Biology subjects

Ersoy, U.

Publications and source records attributed to Ersoy, U..

3 recordsLinked to original sources

bioRxiv2024

Defining the most potent osteoinductive culture conditions for MC3T3-E1 cells reveals no implication of oxidative stress or energy metabolism

MC3T3-E1 preosteoblastic cell line is widely utilised as a reliable in vitro system to assess bone formation. However, the experimental growth conditions for these cells hugely diverge and, particularly, the osteogenic medium (OSM) composition varies in research studies. Therefore, we aimed to define the ideal culture conditions for MC3T3-E1 subclone 4 cells with regards to mineralization capacity and explore if oxidative stress or cellular metabolism processes are implicated. Cells were treated with 9 different combinations of long-lasting ascorbate (Asc) and {beta}-glycerophosphate ({beta}GP) and osteogenesis/calcification were evaluated at 3 different time-points by qPCR, Western blotting and bone nodule staining. Key molecules of oxidative and metabolic pathways were also assessed. It was found that sufficient mineral deposition was achieved only in 150g.mL-1/2mM Asc/{beta}GP combination at 21d in OSM and this was supported by Runx2, Alpl, Bglap and Col1a1 expression levels increase. NOX2 and SOD2 as well as PGC1 and Tfam were also monitored as indicators of redox and metabolic processes, respectively, where no differences were observed. Elevation in OCN protein levels and ALP activity showed that mineralisation comes as a result of these differences. This work defines the most appropriate culture conditions for MC3T3-E1 cells and could be used by other research laboratories in this field.

cell biology↗

bioRxiv2023

Lifelong dietary protein restriction accelerates skeletal muscle loss and reduces muscle fibre size by impairing proteostasis and mitochondrial homeostasis

The early life environment significantly affects the development of age-related skeletal muscle disorders. However, the long-term effects of lactational protein restriction on skeletal muscle are still poorly defined. Our study revealed that male mice nursed by dams fed a low-protein diet during lactation exhibited skeletal muscle growth restriction. This was associated with a dysregulation in the expression levels of genes related to the ribosome, mitochondria and skeletal muscle development. We reported that lifelong protein restriction accelerated loss of type-IIa muscle fibres and reduced muscle fibre size by impairing mitochondrial homeostasis and proteostasis at 18 months of age. However, feeding a normal-protein diet following lactational protein restriction prevented accelerated fibre loss and fibre size reduction in later life. These findings provide novel insight into the mechanisms by which lactational protein restriction hinders skeletal muscle growth and includes evidence that lifelong dietary protein restriction accelerated skeletal muscle loss in later life.

physiology↗

bioRxiv2023

The effect of lactational low protein diet on skeletal muscle during adulthood and ageing in male and female mouse offspring

Sarcopenia is characterised by loss of skeletal muscle mass and function associated with a reduction in muscle fibres. External factors, like exercise and diet, can also influence skeletal muscle mass and contribute to muscle fibre loss. Maternal programming refers to the effect of maternal environmental factors such as nutrition that lead to phenotypic changes in the offspring. Maternal malnutrition has been linked to a reduction in body weight and impaired development of skeletal muscle of the offspring; however, there are no studies that reported the long-term effect of maternal low protein diet on the ageing of skeletal muscles. This study aimed to examine how maternal protein deficiency during lactation affects skeletal muscle development and ageing in the offspring. Pups born from normally fed mothers were lactated by low protein fed mothers. Post-weaning, mice were either maintained on a low protein diet (LPD) or switched to normal protein diet (NPD). Pups born from normally fed mothers and maintained on NPD during lactation and afterwards were used as control. In males, the diet mainly affected the size of the myofibres without major effect on fibre number and led to a reduced grip strength of ageing mice (24 months). Female mice had a lower body and muscle weight at weaning but caught up with control mice at 3 months. During ageing, muscle weight, myofibre number and survival rate of female pups were significantly affected. These findings highlight longitudinal animal research for nutritional programming and the importance of sexual dimorphism in response to challenges. HighlightsO_LIPostnatal low protein diet significantly decreases the survival rate of female but not male mice. C_LIO_LIDuring ageing, female mice fed a low protein diet during lactation have lower muscle weight. C_LIO_LIDuring ageing, female mice fed a low protein diet postnatally maintain their myofibre number. C_LIO_LIMale mice fed a low protein diet postnatally have lower body weight and muscle weight throughout their lifespan. C_LIO_LILow protein diet affects myofibres size of TA muscle of male but not female mice at 3 months of age however this effect is lost during ageing. C_LI

physiology↗