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Effects of L-tryptophan and 1,25(OH)2D3 on the osteogenic differentiation of mesenchymal stem cells isolated from compact bones of broilers and layers

2022 CPRF,Broilers
Poultry birds are vulnerable to bone problems throughout their life or production period due to a rapid growth rate in broilers and an active laying cycle in layers. The calcium-sensing receptor (CaSR) and vitamin D receptor (VDR) are important in calcium and bone metabolism. The objective of this study was to investigate the effect of ligands of CaSR (L-Trp) and VDR [1,25-dihydroxycholecalciferol, 1,25(OH)2D3] on the proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs) isolated from compact bones in 14-day-old Ross 308 chickens and Dekalb pullets. Results showed that the high doses (10 and 15 mM) of L-Trp inhibited proliferation but promoted osteogenic differentiation, and 5 mM L-Trp had a balanced effect between proliferation and osteogenic differentiation in broiler and layer MSCs. Therefore, 5 mM L-Trp was selected for further experiments. The broiler and layer MSCs were then treated with (1) osteogenic medium, osteogenic medium supplemented with (2) 1 nM 1,25(OH)2D3, (3) 2.5 mM Ca2+, (4) 2.5 mM Ca2+ + 5 mM L-Trp, (5) 2.5 mM Ca2+ + 5 mM L-Trp + 1 μM NPS-2143, respectively for 7 days. All data were expressed as mean ± SEM with at least three biological repeats. Statistical significance was determined using one-way ANOVA followed by Dunnett’s multiple-comparison test. P-value < 0.05 was considered statistically significant. The 5 mM L-Trp significantly inhibited the proliferation of broiler and layer MSCs on day 7 (P < 0.05), but 1 nM 1,25(OH)2D3 significantly promoted the proliferation of layer MSCs (P < 0.05). The osteogenic differentiation process in layer MSCs was faster than that in broiler MSCs. Only 2.5 mM Ca2+ + 5 mM L-Trp group significantly increased the mineralization process during osteogenic differentiation in broiler MSCs (P < 0.05). The NPS-2143 supplementation did not block the positive effect of 5 mM L-Trp on ALP activity and the mRNA expression of osteogenic genes. In layer MSCs, Ca2+ alone significantly facilitated mineralization and ALP activity after 7-day osteogenic differentiation (P < 0.05). However, different treatments selectively regulated the mRNA expression of CaSR, VDR, osteogenesis-related genes, β-catenin, and NFATc1 without a consistent trend. Taken together, CaSR and VDR were expressed in boiler and layer MSCs, and L-Trp and 1,25(OH)2D3 can regulate proliferation and osteogenic differentiation in both broiler and layer MSCs depending on the modulator doses and treatment time, which provided evidence that CaSR and VDR could be potential targets to regulate bone formation and remodeling in broilers and layers.
Tags :
1,25(OH)2D3,bone health,L-tryptophan,stem cells
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Presenter

PhD Candidate

University of Manitoba

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