Sıçanlarda polidioksanon ip, hyaluronik asit dolgu ve botulinum toksin-A uygulamalarının deri ve bağ dokusu ile fotoyaşlanma üzerindeki etkilerinin araştırılması
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Dosyalar
Tarih
2020
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Yayıncı
Necmettin Erbakan Üniversitesi Meram Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu projede polidioksanon (PDO) ip, hyaluronik asit (HA) dolgu ve botulinum toksin-A (BTX-A) uygulamaları tek ve kombin kullanım yoluyla değerlendirilmiş olup, deri ve derialtı dokuda oluşturdukları olumlu ya da olumsuz değişimler, kombine uygulamalarında birbirleri üzerindeki sinerjistik ya da antagonistik etkileri, ultraviolet (UV) maruziyetiyle oluşan fotohasar ve fotoyaşlanma süreçlerinde bu etkilerin nasıl değişeceği incelenmiştir. Bu sayede insan derisindeki yaşlanmada seçilecek yönteme karar verilmesi için bakış açısı oluşturulması hedeflenmiştir. Yöntem Projemizde Wistar albino sıçanlar (2-5 aylık, 200-250 g ağırlığında, erkek, 20 adet 10'arlı 2 grup) çalışmaya alınmıştır. Sıçanların bakım ve beslenmeleri Necmettin Erbakan Üniversitesi KONÜDAM Deneysel Tıp Uygulama ve Araştırma Merkezi'nde yapılmıştır. Tüm sıçanların sırt derisi 5 eşit parçaya bölünüp 1-2-3-4-5 olarak numaralandırılmış ve sırt bölgeleri tıraşlandıktan sonra uygun anestezik ajanla genel anesteziye alınmışlardır. 1 nolu alan kontrol alanı olup buraya ip'siz iğne derialtına uygulanıp çıkarılmıştır. 2.alana PDO ip uygulaması deri altında Panniculus carnosus (PC) bölgesine uygulanmıştır. 3.alana HA maddesi intradermal dolgu olarak 0.02 ml uygulanmıştır. 4.alana PDO ip uygulaması ve aynı seansta HA 0.02 ml uygulaması yapılmıştır. 5.alana ise PDO ip ile beraber aynı seansta HA 0.02 ml ve BTX-A 1U olarak uygulanmıştır. İkinci gruptaki 10 sıçana ise aynı işlemler yine tek seferlik uygulanıp 320-400 nm dalga boyunda UVA, 138 mj/cm2 ve 290-320 nm dalga boyu olan UVB 130 mj/cm2 haftada 3 gün olmak üzere 12 hafta boyunca uygulanmıştır. Her 2 gruptaki sıçanların genel durumu haftalık olarak kontrol edilmiştir. Uygulama bölgeleri (ip aksında) ve çevresini de kapsayan doku örnekleri, transvers kesit histolojik değerlendirme ve PCR ekspresyon analizi için 5 mm punch biyopsi işlemi ile başlangıçta, 4. ve 12. haftalarda elde edilmiştir. Histopatolojik olarak epidermis ve dermis kalınlıkları, dermisteki kollagen ve elastinin kalitatif düzeyleri ve dizilimleri, neokollagenez ve yeni elastin sentezi, yabancı cisim granulomları, fibröz kapsül yapısı, histiosit ve dev hücre formasyonu, fibroblast sayıları, inflamatuar hücreler incelenmiş ve kıyaslamalı olarak değerlendirilmiştir. Kollagen 1 (Col1A1), Kollagen-3 (Col3A1), elastin, matriks metalloproteinaz (MMP) 2 ve 9, Nüklear faktör kappa B (NF-κB), interlökin-6 (IL-6) değerleri RT-QPCR ile ölçülmüştür. İstatistiksel hesaplamalarda histopatolojik değerlendirme için SAS University Edition 9.4 programı kullanılarak yapılmış ve RT-qPCR analizi için web tabanlı "RT² Profiler™ PCR Array Data Analysis" programı kullanılmıştır. p<0.05 istatistiksel olarak anlamlı kabul edilmiştir. Bulgular Çalışma 18 sıçanla tamamlanmıştır. Histopatolojik olarak solar elastoz UV alan grupta anlamlı olarak yüksek çıkmıştır. Tüm uygulama alanlarında kollajenin kalitatif düzeyi ve organizasyonu kontrol alanı olan 1. alana kıyasla yüksek bulunmuştur. UV alan deney grubunda UV almayan kontrol gruba kıyasla kollajen histopatolojik olarak daha az ve daha dezorganize saptanmıştır. PCR incelemede Col1A1 ve Col3A1 düzeylerinde UV uygulamasının etkisiyle anlamlı düşüşler gözlenmiştir. İnflamasyon ve vaskülarizasyonda kontrol alana göre materyal uygulanan alanlarda anlamlı artışlar gözlenmiştir. Histopatolojik olarak kapsül yapısı, dev hücre granülomu ve yabancı cisim reaksiyonu da madde uygulanan alanlarda gözlenmiş ve gösterilmiştir. Sonuç Çalışma neticesinde yaşlanma ve fotoyaşlanmada etkili olabilecek PDO ip uygulama, HA dolgu maddesi ve BTX-A'nın deri ve derialtı dokuda meydana getirdikleri reaksiyonlar neticesinde kollajen liflerinin organize düzenlenmesi ve ekspresyonlarını artırması gösterilmiştir. Fotoyaşlanmayı gösteren solar elastozda ve NF-KB ekspresyonunda artış, tip 1 ve tip 3 kollajen düzeylerinde ise azalış anlamlı olarak gösterilmiştir. UV uygulanan grupta (fotoyaşlanma) tek başına dolgu uygulamasının, UV almayan grupta (fizyolojik yaşlanma) ip+dolgu kombine uygulamasının kronik dönemde (12.hafta) antiaging olarak daha effektif sonuçlar getirdiği gözlenmiştir.
This project evaluates polydioxanone (PDO) thread, hyaluronic acid (HA) injection and botulinum toxin type A (BTX-A) practices via single and combined use and analyzes their positive or negative effects on skin and subcutaneous tissue, the synergistic or antagonistic impacts they have on each other during combined application and how these impacts may change during photo damage and photo aging processes caused by UV exposition. It is aimed to create a perspective to make a decision about the method on aging of human skin. Materials and Methods Our project includes Wistar albino rats (2-5 months, weighing 200-250 g, male, and 2 groups consisting of 10). The rats were taken care of and fed in Experimental Medicine Application and Research Centre (KONUDAM) in Necmettin Erbakan University. The skin on the back of all rats were separated into 5 equal parts and given numbers from 1 to 5. After shaving their backs, general anesthesia was induced through a suitable anesthetic agent. Area No. 1 is the control area and thread without needle was applied under the skin and removed from this location. On the Area No. 2, PDO thread application was realized on panniculus carnosus (PC) area of the rats. On the Area No. 3, HA material was applied as 0.02 ml intradermal injection. On the Area No. 4, both PDO thread application and HA 0.02 ml application during the same session were realized. PDO thread and HA 0.02 ml and BTX-A 1U was applied on the Are No. 5 within the same session. The same procedures were applied on 10 rats in the second group only once and then 320-400 nm wavelength UVA 138 mj/cm2 was applied 3 times per week, with combination of 290-320 nm wavelength UVB 130 mj/cm2 for 12 weeks. The general conditions of the rats in both groups were checked on a weekly basis. The tissue samples including the areas of and around the application will initially be obtained during the 4th and 12th weeks through 5 mm punch biopsy for the histological evaluation of transverse section. Histopathologically epidermis and dermis thickness, qualitative levels and alignment of collagen and elastin in dermis, neocollagenesis and new elastin synthesis, foreign body granulomas, fibrosis capsule structure, histiocyte and giant cell formation, fibroblast counts, inflammatory cells were examined and comparatively evaluated. Collagen 1, Collagen 3, elastin, MMP-2, MMP-9, NF-κB, IL-6 gene expression levels were measured. In the statistical analysis, SAS University Edition 9.4 for the histopathological assessments and "RT² Profiler™ PCR Array Data Analysis" for the RT-qPCR analysis were used. p<0.05 value were accepted significant. Results The study was completed with 18 rats. Histopathologically, solar elastosis was significantly higher in the group receiving UV. The qualitative level and organization of collagen in all application areas were found to be higher compared to the control area 1. In the experimental group receiving UV, collagen was less histopathologically less and more disorganized than the non-UV control group. In the PCR examination, significant decreases were observed in Col1A1 and Col3A1 levels due to the effect of UV application. In inflammation and vascularization, significant increases were observed in the areas where the material was applied compared to the control area. Histopathologically, capsule structure, giant cell granuloma and foreign body reaction were also observed and demonstrated in the areas where the substances were applied. Conclusion As a result of the study, it has been shown that PDO thread application, HA filler and BTX-A, which may be effective in aging and photoaging, increase the organized regulation and increase the expression of the collagen fibers. An increase in solar elastosis and NF-KB expression, which shows photoaging, and a decrease in type 1 and type 3 collagen levels were shown to be significant. It was observed that the application of filler alone in the UV treated group (photoaging), and the combination of the application of thread + filler in the non-UV receiving group (physiological aging) brought more effective results in the chronic period (week 12) as antiaging.
This project evaluates polydioxanone (PDO) thread, hyaluronic acid (HA) injection and botulinum toxin type A (BTX-A) practices via single and combined use and analyzes their positive or negative effects on skin and subcutaneous tissue, the synergistic or antagonistic impacts they have on each other during combined application and how these impacts may change during photo damage and photo aging processes caused by UV exposition. It is aimed to create a perspective to make a decision about the method on aging of human skin. Materials and Methods Our project includes Wistar albino rats (2-5 months, weighing 200-250 g, male, and 2 groups consisting of 10). The rats were taken care of and fed in Experimental Medicine Application and Research Centre (KONUDAM) in Necmettin Erbakan University. The skin on the back of all rats were separated into 5 equal parts and given numbers from 1 to 5. After shaving their backs, general anesthesia was induced through a suitable anesthetic agent. Area No. 1 is the control area and thread without needle was applied under the skin and removed from this location. On the Area No. 2, PDO thread application was realized on panniculus carnosus (PC) area of the rats. On the Area No. 3, HA material was applied as 0.02 ml intradermal injection. On the Area No. 4, both PDO thread application and HA 0.02 ml application during the same session were realized. PDO thread and HA 0.02 ml and BTX-A 1U was applied on the Are No. 5 within the same session. The same procedures were applied on 10 rats in the second group only once and then 320-400 nm wavelength UVA 138 mj/cm2 was applied 3 times per week, with combination of 290-320 nm wavelength UVB 130 mj/cm2 for 12 weeks. The general conditions of the rats in both groups were checked on a weekly basis. The tissue samples including the areas of and around the application will initially be obtained during the 4th and 12th weeks through 5 mm punch biopsy for the histological evaluation of transverse section. Histopathologically epidermis and dermis thickness, qualitative levels and alignment of collagen and elastin in dermis, neocollagenesis and new elastin synthesis, foreign body granulomas, fibrosis capsule structure, histiocyte and giant cell formation, fibroblast counts, inflammatory cells were examined and comparatively evaluated. Collagen 1, Collagen 3, elastin, MMP-2, MMP-9, NF-κB, IL-6 gene expression levels were measured. In the statistical analysis, SAS University Edition 9.4 for the histopathological assessments and "RT² Profiler™ PCR Array Data Analysis" for the RT-qPCR analysis were used. p<0.05 value were accepted significant. Results The study was completed with 18 rats. Histopathologically, solar elastosis was significantly higher in the group receiving UV. The qualitative level and organization of collagen in all application areas were found to be higher compared to the control area 1. In the experimental group receiving UV, collagen was less histopathologically less and more disorganized than the non-UV control group. In the PCR examination, significant decreases were observed in Col1A1 and Col3A1 levels due to the effect of UV application. In inflammation and vascularization, significant increases were observed in the areas where the material was applied compared to the control area. Histopathologically, capsule structure, giant cell granuloma and foreign body reaction were also observed and demonstrated in the areas where the substances were applied. Conclusion As a result of the study, it has been shown that PDO thread application, HA filler and BTX-A, which may be effective in aging and photoaging, increase the organized regulation and increase the expression of the collagen fibers. An increase in solar elastosis and NF-KB expression, which shows photoaging, and a decrease in type 1 and type 3 collagen levels were shown to be significant. It was observed that the application of filler alone in the UV treated group (photoaging), and the combination of the application of thread + filler in the non-UV receiving group (physiological aging) brought more effective results in the chronic period (week 12) as antiaging.
Açıklama
Anahtar Kelimeler
Polidioksanon ip, Polydioxanone thread, Botulinum toksin, Botulinum toxins, Tip A dermal dolgular, Type A dermal fillers, Ultraviyole ışınları, Ultraviolet ray, Deri yaşlanması, Skin aging
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Durmaz, K. (2020). Sıçanlarda polidioksanon ip, hyaluronik asit dolgu ve botulinum toksin-A uygulamalarının deri ve bağ dokusu ile fotoyaşlanma üzerindeki etkilerinin araştırılması. (Yayınlanmamış tıpta uzmanlık tezi) Necmettin Erbakan Üniversitesi, Meram Tıp Fakültesi Dahili Tıp Bilimleri Deri ve Zührevi Hastalıklar Anabilim Dalı, Konya.
