## 4 Discussion  
### Assessment of Cytotoxicity According to ISO: 10993-5: 2009  
Quantitative and Qualitative Evaluation  
Cytotoxicity assays performed on the FC and FL membranes at different concentrations showed that almost all evaluated LBMs have an acceptable viability. Only two membranes were below the viability threshold, FL70 (61.13%) and FC30 (62.12%), according to ISO: 10993-5: 2009. The FC30 mixture showed the lowest viability percentage of the LBMs evaluated, reaching 62.12%, while the FC70 sample showed the highest percentage of viability (80.84%). However, due to sample size and \begin{table}
\begin{tabular}{c c c c c c} \hline \hline  & **DFT** & **Contrast** & **Entropy** & **SMA** &
\begin{tabular}{c} **Entropy/DFT** \\ **Relation** \\ \end{tabular} \\ \hline FL30 & 2.56215 \({}^{\text{a,b,c}}\) & 187.93 \({}^{\text{a,b}}\) & 8.9805 \({}^{\text{a,b}}\) & 0.000245 \({}^{\text{a,b}}\) & 3.50506 \\ \hline FC30 & 2.6462 \({}^{\text{a}}\) & 311.902 \({}^{\text{a}}\) & 9.393 \({}^{\text{a}}\) & 0.000569 \({}^{\text{a,b}}\) & 3.54961 \\ \hline FL50 & 2.5922 \({}^{\text{b}}\) & 214.2285 \({}^{\text{a,b}}\) & 9.042 \({}^{\text{b}}\) & 0.000381 \({}^{\text{a,b}}\) & 3.48815 \\ \hline FC50 & 2.5896 \({}^{\text{a,b,c}}\) & 215.95 \({}^{\text{a,b}}\) & 9.1265 \({}^{\text{a,b}}\) & 0.00025 \({}^{\text{a,b}}\) & 3.52428 \\ \hline FL70 & 2.5313 \({}^{\text{c}}\) & 183.5485 \({}^{\text{b}}\) & 8.996 \({}^{\text{b,c}}\) & 0.000182 \({}^{\text{a}}\) & 3.55390 \\ \hline FC70 & 2.5948 \({}^{\text{a,b,c}}\) & 222.6915 \({}^{\text{a,b}}\) & 9.157 \({}^{\text{a,b}}\) & 0.000238 \({}^{\text{b}}\) & 3.52898 \\ \hline \hline \end{tabular} \({}^{\text{(a,b,c)}}\) values (average \(\pm\) standard deviation, \(n=5\)) in a column with the different letters show significant differences (\(p<0.05\)).  
\end{table} 
Table 7: Analysis of texture images of the ESEM micrographs of the LMBs (SMA: second angular momentum). media volume as well as direct contact and lack of evidence of adherence, this would be a preliminary in vitro evaluation. In stereotypically observed micrographs (Figure 2) of the different cytotoxicity evaluations, the fibroblast culture media are observed in a typical arrangement, in the form of a spindle with a distance growth similar to that of the control or blank in the cases of FC samples. The zone of inhibition and minimization of growth was less obvious in the case of FC30 and FC50. For the FL50 and FL70 membranes, a large number of many devitalized cells are observed in the vicinity of the material, as well as small areas of inhibition. In this case, the cell is adopting a contracted form, which would indicate a low adhesion with the scaffolding material. Clear differences are observed among the morphologies adopted by the cells in these samples, leading to the conclusion that cell adhesion depends on the physicochemical characteristics of the surface of the material [43]. Lignocellulosic fibers have hygroscopic properties [44]. This is consistent with many results in which different cell lines showed variable adhesion based on surface wettability [45]. Cells have been considered to generally adhere better to moderately hydrophilic surfaces compared to hydrophobic or hydrophilic ends [46]. The qualitative cytotoxic behavior was classified as mild reactivity since no more than 20% of the cells are round, freely bound and without intracytoplasmic granules, and these do not show changes in morphology (Table 3); a slight inhibition of the growth of occasional lysed cells is observed. LBMs showed considerable swelling due to the absorbed culture medium because of the insert. In the case of the FC30 membrane, there was a deformation and shedding of it. Some results have showed that LBMs do not produce cytotoxic effects in indirect contact with the culture of fibroblast cells in vitro, since no major malformation, degeneration, propagation, or lysis of fibroblast cells was observed according to stereoscopic micrographs [47]. Any cytotoxic effect may be cause for concern.