Determination of Bone Developments of Rat Anterior and Posterior Extremity Bones in Prenatal and Postnatal Period by Double Staining Method

Mustafa ÖZTÜRK, Erdoğan UNUR, Niyazi ACER, Tolga ERTEKİN, Serife ALPA, Mesut MEKER, Yahya TAHTA
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Abstract

Aim: In our study, we aimed to determine the morphological development of the bones of the anterior and posterior extremity by staining the rat fetus and offspring skeletons with the double staining method. Method: In the current study, seven groups three prenatal (16th, 18th, and 20th days) and four postnatal (0th, 3th, 7th and 12th days) were formed from the foetuses and offsprings obtained from 13 pregnant rats. Then, it was stained with double staining method. Anterior and posterior extremity images of the fetuses and offsprings were examined under a stereo microscope, and ossification findings were determined. Total bone and ossification lengths as well as ossification areas were measured using the ImageJ software. Results: The first cartilage destruction in fetuses occurred on the 16th day of pregnancy in the clavicle, scapula, humerus, radius and ulna; It was seen in the femur, tibia and fibula on the 18th day of pregnancy. The first ossification centres were in the clavicle, scapula and humerus on the 18th day of pregnancy; It was seen in the radius, ulna, femur, tibia, fibula and 2-5 metatarsal bones on the 20th day of pregnancy. The secondary ossification centre was seen on the 0th day (birthday) in the scapula and humerus, on the 7th day after birth in the ulna and radius, and on the 12th day after birth in the femur and tibia. In the study, while the ossification rate in 20-day-old foetuses was 48.9% in the humerus, 53.2% in the radius, 55.7% in the ulna, 33.6% in the femur, 43.2% in the tibia, 44.3% in the fibula, it was determined that it reached 69.7% in the humerus, 78.4% in the radius, 73.3% in the ulna, 63.5% in the femur, 75.5% in the tibia, and 69.2% in the fibula on the 12th day after birth. Conclusion: In this study, we revealed the morphological changes of the anterior and posterior extremity bones of fetuses and offsprings in the normal developmental course. We think that these results will shed light on the studies to be conducted on the detection of skeletal anomalies in teratological studies and contribute to a more comprehensive evaluation of the findings to be obtained from the studies to be conducted.
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双染色法测定大鼠产前、产后前后肢体骨发育
目的:采用双染色法对大鼠胎儿和子代骨骼进行染色,观察其前后肢体骨骼的形态发育情况。方法:选取13只妊娠大鼠的胎仔共7组,分别为产前(第16、18、20天)和产后(第0、3、7、12天)。然后用双染色法进行染色。在立体显微镜下检查胎儿和后代的前肢和后肢图像,并确定骨化结果。使用ImageJ软件测量总骨和骨化长度以及骨化面积。结果:胎儿第一次软骨破坏发生在妊娠第16天的锁骨、肩胛骨、肱骨、桡骨和尺骨;妊娠第18天在股骨、胫骨和腓骨可见。妊娠第18天,第一批骨化中心在锁骨、肩胛骨和肱骨;妊娠第20天桡骨、尺骨、股骨、胫骨、腓骨及2 ~ 5跖骨可见。出生后第0天(生日)肩胛骨和肱骨出现继发性骨化中心,出生后第7天尺骨和桡骨出现继发性骨化中心,出生后第12天股骨和胫骨出现继发性骨化中心。本研究中,20日龄胎儿骨化率肱骨为48.9%,桡骨为53.2%,尺骨为55.7%,股骨为33.6%,胫骨为43.2%,腓骨为44.3%,确定在出生后第12天,肱骨骨化率为69.7%,桡骨为78.4%,尺骨为73.3%,股骨为63.5%,胫骨为75.5%,腓骨为69.2%。结论:本研究揭示了胎儿和子代在正常发育过程中四肢前后骨的形态学变化。我们认为,这些结果将有助于对畸形学研究中骨骼异常检测的研究进行阐明,并有助于对即将进行的研究结果进行更全面的评估。
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