In vitro characterization of chitosan scaffolds: influence of composition and deacetylation degree

Abstract

In this study, the influence of degree of deacetylation (DD) and composition on some structural and biological properties of chitosan scaffolds were examined in vitro. 3D chitosan scaffolds of 2% (w/v) and 3% (w/v) composition in different DDs i.e. 75-85% and >85% were prepared by freeze-drying method at -80 degrees C. We noticed that >85% deacetylated chitosan scaffolds of 2% (w/v) composition has a highly interconnected morphological structure having approximately 100 mum pore size with 0.0917 N/mm(2) compression modulus. L929 fibroblastic cells were cultured on chitosan scaffolds in order to evaluate their biocompatibilities. Cell culture studies demonstrated that fibroblastic cell attachment and proliferation is affected by DD. The higher deacetylated chitosan scaffolds strongly supported the attachment and proliferation when compared with the lower deacetylated scaffolds. MTT assay indicated that >85% deacetylated chitosan scaffolds of 2% (w/v) composition, having the highest specific growth rate 0.017 h(-1) of all, was found to be the most suitable for cell culture studies and a potential candidate for tissue engineering with enhanced biostability and good biocompatibility.

Tailoring chitosan/collagen scaffolds for tissue engineering: Effect of composition and different crosslinking agents on scaffold properties

“…Cells grew throughout these scaffolds with the fastest growth rate and doubled the cell population in the lowest time. Similar growth rates and doubling time values were obtained by Seda Tigli et al (2007), who seeded fibroblasts on pure chitosan scaffolds for 5 days. However, they detected a reduction in cell number after 3 days of culture period as well as some difficulties in achieving adequate cell growth throughout the matrix with the smallest pore size.…”

“…Proliferation of MCF-7 cells on the scaffolds was quantitatively assessed with 3-[4,5-dimethylthiazol-2-yl]-diphenyltetrazolium bromide (MTT) (Seda Tigli, Karakecili & Gumusderelioglu, 2007) at different culture periods of up to 5 days. At different culture times, control and inoculated samples were removed from the Petri plates and placed on to 24-well plates containing 1 ml medium per well.…”

“…MTT assay was also applied to the scaffolds without cells and the signal obtained was subtracted from the inoculated scaffold signal. Exponential cell growth was assumed and a specific growth rate () was determined by fitting the following equation to the absorbance data (Seda Tigli et al, 2007):…”

“…It is known that bacterial collagenase (Closrium hisrolyticum) specifically cleaves the peptide bond on the amino side of glycine in collagen sequences (Lee et al, 2001;Weadock, Miller, Keuffel, & Dunn, 1996) while lysozyme hydrolyses and metabolises chitosan molecules, leading to the release of amino-sugars (Seda Tigli et al, 2007). Therefore it may be anticipated that Non-XL Chit enriched compositions should be degraded more in lysozyme containing media while scaffolds with higher collagen content should be more susceptible to the presence of collagenase (Harper & Kang, 1970).…”

In vitro capacity of different grades of chitosan derivatives to induce platelet adhesion and aggregation

“…We used collagen-based chitosan in our study as we were aware of the ability of collagen to initiate platelet aggregation at the site of bleeding tissue [20]. Collagens are crucial for platelet adhesion and subsequent activation on the extracellular matrix of denuded endothelium [21].…”