anti-cancer drug delivery

Despite the tremendous progress in understanding the molecular basis of the disease, cancer still remains one of the leading causes of deaths. Recently, advances in nanotechnology are rapidly enabling the development of novel, multifunctional materials with combined cancer specific targeting, therapeutic and diagnostic functions within a single nanocomplex (NP) that address the shortcomings of traditional disease diagnostic and therapeutic agents. Among the myriad of nanocarriers, magnetic nanoparticles (MNPs) have sparked extensive promise as novel theranostic applications as these MNPs can be directly targeted to the diseased cells with effective therapeutic efficiency. For this, these MNPs should be modified with some highly biocompatible polymers (specially polysaccharides) exhibiting the cancer targeting properties that can strongly interact with receptors expressed on the target cancer to facilitate accurate detection of the specific cancer and enhanced delivery to the target site while reducing unintended tside effects. Over the last few years, many groups have reported hyaluronic acid (HA) as the targeting agent as it directly delivers targeted MNPs to CD44 overexpressed cancer cells. In most of the cases, doxorubicin (DOX) has been used as the anticancer drug as it is largely utilized for treating a broad spectrum of cancers. In our work, we have designed a novel, intravenously injectable, CD44 receptor targeted MNP formulation, where the HA moiety of MNPs facilitate easy detection of cancer cells via receptor specific interactions, DOX can regress the cancer cells with simultaneous imaging efficacy. This theranostic MNPs led to the formation of novel nanoformulation, capable of performing concomitant detection, regression and imaging in in vitro CD44 over expressing cancer cells.

Bioaccessible Organosulfur Compounds in Broccoli Stalks Modulate the Inflammatory Mediators Involved in Inflammatory Bowel Disease.

Inflammatory diseases are strongly associated with global morbidity and mortality. Several mediators are involved in this process, including proinflammatory interleukins and cytokines produced by damaged tissues that, somehow, act as initiators of the autoreactive immune response. Bioactive compounds present in plant-based foods and byproducts have been largely considered active agents with the potential to treat or prevent inflammatory diseases, being a valuable alternative to traditional therapeutic agents used nowadays, which present several side effects. In this regard, the present research uncovers the anti-inflammatory activity of the bioaccessible fraction of broccoli stalks processed, by applying different conditions that render specific concentrations of bioactive sulforaphane (SFN). The raw materials' extracts exhibited significantly different contents of total glucosinolates (GSLs) that ranged between 3993.29 and 12,296.48 mg/kg dry weight (dw), with glucoraphanin as the most abundant one, followed by GI and GE. The indolic GSLs were represented by hydroxy-glucobrassicin, glucobrassicin, methoxy-glucobrassicin, and neo-glucobrassicin, with the two latter as the most abundant. Additionally, SFN and indole-3-carbinol were found in lower concentrations than the corresponding GSL precursors in the raw materials. When exploring the bioaccessibility of these organosulfur compounds, the GSL of all matrices remained at levels lower than the limit of detection, while SFN was the only breakdown product that remained stable and at quantifiable concentrations. The highest concentration of bioaccessible SFN was provided by the high-ITC materials (~4.00 mg/kg dw). The results retrieved on the cytotoxicity of the referred extracts evidenced that the range of supplementation of growth media tested (0.002-430.400 µg of organosulfur compounds/mL) did not display cytotoxic effects on Caco-2 cells. The obtained extracts were assessed based on their capacity to reduce the production of key proinflammatory cytokines (interleukin 6 (IL-6), IL-8, and TNF-α) by the intestinal epithelium. Most of the tested processing conditions provided plant material with significant anti-inflammatory activity and the absence of cytotoxic effects. These data confirm that SFN from broccoli stalks, processed to optimize the bioaccessible concentration of SFN, may be potential therapeutic leads to treat or prevent human intestinal inflammation.

Research Progress on Effects of Ginsenoside Rg2 and Rh1 on Nervous System and Related Mechanisms

Neurological-related disorders are diseases that affect the body’s neurons or peripheral nerve tissue, such as Parkinson’s disease (PD) and Alzheimer’s disease (AD). The development of neurological disorders can cause serious harm to the quality of life and functioning of the patient. The use of traditional therapeutic agents such as dopamine-promoting drugs, anticholinergic drugs, cholinesterase inhibitors, and NMDA receptor antagonists is often accompanied by a series of side effects such as drug resistance, cardiac arrhythmia, liver function abnormalities, and blurred vision. Therefore, there is an urgent need to find a therapeutic drug with a high safety profile and few side effects. Herbal medicines are rich in active ingredients that are natural macromolecules. Ginsenoside is the main active ingredient of ginseng, which has a variety of pharmacological effects and is considered to have potential value in the treatment of human diseases. Modern pharmacological studies have shown that ginsenosides Rg2 and Rh1 have strong pharmacological activities in the nervous system, with protective effects on nerve cells, improved resistance to neuronal injury, modulation of neural activity, resistance to cerebral ischemia/reperfusion injury, improvement of brain damage after eclampsia hemorrhage, improvement of memory and cognitive deficits, treatment of AD and vascular dementia, alleviation of anxiety, pain, and inhibition of ionic-like behavior. In this article, we searched the pharmacological research literature of Rg2 and Rh1 in the field of neurological diseases, summarized the latest research progress of the two ginsenosides, and reviewed the pharmacological effects and mechanisms of Rg2 and Rh1, which provided a new way of thinking for the research of the active ingredients in ginseng anti-neurological diseases and the development of new drugs.

Mitocans induce lipid flip-flop and permeabilize the membrane to signal apoptosis

Pancratistatin (PST) and narciclasine (NRC) are natural therapeutic agents that exhibit specificity toward the mitochondria of cancerous cells and initiate apoptosis. Unlike traditional cancer therapeutic agents, PST and NRC are effective, targeted, and have limited adverse effects on neighboring healthy, noncancerous cells. Currently, the mechanistic pathway of action for PST and NRC remains elusive, which in part inhibits PST and NRC from becoming efficacious therapeutic alternatives. Herein, we use neutron and x-ray scattering in combination with calcein leakage assays to characterize the effects of PST, NRC, and tamoxifen (TAM) on a biomimetic model membrane. We report an increase in lipid flip-flop half-times (t1/2) (≈12.0%, ≈35.1%, and a decrease of ≈45.7%) with 2 mol percent PST, NRC, and TAM respectively. An increase in bilayer thickness (≈6.3%, ≈7.8%, and ≈7.8%) with 2 mol percent PST, NRC, and TAM, respectively, was also observed. Lastly, increases in membrane leakage (≈31.7%, ≈37.0%, and ≈34.4%) with 2 mol percent PST, NRC, and TAM, respectively, were seen. Considering the maintenance of an asymmetric lipid composition across the outer mitochondrial membrane (OMM) is crucial to eukaryotic cellular homeostasis and survival, our results suggest PST and NRC may play a role in disrupting the native distribution of lipids within the OMM. A possible mechanism of action for PST- and NRC-induced mitochondrial apoptosis is proposed via the redistribution of the native OMM lipid organization and through OMM permeabilization.

Perspective of SGLT2i in the Treatment of Abdominal Aortic Aneurysms.

The incidence of abdominal aortic aneurysm (AAA) in the elderly is increasing year by year with high mortality. Current treatment is mainly through surgery or endovascular intervention, which is not sufficient to reduce future risk. Therefore, we still need to find an effective conservative measure as an adjunct therapy or early intervention to prevent AAA progression. Traditional therapeutic agents, such as β-receptor blockers, calcium channel blockers, and statins, have been shown to have limited effects on the growth of AAA. Recently, sodium-glucose cotransport proteins inhibitors (SGLT2is), a new class hypoglycemic drug, have shown outstanding beneficiary effects on cardiovascular diseases by plasma volume reduction, vascular tone regulation, and various unidentified mechanisms. It has been demonstrated that SGLT2i is abundantly expressed in the aorta, and some studies also showed promising results of SGLT2i in treating animal AAA models. This article aims to summarize the recent progress of AAA studies and look forward to the application of SGLT2i in AAA treatment for early intervention or adjunct therapy after surgical repair or stent graft.

Influence of pyrrolizidine alkaloids depletion upon the biological activity of Symphytum officinale L. extracts

Ethnopharmacological relevanceComfrey (Symphytum officinale L., Boraginaceae) root preparations are used as both traditional remedies and therapeutic agents in treating pain and inflammation associated with joint, bone, and muscle ailments. Even though numerous phytochemicals contribute to the beneficial effects of comfrey, the presence of toxic pyrrolizidine alkaloids (PAs) overshadows its uses. Aim of the studyIn this work, different PA-/mucilage-depleted/undepleted comfrey root extracts were subjected to detailed phytochemical characterization and biological evaluation. Materials and methodsThe phytochemical profiling was performed by LC-HRMS/MS. The quantification of PAs and major phenolic compounds was carried out by LC-MS/MS and LC-DAD. Antioxidant and enzyme inhibitory activity was determined by in vitro free radical scavenging, ion reducing, metal chelating, cholinesterase, tyrosinase, amylase, and glucosidase assays. Using an ex vivo model of LPS-stimulated neutrophils, their viability (as measured by flow cytometry) and the release of IL-1β, IL-8, and TNF-α were determined (ELISA assay). Results12 phenolic acids, six PAs, three organic acids, two fatty acids, and two sugars were identified in the obtained comfrey extracts. The PA-depleted materials contained PAs levels below 2 ppm, whereas the removal of mucilage increased the content of rosmarinic acid, globoidnan A, globoidnan B, and rabdosiin. PA-depletion did not significantly affect the antioxidant potential. However, the radical scavenging and metal reducing properties were higher in the mucilage-depleted extracts. Neither PA-depletion nor mucilage-depletion had considerable effects on the in vitro inhibitory activity of cholinesterases, tyrosinase, amylase, and glucosidase or release of ex vivo pro-inflammatory cytokines (e.g., IL-1β, IL-8, and TNF-α) in LPS-stimulated neutrophils. ConclusionsIn light of their superior safety profiles, PA-depleted comfrey extracts can be utilized further in cosmetic and pharmaceutical products.

THE DANGERS OF SILVER NANOPARTICLES TO THE HUMAN BODY AND THE ENVIRONMENT

Nanotechnology is a rapidly growing science of producing and utilizing nano-sized particles that are procedure in nanometers. These nanomaterials are already having an impact on health care for the time being, nanoproducts are used to in various fields. Among the use are silver nanoparticles playa major role in the field the nano revolution and nanomedicine. Nanoparticles with is small size to large surface area (1–100 nm) have potential medical, industrial and agricultural applications. Scientists have carried out significant efforts toward the synthesis of nanoparticles by different means, including physical, chemical and biological methods. These properties can be used to overcome some of the limitations found in traditional therapeutic and diagnostic agents. What makes these materials superior and indispensable a their unique size-dependent properties. Silver nanoparticles have potential antimicrobial activity towards many pathogenic microbes. Along with this antimicrobial activity, silver nanoparticles are showing unacceptable toxic effects on human health and the environment.

Thermosensitive Hydrogel Loaded with Primary Chondrocyte-Derived Exosomes Promotes Cartilage Repair by Regulating Macrophage Polarization in Osteoarthritis.

Intra-articular injection is a classic strategy for the treatment of early osteoarthritis (OA). However, the local delivery of traditional therapeutic agents has limited benefits for alleviating OA. Exosomes, an important type of extracellular nanovesicle, show great potential for suppressing cartilage destruction in OA to replace drugs and stem cell-based administration. In this study, we developed a thermosensitive, injectable hydrogel by in situ crosslinking of Pluronic F-127 and hyaluronic acid, which can be used as a slow-release carrier to durably retain primary chondrocyte-derived exosomes at damaged cartilage sites to effectively magnify their reparative effect. It was found that the hydrogel can sustainedly release exosomes, positively regulate chondrocytes on the proliferation, migration and differentiation, as well as efficiently induce polarization of M1 to M2 macrophages. Intra-articular injection of this exosomes-incorporated hydrogel significantly prevented cartilage destruction by promoting cartilage matrix formation. This strategy also displayed a regenerative immune phenotype characterized by a higher infiltration of CD163+ regenerative M2 macrophages over CD86+ M1 macrophages in synovial and chondral tissue, with a concomitant reduction in pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) and increase in anti-inflammatory cytokine (IL-10) in synovial fluid. Our results demonstrated that local sustained-release primary chondrocyte-derived exosomes may relieve OA by promoting the phenotypic transformation of macrophages from M1 to M2, which suggesting a great potential for the application in OA.

Impact of Pre-Transplant Immunosuppression in CT-ILD Recipients

<h3>Purpose</h3> The use of immunosuppressive biologic agents (BA) in CT-ILD patients has increased. These agents have long half-lives and a high infection risk leading to our hypothesis that these patients will have more infectious complications. This study compares early outcomes in patients who received pre-transplant traditional immunosuppression (MMF, AZA, hydroxychloroquine, cyclophosphamide) (TI), pre-transplant BA (anti-CD20, TNF blockers, IL-6 inhibitors, JAK inhibitors) or no immunosuppression (IS). <h3>Methods</h3> Patients transplanted for ILD between 2/2008-4/2021 were retrospectively divided into 3 groups (grp): (1) TI, (2) BA, and (3) none. Maintenance IS included a calcineurin inhibitor, MMF/AZA, and prednisone. Targeted induction therapy with basiliximab occurred in patients with acute kidney injury post-transplant. Variables examined included baseline characteristics, infection within 6 mo, time to infection, and 1-yr survival. <h3>Results</h3> 161 pts were included in the analysis. 45 received TI, 13 received BA, and 103 received neither. Grps were comparable in age and induction therapy, but more males were present in each grp (p=0.001). Incidence of infection within 6 mo was higher in the TI grp compared to no IS grp (71% vs 46%, p=0.004). Time to initial infection was shorter in the TI grp (86 vs 119 vs 121 d) when compared to the BA and no IS cohorts (p=0.016). There was no difference in length of initial hospital stay or 1-yr survival. <h3>Conclusion</h3> Our single-center experience demonstrates that the utilization of traditional therapeutic immunosuppressive agents when compared to BA or no IS to treat CT-ILD prior to transplantation results in a greater risk of early infection but does not impact survival. Transplantation of patients receiving newer BA does not appear to increase early infection rates, but this may be due to selection bias or low numbers in this analysis. Discontinuing TI well ahead of the transplant may have a protective effect in preventing early post-transplant infection.

A Nanoinformatics Approach to Evaluate the Pharmacological Properties of Nanoparticles for the Treatment of Alzheimer’s Disease

Alzheimer's disease is a destructive nervous system disease which causes structural, biochemical and electrical abnormalities inside the human brain and results due to genetic and various environmental factors. Traditional therapeutic agents of Alzheimer's disease such as tacrine and physostigmine have been found to cause adverse effects to the nervous system and gastrointestinal tract. Nanomaterials like graphene, metals, carbon-nanotubes and metal-oxides are gaining attention as potential drugs against Alzheimer's disease due to their properties such as large surface area, which provide clinical efficiency, targeted drug designing and delivery. Designing new drugs by using experimental approaches is a time-consuming, tedious and laborious process which also requires advanced technologies. This study aims to identify some novel drug candidates against Alzheimer's disease with no or less associated side effects using molecular docking approaches Methods: In this study, we utilized nanoinformatics based approaches for evaluating the interaction properties of various nanomaterials and metal nanoparticles with the drug targets, including TRKB kinase domain, EphA4 and histone deacetylase. Furthermore, the drug-likeness of carbon nanotubes was confirmed through ADME analysis. Carbon nanotubes, either single or double-walled in all the three-configurations, including zigzag, chiral, and armchair forms, are found to interact with the target receptors with varying affinities Conclusion: This study provides novel and clearer insights into the interaction properties and drug suitability of known putative nanoparticles as potential agents for the treatment of Alzheimer's disease.

Anaerobic bacteria mediated ‘smart missile’ targeting tumor hypoxic area enhances the therapeutic outcome of lung cancer

Tumor hypoxia is associated with a negative impact on the clinical treatment of lung cancer. Traditional therapeutic agents cannot be efficiently transported to tumor tissues, resulting in a reduced efficacy and serious toxic side effects. Therefore, the aim of this study is to propose a novel drug delivery strategy to solve this problem. The anaerobic Bifidobacterium infantis (B. infantis, Bi) is implanted in the hypoxic area of the tumor as the pre-implanted target. The bacterial antibody decorated nano-drug missile (Ab-DOX-s-s-NPS) actively targets the interior of tumor tissue, and enriches therapeutic drug doxorubicin into tumor hypoxic area through bacterial recruitment. The study confirms that the increased concentration of Ab-DOX-s-s-NPS in tumor tissues through the recruitment of B. infantis plays an excellent anti-tumor effect, while the systemic side effects are significantly reduced, suggesting the good therapeutic effect of the B. infantis-mediated drug delivery strategy against lung cancer. Therefore, this work provides an elegant platform for the in vivo delivery of chemotherapeutic drugs to the hypoxic area of the tumor, representing a novel strategy potentially effective in the treatment of solid tumors.

Supramolecular Nanomedicines of In-Situ Self-Assembling Peptides.

Nanomedicines provide distinct clinical advantages over traditional monomolecular therapeutic and diagnostic agents. Supramolecular nanomedicines made from in-situ self-assembling peptides have emerged as a promising strategy in designing and fabricating nanomedicines. In-situ self-assambly (SA) allows the combination of nanomedicines approach with prodrug approach, which exhibited both advantages of these strategies while addressed the problems of both and thus receiving more and more research attention. In this review, we summarized recently designed supramolecular nanomedicines of in-situ SA peptides in the manner of applications and design principles, and the interaction between the materials and biological environments was also discussed.

The use of Clostridium in cancer therapy: a promising way

In recent decades, the traditional cancer therapeutic agents such as surgery, radiotherapy, and chemotherapy have failed. One of the main reasons for repeated failures in the field of solid tumors is the hypoxic/necrotic regions within the tumors. Recently, looking for new therapeutic approaches, using bacteria as a therapeutic agent for cancer attract attention. In the case of solid tumors, several strains of anaerobic bacteria have been shown to selectively colonize, which made them the suitable agent for selective targeting and destruction. Interestingly to know that Clostridium species as an anaerobic bacterium has shown the high ability for selective internalization and colonization in the hypoxic and necrotic areas of the tumor which allow them to use as a target delivery for therapeutics agent and significant oncolytic. Furthermore, Clostridium can be modified to use as a producer of prodrug-converting enzymes or cytokines that allow the production of specific proteins or toxic agents within the tumors resulting in tumor destruction. The present study is a comprehensive review of the progress and current status of Clostridia as a promising agent for anticancer treatment.

When fat meets the engine: implications of dietary rumenic acid on myosin-targeting therapies in heart failure.

When fat meets the engine: implications of dietary rumenic acid on myosin-targeting therapies in heart failure.

Aqueous extract of Chrysanthemum morifolium Ramat. inhibits RANKL-induced osteoclast differentiation by suppressing the c-fos/NFATc1 pathway

ObjectiveThe flower of chrysanthemum, used worldwide as a medicinal and edible product, has shown various bioactivities, such as anti-inflammatory, antioxidant, anti-tumorigenic, and hepatoprotective activities, as well as cardiovascular protection. However, the effect of Chrysanthemum morifolium Ramat. on the regulation of osteoclast differentiation has not yet been reported. In this study, we aimed to investigate the inhibitory effect of Chrysanthemum morifolium Ramat. water extract (CME) on RANKL-induced osteoclast differentiation in mouse bone marrow-derived macrophages (BMMs). Study designBone marrow-derived macrophages (BMMs) isolated from the C57BL/6 J mice. The viability of BMMs was detected with MTT assays. Inhibitory effects of CME on osteoclast differentiation and bone resorption was measured by TRAP staining and Pit assay. Osteoclast differentiation-associated gene expression were assessed by Real-time quantitative polymerase chain reaction. Intracellular signaling molecules was assessed by western blot. ResultsCME significantly inhibited osteoclast differentiation in BMMs without cytotoxicity, besides inhibiting MAPK/c-fos and PLCγ2/CREB activation. The inhibitory effects of CME on differentiation-related signaling molecules resulted in significant repression of NFATc1 expression, which is a key transcription factor in osteoclast differentiation, fusion, and activation. ConclusionOur results confirmed the inhibition of RANKL-induced PLCγ2/CREB/c-fos/NFATc1 activation by CME during osteoclast differentiation. The findings collectively suggested CME as a traditional therapeutic agent for osteoporosis, RA, and periodontitis.

Standardized aqueous stem bark extract of Gmelina arborea roxb. possesses nephroprotection against adriamycin-induced nephrotoxicity in Wistar rats

Nephrotoxicity is a major limitation of adriamycin (ADR) chemotherapy. We hypothesized that administration of standardized aqueous bark extract of Gmelina arborea Roxb. (GA) (Family; Verbenaceae), a traditional therapeutic agent, may reduce the nephrotoxicity caused by ADR in Wistar rats. The dose-dependent nephroprotective activity of the standardized GA extract was investigated in ADR-induced (20 mg/kg, ip) nephrotoxicity in male Wistar rats (n = 6/group). The lyophilized powder of the aqueous refluxed (4 h) GA extract was administered at 100, 300 and 500 mg/kg doses orally for three consecutive days. Fosinopril sodium (0.09 mg/kg) was used as the positive control. Assessment of biochemical parameters on serum, urine and histopathology on H and E stained kidney sections were done at the end of the intervention. The treatment with GA and fosinopril decreased the elevation of serum creatinine, blood urea nitrogen, cystatin C, β2-microglobulin and loss of total protein in urine in nephrotoxic rats in a dose-dependent manner (p < 0.05). In contrast, serum concentrations of albumin and total protein were increased significantly (p < 0.05). H and E stained kidney sections showed an attenuation of renal parenchymal injury following the treatment. The aqueous extract of GA demonstrated antioxidant potential in vitro. Present findings conclude that the standardized aqueous extract of GA stem bark exerted a dose-dependent protection against ADR-induced nephrotoxicity in vivo and may be a promising adjunct in ADR chemotherapy.

Recent advances in the management of non-infectious posterior uveitis.

To review the current regimens and novel therapeutic modalities in various stages of research and development for the management of non-infectious posterior uveitis (NIPU). We performed a thorough review of current literature using PubMed, Google Scholar and Clinicaltrials.gov to identify the published literature about the available therapeutics and novel drugs/therapies in different stages of clinical trials. The current management regimen for non-infectious posterior uveitis includes corticosteroids, immunomodulatory therapies and anti-metabolites. However, NIPU requires long-term management for efficacious remission of the disease and to prevent disease relapse. Long-term safety issues associated with steroids have led to efforts to develop novel therapeutic agents including biological response modulators and immunosuppressants. The current therapeutic agents in various stages of development include calcineurin inhibitors, biologic response modifiers and a more a comprehensive modalities like ocular gene therapy as well as novel drug delivery mechanisms for higher bioavailability to the target tissues, with minimal systemic effects. Novel efficacious therapeutic modalities under development will help overcome the challenges associated with the traditional therapeutic agents.

In-vitro antiulcer activities of petal extract of Crocus sativus var. cashmerianus

Medicinal plants have been known for millennia as a rich source of traditional therapeutic agents for the prevention of diseases and ailments. The aim of the present study was performed to evaluate the antiulcer activities of hydro-alcoholic extracts of petals of Crocus sativus var. Cashmerianus by in-vitro methods viz. acid neutralizing capacity and H+/K+ - ATPase inhibition activity. In acid neutralizing capacity method, the petals extract significantly reduced acidity to 6.10 at a concentration of 1000 mg/ml as compared to 11.90 with standard 500 mg/ml of Aluminium hydroxide + Magnesium hydroxide combination. However, H+/K+ - ATPase inhibition activity method, petals extract showed maximum percentage inhibition of 70.31 % at the concentration 400µg/ml as compared to 73.82 % with a similar dose of standard Omeprazole. The IC 50 value of petals extract of C. sativus var. cashmerianus is shown 100 µg/ml in comparison with standard omeprazole of 82.5 µg/ml. The study reveals that the petals extract of C. sativus var. cashmerianus may contain compounds possessing acid neutralize and enzyme inhibition activities, thus it can be used as an alternative medicine for gastrointestinal disorders.

Inhibitory effect of Chaenomelis Fructus ethanol extract on receptor activator of nuclear factor-kappa B ligand-mediated osteoclastogenesis

The fruit of Chaenomeles sinensis (Thouin) Koehne (Chaenomelis Fructus) known as “Mo-Gua” in Korea has been commonly used in traditional medicine to treat inflammatory diseases, such as sore throat. However, its effect on bone metabolism has not been elucidated yet. Here, we examined the effect of Chaenomelis Fructus ethanol extract (CFE) on receptor activator of nuclear factor (NF)-κB ligand (RANKL)-mediated osteoclast differentiation and formation. CF-E considerably inhibited osteoclast differentiation and tartrate-resistant acid phosphatase-positive multinuclear cell formation from bone marrow-derived macrophages and osteoclast precursor cells in a dose-dependent manner. In addition, the formation of actin rings and resorption pits were significantly suppressed in CF-E-treated osteoclasts as compared with the findings in non-treated control cells. Consistent with these phenotypic inhibitory results, the expressions of osteoclast differentiation marker genes (Acp5, Atp6v0d2 , Oscar, CtsK, and Tm7sf4) and Nfatc1 , a pivotal transcription factor for osteoclastogenesis, were markedly decreased by CF-E treatment. The inhibitory effect of CF-E on RANKL-induced osteoclastogenesis was associated with the suppression of NFATc1 expression, not by regulation of mitogen-activated protein kinases and NF-κB activation but by the inactivation of phospholipase C gamma 1 and 2. These results indicate that CF-E has an inhibitory effect on osteoclast differentiation and formation, and they suggest the possibility of CF-E as a traditional therapeutic agent against bone-resorptive diseases, such as osteoporosis, rheumatoid arthritis, and periodontitis.

Inhibitory Effect of Rosae Multiflorae Fructus Extracts on the Receptor Activator of NF-κB Ligand-Induced Osteoclastogenesis through Modulation of P38- and Ca2+-Mediated Nuclear Factor of Activated T-Cells Cytoplasmic 1 Expression

BackgroundRosae Multiflorae fructus (RMF), known to have anti-inflammatory and antioxidant properties, has been used as a traditional remedy for inflammatory diseases such as arthritis in Eastern Asia. However, its effect on osteoclasts, which play a crucial role in resorptive inflammatory bone diseases, is yet to be elucidated.MethodsThe effect of extract of RMF (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-mediated osteoclastogenesis was examined by tartrate-resistant acid phosphatase (TRAP) staining, real-time polymerase chain reaction and western blot analysis. In addition, RANKL-induced Ca2+-oscillation was also investigated.ResultsRMF-E remarkably inhibited TRAP+-osteoclast and resorptive pit formation in a dose-dependent manner. In addition, the expression of c-Fos and nuclear factor of activated T-cells cytoplasmic, known as pivotal transcription factors for osteoclast formation in vitro and in vivo, and that of the osteoclast differentiation markers such as Acp5, Oscar, CtsK, Atp6v0d2, Tm7sf4, and Nfatc1 were significantly decreased by RMF-E treatment during osteoclastogenesis. The inhibitory effect of RMF-E on RANKL-induced osteoclastogenesis was caused by the suppression of p38 mitogen-activated protein kinase activation, and RANKL-induced Ca2+-oscillation removal via inactivation of Bruton's tyrosine kinase (BTK), and subsequently phospholipase C-γ2.Conclusions RMF-E negatively regulates osteoclast differentiation and formation. These findings suggest the possibility of RMF-E as a traditional therapeutic agent against osteoclast-related bone disorders such as osteoporosis, rheumatoid arthritis, and periodontitis.