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The establishment of non-invasive analytical tools for assessing the in-situ use of biomaterials for surgical implants or scaffolds in tissue engineering and polymer-based therapies is fundamental. This study established a method for fluorescent tracking of the degradation of a chitosan membrane scaffold for use in vitro in bioreactors and ultimately in vivo. The basis of this tracking system is a fluorescence emitting biomaterial obtained by covalent binding of the fluorophore tetramethylrhodamine isothiocyanate (TRITC) onto the backbone of chitosan. Using confocal microscopy, this study quantitated the reductions in fluorescence intensity of the membrane and correlated these decreases with weight loss during polymer breakdown, thereby providing a technique for non-destructively assessing the extent of degradation of chitosan materials over time in vitro. Using multispectral imaging in a mouse model, the study assessed the degradation profile of the fluorophore-labeled biomaterial in vivo in real time and identified the dispersing pathway of the chitosan membrane degradation products in vivo. The results revealed that TRITC conjugated chitosan was biocompatible and supported bone cell growth. The changes in fluorescence intensity correlated well with weight loss up to 16 weeks of in vitro culture and could be monitored over two weeks in vivo.

The state of the art approaches for tailoring the degradation of chitosan scaffolds are based on altering the chemical structure of the polymer. Nevertheless, such alterations may lead to changes in other properties of scaffolds, such as the ability to promote cell adhesion. The aim of this study was to investigate the influence of physical parameters such as porosity and fibre diameter on the degradation of chitosan fibre-mesh scaffolds, as a possible way of tailoring the degradation of such scaffolds. Four sets of scaffolds with distinct fibre diameter and porosity were produced and their response to degradation and cell adhesion was studied. The degradation study was carried out at 37degrees C in a lysozyme solution for five weeks. The extent of degradation was expressed as percentage of weight loss of the dried scaffolds after lysozyme treatment. Cell adhesion was assessed by Confocal Microscopy. The results have shown that the scaffolds with higher porosity degrade faster and that, within the same range of porosity, the fibres with smaller diameter degrade slightly faster. Furthermore, the morphological differences between the scaffolds did not affect the degree of cell adhesion, and the cells were observed throughout the thickness of all four types of scaffolds.

Antimicrobial properties of gallium(III)-and iron(III)-loaded polysaccharides affecting the growth of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, in vitro. ACS Applied Bio Materials, 3(11), pp. 7589-7597.

Scaffolds based on aligned and non-aligned poly (L-lactic acid) (PLLA)/polycaprolactone (PCL) fibers obtained by electrospinning, associated to electrosprayed hydroxyapatite (HA) for tissue engineering applications were developed and their performance was compared in terms of their morphology and biological and mechanical behaviors. The morphological results assessed by scanning electron microscopy showed a mesh of PLLA/PCL fibers (random and perfectly aligned) associated with aggregates of nanophased HA. Fourier transform infrared spectrometry confirmed the homogeneity in the blends and the presence of nanoHA in the scaffold. As a result of fiber alignment a 15-fold increase in Young’s Modulus and an 8-fold increase in tensile strength were observed when compared to non-aligned fibers. In PLLA/PCL/HA scaffolds, the introduction of nanoHA caused a remarkable improvement of the mechanical strength of this material acting as a reinforcement, enhancing the response of these constructs to tensile stress. In vitro testing was evaluated using osteoblast (MC3T3-E1) cells. The results showed that both fibrous scaffolds were able to support osteoblast cell adhesion and proliferation and that fiber alignment induced increased cellular metabolic activity. In addition, the adhesion and proliferation of Staphylococcus aureus were evaluated and a lower number of colony forming units (CFUs) was obtained in the scaffolds with aligned fibers.