Turkiye Klinikleri Journal of Dental Sciences

.: ORIGINAL RESEARCH
Travma Nedeniyle Kırılan Daimî Anterior Dişlerin Pulpasından Mezenkimal Kök Hücre İzolasyonu ve Karakterizasyonu
Mesenchymal Stem Cell Isolation and Characterization of Dental Pulp from Fractured Permanent Anterior Teeth Due to Trauma
Şerife Buket BOZKURTa, Sema Sezgin HAKKIb
aHacettepe Üniversitesi Diş Hekimliği Fakültesi, Ar-Ge Laboratuvarı, Ankara, TÜRKİYE
bSelçuk Üniversitesi Diş Hekimliği Fakültesi, Periodontoloji ABD, Konya, TÜRKİYE
Turkiye Klinikleri J Dental Sci. 2021;27(4):600-7
doi: 10.5336/dentalsci.2020-79168
Article Language: TR
Full Text
ÖZET
 Amaç: İnsan dental pulpa mezenkimal kök hücrelerin (İDPMKH) sağlıklı pulpa dokularından izole edildiğine dair çok sayıda araştırma olmakla birlikte, travma nedeniyle kırılan dişlerden İDPMKH'lerin izolasyonuna ilişkin çalışma çok sınırlı sayıdadır. Bu çalışmada, travma nedeniyle dişleri kırılmış hastanın kanal tedavisi sırasında elde edilen pulpa dokularından, mezenkimal kök hücre (MKH) izolasyonu yapılıp yapılamayacağı test edildi. Gereç ve Yöntemler: Beş gün önce düştüğü için üst çene santral dişleri (#11, #21) kırılan 15 yaşındaki kadın hasta, ağrı şikâyetiyle pedodonti kliniğine başvurdu. Kırığı görmek, izolasyonu sağlamak ve düzgün bir şekilde restore etmek için krestal alveoler kemiğe osteotomi yapılması için flep cerrahisine karar verildi. Dişler, kanal tedavisi yapılmadan restore edilemeyeceği için pulpa dokusu ekstirpe edildi. Avasküler görünümlü pulpa dokuları eksplant yöntemiyle kültür ortamına alındı ve hücre büyümesi gözlemlendi. Hücrelerin 3. pasajından sonra RNA izole edildi ve MKH'ye özgü hücre yüzey belirteçleri gerçek zamanlı polimeraz zincir reaksiyonu [real-time polymerase chain reaction (RT-PCR)] ile çalışıldı. Ayrıca hematopoietik kök hücre yüzey belirteçleri doğrulama için değerlendirildi. Bulgular: RT-PCR sonuçları, hücrelerin MKH spesifik yüzey belirteçlerini (CD73, CD90, CD105) eksprese ettiğini ve hematopoietik kök hücre yüzey belirteçlerinin (CD34, CD45), ekspresyonunun çok düşük düzeyde olduğunu gösterdi. Sonuç: Travma görmüş kırılmış dişlerden MKH izolasyonunun yapılabilmesi, otolog dental pulpa MKH izole edilerek, hücre temelli kanal tedavilerinin mümkün olabileceğini düşündürmektedir. Bu yaklaşım da diş hekimliğinde travma hastalarının tedavisinde pulpanın revaskülarizasyonu ve rejenerasyonu açısından umut vaat edici görünmektedir.

Anahtar Kelimeler: Mezenkimal kök hücreler; diş pulpası; diş kırıkları
ABSTRACT
 Objective: Although many researches have been reported regarding isolation of human dental pulp mesenchymal stem cells (HDPMSCs) from healthy pulp tissues, there are limited studies on isolation of HDPMSCs from the fractured teeth due to trauma. In this study, it was tested whether mesenchymal stem cell (MSC) could be isolated from pulp tissues that were obtained during the canal treatment of fractured teeth due to trauma. Material and Methods: A 15- year-old woman who had fractured maxillary central incisor teeth (#11, #21) due to falling five days ago admitted to pediatric dentistry with pain. It was decided a flap surgery to do osteotomy for crestal alveolar bone to view fracture and to provide isolation and restore properly. Since the teeth can not be restored without root canal treatment, pulp tissue was extirpated. The avascular-like pulp tissues were cultured using explantation method and cell growth was observed. After third passage, RNA was isolated and cell surface markers specific for MSC were studied by real-time polymerase chain reaction (RT-PCR). Also, hematopoietic stem cell surface markers were evaluated for confirmation. Results: RT-PCR analysis displayed that cells expressed specific cell surface markers of MSC (CD73, CD90, CD105) and expression of hematopoietic stem cell surface markers (CD34,CD45) of cells were determined low levels. Conclusion: Possibility of the MSCs isolation from fractured teeth suggested that cell based canal treatments can be possible by isolating autologous dental pulp MSCs in patient. This approach seems promising for pulp revascularization and regeneration in the treatment of trauma patients in dentistry.

Keywords: Mesenchymal stem cell; dental pulp; tooth fractures
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