## 3 Results
The B-antiseptic released from BC displayed significantly higher antimicrobial activity against all analyzed microorganisms in comparison to PO and P (Kruskal-Wallis test with post-hoc [PERSON]'s analysis, \(p<0.0001\), Table 2). Release of PO and P from BC also inhibited microbial growth, however there were no statistically significant differences between inhibition zone sizes formed in result of activity of these two antiseptics ([PERSON] test with post-hoc [PERSON]'s analysis, \(p>0.0001\), Table 2). _P. aeruginosa_ was most resistant to all applied antiseptics among tested species, while _A. actinomycetemcomitans_ was the most sensitive. Very high _C. albicans_ growth inhibition zones were observed in the setting of antiseptic B (Table 2). These analyses, with results presented above, were designed to investigate whether antiseptic agents were released from BC carriers to demonstrate antimicrobial activity. Next, another experiment was performed, namely the culturing biofilms of tested pathogens on discs made of HA, which is the main inorganic building component of bone and teeth. The diameter of the customized HA discs was 12 mm; its surface is presented in Figure 6A. The ability of microorganisms to form multilayer biofilms on the substrate was presented in earlier work of ours [33; 34; 35]; herein, we show it in Figure 6B with _S. aureus_ as an exemplar. Next, the activity of antiseptic BC carriers was analyzed in in vitro settings imitating the environment of the oral cavity, as presented in Section 2.12. Over the contact time of 24 h, all antiseptics were able to completely eradicate the biofilm-forming cells from the HA surface. Results of the antiseptic efficiency for shorter contact times (e.g., 1 h and 30 min) are presented in Figures 7 and 8, respectively. \begin{table}
\begin{tabular}{c c c c c c c} \hline \hline  & & & & & & \\ \hline \hline
**Heading** & S. _ aureus_ & _E. faecalis_ & _P. aeruginosa_ & _C. albicans_ & _Str. mutans_ & _A.actinomycetemcomitans_ \\ \hline B & \(25\pm 1\) & \(24\pm 1\) & \(9\pm 2\) & \(28\pm 3.5\) & \(23\pm 3\) & \(34\pm 4.5\) \\ PO & \(6\pm 0.5\) & \(5\pm 1\) & \(2\pm 1\) & \(4\pm 0.3\) & \(11\pm 1\) & \(10\pm 1.5\) \\ P & \(5\pm 1\) & \(3\pm 0.5\) & \(2\pm 0.5\) & \(5\pm 1\) & \(10\pm 1.5\) & \(9\pm 2\) \\ \hline \hline \end{tabular}
\end{table} 
Table 2: Average microbial growth inhibition zone results of the antimicrobial activity of antiseptics released from BC carriers. Figure 6: _Cont._  
Figure 6: Surface of HA disc native (**A**) 1000\(\times\) magnification; and (**B**) covered with biofilm of _S. aureus_ at 500\(\times\) magnification with the inset (**b\({}^{\prime}\)**) at 20,000\(\times\) magnification. Red arrows indicate: a—top layer of the biofilm, b—cross-section of the biofilm from the top layer to the basal layer, and c—surface of the biofilm growth. Blue arrow indicates a magnified picture of the biofilm cross-section showing numerous layers of staphylococcal cells within the biofilm structure. Images were taken on an EVO MA microscope. Figure 7: Percentage reduction of biofilm-forming cells from HA discs after 1 h contact time. B, PO, and P stand for Braunol, Pront Oral, and Prontosan, respectively. Aa, Ef, Sa, Pa, and Ca stand for _A. actinomycetemcomitans, E. faecalis, P. aeruginosa,_ and _C. albicans_, respectively. As shown in Figures 7 and 8, the resistance levels of all pathogens against B, PO, and P were grouped into two clearly distinct sets. One of them clustered _S. aureus_, _E. faecalis_, and _A. actinomycetemcomitans_ together, while another one clustered _C. albicans_ and _P. aeruginosa_ together. In the case of the first group, antiseptics displayed a high (\(>\)70%) antibiofilm efficacy, while in the case of the latter group, the analogical cut-off point was established at 60% and its lowest values were at 23% (_C. albicans_ biofilm versus P with a 30 min contact time, Figure 8). Differences observed between the level of eradication of _S. aureus_, _E. faecalis_, and _A. actinomycetemcomitans_ versus _C. albicans_ and _P.