International Journal of Biological Macromolecules
Structural features and bioactivities of the chitosan
Introduction
Chitosan, a deacetylated form of chitin, is a natural antimicrobial compound (Fig. 1). On the one hand it can be obtained from crustacean shells (crabs, shrimp and crayfishes) either by chemical or microbiological processes and on the other hand it can be produced by some fungi (Aspergillus niger, Mucor rouxii, Penecillium notatum) [1], [2], [3], [4], [5]. Chitosan has one primary amino and two free hydroxyl groups for each C6 building unit. Due to the easy availability of free amino groups in chitosan, it carries a positive charge and thus in turn reacts with many negatively charged surfaces/polymers and also undergoes chelation with metal ions [6] like cobalt [7]. Thus, it is utilized for separation of metals.
Chitosan is currently receiving a great deal of attention for medical and pharmaceutical applications. Indeed, chitosan is known for its biocompatibility allowing its use in various medical applications such as topical ocular application [8], implantation [9] or injection [10]. Moreover, chitosan is metabolized by certain human enzymes, e.g. lysozyme, and can be considered as biodegradable [11], [12]. In addition, it has been reported that chitosan acts as a penetration enhancer by opening epithelial tight-junctions [13], [14]. Due to its positive charges at physiological pH, chitosan is also bioadhesive, which increases retention at the site of application [15], [16]. Chitosan also promotes wound-healing [17], [18] and has bacteriostatic effects [19], [20].
Renal ischemia is associated mostly with acute renal failure. Moreover, transplantation of any organ necessarily involves an unavoidable period of ischemia. Clinical and experimental studies have provided evidence that ischemic cell injury is mediated by ROS [21], [22]. Ischemic reperfusion injury (IRI) is a key factor to affect early function of transplanted kidneys. It don’t only cause the delay in restoration of normal graft function, but also promote acute rejection through changing immune mechanism. In addition, IRI, as a main non-antigen-dependent factor, affects long-term survival and function of transplanted organs and plays a important part in occurance and development of loss of chronic graft function. ROS per se have also been shown to compromise renal function, depress glomerular filtration, impair glomerular sieving function [23], [24], [25], and induce apoptosis in renal cells [26]. Antioxidants improve organ functions [27] and attenuate apoptosis [28], [29].
In the present study, the chemical structure of chitosan was examined. In addition, effect of chitosan on renal tubular epithelial cell apoptosis was investigated.
Section snippets
Nitric oxide radical inhibition assay
Nitric oxide radical scavenging can be estimated by the use of Griess Illosvoy reaction [30]. In this study, the Griess-Illosvoy reagent was modified by using naphthyl ethylene diamine dihydrochloride (0.1% w/v) instead of 1-napthylamine (5%). The reaction mixture (3 ml) containing sodium nitroprusside (10 mM, 2 ml), phosphate buffer saline (0.5 ml) and chitosan (0.5–2.0 mg) was incubated at 25 °C for 150 min. After incubation, 0.5 ml of the reaction mixture was mixed with 1 ml of sulfanilic acid
Result and discussion
The FT-IR spectra (3500–600 cm−1) showed some different bands of chitosan. The absorption at 3439 cm−1 is stretching vibration of –OH and –NH2 bonds, indicating the association of the hydrogen-bond between them. The absorptions of approximately 1659, 1379 and 1321 cm−1 are characteristic of the tertiary amine bond. The absorption bond between 2920 and 2873 cm−1 is due to C–H stretch; amide II peak to N–H and C–N torsional vibrations, while amide III peak is associated to CH2 residual groups from
Acknowledgement
This study was supported by Science research fund from health bureau of ChongQing city (2010-2-042): Role and mechanism of loss of equilibrium of Th17/Treg cell subset in acute rejection in renal transplantation and Science research fund from Chongqing Medical University (XBYB2008001): Proteomics analysis in rat renal allograft rejection.
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