The Efficacy of Botulinum Toxin A Injection for Gastrocnemius Hypertrophy: A Prospective, Randomized, Double-blinded Controlled Trial.

Background: Many individuals hold an interest in aesthetic appeal, with one aspect of physical attractiveness being the alluring contour of the lower leg. Utilizing botulinum toxin A (BTX-A) injections offers several advantages, including a short procedure time, low pain, and a speedy recovery. With a demand for high-level evidence regarding the effectiveness of BTX-A injections for correction of lower leg contour, we evaluated the safety and efficacy of BTX-A injection for improvement of gastrocnemius muscle hypertrophy. Methods: We conducted a prospective, randomized, and controlled clinical trial to evaluate whether the injection of BTX-A into the gastrocnemius muscle could decrease muscular hypertrophy. The patients were randomized into a low-dose injection (60 units) group and a high-dose injection group (100 units) for each leg. Demographics, clinical outcome, and satisfaction score were compared between the two groups. Results: A total of 20 patients and 40 legs were enrolled in this study. Clinical and surgical demographics were similar between the two groups. BTX-A injection showed a significant decrease in the circumference of the calf after 8 weeks (preinjection: 36.35 ±â€…0.63 cm versus postinjection: 35.87 ±â€…0.61 cm; P = 0.03). However, no significant difference was observed between the low- and the high-dose group (-0.52 ±â€…0.74 cm versus -0.44 ±â€…1.04 cm, P = 0.78). Conclusions: BTX-A injection can be a good noninvasive method for the correction of hypertrophic gastrocnemius muscles. This study supports the use of BTX-A injections in patients unsatisfied with lower leg hypertrophy.

The Double Mediating Effect of Family Support and Family Relationship Satisfaction on Self-Compassion and Meaning in Life among Korean Baby Boomers.

This study identified the relationship between self-compassion and meaning in life among Korean baby boomers and examined the double mediating effect of family support and family relationship satisfaction on this relationship. For this purpose, data were collected from 400 baby boomers (born between 1955-1963) using the self-compassion, meaning in life, family support, and family relationship satisfaction scales. PROCESS Macro 3.5 Model 6 was used to analyze the double mediating effects. The results revealed that first, there was a significant correlation between the self-compassion, meaning in life, family support, and family relationship satisfaction of this study. Second, in the relationship between self-compassion and the meaning in life, family support, and family relationship satisfaction were found to have a partial mediating effect and a double mediating effect. The implications and limitations of these findings are also discussed.

Comparison of physicochemical properties and antioxidant activities of fermented soybean-based red pepper paste, Gochujang, prepared with five different red pepper (Capsicum annuum L.) varieties.

The purposes of this study were to identify physicochemical properties and evaluate bioactive compound levels and antioxidant characteristics at 30 day intervals during the 90 days of fermentation of gochujang fortified with five different varieties of red pepper: Juktoma pepper (RP1), facing heaven pepper (RP2), Thai chili pepper (RP3), bird's eye pepper (RP4), and red bell pepper (RP5). Physicochemicals properties, including reducing sugar, capsaicin, pH, ß-carotene, and color parameters, of gochujang were evaluated. Antioxidant compounds of total polyphenols and total flavonoids were analyzed with antioxidant activities of DPPH and FRAP assays. The results showed that gochujangs (GRP1, GRP5) fortified with RP1, and RP5, had consistently higher values of reducing sugars, total polyphenols, and total flavonoids with antioxidant activities, but lower values of capsaicin, pH, ß-carotene, and color parameters as compared to GRP2, GRP3, GRP4 during 90 days of fermentation. GRP5 especially had the highest reducing sugar, amino acid contents, total polyphenols, and total flavonoids with antioxidant activities and the lowest value of capsaicin during the 90 days of fermentation.

Physicochemical and antioxidant properties of black garlic.

Black garlic (BG) is a processed garlic product prepared by heat treatment of whole garlic bulbs (Allium sativum L.) at high temperature under high humidity for several days, resulting in black cloves with a sweet taste. BG has recently been introduced to the Korean market as a product beneficial to health. To clarify how BG changes during the 35 day aging period, the physicochemical characteristics, antioxidant contents, and antioxidant activities were evaluated under controlled conditions of 70 °C and 90% relative humidity. Reducing sugar and total acidity of BG increased during the aging period, whereas pH decreased from pH 6.33 to 3.74. Lightness and yellowness values of BG radically decreased during the aging period, whereas redness values increased significantly. Antioxidant components, including the total polyphenol and total flavonoids contents of BG, increased significantly until the 21st day of aging (p < 0.05) and correspondingly, the antioxidant activities of BG, measured by DPPH, ABTS, FRAP, and reducing power assays, were highest on the 21st day of aging. These results indicate that BG can be considered to not only possess antioxidant properties during the aging period, but also to reach its optimal antioxidant properties at the 21st day of aging.

Actein isolated from black cohosh promotes the function of osteoblastic MC3T3-E1 cells.

Actein, isolated from black cohosh, was subjected to in vitro experiments to investigate its functional bioactivities in osteoblastic MC3T3-E1 cells. Actein caused a significant elevation of alkaline phosphatase activity, collagen synthesis, osteocalcin production, mineralization, and glutathione content in the cells, suggesting that actein has a stimulatory effect on osteoblastic bone formation or has potential activity against osteoporosis. We investigated the protective effects of actein on mitochondrial electron transport inhibitor, antimycin A induced toxicity in osteoblastic MC3T3-E1 cells. Exposure of MC3T3-E1 cells to antimycin A caused significant decrease in cell viability and mineralization. However, pretreatment with actein prior to antimycin A exposure significantly reduced antimycin A-induced cell damage by preventing mitochondrial membrane potential dissipation, complex IV inactivation, cardiolipin oxidation, ROS release, and nitrotyrosine increase, suggesting that actein may be useful for protecting mitochondria against a burst of oxidative stress. In addition, actein increased the phosphorylation of CREB (cAMP-response element-binding protein) inhibited by antimycin A and decreased the production of TNF-α induced by antimycin A. These findings suggest that actein could prevent oxidative damage to osteoblasts in osteoporotic patients.

The protective effects of Achyranthes bidentata root extract on the antimycin A induced damage of osteoblastic MC3T3-E1 cells.

Achyranthes bidentata (A. bidentata) Blume is a medicinal herb with the property of strengthening bones and muscles and ensuring proper downward flow of blood in terms of the therapeutic theory of traditional medicine. In the present study, the effect of A. bidentata root extract (AE) on osteoblast function was investigated in osteoblastic MC3T3-E1 cells. AE caused a significant elevation of alkaline phosphatase activity, collagen synthesis, osteocalcin production, and mineralization in the cells (P < 0.05). AE also decreased the production of TNF-α, IL-6, and RANKL induced by antimycin A, mitochondrial electron transport inhibitor. Exposure of MC3T3-E1 cells to antimycin A caused significant reduction of cell viability and mineralization. However, pretreatment with AE prior to antimycin A exposure significantly reduced antimycin A-induced cell damage by preventing mitochondrial membrane potential dissipation, ATP loss, ROS release, and nitrotyrosine increase, suggesting that AE may be useful for protecting mitochondria against a burst of oxidative stress. Moreover, AE increased the phosphorylation of cAMP-response element-binding protein inhibited by antimycin A. Our study demonstrates that A. bidentata could significantly prevent osteoblast damage in aged patients.

Liquiritigenin restores osteoblast damage through regulating oxidative stress and mitochondrial dysfunction.

We investigated the protective effect of liquiritigenin, one of the flavonoids present in Glycyrrhizae radix, against antimycin A-induced mitochondrial dysfunction in MC3T3-E1 osteoblast cells. Osteoblastic MC3T3-E1 cells were pre-incubated with liquiritigenin before treatment with antimycin A, and markers of mitochondrial function and oxidative damage were examined. In addition, the effects of liquiritigenin on the activation of phosphoinositide 3-kinase (PI3K) were examined in MC3T3-E1 cells. Liquiritigenin protected MC3T3-E1 cells from antimycin A-induced cell death. However, the PI3K inhibitor, LY294002, significantly attenuated liquiritigenin-mediated cell survival, indicating the involvement of PI3K in the cytoprotective effect of liquiritigenin. Pretreatment with liquiritigenin prior to antimycin A exposure significantly reduced antimycin A-induced PI3K inactivation, mitochondrial membrane potential dissipation, complex IV inactivation, and ATP loss. Liquiritigenin also reduced mitochondrial superoxide generation, nitrotyrosine production, and cardiolipin peroxidation during mitochondrial complex inhibition with antimycin A. Taken together, the results of this study show that modulation of PI3K, antioxidant effects, and the attenuation of mitochondrial dysfunction by liquiritigenin represent an important mechanism for its protection of osteoblasts against cytotoxicity resulting from mitochondrial oxidative stress.

Xanthohumol modulates the expression of osteoclast-specific genes during osteoclastogenesis in RAW264.7 cells.

RANKL has been shown to play a critical role in osteoclast formation and bone resorption. Thus, agents that suppress RANKL signaling have a potential to suppress bone loss. In this study, we examined the ability of xanthohumol, a structurally simple prenylated chalcone, to suppress RANKL signaling during osteoclastogenesis in RAW264.7 cells. Xanthohumol markedly inhibited RANKL-induced TRAP activity, multinucleated osteoclasts formation, and resorption-pit formation. In experiments to elucidate its mechanism of action, xanthohumol was found to suppress RANKL-induced expression of TRAF6, GAB2, ERK, c-Src, PI3K, and Akt genes. Moreover, RANKL-induced expressions of c-Fos and NFATc1, which are crucial transcription factors for osteoclastogenesis, were reduced by treatment with xanthohumol. Xanthohumol also inhibited RANKL-induced expression of bone-resorption related osteoclast-specific genes (carbonic anhydrase II, TCIRG, CLCN7, OSTM1, cathepsin K, and MMP-9). These data demonstrate that xanthohumol inhibits osteoclastogenesis by modulating RANKL signaling and may be useful for the prevention of bone-destructive diseases such as osteoporosis, arthritis and periodontitis.

Effect of zinc oxide nanoparticles on the function of MC3T3-E1 osteoblastic cells.

Zinc oxide nanoparticles (ZnO NPs) can be ingested directly when used in food, food packaging, drug delivery, and cosmetics. This study evaluated the cellular effects of ZnO NPs (50 and 100 nm diameter particle sizes) on the function of osteoblastic MC3T3-E1 cells. ZnO NPs showed cytotoxicity at concentrations of above 50 µg/ml, and there was no significant effect of the size on the cytotoxicity of ZnO NPs. Within the testing concentrations of 0.01~1 µg/ml, which did not cause a marked drop in cell viability, ZnO NPs (0.1 µg/ml) caused a significant elevation of alkaline phosphatase activity, collagen synthesis, mineralization, and osteocalcin content in the cells (P < 0.05). Moreover, pretreatment with ZnO NPs (0.01~1 µg/ml) significantly reduced antimycin A-induced cell damage by preventing mitochondrial membrane potential dissipation, complex IV inactivation, and ATP loss. Measurement of reactive oxygen species (ROS) indicated decrease in ROS level upon exposure to ZnO nanoparticles (0.01 µg/ml). Hence, our study indicated that ZnO nanoparticles can have protective effects on osteoblasts at low concentrations where there are little or no observable cytotoxic effects.

Paeoniflorin isolated from Paeonia lactiflora attenuates osteoblast cytotoxicity induced by antimycin A.

The protective effects of paeoniflorin isolated from Paeonia lactiflora against pharmacological inhibition of the respiratory chain were studied using osteoblastic MC3T3-E1 cells. Here we show that paeoniflorin decreases cell death induced by antimycin A, an inhibitor of mitochondrial complex III. Paeoniflorin restored antimycin A-induced inactivation of phosphoinositide 3-kinase (PI3K) and thioredoxin reductase, suggesting that PI3K and thioredoxin reductase may be involved in paeoniflorin-induced cytoprotective responses. We also examined the effect of paeoniflorin on mitochondrial dysfunction and oxidative stress induced by antimycin A. Paeoniflorin inhibited mitochondrial membrane potential dissipation, ATP loss, inactivation of complexes I and IV, cytochrome c release, and cardiolipin oxidation induced by antimycin A. In addition, paeoniflorin prevented antimycin A-induced ROS release and nitrotyrosine increase. These results imply that paeoniflorin protects osteoblasts from antimycin A-induced cell death via improved mitochondrial function.

Gold nanoparticles attenuates antimycin A-induced mitochondrial dysfunction in MC3T3-E1 osteoblastic cells.

Gold nanoparticles have shown promising biological applications due to their unique properties. Understanding the interaction mechanisms between nanomaterials and biological cells is important for the control and manipulation of these interactions for biomedical applications. In the present study, we investigated the effects of gold nanoparticles on the differentiation of osteoblastic MC3T3-E1 cells and antimycin A-induced mitochondrial dysfunction. The results showed that gold nanoparticles (5, 10, and 20 nm) caused a significant elevation of cell growth, alkaline phosphatase activity, collagen synthesis, and osteocalcin content in the cells (P < 0.05). Moreover, pretreatment with gold nanoparticles prior to antimycin A exposure significantly reduced antimycin A-induced cell damage by preventing mitochondrial membrane potential dissipation, complex IV inactivation, ATP loss, cytochrome c release, cardiolipin peroxidation, and reactive oxygen species generation. Taken together, our study indicated that gold nanoparticles may improve the differentiation and have protective effects on mitochondrial dysfunction of osteoblastic cells.

Protective effect of albiflorin against oxidative-stress-mediated toxicity in osteoblast-like MC3T3-E1 cells.

Albiflorin isolated from Paeoniae Radix was investigated for its ability to protect against antimycin A-induced osteoblast toxicity in the MC3T3-E1 cell line. MC3T3-E1 cells showed significantly reduced viability, increased apoptosis and lactate dehydrogenase release, elevated ROS/RNS levels, and decreased mitochondrial function after exposure to antimycin A. Pretreatment with albiflorin reversed the loss of cell viability in antimycin A-treated cultures. Similarly, pretreatment with albiflorin before antimycin A resulted in decreased apoptosis and lactate dehydrogenase release, decreased ROS/RNS levels, and increased mitochondrial function compared to antimycin A-treated cultures. In addition, albiflorin increased the mineralization reduced by antimycin A. Albiflorin reduced antimycin A-induced mitochondrial cytochrome c loss and cardiolipin peroxidation, conferring protection against ROS. These results confirmed the crucial role of cytochrome c and cardiolipin in the underlying mechanistic action of albiflorin. Therefore, the results suggest that albiflorin enhances mitochondrial function to suppress antimycin A-induced oxidative damage via the preservation of cytochrome c and cardiolipin. All of these data indicate that albiflorin may reduce or prevent osteoblast degeneration in osteoporosis.

Sciadopitysin protects osteoblast function via its antioxidant activity in MC3T3-E1 cells.

Age-related osteoblast dysfunction is the main cause of age-related bone loss in both men and women. In the present study, the effect of sciadopitysin, a type of biflavonoids, on osteoblast function was investigated in osteoblastic MC3T3-E1 cells. Sciadopitysin caused a significant elevation of alkaline phosphatase activity, collagen synthesis, osteocalcin production, mineralization, and glutathione content in the cells (P<0.05). Sciadopitysin also decreased the production of tumor necrosis factor-a (TNF-α) induced by antimycin A, a mitochondrial electron transport inhibitor. We investigated the protective effects of sciadopitysin on antimycin A-induced toxicity in osteoblastic MC3T3-E1 cells. Exposure of MC3T3-E1 cells to antimycin A caused a significant reduction in osteoblast dysfunction. However, pretreatment with sciadopitysin prior to antimycin A exposure significantly reduced antimycin A-induced cell damage by preventing mitochondrial membrane potential dissipation, adenosine triphosphate (ATP) loss, reactive oxygen species (ROS) release, and nitrotyrosine increase, suggesting that sciadopitysin may be useful for protecting mitochondria against a burst of oxidative stress. Moreover, sciadopitysin increased phosphorylation of cAMP-response element-binding protein (CREB) inhibited by antimycin A. Our results demonstrate that sciadopitysin may reduce or prevent osteoblasts degeneration.

Pinacidil stimulates osteoblast function in osteoblastic MC3T3-E1 cells.

This study examined the effect of pinacidil, a nonselective adenosine triphosphate-sensitive potassium channel opener, on the function of osteoblastic MC3T3-E1 cells. Pinacidil caused a significant elevation of collagen synthesis, alkaline phosphatase activity, osteocalcin synthesis and mineralization in the cells (p < 0.05). Pinacidil significantly decreased the production of osteoclast differentiation inducing factors such as TNF-α, IL-6 and receptor activator of nuclear factor-κB ligand in the presence of antimycin A, which inhibits mitochondrial electron transport. Moreover, pinacidil prevented antimycin A-induced reactive oxygen species and nitrotyrosine production. These results demonstrate that pinacidil may have positive effects on skeletal structure.

Effect of magnolol on the function of osteoblastic MC3T3-E1 cells.

OBJECTIVES: In the present study, the ability of magnolol, a hydroxylated biphenyl compound isolated from Magnolia officinalis, to stimulate osteoblast function and inhibit the release of bone-resorbing mediators was investigated in osteoblastic MC3T3-E1 cells. METHODS: Osteoblast function was measured by cell growth, alkaline phosphatase activity, collagen synthesis, and mineralization. Glutathione content was also measured in the cells. Bone-resorbing cytokines, receptor activator of nuclear factor-κB ligand (RANKL), TNF-α, and IL-6 were measured with an enzyme immunoassay system. RESULTS: Magnolol caused a significant elevation of cell growth, alkaline phosphatase activity, collagen synthesis, mineralization, and glutathione content in the cells (P < 0.05). Skeletal turnover is orchestrated by a complex network of regulatory factors. Among cytokines, RANKL, TNF-α, and IL-6 were found to be key osteoclastogenetic molecules produced by osteoblasts. Magnolol significantly (P < 0.05) decreased the production of osteoclast differentiation inducing factors such as RANKL, TNF-α, and IL-6 in the presence of antimycin A, which inhibits mitochondrial electron transport and has been used as an ROS generator. CONCLUSION: Magnolol might be a candidate as an agent for the prevention of bone disorders such as osteoporosis.

Protective effect of apocynin on antimycin A-induced cell damage in osteoblastic MC3T3-E1 cells.

Apocynin is a naturally occurring methoxy-substituted catechol, experimentally used as an inhibitor of NADPH-oxidase. In the present study, we investigated the protective effects of apocynin on antimycin A (AMA)-induced toxicicy in osteoblastic MC3T3-E1 cells. Exposure of MC3T3-E1 cells to AMA caused significant cell viability loss, as well as mitochondrial membrane potential (MMP) dissipation, complex IV inactivation, ATP loss, intracellular calcium ([Ca2+]i) elevation and oxidative stress. Pretreatment with apocynin prior to AMA exposure significantly reduced AMA-induced cell damage by preventing MMP dissipation, complex IV inactivation, ATP loss, [Ca2+]i elevation and oxidative stress. These results suggest that apocynin has a protective effect against AMA-induced cell damage by its antioxidant effects and the attenuation of mitochondrial dysfunction. Apocynin also induced the activation of PI3K (phosphoinositide 3-kinase), Akt (protein kinase B) and CREB (cAMP-response element-binding protein) inhibited by AMA. All these data indicate that apocynin may reduce or prevent osteoblasts degeneration in osteoporosis or other degenerative disorders.

Effect of pollen from Typha angustata on hydrogen peroxide induced toxicity in osteoblastic MC3T3-E1 cells.

BACKGROUND: Typha angustata is a traditional Chinese medicine, commonly used for a variety of clinical disorders, including atherosclerosis and wound healing. In the present study, the protective effects of T. angustata pollen extract (TE) on the response of osteoblast to oxidative stress were evaluated. MATERIALS AND METHODS: Osteoblastic MC3T3-E1 cells were incubated with H(2)O(2) and/or TE, and markers of osteoblast function and oxidative damage were examined. RESULTS: TE treatment significantly (P < 0.05) reversed the cytotoxic effect of H(2)O(2) and this effect was blocked by ICI182780, suggesting that TE's effect might be partly involved in estrogen action. TE significantly (P < 0.05) increased collagen content, alkaline phosphatase activity, calcium deposition of osteoblasts in the presence of H(2)O(2) and these effects were blocked by rottlerin and PD98059, suggesting that the induction of differentiation by TE is associated with increased activation of protein kinase C and ERK. Moreover, H(2)O(2)-induced reduction of osteocalcin was significantly recovered in the presence of TE. Pretreatment with TE also decreased the increase in receptor activator of nuclear factor-kB ligand, malondialdehyde, and protein carbonyl induced by H(2)O(2). CONCLUSION: These results suggest that the pollen of T. angustata may be useful for the protection of H(2)O(2)-induced oxidative damage and dysfunction in osteoblasts.

Apocynin stimulates osteoblast differentiation and inhibits bone-resorbing mediators in MC3T3-E1 cells.

Apocynin is a naturally occurring methoxy-substituted catechol, experimentally used as an inhibitor of NADPH-oxidase. In the present study, the effect of apocynin on the function of osteoblastic MC3T3-E1 cells was studied. Apocynin caused a significant elevation of alkaline phosphatase (ALP) activity, collagen content, and mineralization in the cells (P<0.05). Antimycin A (AMA), which inhibits complex III of the electron transport system, has been used as a reactive oxygen species (ROS) generator in biological systems. We exposed cultured osteoblastic MC3T3-E1 cells to AMA with or without pretreatment with apocynin. Apocynin significantly (P<0.05) increased cell survival, calcium deposition, and osteoprotegerin release and decreased the production of ROS and osteoclast differentiation inducing factors such as TNF-α, IL-6, and receptor activator of nuclear factor-kB ligand (RANKL) in the presence of AMA. These results demonstrate that apocynin can protect osteoblasts from mitochondrial dysfunction-induced toxicity and may have positive effects on skeletal structure.

Mitochondrial defects and cytotoxicity by antimycin A on cultured osteoblastic MC3T3-E1 cells.

Antimycin A (AMA), which inhibits complex III of the electron transport system, has been used as a reactive oxygen species (ROS) generator in biological systems. We investigated the effects of AMA on various parameters related to mitochondrial function in osteoblastic MC3T3-E1 cells. Here, we show that AMA-induced cell death was accompanied by the loss of ATP, complex I and IV activities, and mitochondrial membrane potential. Moreover, AMA stimulated oxidative stress and induced cytochrome c release from mitochondria in osteoblasts. Our data support AMA-induced death in osteoblasts via a mitochondria-dependent pathway. These biochemical changes in mitochondria were effectively prevented upon pre-treatment with ROS scavengers, indicating that ROS plays a critical role as an upstream controller in the AMA-induced cell dysfunction.

Involvement of PI3K/Akt/CREB and redox changes in mitochondrial defect of osteoblastic MC3T3-E1 cells.

Antimycin A (AMA) is an inhibitor of mitochondrial electron transport via its binding to complex III. In the present study, the mechanisms involved in AMA-induced cell damage were investigated. Treatment of osteoblastic MC3T3-E1 cells with AMA decreased adenosine 3',5'-cyclic monophosphate (cAMP) level, activities of phosphoinositide 3-kinase (PI3K) and Akt (protein kinase B), and phosphorylated CREB (cAMP-response element-binding protein). To examine whether AMA-induced cell damage involves altered metabolism of pyridine nucleotides, the levels of NAD(+), NADH, NADP(+), and NADPH were measured. Treatment with AMA significantly decreased the levels of NAD(+) and NADPH. Moreover, the activities of aconitase and thioredoxin reductase were decreased by AMA treatment. These results suggest that PI3K/Akt/CREB pathway and pyridine nucleotide (NAD(+) and NADPH) are related to mitochondria function of osteoblasts.