Long-term Biocompatibility Evaluation of Porous Biomaterials

The biocompatibility of the synthesized porous materials (CS, CS_Gel, HAp(ESM)_CS_Gel_BA) was additionally evaluated with the help of fluorescence microscopy, using a cell tracker for long-term tracing of living cells, called RED CMTPX fluorophore (Thermo Fischer Scientific, Massachusetts, MA, USA). The synthesized materials were added in the respective cell cultures along with the CMTPX tracker. After 5 days, the morphology and viability of the AFSCs were evaluated. The CMTPX fluorophore was added in the culture medium at a final concentration of 5 μM. The mediums were incubated for 30 min so the dye penetrated the cells completely. After the incubation period, the cells were washed with PBS (phosphate buffer saline) and analyzed by fluorescent microscopy. The photomicrographs were taken with Olympus CKX 41 digital camera driven by CellSense Entry software (Olympus, Tokyo, Japan) [41 (link)].