Isolation of polysaccharides alpha- and beta-glucans of Pleurotus ostreatus and evaluation of their biocompatibility after surface coating

Year 2019, Volume: 10 Issue: 1, 56 - 69, 25.04.2019

Leyla Kayiş Gökhan Duruksu

Abstract

Glucans, which are polysaccharides in the
fungal cell wall, are formed by glucose monomers linked by O-glycosidic bonds.
Besides of their medical applications, one of the most promising features of
glucans is the stimulatory effect on the immune system of beta-glucans, in
which glucose units are linked by
beta-1,3 and -1,6 glycosidic bonds. In this study, it was aimed to investigate
the effect of alpha- and beta-glucans on the cell viability and proliferation,
which were derived from Pleurotus
ostreatus by hot water/caustic extraction. Total glucan was quantified by
the phenol-sulphuric acid method, and
alpha/beta-glucan purity was determined by the enzymatic
method. After coating the surface of culture flask
with the glucans, their effects on mesenchymal stem cells were examined by
toxicity and cell proliferation assays. Alpha- and beta-glucans from P. ostreatus
were obtained with a purity of 97.87%
±1.45 and 97.79% ±0.28, respectively. In cell adhesion analysis, alpha-glucans
were found to support the attachment (88.10-95.75%), while the adhesion ratio
on beta-glucan-coated surfaces decreased to 61.26% ±8.04. The cells on
alpha-glucan-coated surfaces were proliferated 1.44 times faster than those on
beta-glucan, and 1.56 times faster than those on the uncoated surface. As a result, alpha- and beta-glucan, obtained by
hot water/caustic extraction processes, have been shown to have no toxic
effects on the cells, and both glucans were demonstrated to be biocompatible
biomaterials by in vitro tests. However, the cell adhesion on beta-glucan
coated surfaces was observed to be low. Alpha-glucan was shown to promote both
cell adhesion and proliferation. Therefore, alpha-glucan may have potential
applications in cell culture and tissue engineering studies.

Keywords

Biomaterials, Fungi, Polysaccharides, Glucan, Surface Coating

Pleurotus ostreatus alfa- ve beta- glukan polisakkaritlerinin izolasyonu ve biyouyumluluklarının yüzey kaplaması sonrasında incelenmesi

Abstract

Glukanlar mantarların hücre duvarında bulunan
O-glikozidik bağlar ile bağlanmış glikoz monomerlerinin oluşturduğu
polisakkaritlerdir. Glukanların birçok tıbbi uygulamalarının yanı sıra en öne
çıkan özelliklerinden biri ise beta-1,3 ve -1,6 glikozidik bağlarıyla birleşmiş
glikoz zincirinin oluşturduğu beta-glukanların bağışıklık sistemi üzerindeki
uyarıcı etkisidir. Bu çalışmada Pleurotus
ostreatus 'tan (istiridye mantarı) sıcak su ve sıcak kostik çözelti ile
ekstraksiyonu sonrasında alfa- ve beta-glukan izolasyonu gerçekleştirilerek
elde edilen polisakkaritlerin hücre canlılığı ve çoğalması üzerindeki
etkilerinin incelenmesi amaçlanmıştır. Toplam glukan miktarları fenol-sülfürik
asit metoduyla ve alfa/beta-glukan saflıkları enzimatik yöntemle analiz edilmiştir.
Kültür kapları glukanla kaplandıktan sonra toksisite ve hücre çoğalma
analizleriyle mezenkimal kök hücreler üzerindeki etkileri incelenmiştir.
İşlemler sonucunda %97.87 ±1.45 saflıkta alfa-glukan ve %97.79 ±0.28 saflıkta
beta-glukan P. ostreatus 'tan elde
edilmiştir. Hücre tutunma analizinde alfa-glukanların tutunmayı desteklediği
(%88.10-95.75) gözlemlenmişken beta-glukan kaplı yüzeylerde tutunma oranının
%61.26 ±8.04 değerine düştüğü belirlenmiştir. Alfa-glukan kaplı yüzey
üzerindeki hücreler beta-glukan kaplı yüzey üzerindeki hücrelere göre 1.44 kat,
kaplı olmayan yüzey üzerindeki hücrelere göre 1.56 kat daha hızlı
çoğalmışlardır. Sonuç olarak sıcak su/kostik işlemleriyle elde edilen alfa- ve
beta-glukanların hücreler üzerinde toksik etkisinin olmadığı ve her iki
glukanın biyouyumlu bir malzeme olduğu in vitro testlerle gösterilmiştir. Ancak
beta-glukan kaplı yüzeylerde hücre tutunması düşük olduğu gözlemlenmiştir.
Alfa-glukanın hücre tutunmasını ve çoğalmasını desteklediği görülmüştür. Bu
özellikleri ile hücre kültürü ve doku mühendisliği çalışmalarında alfa-glukanın
potansiyel uygulama alanlarının olabileceği ortaya konmuştur.

Keywords

Biyomalzeme, Mantar, Polisakkarit, Glukan, Yüzey Kaplama

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