Dr LIMY WONG1,2, Dr JENNY YY OOI2, Mr YUNG SHING TSANG2, Professor LAWRENCE P MCMAHON1,2
1Eastern Health, Box Hill, Australia, 2Monash University Eastern Health Clinical School, Box Hill, Australia
Biography:
Dr Limy Wong is a Nephrologist and Postdoctoral Fellow at Eastern Health. She graduated with First Class Honours in Medicine from the Royal College of Surgeons in Ireland and completed Nephrology specialist training in Ireland and the UK. She completed a Wellcome Trust Clinical PhD Fellowship at the University of Cambridge, studying the genetics of ANCA-associated vasculitis. She was awarded the 2021 and 2022 RACP Jacquot Research Establishment Fellowship, which support her current research focus in understanding the biological basis of sarcopenia in patients with chronic kidney disease and to determine the relationship between sarcopenia and renal osteodystrophy.
Aim:
This study aimed to investigate the biochemical interactions between osteoblasts and myoblasts using an in vitro co-culture system that more accurately simulates the physiological environment.
Background:
Chronic kidney disease (CKD) leads to significant musculoskeletal decline, characterised by the concurrent loss of bone and skeletal muscle mass—sarcopenia. Increasing research reveals that bone and muscle engage in biochemical and molecular crosstalk beyond simple mechanical interaction. Much of the current understanding stems from simplistic monolayer cell models, which fail to reflect the complexity of in vivo environments, especially in the presence of uraemic toxins. A deeper understanding of this bone-muscle communication could yield novel therapeutic targets for CKD-related musculoskeletal deterioration.
Methods:
Normal human osteoblasts (NHost) and human skeletal muscle myoblasts (HSMM) were cultured separately and together using Transwell plates. Osteoblasts were differentiated over 14 and 21 days, while myoblasts were induced to form myotubes over 7 days. To replicate uraemic conditions, myoblasts were treated with indoxyl sulphate (IS), a uraemic toxin, before co-culture. Myoblast proliferation was assessed using BrdU assay, and differentiation was evaluated via myotube fusion index and diameter. Proteomic analysis of conditioned media was performed using liquid chromatography-mass spectrometry (LC-MS).
Results:
Enhanced myoblast proliferation was observed in co-culture compared to monoculture, even in the presence of IS, although myogenic differentiation remained unchanged. LC-MS identified 29 unique proteins in the co-culture media, including potential osteoblast-derived osteokines tenascin-C (TNC), glypican-1 (GPC1), and collagen type XI alpha 2 (COL11A2), suggesting a paracrine effect.
Conclusion:
Osteoblasts promote myoblast proliferation through potential secreted factors, highlighting the utility of co-culture models to study bone-muscle interactions and their therapeutic implications for CKD patients.
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