Sıçan Ön ve Arka Ekstremite Kemiklerinin Prenatal ve Postnatal Dönemdeki Kemik Gelişimlerinin İkili Boyama Yöntemi ile Belirlenmesi

Year 2023, Volume: 7 Issue: 2, 190 - 199, 29.10.2023

Mustafa Öztürk , Erdoğan Unur , Niyazi Acer , Tolga Ertekin , Serife Alpa , Mesut Meker , Yahya Tahta

https://doi.org/10.30565/medalanya.1262518

Abstract

Amaç: Çalışmamızda sıçan fetus ve yenidoğan iskeletinin ikili boyama yöntemi ile boyanarak, ön ve arka ekstremiteye ait kemiklerin morfolojik gelişimlerinin tespitini amaçladık.

Yöntem: Bu çalışmada 13 adet gebe sıçandan elde edilen fetüs ve yavrulardan 3'ü prenatal (16.,18. ve 20. gün) ve 4'ü postnatal (0, 3.,7. ve 12.gün) olmak üzere 7 grup oluşturuldu. Daha sonra ikili boyama yöntemiyle boyandı. Fetüs ve yavruların ön ve arka ekstremite görüntüleri stereo mikroskop altında incelenerek ossifikasyon bulguları belirlendi. ImageJ yazılımı kullanılarak toplam kemik ve ossifikasyon uzunlukları ile ossifikasyon alanları ölçüldü.

Bulgular: Fetüslerdeki ilk kıkırdak yıkımı clavicula, scapula, humerus, radius ve ulna’da gebeliğin 16. gününde; femur, tibia ve fibula’da ise gebeliğin 18. gününde görüldü. İlk kemikleşme merkezi clavicula, scapula ve humerusta gebeliğin 18. gününde; radius, ulna, femur, tibia, fibula ve 2-5 metatarsal kemiklerde gebeliğin 20. gününde görüldü. İkincil kemikleşme merkezi scapula ve humerusta 0. günde (doğum günü), ulna ve radiusta doğumdan sonra 7. günde, femur ve tibiada doğumdan sonra 12. günde görüldü. Çalışmada 20 günlük fetuslerde kemikleşme oranı humerus’ta %48.9, radius’ta %53.2, ulna’da %55.7, femur‘da %33.6, tibia’da %43.2, fibula’da %44.3’ken, doğumdan sonraki 12. günde humerus’ta %69.7, radius’ta %78.4, ulna’da %73.3, femur’da %63.5, tibia’da %75.5, fibula’da %69.2’e ulaştığı tespit edildi

Sonuç: Bu çalışmada, fetüslere ve yenidoğan yavrulara ait ön ve arka ekstremite kemiklerinin normal gelişim seyrindeki morfolojik değişimlerini ortaya koyduk. Bu sonuçların teratolojik çalışmalarda iskelet anomalilerinin (malformasyon, varyasyon ve diğer anomaliler) tespitine yönelik yapılacak çalışmalara ışık tutacağını ve yapılacak çalışmalardan elde edilecek bulguların daha kapsamlı değerlendirilmesine katkı sunacağını düşünmekteyiz.

Keywords

Sıçan, İkili Boyama, Kemikleşme, Kemik Gelişimi, Image J, Toksikoloji.

Supporting Institution

Erciyes Üniversitesi Bilimsel Araştırma Projeleri Birimi

Project Number

TSY-11-3723

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

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.

Keywords

Rat, Double Staining, Ossification, Bone Development, Image J, Toxicology.

Project Number

TSY-11-3723

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