Abstract 1153 Sea cucumber's Role in Bone Health: Exploring Its Impact on Osteogenic Differentiation for Osteoporosis Supplementation

The sea cucumber, Holothuria scabra, or sandfish, is the marine crustaceans that wildly distributed abundant throughout the Indo-Pacific region. Due to a commercially high values species in Asian countries, the previous research also focused on the bioactive compounds, nutritional components and growth factor gene expression. According to the regeneration capacity of sea cucumber, our group studied on the effect of H. scabra extract on human placenta derived mesenchymal stem cells (MSCs) and we found that sea cucumber extract could exert the proliferation rate of MSCs. Additionally, we investigated the tissue expression in sea cucumber by transcriptomic analysis and explored that H. scabra contained a numerous of growth factor genes including TGF-β superfamily, Neurotrophin (NT) and Fibroblast growth factor (FGF) mRNA. Osteoporosis is a major public health problem worldwide, with more risk for bone fractures. Clinical treatments for osteoporosis are limited due to strong adverse events and drug instability. Alternative medical therapeutic agents for bone degeneration mainly focus on natural products with less adverse effects. This study aimed to investigate the effect of IW extracted from H. scabra on proliferation, extracellular matrix maturation, and mineralization of MC3T3-E1 preosteoblasts and and downstream signaling pathway during osteogenesis illustrated by transcriptomic analysis. The MC3T3-E1 cells were seeded and grown in the plates for 3 days prior to switching to the osteogenic differentiation medium as the culture medium. In the treatment groups, culture media were added with different concentrations of IW extract from H. scabra (0.1, 0.5, 1, and 5 μg/ml) for 21 consecutive days. On day 7, 14, and 21, cells were harvested and quantified for the alkaline phosphatase (ALP) activity and alizarin red S (ARS) staining. The transcriptome analysis was performed after total RNA extraction. The results showed that the optimal concentration of crude IW extract of H. scabra which did not decrease cell viability in long-term exposure was less than 10 μg/ml. On days 14 and 21 of differentiation, ALP levels were significantly enhanced by 1 and 5 μg/ml of IW extract in a dose-dependent manner when compared with the control group. On day 14 of osteogenic differentiation, transcriptional changes compared to the control osteogenic induction medium and osteogenic induction medium containing 5 μg/ml of IW extract were potentially related in four signaling pathways: TGF-β, PI3K-Akt, MAPK, and Wnt signaling pathways. The genes related to the bone formation process were upregulated, including bone morphogenetic protein 8a (Bmp8a), Bmp4, inhibitor of DNA binding 3 (Id3), Id2, collagen, type I, alpha 1 (Col1a1), fibroblast growth factor 2 (Fgf2), fibroblast growth factor receptor 2 (Fgfr2), Axin2, low density lipoprotein receptor-related protein 5 (Lrp5), and frizzled class receptor 1 (Fzd). The study suggests that IW extracted from H. scabra can enhance osteogenic differentiation and mineralization of MC3T3-E1 preosteoblasts through TGF-β, PI3K-Akt, MAPK, and Wnt signaling pathways, which may be used as an alternative naturally derived supplement for treating ostoporosis with minimal side effects. This work was supported by Thammasat University Research Unit in Biotechnology and its application of aquatic animals, the Research Fund of Chulabhorn International College of Medicine, Thammasat University (Grant No. F 4/2562) and the financial support provided by Thai Science Research and Innovation Fundamental Fund (TUFF 96/2566).