Emrah YÜCESAN, Harun BASOGLU, Beyza GÖNCÜ, Nur ÖZTEN KANDAŞ, Yeliz Emine ERSOY, Fahri AKBAŞ, Erhan AYŞAN

Mikroenkapsüle edilen paratiroid hücrelerinin in-vitro optimizasyonu

Amaç: Enkapsülasyon, bir maddenin ya da bir karışımın diğer bir malzeme ya da sistemle kaplandığı ya da içine sıkıştırıldığı tekniktir. Mikroenkapsülasyon tekniği, tıp dahil olmak üzere pek çok farklı disiplin tarafından uygulanmaya çalışılan ve halen en uygun enkapsülasyon tekniğinin bulunması için araştırmaların yürütüldüğü bir tekniktir. Mikroenkapsülasyon tekniğinde birbirinden farklı maddeler uygulanabilir. Bu maddeler elde edilme kaynaklarına göre doğal ya da sentetiktir. Mikroenkapsülasyon tekniği tıp pratiğinde özellikle pankreas beta hücrelerinin, Tip 1 Diabetes Mellitus (DM) hastalarına verilmesinde kullanılmaktadır. Mikroenkapsülasyonun uygulandığı bir başka durum ise hipoparatiroidi tanılı hastalara paratiroid hücrelerinin verilmesidir. Paratiroid hücrelerinin, mikroenkapsülasyon işleminde kullanılabilebilmesinin en önemli özelliklerinden biri, hücrelerin görece homojen yapıda olmaları, eksikliğinin organizmada doğrudan gözlemlenebilir semptomlara neden olması dolayısıyla oluşacak yanıtın hızlıca belirlenebilmesidir. Yöntemler: Çalışmamızda paratiroidhiperplazisi tanısı almış insandan elde edilen paratiroid dokularından izole edilen hücreler, %2'lik ultra saf aljinat ile muamele edildi. İşlem sırasında her bir kapsül sayısı hesaplandı ve kapsül başına 5X106, 10X106, 20X106 ve 50X106 hücre eklendi. Sonrasında 75 gün boyunca içlerinde paratiroid hücresi bulunan kapsüllerin morfolojik özellikleri ve parathormonsekresyon yeteneklerinin zaman bağlı değişimi gözlemlendi. Bulgular: 75 gün sonrasında tüm gruplar için korele olarak parathormon düzeyinde düşüş tespit edildi. Kapsüler formasyonda ciddi bir bozulmanın görülmediği grup, 20X106 hücre uygulanan grup oldu. Sonuç: Çalışmamızda model olarak değerlendirdiğimiz paratiroid hücrelerinin kapsülasyonu için en uygun koşulları (kapsülasyonda kullanılacak madde, destekleyici tampon, uygulanan maddelerin yüzdeleri, miktarları) belirlendi. Ayrıca günlere bağlı olarak kapsüllerdeki bozulmalar da tespit edildi. İlerleyen çalışmalarda optimize ettiğimiz koşulların hipoparatiroidizm tanılı hasta gruplarında denenmesi ve sonuçların burada sunduğumuz in vitro deney sonuçları ile karşılaştırılması gerekmektedir.

In-vitro optimization of microencapsulated parathyroid cells

Objective: Encapsulation is a process that a mixture combined and coated with another material or system. Microencapsulation has been the most frequently used technique for several different disciplines and it is still a common technique in which researches are carried out to detect the most appropriate encapsulation technique. Different substances can be applied for this technique. Applied materials have positive and negative potencies. Microencapsulation technique is used in medicine, particularly pancreatic ?-cells are given for treating Type 1 Diabetes Mellitus (DM) patients. Another research field of technique have been investigated for the treatment of the patients which diagnosed with permanent hypoparathyroidsm with the advantage of via capsulated parathyroid cell transplantation and as a result the improvement of parathormone levels also amelioration of the clinical symptoms who prevents the graft versus host reaction to the transplanted cells as a consequences of capsulation process. The most important characteristics of parathyroid cells includes relatively a homogenous population and in the absence, symptoms are directly observable therefore microencapsulation process made them useful for determination of results. Methods: In this study isolated parathyroid tissue cells, which were mixed with 2% ultrapure alginate, from a patient diagnosed with parathyroid hyperplasia were used. During this process capsule quantity for each cell/capsule groups of 5X106, 10X106, 20X106and 50X106 were calculated. Afterwards each capsulated parathyroid cell groups were evaluated with their altered morphological features and parathormone release ability for 75 days. Results: Throughout 75 days, gradual decrease in parathormoneleves for each group was evaluated. In particular 20X106 cells with capsulated group were stable for capsulary formation. Conclusion: In our study, the optimal conditions (substance for capsulation, supporting medium, percentage and amounts of material) for encapsulation of parathyroid cells (as a model) were determined. In addition, deterioration in the capsules was detected depending on the days. It is necessary for further investigations to test the conditions we optimized. Similar conditions should be evaluate with the patients have hypoparathyroidism and compare with the in vitro test results we provide here.

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