Stimulatory effects of creatine on metabolic activity, differentiation and mineralization of primary osteoblast-like cells in monolayer and micromass cell cultures

Abstract

The effects of creatine (Cr) supplementation on primary rat osteoblast-like cells cultured as monolayer and micromass were investigated. Cr was added to the medium at concentrations of either 10 or 20 mM. At various time points, the cell cultures were analyzed morphologically, metabolically and biochemically.The degree of differentiation of primary osteoblast-like cell cultures was higher in micromass cultures compared to monolayer cultures, as judged by alkaline phosphatase (ALP) activity and extent of mineralization. In both culture systems, Cr supplementation showed positive effects, which were dependent on the organizational level of the osteoblast-like cells in such a way that the cells in monolayer culture showed significantly increased metabolic activity, ALP activity and mineralization in the presence of Cr than without the supplement. In micromass cultures, Cr also significantly enhanced ALP activity and mineralization, without affecting metabolic activity. The effect of Cr on ALP activity was more pronounced at higher concentrations of Cr, but 20 mM Cr already showed some adverse effects on cell viability. In conclusion, chemically pure Cr added to low serum cell culture medium has a stimulatory effect on metabolic activity, differentiation and mineralization of osteoblast-like cells indicating that Cr supplementation could also be used as a potential clinical intervention to stimulate cell growth, differentiation and mineralization during bone repair in vivo.

Influência da suplementação de creatina sobre a massa óssea de ratos espontaneamente hipertensos

“…11,12 In fact, a study has shown that incubating Cr in a culture medium with primary osteoblasts has stimulating effects on the differentiation, metabolic activity, and bone mineralization, elevating the phosphorylcreatine/Cr ratio and preserving the ultrastructure and mitochondrial function of osteoblasts. 8 In vivo evidence 13,14 has corroborated those findings. Supplementation with Cr can increase the BMD and cause benefi cial biomechanical adaptations in the femur of healthy rats.…”

“…7 The energy need of bone cells to survive, proliferate, differentiate, and synthesize extracellular matrix is known to be high. 8 Evidence has shown that part of the energy required for those processes originates from creatine (Cr; α-methyl guanidine-acetic acid), which plays a central role in maintaining Rev Bras Reumatol 2012;52(3):453-461 ATP and ADP levels in several tissues, such as skeletal muscle, brain, testicles, cartilage, and bone (for a recent and comprehensive review, see Wallimann et al 9 ). The hypothesis that Cr could play an important role in bone metabolism was fi rst suggested based on the identifi cation of creatine kinase isoforms (CK), enzyme responsible for the reversible reaction as follow: phosphocreatine + ADP + H + creatine + ATP in the bone.…”

Información sobre la creatina durante el siglo XXI en España = Information about creatine during the 21st century in Spain

“…Sin embargo, evidencias posteriores demostraron que la creatina induce la expresión de genes y proteínas relacionados con la hipertrofia (Deldique, 2008;Safdar, 2008), así como la activación de las células satélite (Olsen, 2006). Además, como resultado de este aumento de masa muscular, se produce un incremento en la densidad mineral ósea (Chilibeck, 2005;Gerber, 2005). Recientemente, el interés suscitado por la creatina, se ha trasladado al ámbito clínico, en el cual sus beneficios han sido reconocidos en el tratamiento de enfermedades caracterizadas por la pérdida de masa muscular, la disminución de la densidad ósea, problemas articulares, trastornos del sistema nervioso central y trastornos metabólicos (Gualano, 2010).…”

Efeitos do exercício físico associado à suplementação de creatina na massa óssea de ratas ovariectomizadas

“…De fato, o osso é um tecido que requer grande aporte energético para realizar os processos de diferenciação, proliferação e síntese de matriz óssea (Gerber et al, 2005). Esses processos decorrem da energia proveniente da glicólise, oxidação fosforilativa e do sistema ATP⁄PCr (Wallimann et al, 1994).…”

“…Os autores sugerem que a oferta de Cr tenha sido responsável por salientar a provisão de energia no tecido ósseo através do sistema ATP⁄PCr, contribuindo para a diferenciação das células ósseas, favorecendo, portanto, a formação óssea (Antolic et al, 2007;Gerber et al, 2005). Em consonância com esse achado, um recente estudo demonstrou que a suplementação de creatina foi capaz de aumentar o CMO (i.e.…”