# Examination of the Quality of Particulate and Filtered
## 3 Results
### Microbiological Analysis  
#### 3.3.1 Distribution of Gram Staining Forms and Morphology  
Bone chip samples from all four groups contained viable microorganisms, even after Clindamycin pretreatment (Figure 5). In total 259 morphologically different microorganisms were observed. The bacterial species predominantly found belonged to the families of _Streptococcaceae_ and _Staphylococcaceae_ (95.7%). Rod-shaped microorganisms were found in lower percentage (3.9%). Yeast could only be isolated from one bone chip sample (0.4%).  
\begin{table}
\begin{tabular}{c c c c c} \hline \hline
**Patient Group** & **Number of Samples** & **Outgrown Samples** & **Outgrown Time [days]** & **SEM** & **Outgrowth Probability [\%]** \\ \hline Cortical w/antibiotic pretreatment & 15 & 13 & 12.85 \(\pm\) 3.24 & 0.90 & 86.67 \\ \hline Cortical w/o antibiotic pre-treatment & 30 & 21 & 15.81 \(\pm\) 5.32 & 1.16 & 70.00 \\ \hline Cortical-cancellous w/antibiotic pretreatment & 20 & 17 & 13.47 \(\pm\) 5.25 & 1.27 & 85.00 \\ \hline Cortical-cancellous w/o antibiotic pretreatment & 19 & 14 & 7.00 \(\pm\) 2.18 * & 0.58 & 73.68 \\ \hline \hline \end{tabular}
\end{table}
Table 2: Outgrowth probability and time of the bone chip samples with respect to the patient group.  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**Patient Group** & **Number of Samples** & **Outgrown Samples** & **Outgrown Probability [\%]** \\ \hline Cortical w/\& w/o antibiotic pretreatment & 45 & 34 & 75.56 \\ Cortical-cancellous w/\& w/o antibiotic pretreatment & 39 & 31 & 79.49 \\ Cortical and cortical-cancellous w/antibiotic pretreatment & 35 & 30 & 85.71 * \\ Cortical and cortical-cancellous w/o antibiotic pretreatment & 49 & 35 & 71.43 \\ \hline \hline \end{tabular}
\end{table}
Table 3: Outgrowth probability of the bone chip samples after regrouping of the patient groups.  
Figure 4: In vitro osteogenic differentiation. (**A**) Alizarin Red S staining after 28 days in vitro osteogenic differentiation of BDSCs for the patient groups cortical bone chips with antibiotic pre-treatment (group I), cortical bone chips without antibiotic pre-treatment (group II), cortical-cancellous bone chips with antibiotic pre-treatment (group III) and cortical-cancellous bone chips without antibiotic pre-treatment (group IV) as indicated. Main frame: osteogenic induced BDSCs, insert: non-induced BDSCs served as negative control, magnification 40\(\times\). (**B**) RT-PCR for the osteogenic markers osteonectin (ON) and bone morphogenetic protein 2 (BMP-2) after 4 weeks osteogenic differentiation of BDSCs. \(\beta\)-actin served as internal control, H\({}_{2}\)O served as negative control. The pictures shown are representative for outgrowth bone chip samples of the respective group. C: non-induced BDSCs, O: osteogenic induced BDSCs.  
Figure 3: Characterization of bone chip-derived stem cells (BDSCs). (**A**) Isolated dental follicle (**B**) Bone material in grooves of the drill. (**C**) Particulate jawbone material in a syringe. Size bar equals 5 mm. (**D**) Cells proliferating out of the bone chip material. (**E**) RT-PCR for the mesenchymal stem cell markers CD73, CD90, and CD105 in BDSCs for the patient groups: cortical bone chips with antibiotic pre-treatment (group I), cortical bone chips without antibiotic pre-treatment (group III) and cortical-cancellous bone chips without antibiotic pre-treatment (group IV) as indicated. \(\beta\)-Actin served as internal expression control, H\({}_{2}\)O served as negative control. The pictures shown are representative for outgrowth bone chip samples of the respective group.  
In the bone chip samples that showed cell outgrowth, 152 different microorganism forms were detected. 96.1% of those were gram-positive cocci and 3.9% were gram-positive rods. From the bone chip samples for which no cell outgrowth could be observed, 107 diverse shaped microorganisms could be isolated. Among them 95.3% were gram-positive cocci, 3.7% were gram-positive rods and 1 microorganism could be identified as yeast.