## 4 Discussion
### Biocompatibility and Pharmaceutical Activity  
Toxicity is among the one most common issues related to the fabrication of biomimetrics, particularly if they are designed to be left in situ for long periods [38]. Cell cultures with human fibroblasts were designed to evaluate the potential cytotoxicity of VitE 0.1 and VitE 1 scaffolds. Cytotoxicity evaluation usually takes 48 h maximum; in this case, both the proliferation through the scaffolds and the possible cell toxicity over long periods of time were taken into consideration. Excellent biocompatibility was overall demonstrated, and in fact, no signs of toxicity were evidenced over the 28 days of cultures in any of the tested scaffolds. Interestingly, the cells started to adhere to the 3D scaffold surfaces with a little delay in the VitE 1 scaffolds, and, in general, all of the scaffold types seemed to adhere more and more over time on the substrate (Figure 7). A short delay in terms of attachment followed by boosted proliferation was also already observed and measured in the previously work referenced [27].  
VitE, apart from showing no toxicity, was demonstrated to be a good radical scavenger. In fact, a discrete antioxidant charge was observed after 5 min in all of the samples. In particular, VitE 1 increased its antioxidant activity over 47 min of analysis, while VitE 0.1 activity remained constant. The idea that the wider reservoir of VitE in the scaffolds might lead to a more prolonged antioxidant activity should not be excluded. Moreover, organic solvents were used in this experimental model to accelerate the reaction and to measure it in a reliable way; however, the scaffolds would not be applied in a similar environment. Indeed, VitE quickly dissolves in organic solvents, favoring its release from the scaffolds. In accordance with dissolution in ethanol, it is quite probable that in less than one hour, only a small amount of the drug would have been released, suggesting that once applied in situ, being that the release much slower, the antioxidant activity could be yielded in a lower amount and over a longer time. The demonstrated antioxidant activity of the VitE scaffolds might create a favorable environment for the development of re-growing tissue in wounds.  
The test was conducted using scaffolds as large as antibiogram disks and clearly showed that no bacteria grew under the scaffold area. Inhibition ring measurements were only 2 mm wider than the diameter of the disks, probably due to the poor solubility of VitE in the culture medium. Indeed, the VitE scaffolds were not conceived as systems intended to freely release the compound but rather were intended as systems incorporating it. This approach is compatible with intrinsic chitosan scaffold degradability over long periods. Since chitosan is the agent that is supposed to provide antimicrobial activity and since its biodegradation velocity due to enzymes present in vivo is too complicated to be generally defined [39], its actual antimicrobial activity should be measured in an environment that is more representative of chronic wounds. Good signs of antimicrobial potential for the chitosan-based scaffolds containing VitE were still evident from the experimental results.