Publication: 3B yazıcı teknoloji ile antimikrobiyal nar ekstraktlı dental membran üretimi
Abstract
Geçmişten günümüze bitkisel kaynaklar kullanılarak tedavi yöntemleri, daha pratik olması ve kullanım kolaylığı gibi sebeplerden dolayı sıkça tercih edilmiştir ve halen bitkisel tedavi yöntemleri varlığını korumaktadır. Aynı şekilde son zamanlarda 3 boyutlu yazıcılar kullanılarak yeni tedavi edici yapıların üretimi gerçekleşmektedir. Bu biyoyazıcılardan üretilen yapılar hasar görmüş dokuyu korumak, tedavi etmek ve desteklemek amacıyla hücrelerin büyümesine olanak sağlayarak tedavi etmeye yarayacak yapıları kolaylıkla istenilen tasarımda üretmeyi mümkün kılması bakımından bu çalışmada tercih edilmiştir. Üretilen iskelelere kullanımı çok eskilere dayanan antimikrobiyal özelliğe sahip olan Punica granatum L. (Punicaceae) olarak isimlendirilen nar kabuğundan ve nar tanelerinden ekstreler eklenerek, iskelelerin dental membrane olarak kullanılması amacıyla üretim gerçekleşmiştir. Nar kabuğundan ve nar tanelerinden ayrı proseslerde elde edilen ekstreler 1:1 oranında karışım haline getirilerek iskele yapılarında kullanılmıştır. Yapıyı güçlendirebilmek amacıyla Polivinil alkol (PVA) polimeri ana polimer olarak kullanıldı ve nişasta eklenmesiyle yapının biyouyumluluğuna etkisi incelenmiştir. Üretimi gerçekleştirilen tüm iskelelerin morfolojik yapısı taramalı elektron mikroskopu ve optik mikroskop ile incelenmiştir. İskelelerin kimyasal bağ etkileşim analizleri için fourier dönüşümlü kızılötesi spektroskopisi kullanılarak incelendi ve termal analizleri için diferansiyel taramalı kalorimetre kullanılmıştır. İskelelerin mekanik dayanımları çekme testiyle incelendi ve ilaç salım özellikleri vücut ortamını taklit edilerek fosfat tampon çözeltisi içerisinde farklı zaman aralıklarında UV- Spektrofotometre ile analiz edildi. İskeleler üzerinde insan diş eti fibroblast hücresi kullanılarak iskelelerin sitotoksisitesi ve iskeleler üzerinde hücre canlılığı MTT analizi ve floresan boyama kullanılarak test edilmiştir. Ekstresi yapılan ve iskelelerin içeriğinde kullanılan nar kabuğu ve tanelerinin ekstresi yüksek basınçlı sıvı kromatografisi kullanılarak biyolojik içeriği analiz edilmiştir. Aynı şekilde nar ekstre karışımını içeren iskelelerin antimikrobiyal etkisi S. aureus (ATCC® 6538™), C. albicans (ATCC® 5314™), ve E. faecalis (ATCC® 29212™) mikroorganizmaları üzerinde test edilmiştir. Tüm analizler sonucu %3 nar ekstre karışımını ve %2 nişasta içeren iskelelerin diğer iskelelere nazaran daha iyi özellikler gösterdiği gözlemlenmiştir.
Natural herbal based products are always preferred and have been used since the dawn of time, due to the ease and practicality of use, and they are still preferred nowadays. Similarly, 3D printing method can be applied for the production of scaffolds that could be used in the treatment of different diseases. 3D printing technique, is an easy way to produce scaffolds which could help effected tissue healing and provide the necessary support to the damaged structures for faster recovery as well as protect, treat and provide the required environment for cell growth. For those reasons 3D bioprinting method was chosen for this study. Pomegranate is well known folk treatment substance. Pomegranate peel and seed extract was obtained and added into the scaffolds due to its high antimicrobial activity to be used in dental membrane scaffolds. Pomegranate peel and seeds extracts were applied in separate processes and mixed at 1:1 ratio following which they were added into the scaffolds. In order to increase the mechanical strength of scaffolds, Poly vinyl alcohol (PVA) polymer was used as the main ingredient and the effect of starch addition on the biocompatibility of the structure was investigated. The morphological structure of all fabricated scaffolds were examined using scanning electron microscope and optical microscope. The scaffolds chemical bond interactions were examined using Fourier transform infrared spectroscopy and differential scanning calorimetry was used for thermal analysis of each scaffolds. The mechanical strength of the scaffolds was examined using mechanical tensile strength testing method and drug release kinetics were analyzed using UV-Spectrophotometer at different time intervals in phosphate buffer solution which helps simulating the body environment. The cytotoxicity analysis of the scaffolds on human gingival fibroblast cells and the cell viability on the scaffolds were tested using MTT analysis and fluorescent staining technique. The biological content of the extracted pomegranate peel and seeds, used in the contents of the scaffolds, was analyzed using high pressure liquid chromatography. Likewise, the antimicrobial effect of scaffolds containing the pomegranate extract mixture was tested on S. aureus (ATCC® 6538™), C. albicans (ATCC® 5314™), and E. faecalis (ATCC® 29212™) microorganisms. As a result of all analyzes, it was observed that the scaffold containing 3% pomegranate extract mixture and 2% starch showed the best properties compared to other scaffolds.
Natural herbal based products are always preferred and have been used since the dawn of time, due to the ease and practicality of use, and they are still preferred nowadays. Similarly, 3D printing method can be applied for the production of scaffolds that could be used in the treatment of different diseases. 3D printing technique, is an easy way to produce scaffolds which could help effected tissue healing and provide the necessary support to the damaged structures for faster recovery as well as protect, treat and provide the required environment for cell growth. For those reasons 3D bioprinting method was chosen for this study. Pomegranate is well known folk treatment substance. Pomegranate peel and seed extract was obtained and added into the scaffolds due to its high antimicrobial activity to be used in dental membrane scaffolds. Pomegranate peel and seeds extracts were applied in separate processes and mixed at 1:1 ratio following which they were added into the scaffolds. In order to increase the mechanical strength of scaffolds, Poly vinyl alcohol (PVA) polymer was used as the main ingredient and the effect of starch addition on the biocompatibility of the structure was investigated. The morphological structure of all fabricated scaffolds were examined using scanning electron microscope and optical microscope. The scaffolds chemical bond interactions were examined using Fourier transform infrared spectroscopy and differential scanning calorimetry was used for thermal analysis of each scaffolds. The mechanical strength of the scaffolds was examined using mechanical tensile strength testing method and drug release kinetics were analyzed using UV-Spectrophotometer at different time intervals in phosphate buffer solution which helps simulating the body environment. The cytotoxicity analysis of the scaffolds on human gingival fibroblast cells and the cell viability on the scaffolds were tested using MTT analysis and fluorescent staining technique. The biological content of the extracted pomegranate peel and seeds, used in the contents of the scaffolds, was analyzed using high pressure liquid chromatography. Likewise, the antimicrobial effect of scaffolds containing the pomegranate extract mixture was tested on S. aureus (ATCC® 6538™), C. albicans (ATCC® 5314™), and E. faecalis (ATCC® 29212™) microorganisms. As a result of all analyzes, it was observed that the scaffold containing 3% pomegranate extract mixture and 2% starch showed the best properties compared to other scaffolds.