DIHYDROTESTOSTERONE DOWNREGULATES BONE RESORPTION ACTIVITY OF OSTEOCLASTS IN DOSE DEPENDENT MANNER: AN IN VITRO MODEL USING RAW 264.7 CELLS

Objective: Numerous studies have evidenced the bone regulatory potential of dihydrotestosterone in androgen-deficient osteoporosis. The present study was thus aimed to explore the translational mechanism of dihydrotestosterone to down-regulate the bone resorption activity of osteoclasts using RAW 264.7 cells as in vitro model.Methods: Prior to analyze the efficacy of dihydrotestosterone (5α-DHT) to alleviate osteoclastic differentiation, their cell viability and cell proliferative ability was assessed using lactate dehydrogenase (LDH) and MTS assays. The osteoclastic differentiation capacity of dihydrotestosterone was evaluated by measuring TRAP activity and the expression of bone resorption-related proteins such as matrix metallopeptidase-9 (MMP-9), cathepsin-K, tartrate-resistant acid phosphatase (TRAP) and NFATc1. Moreover, the effects of dihydrotestosterone were also evaluated on superoxide (free radicals) generation and superoxide dismutase (SOD) activity in RANKL-induced osteoclasts.Results: Dihydrotestosterone showed no toxicity towards RAW 264.7 cells and significantly enhanced their proliferation and growth rates in a dose-dependent fashion. It was also observed that dihydrotestosterone exhibits a remarkable inhibitory effect on differentiation, maturation and activation of osteoclasts. The marked inhibition of differentiation and activation of osteoclasts caused by 5α-DHT was due to down-regulation of the expression of MMP-9, cathepsin-K, TRAP, NFATc1, generation of superoxide and up-regulation of SOD activity in the RAW 264.7 cells.Conclusion: Resulting data provided substantially in vitroevidence for the pronounced anti-osteoclastogenetic activity of dihydrotestosterone and its therapeutic value in treating osteoporosis and other bone-erosive disorders.Â

Dihydrotestosterone, a robust promoter of osteoblastic proliferation and differentiation: understanding of time-mannered and dose-dependent control of bone forming cells

Objective(s): The present study was aimed to evaluate the time-mannered and dose-dependent effects of 5α-dihydrotestosterone (5α-DHT) on the proliferation and differentiation of bone forming cells using MC3T3-E1 cells. Materials and Methods: Cell proliferation was analyzed using MTS and phase contrast microscopic assays. Osteogenic differentiation was assessed through a series of in vitro experiments including crystal violet staining, alkaline phosphatase (ALP) activity, and Van Gieson (VG) staining. Taken together, the efficiency of bone mineralization was examined by using alizarin red s (ARS) staining, Von Kossa staining, scanning electron microscopy (SEM) and energy dispersive x-ray (EDX) analysis. Results: The resulting data revealed that 5α-DHT exhibits promising potential particularly at a dose of 0.1 ng/ml, in promoting the growth of MC3T3-E1 cells compared to the control group (CN). Moreover, a significantly higher ALP activity was evident in the experimental group treated with 5α-DHT compared to the CN group at various time intervals. MC3T3-E1 cells treated with 5α-DHT also expressed a remarkably higher collagen deposition and mineralization (calcium and phosphate contents) compared to the CN group at various time intervals. Conclusion: Conclusively, we suggest that 5α-DHT exhibits outstanding potential of promoting proliferation and differentiation in osteoblasts which could be the in vitro basis for the efficacy of 5α-DHT in the treatment of androgen-deficient male osteoporosis

Exploring dynamic biomedical algorithm of Eurycoma longifolia Jack and its bioactive phytochemicals: A review of pharmacokinetic and pharmacodynamic implications and future prospects

Eurycoma longifolia Jack (E. longifolia) is a well-recognized traditional herbal medicine that offers a wide dynamic range of biomedical applications including anti-osteoporotic, anticancer, anti-proliferative, anti-malarial, antimicrobial, antioxidant, aphrodisiac, anti-inflammatory, anxiolytic, anti-diabetic, anti-rheumatism and anti-ulcer properties. This review aims to overview the pharmacokinetic and a pharmacodynamic algorithm of E. longifolia and its bioactive components. Analysis of pharmacokinetic profile revealed that E. longifolia exhibit higher bioavailability, high volume of distribution, slow elimination rate, and does not show inhibitory effects on cytochrome P450 isoenzymes. E. longifolia has been used, alone or in combination with other pharmacological agents, in the form of crude extracts, standard extracts, or decoctions of different plant parts (i.e., herbs, shrubs, stem, leaves, and roots) for the treatment of various ailments in animals and humans. Among various bioactive constituents, eurycomanone has been found to be the most remarkable, super-stable, versatile, and most potent phytochemical (isolated or extracted from root extracts) against various types of animals and human diseases. Based on its well-established pharmacokinetic and pharmacodynamic profiles, we suggested that E. longifolia can be a well-accepted complementary and alternative medicine for the treatment of different types of human ailments

Exploring molecular mechanism of bone-forming capacity of Eurycoma longifolia: Evidence of enhanced expression of bone-related biomarkers

Background: Among the numerous well-documented medicinal herbs, Eurycoma longifolia (EL) has gained remarkable recognition due to its promising efficacy of stimulating bone formation in androgen-deficient osteoporosis. Though numerous animal studies have explored the bone-forming capacity of EL, the exact mechanism was yet to be explored. Objective(s): The present study was aimed to investigate the mechanism of bone-forming capacity of EL using MC3T3-E1 as an in vitro osteoblastic model. Materials and methods: The cell differentiation capacity of EL was investigated by evaluating cell growth, alkaline phosphatase (ALP) activity, collagen deposition and mineralization. Taken together, time-mannered expression of bone-related mediators which include bone morphogenic protein-2 (BMP-2), ALP, runt-related transcription factor-2 (Runx-2), osteocalcin (OCN), type I collagen, osteopontin (OPN), transforming growth factor-β1 (TGF-β1) and androgen receptor (AR) were measured to comprehend bone-forming mechanism of EL. Results: Results demonstrated a superior cell differentiation efficacy of EL (particularly at a dose of 25 μg/mL) that was evidenced by dramatically increased cell growth, higher ALP activity, collagen deposition and mineralization compared to the testosterone. Results analysis of the bone-related protein biomarkers indicated that the expression of these mediators was well-regulated in EL-treated cell cultures compared to the control groups. These findings revealed potential molecular mechanism of EL for the prevention and treatment of male osteoporosis. Conclusion: The resulting data suggested that EL exhibited superior efficacy in stimulating bone formation via up-regulating the expression of various mitogenic proteins and thus can be considered as a potential natural alternative therapy for the treatment of osteoporosis. Keywords: Eurycoma longifolia, Osteoporosis, Bone formation, Molecular mechanis

EFFECT OF EURYCOMA LONGIFOLIA ON SEXUAL BEHAVIOR IN SEXUALLY DYSFUNCTIONAL MALE: A SYSTEMATIC REVIEW

Objective: This systematic review was conducted to summarize and evaluate the effect of Eurycoma longifolia (EL) on sexual behavior in the sexually dysfunctional male.Methods: Ovid Medline, Wiley Online Library, Scopus, BioMed Central, Hindawi and Google Scholar databases were searched for relevant studies published from 1995 to 2016 and search was limited to relevant studies published in English. Studies assessing aphrodisiac properties of EL on male sexual behavior in sexually dysfunctional animal models or human were included.Results: Among the 155 studies identified in the literature search, a total of 6 eligible articles (5 animal studies and one human study) were selected and reviewed. All studies reported positive aphrodisiac activities and supported the use of the plant as an aphrodisiac.Conclusion: This systematic review highlighted the effect of EL extract as an aphrodisiac agent to improve sexual behavior in the sexually dysfunctional male. Further studies are required to determine the specific mechanisms of action and identification of the bioactive components of EL responsible for its potential efficacy

Mechanistic Insight Into the Efficient Osteogenic Potential of Dihydrotestosterone: Exploring Sequential Expression of Bone-Related Protein Biomarkers

Numerous in vitro, in vivo and clinical studies have evidenced the outstanding potential of dihydrotestosterone (DHT) in the treatment of male osteoporosis. Despite of promising clinical efficacy of DHT in regulating the skeletal growth and homeostasis, the exact molecular and translational mechanism is yet to be explored. This study was aimed to investigate the bone-forming molecular mechanism of DHT using MC3T3-E1 cell line as in vitro model. The mechanism of bone-forming ability of DHT was assessed by evaluating the time-mannered expression of bone-related biomarkers such as bone morphogenic protein-2 (BMP-2), alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx-2), osteocalcin (OCN), type I collagen, osteopontin (OPN), transforming growth factor-β1 (TGF-β1) and androgen receptor (AR). Results demonstrated a remarkable efficacy of DHT (at a dose of 0.1 ng/mL) in promoting the expression of these vital bone-forming mediators. The resulting analysis revealed that the DHT-0.1 group showed higher expression of BMP-2 (106±9 pg/mL), ALP (381±16 pg/mL), Runx-2 (664±32 pg/mL), OCN (2265±111 pg/mL), type I collagen (276±16 pg/mL), TGF-β1 (81±7 pg/mL) and AR (411±21 pg/mL) compared to the control (CN) and other DHT-treated groups. These findings provide an in vitro evidence for the bone-forming capacity of DHT and its therapeutic significance for the treatment of male osteoporosis

OFF-FARM EMPLOYMENT IN SOUTHERN SHAN STATE

This research highlight presents key features of off-farm employment in Myanmar’s Southern Shan State. Analysis is based on the Shan Household Agriculture and Rural Economy Survey (SHARES), which collected detailed information on livelihoods and agriculture from 1562 households in nine townships. In this study, off-farm employment is defined as any income generating activity that does not take place on the household’s farm. Off-farm employment can therefore include both agriculture and non-agriculture related activities. We distinguish four main categories of off-farm employment: casual wage employment, salaried employment, non-farm enterprises, and natural resource extraction. Findings related to migration are presented in a separate research highlight. Below, we present the research findings in five sub-sections. The first provides a general overview of off-farm employment, and is followed by subsections presenting results on each of the four main categories of off-farm employment listed above

EURYCOMA LONGIFOLIA, A MALAYSIAN MEDICINAL HERB, SIGNIFICANTLY UPREGULATES PROLIFERATION AND DIFFERENTIATION IN PRE-OSTEOBLASTS (MC3T3-E1): AN IN VITRO MODEL

Objective: Eurycoma longifolia (EL), a well-recognized Malaysian medicinal herb, has gained widespread popularity due to its ability to protect against bone calcium loss in androgen-deficient osteoporosis. Nevertheless numerous animal studies have proved the bone protective effect of EL; however, the exact mechanism is not well-explained yet. Thus, the present study was aimed to explore the in vitro basis of bone protective effects of EL by using mouse pre-osteoblast cell line (MC3T3-E1).Methods: The cytotoxicity and proliferative potential of EL were evaluated by lactate dehydrogenase (LDH) and cell counting methods. Despite cell growth, the ability of EL to promote osteogenic differentiation of bone-forming cells was assessed by quantifying collagen (early differentiation marker) and calcium (late differentiation marker) in EL-treated bone forming cells.Results: Resulting data obtained from dose optimization study revealed that EL at 5 to 50 µg/ml concentration showed marked effects in significantly promoting cell growth in MC3T3-E1 cells. As such, resulting data also demonstrated the superior potential of EL in up regulating collagen synthesis and mineralization (calcium deposition) in MC3T3-E1 cells at 25 µg/ml, in comparison to untreated (negative control) and dihydrotestosterone (5α-DHT)-treated cells (positive control).Conclusion: These pronounced effects of EL on osteoblasts provide an in vitro basis for the bone protective potential of EL and thus can be considered as an alternative regimen for the treatment of androgen-deficient male osteoporosis.Â