Ex Vivo Produced Oral Mucosa Equivalent by Using the Direct Explant Cell Culture Technique

Abstract

Objective: The aim of this study is the histological and immunohistochemical evaluation of ex vivo produced oral mucosal equivalents using keratinocytes cultured by direct explant technique. Material and Methods: Oral mucosa tissue samples were obtained from the keratinized gingival tissues of 14 healthy human subjects. Human oral mucosa keratinocytes from an oral mucosa biopsy specimen were dissociated by the explant technique. Once a sufficient population of keratinocytes was reached, they were seeded onto the type IV collagen coated “AlloDerm” and taken for histological and immunohistochemical examinations at 11 days postseeding of the keratinocytes on the cadaveric human dermal matrix. Results: Histopathologically and immunohistochemically, 12 out of 14 successful ex vivo produced oral mucosa equivalents (EVPOME) that consisted of a stratified epidermis on a dermal matrix have been developed with keratinocytes cultured by the explant technique. Conclusion: The technical handling involved in the direct explant method at the beginning of the process has fewer steps than the enzymatic method and use of the direct explant technique protocol for culturing of human oral mucosa keratinocyte may be more adequate for EVPOME production. Turkish Anahtar Kelimeler: Ağız mukozası, keratinosit, epidermis, dermal matriks Amaç: Bu çalışmanın amacı, direkt eksplant teknik kullanılarak, vücut dışında ağız mukozası keratinositlerinden üretilen ağız mukozası eşdeğerlerinin, histolojik ve immünohistokimyasal olarak incelenmesidir. Gereç ve yöntem: Ağız mukozası örnekleri, 14 sağlıklı insanın keratinize dişeti dokularından elde edilmiştir. İnsan ağız mukozası keratinositleri, ağız mukozası örneklerinden, eksplant teknik ile ayrıştırılmıştır. Yeterli çoğunluğa ulaştıktan sonra keratinositler, tip IV kollajen kaplı “Alloderm” üzerine ekilmiş ve kadavra kaynaklı insan dermal matriksi üzerine ekimlerinden 11 gün sonra histolojik ve immünokimyasal incelemeye tabi tutulmuştur. Bulgular: Histopatolojik ve immünohistokimyasal olarak, 14 örnekten 12sinde, dermal matriks üzerinde, eksplant teknik ile oluşan keratinositlerden elde edilen çok katlı yassı epitelden oluşan vücut dışında üretilmiş ağız mukozası eşdeğeri başarılı bir şekilde oluşturulmuştur. Sonuç: Enzimatik tekniğe göre direct eksplant yöntem, sürecin başında daha az sayıda basamak gerektirmekte ve vücut dışında üretilmiş ağız mukozası eşdeğeri üretimi için ağız mukozası keratinositlerinin elde edilmesinde daha uygun bir yöntem olarak öne çıkmaktadır.

Keywords

• Oral mucosa, keratinocyte, epidermis, dermal matrix

Ex Vivo Produced Oral Mucosa Equivalent by Using the Direct Explant Cell Culture Technique

Öz

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