# Adipose Stem Cell Response to Borophosphate Bioactive Glass
## 3 Results  
### Glass Properties  
Table 3 displays the pH as reported by [PERSON] et al. [39], recorded 24 h post the addition of 3 mg/mL of the three glasses from the current study into DI water at 37\({}^{\circ}\)C. The borate-free X0 glass yielded an acidic solution (pH \(\sim\) 2.6); the phosphate free X60 glass produced an alkaline solution (pH \(\sim\) 8.5), and the X40 borophosphate glass produced a pH-neutral solution (pH \(\sim\) 7). These same glass particles were introduced to ASCs under standard culture conditions, and both the pH and the released ions were measured (Table 3). After 24 h, the pH values of the CCM with the acidic X0 glass and with the pH-neutral X40 glass particles showed no significant differences from the baseline CCM values. However, the CCM pH with the alkaline X60 glass particles exhibited a slight increase. The difference in the pH values from the water experiments and the CCM experiments reflects the buffering effects of the latter. \begin{table}
\begin{tabular}{c c c c c c c c c} \hline \hline \multirow{2}{*}{**Sample**} & \multirow{2}{*}{**Time (h)**} & \multicolumn{3}{c}{**pH**} & \multicolumn{3}{c}{**Ions Released**} \\ \cline{3-8}  & & & \multicolumn{2}{c}{**P**} & \multicolumn{3}{c}{**B**} & \multicolumn{2}{c}{**Ca**} \\ \cline{3-8}  & & & **DPm** & **\%** & **ppm** & **\%** & **ppm** & **\%** \\ \hline \multirow{3}{*}{CCM} & 5 & ND & 7.32 & 34.80 \(\pm\) 1.45 & ND & ND & ND & 65.82 \(\pm\) 2.68 & ND \\ \cline{2-8}  & 24 & ND & 7.53 & 36.65 \(\pm\) 0.88 & ND & ND & ND & 68.41 \(\pm\) 1.07 & ND \\ \hline \multirow{3}{*}{X0} & 5 & ND & 7.23 & 72.04 \(\pm\) 0.47 & 4 & ND & ND & 68.84 \(\pm\) 2.67 & 1 \\ \cline{2-8}  & 24 & 2.6 & 7.49 & 197.03 \(\pm\) 5.91 & 19 & ND & ND & 101.28 \(\pm\) 1.64 & 15 \\ \hline \multirow{3}{*}{X40} & 5 & ND & 7.25 & 121.06 \(\pm\) 3.61 & 22 & 126.10 \(\pm\) 1.46 & 46 & 82.36 \(\pm\) 1.37 & 5 \\ \cline{2-8}  & 24 & 7.1 & 7.39 & 176.01 \(\pm\) 4.24 & 36 & 217.40 \(\pm\) 1.06 & 80 & 73.26 \(\pm\) 1.11 & 2 \\ \hline \multirow{3}{*}{X60} & 5 & ND & 7.86 & 23.27 \(\pm\) 0.30 & ND & 249.43 \(\pm\) 8.00 & 50 & 173.16 \(\pm\) 3.79 & 29 \\ \cline{2-8}  & 24 & 8.6 & 7.67 & 24.96 \(\pm\) 0.18 & ND & 301.26 \(\pm\) 6.03 & 61 & 201.43 \(\pm\) 5.03 & 36 \\ \hline \hline \end{tabular}
\end{table} 
Table 3: The pH and ion concentration of the cell culture media were measured after interactions with 2.5 mg/mL BPBGs for the specified durations. The pH values for deionized water (DI) correspond to 3 mg/mL of each glass after 24 h, as reported previously [39]. The percent refers to the proportions of ions released from each glass into the media. ND: not detected. Table 3 also shows the respective concentrations of calcium, borate, and phosphate species in the CCM solutions after 5 h and 24 h with the glass particles. Borate ions were not detected in either the CCM baseline or the CCM after reacting with the X0 particles. However, significant concentrations of borate ions were found in the CCM solutions containing X40 and X60 particles. Approximately 80% of the borate ions were released from the X40 glass, while \(\sim\)61% of borate ions were released from the X60 glass after 24 h (Table 3). Similar calculations were performed for the phosphate and calcium concentrations in solution, after normalizing for their respective concentrations in the baseline CCM. These values, however, are likely affected by the precipitation of calcium phosphate phases on the surfaces of the reacting glass particles [39]. The calcium ion concentrations of the CCM after 24 h with X0 and X40 glasses were slightly greater than the baseline concentrations, but the CCM with X60 particles had much greater concentrations of calcium ions. For both X0 and X40 glasses, the phosphate content of the CCM solutions increased over the baseline values at both timepoints evaluated. For X60, the phosphate content of the CCM solution decreased from about 36 ppm to 24 ppm after reacting with the X60 glass particles.