In-vitro and in-vivo degradation studies of freeze gelated porous chitosan composite scaffolds for tissue engineering applications

Tissue engineering approaches have been adapted to reconstruct and restore functionality of impaired tissue for decades. Porous biomimetic composite scaffolds of Chitosan (CH) with hydroxyapatite (HA) for bone regeneration have also been extensively studied in the past. These porous scaffolds play a critical role in providing successful regeneration by acting as a three-dimensional template for delivering nutrients and metabolites and the removal of waste by products. The aim of the current study was to investigate in-vitro and in-vivo degradation rates of porous freeze gelated chitosan (CH) and CH hydroxyapatite scaffolds by scanning electron microscopy (SEM) to observe for morphological changes, Fourier Transform Infrared Spectroscopy (FTIR) in conjunction with photo-acoustic sampling (PAS) accessory for the analysis of chemical changes, pH analysis and UV–Vis spectroscopy of degraded supernatant. SEM results showed significant alterations in the surface morphology. FTIR-PAS spectra showed changes in the finger print region and glycosidic bonds showed signs of breakage. pH values and UV–Vis spectroscopy of the degraded supernatant were indicative of CH bonds scission in neat samples. HA incorporated specimens showed more stability. Histological sections performed after in-vivo implantation also showed greater cellular infiltration and delayed degradation profiles by HA loaded samples. Within 30 days of implantation, neat CH scaffolds showed complete in-vivo biodegradation. The current findings show the advantage of adding hydroxyapatite to porous templates which enhances hard tissue regeneration. In addition, it allows easy and cost effective fabrication of bioactive composite scaffolds

Ultrasound irradiation in the production of ethanol from biomass

Ethanol produced from renewable biomass, such as lignocellulosic feedstock, is one of the alternative energy resources that can be environmentally friendly. However, physical and chemical barriers caused by the close association of the main components of lignocellulosic biomass, as well as starch, hinder the hydrolysis of cellulose and hemicellulose in lignocellulose as well as amylase and amylopectin in starch to fermentable sugars. One of the main goals of pretreatment for enzymatic hydrolysis is to increase the enzyme accessibility for improving digestibility of cellulose and starch. Ultrasound irradiation applied to cellulosic materials and starch-based feedstock was found to enhance the efficiency of hydrolysis and subsequently increase the sugar yield. Prior research conducted on applying ultrasonic technology for cellulose and starch pretreatment has considered a variety of effects on physical and chemical characteristics, hydrolysis efficiency and ethanol yield. This paper reviews the application of ultrasound irradiation to cellulose and starch prior to and during hydrolysis in terms of sugar and ethanol yields. It also addresses characteristics such as accessibility, crystallinity, degree of polymerization, morphological structure, swelling power, particle size and viscosity as influenced by ultrasonic treatment. © 2014 Elsevier Ltd

Preparation of gold nanoparticles stabilized by chitosan using irradiation and sonication methods

Gold nanoparticles (AuNPs) were synthesized, in the absence of any reducing agent, using ionizing radiation or ultrasound in aqueous solutions of chloroauric acid (HAuCl4). Chitosan (average molecular weight 158 kDa, degree of deacetylation 90 %) was used as a stabilizing agent. Both techniques yielded AuNPs which were stable in solution at RT for at least 3 months after synthesis. UV-Vis spectroscopy was used to follow substrate decay, nanoparticles formation, size of the gold core and particles stability. Hydrodynamic radii and polydispersion of the chitozan-stabilized AuNPs (i.e. the whole core-shell nanoparticles) were determined by dynamic light scattering. Zeta potential measurements were performed to assess the surface charge and stability of the particles. Influence of synthesis parameters and presence of isopropanol on the formation and properties of the products have been described and reaction mechanisms have been discussed. Radiation and sonochemical methods are demonstrated to be very efficient, fast and easy-to-control methods of synthesizing gold nanoparticles, leaving behind no unreacted reducing agent or unwanted side products, while stabilization by chitosan provides AuNPs with excellent stability and long shelf life

Procedure for determination of the molecular weight of chitosan by viscometry

The aim of this publication is to facilitate the estimation of chitosan molecular weight (MW) in laboratories with no access to sophisticated analytical instruments, by applying the easily accessible and economical capillary viscometry. The procedure of viscosity-average molecular weight (Mv) determination is described in details. The examples provided encompass testing of the experimental procedure for determination of the Mv of chitosan with a low-molecular weight of 7.7 kDa and 88 kDa, after verification with a high-molecular weight polymer (477 kDa). The experimental work demonstrated the importance of the initial concentration of low-MW chitosan for the accurate determination of intrinsic viscosity and, as a consequence, the viscosity-average molecular weight

Determination of degree of deacetylation of chitosan - comparision of methods

Degree of deacetylation (DD) is one of the main parameters characterizing chitosan. The most precise measurements of DD require sophisticated equipment (NMR spectrometer), not available at many laboratories worldwide working on chitosan. There is a need for low-cost, simple, yet sturdy and reliable methods and procedures for DD determination. The aim of this work was to test and compare – on the same set of chitosan samples - a few of the existing analytical techniques and provide recommendation for selecting DD determination methods. Tests were performed on four chitosans of nominal DD in the range 70 - 95%. Three different methods of titration, two different methods of spectroscopy UV / Vis and infrared spectroscopy using various calculation approaches were used. The results are summarized and compared with values obtained by 1H NMR, considered as the reference method. Moreover, evaluation of the ease of performance and availability of reagents in the given methods was performed. On that basis, recommendations for selection of DD determination methods were formulated

Synthesis of chitosan and carboxymethyl chitosan hydrogels by electron beam irradiation

Solutions of chitosan (CS) and carboxymethyl chitosan (CMCS) were subjected to irradiation by electron beam in presence of a crosslinking agent (poly(ethylene glycol) diacrylate - PEGDA) in order to produce carboxymethyl chitosan- and chitosan-based hydrogels. PEGDA macromonomer itself undergoes simultaneous polymerization and crosslinking either in neutral water or in acidic medium. Acidic solutions of chitosan of 0.5, 1 and 2% can be effectively crosslinked with PEGDA to form a gel. Although CMCS undergoes radiation-initiated crosslinking only at high concentration in water (over 10%), the presence of PEGDA in solution facilitated hydrogel formation even at lower concentration of CMCS. Formation of CS and CMCS hydrogels required irradiation doses lower than those needed for sterilization, i.e. 25 kGy, in some cases even as low as 200 Gy. Sol-gel analysis revealed relatively high gel fraction of obtained hydrogels, up to 80%, and good swelling ability. Both parameters can be easily controlled by composition of the initial solution and irradiation dose. Possible mechanisms of crosslinking reactions were proposed, involving addition of the polysaccharide macroradicals to a terminal double bond of PEGDA