Basit yıkama yöntemi ile hazırlanmış semen örneklerinde trombositten zengin plazmanın (TZP) etkisinin değerlendirilmesi
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Dosyalar
Tarih
2019
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Necmettin Erbakan Üniversitesi Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
AMAÇ: Trombositten Zengin Plazma (TZP)'nın içinde bulunan büyüme faktörlerinin sperm parametrelerine etkisinin olup olmadığını araştırmaktır. TZP, kandan elde edilen yüksek konsantrasyonda trombosit içeren plazma olarak karşımıza çıkmaktadır ve hâlihazırda birçok alandaki araştırmalarda ve tedavilerde kullanımı mevcuttur. TZP'de temel yaklaşım, trombositleri patlatma aşamasından sonra büyüme faktörlerini ve sitokinleri açığa çıkarıp proliferasyonu, yenilenmeyi ve farklılaşmayı uyararak hücresel çoğalma, kollajen üretimi, hyaluronik asit üretimi, epidermal hücre büyümesi, anjiyogenez gibi sistemleri harekete geçirerek farklı tipteki tedavilerde kullanılabilir yapmaktır. Bizim asıl amacımız ise semen örneklerinde kullanmak için hazırlayacağımız TZP ile spermi çift yıkama yapmak, daha sonrasında ise TZP 'nin bazı semen parametrelerine (motilite, sayı) olan etkisine bakmaktır. Ayrıca, TZP'nin sperm hazırlama tekniklerinden biri olan basit yıkama işleminde kullanılan medyumlara ek bir alternatif olması beklenmektedir. MATERYAL- METOD: Gönüllü onam formlarıyla bilgilendirilmiş 10 gönüllünün her birinden 10 cc kan alındı. Gönüllülerden alınan bu kanlardan TZP elde edilerek çalışmamıza dâhil edildi. Gönüllü onam formları ile bilgilendirilmiş ve spermiyograma gelen 20 hastadan azospermi grupları hariç diğer tüm gruplar çalışmaya dâhil edildi. Alınan numunelerinin spermiyogram sonuçlarına bakıldıktan sonra atık hedefli çalışmamıza geçildi. Likefiye olan semen kontrol ve deney grubu olmak üzere eşit hacimde 2 grupta randomize edildi. Kontrol grubu PBS ile rutinde olan basit yıkama metoduyla iki kere yıkandı. Deney grubunda ise semen örnekleri TZP ile muamele edilerek iki kere yıkandı ve her iki grubun 15. , 30. , 45. dk. ve son olarak 2. santrifüj sonrası sonuçlarına bakılarak karşılaştırmalar yapıldı ve istatistiksel olarak değerlendirildi. BULGULAR: Çalışmamıza yaşları 18-50 arasında olan 20 gönüllü hasta katıldı. Spermiyograma gelen ve azospermi hariç tüm hastalar çalışmamıza dâhil edildi. Spermiyogram analizleri yapıldıktan sonra azospermi olduğu anlaşılan spermler çalışma dışı bırakılıp yeniden gönüllü formlarıyla bilgilendirilen hastalardan tekrardan örnekler alınıp çalışmaya devam edildi. Deney ve Kontrol gruplarımızda basit yıkama yapıldığı ve yıkama medyumları da 1/1 kullanıldığı için konsantrasyon analizimizde bir değişiklik gözlemlenmedi. TZP grubundaki+4 motilite analizinde grup etkisi p=0.0004, zaman etkisi p<0.0001, grup-zaman etkisinin p değeri 0.02 olduğu ve tüm bu değerlerin 0.05'den küçük olduğu dikkate alındığında analizin bu kısmı anlamlı bulundu. +3 motilite analizinde grup etkisi p<0.0001, zaman etkisi p<0.0001, grup-zaman etkisinin p<0.0001 olduğu ve tüm bu değerlerin 0.05'den küçük olduğu dikkate alındığında analizin bu kısmı anlamlı bulundu. +2 motilite analizinde grup etkisi p=0.04 olduğu için sonuç anlamlı, zaman etkisi p=0.1, grup-zaman etkisinin p=0.4 olduğu ve tüm bu değerlerin 0.05'den büyük olduğu dikkate alındığında analizin bu kısmı anlamlı bulunmadı. +1 motilite analizinde grup, zaman ve grup-zaman etkisi p<0.0001 olduğu için anlamlı bulunmuştur. +3 ve +4 motilite değerlerinin toplam analizinde ise grup, zaman, grup-zaman etkisi p<0.0001 olduğu yani p<0.05 olduğu için sonuç anlamlı bulunmuştur. SONUÇ: Çalışmamızda azospermi hariç çalışmaya dâhil edilen gruplarda ilk 15. dakikada istatistiksel olarak anlamlı kabul edilen motilite artışı gözlemlenmiştir. TZP'nin ilerleyen zaman diliminde yani çalışma sürelerimiz olan 30 ve 45.dk.'larda etkisini kaybederek spermin motilitesinde bir azalma gösterilmiştir. 45.dk sonrası yapılan 2. santrifüj sonrası TZP ile yeniden muameleden sonra ölçüme bakıldığında ilk 15.dk kadar olmasa da yine anlamlı bir artma meydana gelmiştir.
OBJECTIVE: The aim of this study was to investigate the effect of growth factors on sperm parameters in platelet-rich plasma (PRP). PRP is a high concentration platelet-containing plasma obtained from blood and it is currently used in many research and treatment areas. The basic approach in PRP is making it usable in different types of treatments by activating systems such as the proliferation that occurred by extracting growth factors and cytokines after the blasting step of platelets, cellular proliferation, stimulating proliferation, collagen production, hyaluronic acid production, epidermal cell growth, angiogenesis and more which obtained by inducing regeneration and differentiation. Our main goal is looking for some effects of PRP on some semen parameters (motility, concentration) after double washing the sperm with the PRP which will be prepared to use in semen samples. Moreover, TZP is expected to be an additional alternative to the mediums used in the simple washing process, which is one of the techniques of sperm preparation. MATERIAL AND METHODS: 10 cc blood was collected from each of 10 volunteers who were informed by their volunteer consent forms. PRP was obtained from the blood samples taken from volunteers and included in our study. All the groups were included in the study except the azoospermia groups among 20 patients who were informed by their consent forms and were given the spermiogram. After looking at the spermiogram results of the samples taken, our waste-targeted study started. Semen was randomized into two groups of equal volume, control and experimental group. The control group was washed twice with PBS using a simple washing method. In the experimental group, semen samples were washed twice by treating it with PRP in each 15th, 30th, 45th minutes. Finally, after the second centrifugation, the results were compared and evaluated statistically. FINDINGS: Twenty volunteer patients ages between 18-50 participated in our study. All patients were included in our study except azoospermias. After the analysis of the spermiogram, the patients who were found to have azoospermia were excluded from the study. No change was observed in our concentration analysis since 1/1 of the wash media was used and simple washing was performed in our experiment and control groups. In the + 4 motility analysis of the PRP group, the effect of group was p = 0.0004, the time effect was p <0.0001, the p-value of group-time effect was 0.02 and because all of these values were less than 0.05, this part of the analysis was found significant. In the +3 motility analysis, the effect of the group was p <0.0001, the time effect was p <0.0001, the group-time effect was p <0.0001 and also this part was found significant because all these values were less than 0.05. In the +2 motility analysis, since the effect of the group was p = 0.04, the result was significant, the time effect was p = 0.1, the group-time effect was p = 0.4 and all of these values were greater than 0.05 so this part of the analysis was not considered significant. In the +1 motility analysis, the group was found to be significant since the time and group-time effect were p <0.0001. In the total analysis of +3 and +4 motility values, the result was significant since the group, time and group-time effects were p <0.0001 which are p <0.05. CONCLUSION: In our study, the increase of the motility is observed and accepted as statistically significant at the first 15th minute for each group that included to the study, except for azoospermia. A decrease in sperm motility has shown in the 30th and 45th minutes of the treatment period of the PRP. After the second centrifugation, which is done after 45 minutes, samples are re-treated with PRP and an important increase in sperm motility was observed again but not as much as it was in the first 15 minutes of our study.
OBJECTIVE: The aim of this study was to investigate the effect of growth factors on sperm parameters in platelet-rich plasma (PRP). PRP is a high concentration platelet-containing plasma obtained from blood and it is currently used in many research and treatment areas. The basic approach in PRP is making it usable in different types of treatments by activating systems such as the proliferation that occurred by extracting growth factors and cytokines after the blasting step of platelets, cellular proliferation, stimulating proliferation, collagen production, hyaluronic acid production, epidermal cell growth, angiogenesis and more which obtained by inducing regeneration and differentiation. Our main goal is looking for some effects of PRP on some semen parameters (motility, concentration) after double washing the sperm with the PRP which will be prepared to use in semen samples. Moreover, TZP is expected to be an additional alternative to the mediums used in the simple washing process, which is one of the techniques of sperm preparation. MATERIAL AND METHODS: 10 cc blood was collected from each of 10 volunteers who were informed by their volunteer consent forms. PRP was obtained from the blood samples taken from volunteers and included in our study. All the groups were included in the study except the azoospermia groups among 20 patients who were informed by their consent forms and were given the spermiogram. After looking at the spermiogram results of the samples taken, our waste-targeted study started. Semen was randomized into two groups of equal volume, control and experimental group. The control group was washed twice with PBS using a simple washing method. In the experimental group, semen samples were washed twice by treating it with PRP in each 15th, 30th, 45th minutes. Finally, after the second centrifugation, the results were compared and evaluated statistically. FINDINGS: Twenty volunteer patients ages between 18-50 participated in our study. All patients were included in our study except azoospermias. After the analysis of the spermiogram, the patients who were found to have azoospermia were excluded from the study. No change was observed in our concentration analysis since 1/1 of the wash media was used and simple washing was performed in our experiment and control groups. In the + 4 motility analysis of the PRP group, the effect of group was p = 0.0004, the time effect was p <0.0001, the p-value of group-time effect was 0.02 and because all of these values were less than 0.05, this part of the analysis was found significant. In the +3 motility analysis, the effect of the group was p <0.0001, the time effect was p <0.0001, the group-time effect was p <0.0001 and also this part was found significant because all these values were less than 0.05. In the +2 motility analysis, since the effect of the group was p = 0.04, the result was significant, the time effect was p = 0.1, the group-time effect was p = 0.4 and all of these values were greater than 0.05 so this part of the analysis was not considered significant. In the +1 motility analysis, the group was found to be significant since the time and group-time effect were p <0.0001. In the total analysis of +3 and +4 motility values, the result was significant since the group, time and group-time effects were p <0.0001 which are p <0.05. CONCLUSION: In our study, the increase of the motility is observed and accepted as statistically significant at the first 15th minute for each group that included to the study, except for azoospermia. A decrease in sperm motility has shown in the 30th and 45th minutes of the treatment period of the PRP. After the second centrifugation, which is done after 45 minutes, samples are re-treated with PRP and an important increase in sperm motility was observed again but not as much as it was in the first 15 minutes of our study.
Açıklama
Yüksek Lisans Tezi
Anahtar Kelimeler
Sperm Motilite ve Morfoloji, Sperm Yıkama, Trombosit, Trombositten Zengin Plazma (TZP), Sperm Motility and Morphology, Sperm Washing, Platelet, Platelet Rich Plasma (PRP)
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Duran, M. (2019). Basit yıkama yöntemi ile hazırlanmış semen örneklerinde trombositten zengin plazmanın (TZP) etkisinin değerlendirilmesi. (Yayınlanmamış Yüksek Lisans Tezi). Necmettin Erbakan Üniversitesi, Sağlık Bilimleri Enstitüsü Histoloji Ve Embriyoloji Anabilim Dalı, Konya
