Radyoterapinin cilt komplikasyonlarının önlenmesinde yeni bir tedavi önerisi; wharton jeli mezenkimal kök hücre ve conditioning medium etkinliğinin karşılaştırılması
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Dosyalar
Tarih
2020
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi, Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Kanser günümüzde sıkılığı giderek artan, mortalite ve morbiditeye yol açan önemli bir hastalıktır. Uluslararası Kanser Araştırmaları Ajansına göre geçtiğimiz günlerde dünya çapında 7,6 milyon ölümün 1,27 milyonu yeni kanserden kaynaklandığını tahmin etti (Ferlay, Bray et al. 2004). Onkolojik tedavi gören hastaların %80'i tedavinin bir bölümünde radyoterapi görmektedir. Onkolojik tedavinin başarısından bağımsız olarak radyoterapi alan hastaların hemen hemen tamamında radyoterapiye bağlı cilt reaksiyonları gözlenmekte, akut ve bunu takip eden süreçte kronik radyodermatit görülmektedir. Radyodermatit günümüzde çeşitli farmakolojik yöntemlerle çözüme ulaştırılmaya çalışılsa da etkin bir tedavisi olmayan, radyoterapiye bağlı dermal hücre ve fonksiyonel matriks kaybının önlenemediği önemli bir sorundur. Tedavi sürecini kısıtlayan ve hayat kalitesini düşüren en önemli etmendir. Günümüzde meydana gelen bu dermal hücre eksikliği ve matriksin yerine koyulmasında kök hücre bazlı çalışmalar öne çıkmaktadır. Bu çalışmada daha önce yapılmış kök hücre çalışmaları baz alınarak; radyoterapi sonucu gelişmiş cilt hasarının önlenmesinde Wharton Jeli kaynaklı mezenkimal kök hücre, bu hücrelerden elde edilen Conditioning Medium'un etkinliği araştırılmış ve klinik pratikte yer edinmiş olan yağ enjeksiyonu ile karşılaştırılmıştır. Ratların sırt bölgelerinde 4x2 cmlik alana 2mm derinliğe ulaşacak şekilde toplamda 30 Gy radyoterapi uygulanmasını takiben; 30., 37. ve 45. günlerde 1. gruba Wharton Jeli kaynaklı kök hücre, 2. gruba Conditioning Medium uygulaması yapıldı. 3. grup olan yağ enjeksiyonu grubuna 30. günde otolog yağ enjeksiyonu yapıldı. 60 günde cilt örnekleri alınarak deney sonlandırıldı. Elde edilen örnekler makroksopik olarak National Cancer Institute'ün radyodermatit sınıflamasıyla değerlendirildi. Mikroskopik olarak da epidermis-dermis kalınlığı, cilt eki sayısı, HE boyamasında inflamasyon düzeyi ile değerlendirildi. İmmünhistokimyasal değerlendirmede anti-VEGF boyaması ile vaskülarizasyon değerlendirilirken, anti-TGF boyaması ile cildin fibrozisi değerlendirildi. Dermisteki kollojenin maturitesi ise Picro Sirius Red boyası ile değerlendirildi. Çalışmamızda WJ-MSC tedavisi alan grup makroskobik bakıda en iyi skorları alan gruptur. Bu grupta makroskobik yara iyileşmesi daha iyi, alopesi miktarı daha az olarak saptanmıştır. 1 rat dışında (Grade 2) tüm ratlar grade 1 olarak belirlenmiştir. Bu durumun mikroskopiye yansıması da benzer şekildedir. WJ-MSC grubunda epidermis (ortalama 32.8μm) ve dermisin (ortalama 1147.40μm) daha kalın olarak saptandığı ve korunduğu gözlenmiştir (kontrol grubunda epidermis 16.22 μm, dermis 759 μm). Cilt eki sayısının en fazla olduğu, kıl köklerinin en fazla korunduğu grup; WJ-MSC grubudur (mikroskopik alanda ortalama 4.17; kontrol grubunda 1,67). WJ-MSC grubunda epidermis ve dermis normal cilde en yakın, cilt ekleri en fazla ve sağlam olarak saptanmıştır. Çalışmamızda literatürle uyumlu olarak WJ-MSC'nin radyoterapide meydana gelen ağır inflamatuar süreci minimalize ederek yara iyileşmesini arttırdığı saptanmıştır. Yağ grefti grubunda ise greft kaybı sonucu gelişen yağ nekrozu sebebiyle akut inflamasyon baskın şekilde devam etmektedir. Bu veriden hareketle cilt rejenerasyonu amacı ile WJ-MSC tedavisinin yağ greftine oranla çok daha iyi olacağı sonucuna ulaşılmıştır. WJ-MSC grubunda dermal fibrozis minimal düzeyde (grade 1) gelişirken yağ enjeksiyonu ve Sham grubunda grade 3 olarak saptanmıştır. WJ-MSC hem dermis kalınlığının fazla olması hem de skar indeksine göre matür kollojen içermesi dermis miktarının ve fonksiyonunun korunduğu ve kronik dönemde gelişen fibrozisi engellendiği sonucunu bize vermektedir. CM grubu ile WJ-MSC grubunun sonuçları benzer şekilde diğer gruplara göre belirgin düzeyde üstün olarak saptanmıştır. CM grubu makroskopik, mikroskopik (epidermis, dermis kalınlığı, mikroskopik bakıda cilt eki sayısı) parametrelerde; WJ-MSC grubunun ardından en yüksek skorları alan grup ve istatistiksel olarak WJ-MSC grubu ile aynı kümede olarak belirlenmiştir. Çalışmamızın literatürdeki çalışmalara üstünlüğü bu etkinin WJ-MSC'nin etkisi ile karşılaştırılabilinmesidir. Bu karşılaştırılmada WJ-MSC grubunun akut inflamasyon skormasında anti-inflamatuar etkisinin CM grubunda gözlenmediği ve CM grubunun inflamasyon skorunun daha yüksek olduğu saptanmıştır (WJ-MSC grubunda 1; CM grubunda 2). WJ-MSC'lerin klinik uygulamada kendine yer edinmiş olan otolog yağ enjeksiyonu etkilerinden üstün olup olmadığı, klinik pratiğe uygun olup olmadığı karşılaştırmak açısıdan önemli bir parametredir. Yağ enjeksiyonu grubunun makroskobik skorlamasının kontrol ve Sham grubuna göre daha iyi olduğu saptanmıştır. Fakat bu grup WJ-MSC ve CM grubu ile karşılaştırıldığında makroskobik skorlarının WJ-MSC ve CM grubunda daha iyi olduğu görülmüştür (WJ-MSC için en çok grade 1, CM için yarı yarıya grade 1 ve 2, yağ enjeksiyonunda en çok grade 2). Mikroskopik bakıda ise yağ enjeksiyonunun epidermis ve dermis kalınlığı üzerine istatistiksel olarak anlamlı bir etkisi olmadığı gözlenmiştir (sig.: 0.000 < p-value: 0.05).Yağ enjeksiyonu grubunda greftlerde yağ nekrozu ve nekroze alanların çevresinde en çok grade 4 olmak üzere inflamatuar alanlar dikkati çekmektedir. Yağ enjeksiyonu grubudaki örneklerde kronik inflamatuar süreç belirteci olan anti TGF-β boyamasının a en yüksek fibrozis skoru olan grade 3 olarak skorlanması; yağ enjeksiyonunun radyodermatitte gelişen akut ve kronik inflamasyonu baskılamadığı; hatta nekroze yağ grefti alanları sebebiyle akut inflamasyonu arttırdığı saptanmıştır. Bu sonuçtan hareketle radyodermatit tablosunda WJ-MSC ve CM tedavisinin cilt bütünlüğünü korumada yağ enjeksiyonuna göre çok daha etkili bir yöntem olduğu saptanmıştır. Wharton jeli mezenkimal kök hücre ve Conditioning medium'u radyasyona bağlı cilt yaralanmasında onkolojik olarak güvenilir, dermis ve epidermis yapılarını rejenere ederken inflamasyonu azaltması ile etkili bir tedavi yöntemidir.
A New Treatment for the Prevention of Radiotherapy Skin Complications; Comparison of Wharton's Jelly Mesenchymal Stem Cell and Conditioning Medium Efficacy Cancer is an important disease with increasing frequency, leading to mortality and morbidity. According to the International Agency for Research on Cancer, it recently estimated that 1.27 million of 7.6 million deaths worldwide were caused by new cancer (Ferlay, Bray et al. 2004). 80% of patients receiving oncological treatment receive radiotherapy in some part of the treatment. Regardless of the success of oncological treatment, almost all patients who receive radiotherapy have skin reactions due to radiotherapy, and acute and chronic radiodermatitis is observed in the following period. Radiodermatitis is an important problem that has no effective treatment and the loss of dermal cells and functional matrix due to radiotherapy cannot be prevented, even though various pharmacological methods are tried to be solved today. It is the most important factor that restricts the treatment process and reduces the quality of life. Stem cell-based studies come to the fore in this dermal cell deficiency and replacement of the matrix. In this study, based on previous stem cell studies; The efficacy of Wharton's Jelly-derived mesenchymal stem cells and Conditioning Medium obtained from these cells in preventing skin damage developed as a result of radiotherapy have been investigated and compared with the otolog fat injection that has taken place in clinical practice. After a total of 30 Gy radiotherapy was applied to reach a depth of 2mm in an area of 4x2 cm on the back of the rats. On the 30th, 37th and 45th days, Wharton's Jelly derived stem cells were applied to the first group and Conditioning Medium was applied to the second group. Autologous fat injection was applied on the 30th day to the third group, the fat injection group. Skin samples were taken in 60 days and the experiment was terminated. The samples obtained were evaluated macroxopically with the National Cancer Institute's radioodermatitis classification. Microscopically, epidermis dermis thickness, number of skin appendages, and inflammation level in HE staining were evaluated. While vascularization was evaluated with anti-VEGF staining in immunohistochemical evaluation, skin fibrosis was evaluated with anti-TGF staining. The maturity of the collagen in the dermis was evaluated with Picro Sirius Red dye. In our study, the group that received WJ-MSC treatment was the group with the best scores in macroscopic examination. Macroscopic wound healing was better and the amount of alopecia was lower in this group. All rats except 1 rat (Grade 2) were determined as Grade 1. The reflection of this situation on microscopy is similarly. In the WJ-MSC group, it was observed that epidermis (mean 32.8μm) and dermis (mean 1147.40μm) were detected and protected thicker (epidermis 16.22 μm, dermis 759 μm in control group). The group with the highest number of skin attachments and the most preserved hair follicles; WJ-MSC group (mean 4.17 in microscopic field; 1.67 in control group). In the WJ-MSC group, epidermis and dermis were found to be closest to normal skin, skin appendages were the most and intact. In our study, in accordance with the literature, it was determined that WJ-MSC increased wound healing by minimizing the severe inflammatory process that occurs in radiotherapy. In the fat graft group, acute inflammation continues predominantly due to fat necrosis as a result of graft loss. Based on this data, it was concluded that WJ-MSC treatment would be much better than fat graft for skin regeneration. While dermal fibrosis developed minimally (grade 1) in the WJ-MSC group, it was found to be grade 3 in the fat injection and Sham group. WJ-MSC both having a large dermis thickness and containing mature collagen according to the scar index gives us the conclusion that the amount and function of the dermis are preserved and fibrosis developing in the chronic period is prevented. The results of the CM group and the WJ-MSC group were similarly significantly superior to the other groups. CM group in macroscopic, microscopic (epidermis, dermis thickness, number of skin appendages in microscopic examination) parameters; The group with the highest scores after the WJ-MSC group and was statistically determined to be in the same group with the WJ-MSC group. The advantage of our study over the studies in the literature is that this effect can be compared with the effect of WJ-MSC. In this comparison, it was found that the anti-inflammatory effect was not observed in the acute inflammation score of the WJ-MSC group, and the inflammation score of the CM group was higher (1 in the WJ-MSC group; 2 in the CM group). Whether WJ-MSCs are superior to autologous fat injection effects that have taken place in clinical practice is an important parameter in terms of comparing whether they are suitable for clinical practice or not. It was determined that macroscopic scoring of the fat injection group was better than the control and Sham groups. However, when this group was compared with the WJ-MSC and CM groups, it was observed that the macroscopic scores were better in the WJ-MSC and CM groups (most grade 1 for WJ-MSC, half grade 1 and 2 for CM, grade 2 for fat injection most). In the microscopic examination, it was observed that the fat injection did not have a statistically significant effect on the epidermis and dermis thickness (sig: 0.000.
A New Treatment for the Prevention of Radiotherapy Skin Complications; Comparison of Wharton's Jelly Mesenchymal Stem Cell and Conditioning Medium Efficacy Cancer is an important disease with increasing frequency, leading to mortality and morbidity. According to the International Agency for Research on Cancer, it recently estimated that 1.27 million of 7.6 million deaths worldwide were caused by new cancer (Ferlay, Bray et al. 2004). 80% of patients receiving oncological treatment receive radiotherapy in some part of the treatment. Regardless of the success of oncological treatment, almost all patients who receive radiotherapy have skin reactions due to radiotherapy, and acute and chronic radiodermatitis is observed in the following period. Radiodermatitis is an important problem that has no effective treatment and the loss of dermal cells and functional matrix due to radiotherapy cannot be prevented, even though various pharmacological methods are tried to be solved today. It is the most important factor that restricts the treatment process and reduces the quality of life. Stem cell-based studies come to the fore in this dermal cell deficiency and replacement of the matrix. In this study, based on previous stem cell studies; The efficacy of Wharton's Jelly-derived mesenchymal stem cells and Conditioning Medium obtained from these cells in preventing skin damage developed as a result of radiotherapy have been investigated and compared with the otolog fat injection that has taken place in clinical practice. After a total of 30 Gy radiotherapy was applied to reach a depth of 2mm in an area of 4x2 cm on the back of the rats. On the 30th, 37th and 45th days, Wharton's Jelly derived stem cells were applied to the first group and Conditioning Medium was applied to the second group. Autologous fat injection was applied on the 30th day to the third group, the fat injection group. Skin samples were taken in 60 days and the experiment was terminated. The samples obtained were evaluated macroxopically with the National Cancer Institute's radioodermatitis classification. Microscopically, epidermis dermis thickness, number of skin appendages, and inflammation level in HE staining were evaluated. While vascularization was evaluated with anti-VEGF staining in immunohistochemical evaluation, skin fibrosis was evaluated with anti-TGF staining. The maturity of the collagen in the dermis was evaluated with Picro Sirius Red dye. In our study, the group that received WJ-MSC treatment was the group with the best scores in macroscopic examination. Macroscopic wound healing was better and the amount of alopecia was lower in this group. All rats except 1 rat (Grade 2) were determined as Grade 1. The reflection of this situation on microscopy is similarly. In the WJ-MSC group, it was observed that epidermis (mean 32.8μm) and dermis (mean 1147.40μm) were detected and protected thicker (epidermis 16.22 μm, dermis 759 μm in control group). The group with the highest number of skin attachments and the most preserved hair follicles; WJ-MSC group (mean 4.17 in microscopic field; 1.67 in control group). In the WJ-MSC group, epidermis and dermis were found to be closest to normal skin, skin appendages were the most and intact. In our study, in accordance with the literature, it was determined that WJ-MSC increased wound healing by minimizing the severe inflammatory process that occurs in radiotherapy. In the fat graft group, acute inflammation continues predominantly due to fat necrosis as a result of graft loss. Based on this data, it was concluded that WJ-MSC treatment would be much better than fat graft for skin regeneration. While dermal fibrosis developed minimally (grade 1) in the WJ-MSC group, it was found to be grade 3 in the fat injection and Sham group. WJ-MSC both having a large dermis thickness and containing mature collagen according to the scar index gives us the conclusion that the amount and function of the dermis are preserved and fibrosis developing in the chronic period is prevented. The results of the CM group and the WJ-MSC group were similarly significantly superior to the other groups. CM group in macroscopic, microscopic (epidermis, dermis thickness, number of skin appendages in microscopic examination) parameters; The group with the highest scores after the WJ-MSC group and was statistically determined to be in the same group with the WJ-MSC group. The advantage of our study over the studies in the literature is that this effect can be compared with the effect of WJ-MSC. In this comparison, it was found that the anti-inflammatory effect was not observed in the acute inflammation score of the WJ-MSC group, and the inflammation score of the CM group was higher (1 in the WJ-MSC group; 2 in the CM group). Whether WJ-MSCs are superior to autologous fat injection effects that have taken place in clinical practice is an important parameter in terms of comparing whether they are suitable for clinical practice or not. It was determined that macroscopic scoring of the fat injection group was better than the control and Sham groups. However, when this group was compared with the WJ-MSC and CM groups, it was observed that the macroscopic scores were better in the WJ-MSC and CM groups (most grade 1 for WJ-MSC, half grade 1 and 2 for CM, grade 2 for fat injection most). In the microscopic examination, it was observed that the fat injection did not have a statistically significant effect on the epidermis and dermis thickness (sig: 0.000.
Açıklama
Anahtar Kelimeler
Kök Hücre, Wharton Jeli, Conditioning Medium, Radyoterapi, Radyodermatit, Stem Cell, Wharton‘s Jelly, Radiotherapy, Radiodermatitis
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Bilirer, A. (2020). Radyoterapinin cilt komplikasyonlarının önlenmesinde yeni bir tedavi önerisi; wharton jeli mezenkimal kök hücre ve conditioning medium etkinliğinin karşılaştırılması. (Uzmanlık Tezi). Selçuk Üniversitesi, Tıp Fakültesi, Konya.
