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Establishment and evaluation of the primary cultured tibial osteoblast model of broiler chicks

CAO Su-mei, LI Ting-ting, SHAO Yu-xin, ZHAO Yu-zhen, ZHANG Li-yang, LU Lin, ZHANG Ri-jun, HOU Shui-sheng, LIAO Xiu-dong, LUO Xu-gang, WANG Run-lian
2023, 22 (2): 551-558.   DOI: 10.1016/j.jia.2022.08.051
Abstract225)      PDF in ScienceDirect      

Osteoblasts are considered as a major factor contributing to bone development and mineralization, however, few studies have been done to establish and evaluate the primary cultured tibial osteoblast model of broiler chicks.  Therefore, in the present study, two experiments were conducted to establish and evaluate the primary cultured tibial osteoblast model of broiler chicks.  In experiment 1, osteoblasts were isolated from the tibia of one-day-old Arbor Acre male broiler chicks using the explant method and identified through the cell morphology, alkaline phosphatase (ALP) and alizarin red staining.  Experiment 2 was carried out to evaluate the vitality and mineralization of primary cultured tibial osteoblasts of broilers on days 4, 8, 12, 16, 20, 24, 28 and 32 after incubation, respectively.  The results from experiment 1 demonstrated that primary cultured tibial osteoblasts of broilers showed a spindle-shaped, triangular or polygonal morphology.  More than 95% of the cells were stained blue-black after ALP staining, and mineralized nodules were formed after 4 days of continuous incubation.  in experiment 2, lactate dehydrogenase (LDH) activity stayed at a relatively stabilized level although incubation time affected (P=0.0012) it during the whole culture period.  Additionally, incubation time affected (P≤0.0001) the number and proportion of the area of mineralized nodules.  They increased linearly and quadratically (P<0.04) with the increase of incubation time, and remained at a stabilized level from 24 to 32 days of incubation.  The estimates of the optimal incubation time were 17 and 26 days based on the best fitted broken-line or quadratic models (P<0.0001) of the number and proportion of the area of mineralized nodules, respectively.  These results indicate that the primary cultured tibial osteoblast model of broilers has been established successfully by the explant method, and it showed typical osteoblast morphology and characteristics of ALP activity and mineralization, and could maintain a relatively stabilized vitality from 4 to 32 days of incubation; and the optimal incubation time of primary tibial osteoblasts was 17 to 26 days.  Therefore, it could be used to further study the underlying mechanisms of bone development and mineralization of broiler chicks.

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Kinetics of selenium absorption in ligated small intestinal loops of chicks
LIU Guo-qing, ZHANG Shu-min, AN Zhi-min, FENG Yan-zhong, DONG Xue-yu, LI Su-fen, LU Lin, ZHANG Li-yang, WANG Run-lian, LUO Xu-gang, LIAO Xiu-dong
2020, 19 (8): 2095-2102.   DOI: 10.1016/S2095-3119(20)63194-X
Abstract135)      PDF in ScienceDirect      
Selenium (Se) is an essential trace element that has a large number of biological functions for broilers.  However, the absorption kinetics of Se from sodium selenite in the small intestine of broilers remains unclear.  Therefore, two experiments were conducted with 28-d-old commercial male broilers to study the kinetics of Se absorption in ligated small intestinal segments.  In experiment 1, the Se absorption in duodenal, jejunal, and ileal segments at different post-perfusion time points (0, 20, 40, 60, 80, 100 and 120 min) were compared.  In experiment 2, a kinetic study of Se absorption was conducted with the duodenal, jejunal, and ileal loops perfused with solutions containing 0, 0.0375, 0.075, 0.15, 0.30, or 0.60 μg mL–1 of Se as sodium selenite, and Se contents in perfusates were determined at 100 min after perfusion.  The results from experiment 1 showed that the Se absorption increased in an asymptotic response (P<0.0001) to post-perfusion time within 120 min in all the small intestinal segments, but increased linearly (P<0.0001) at less than 100 min after perfusion in duodenal and ileal segments, while more than 96.0% of the maximum Se absorption occurred at 100 min after perfusion in each small intestinal segment.  In experiment 2, there was no difference (P>0.05) in the Se absorption rate among different ligated small intestinal segments perfused with solutions containing 0.0375–0.15 μg mL–1 of Se, however, the Se absorption rate was higher (P<0.05) in the jejunum than that in the duodenum perfused with solutions containing 0.30–0.60 μg mL–1 of Se.  The kinetic curves of Se absorption demonstrated that the Se absorption was a saturated carrier-mediated process in the duodenum, and the maximum absorption rate was 1 271 pg min–1 cm–1; whereas the Se absorptions were a non-saturated diffusion process in the jejunum and ileum, and the diffusive constants were 2 107 and 1 777 cm2 min–1, respectively.  The results from the present study indicate that the jejunum is the main Se absorption site, and the Se absorption is a saturated carrier-mediated process in the duodenum, but a non-saturated diffusion process in the jejunum and ileum of broilers.
 
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