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Söhling, et al.
A

B

Figure 5. Osteogenic activity of the composite material. (A) MSCs were cultured in the presence of PLA and PLA/BG 20%, respectively.
Calcium deposition was visualized by Alizarin Red staining. The morphometric analysis of alizarin staining is depicted in (B).

suppression after its release, calcium was immediately shows expressed proteins and their main functional
complexed after release by the selective calcium chelator assignment.
EGTA, proportionally added to the medium. MSCs were
incubated for 72 h with control medium, with medium 4. Discussion
conditioned 72 h with BG20 or medium conditioned In this study, a 3D printable biomaterial made of PLA
72 h with BG20 and EGTA. IL-6 gene expression was and a BG content of up to 20% was investigated for
evaluated as a readout parameter. Significant inhibition of cytocompatibility and immunoreactive potential.
IL-6 gene expression by BG20 conditioned medium was Pooled human MSCs were cultured on bilayer printed
recorded. This suppression was almost completely absent disk-shaped test specimens, provided with increasing
by the addition of EGTA (Figure 8). BG fractions, and gene expression was determined.

3.6. Immune Stimulating potential of BG20 Furthermore, osteogenicity and inflammatory potential
of the material were analyzed. Osteogenic genes
Immune stimulating potential of BG20 was determined underwent only slight changes in activity, with only ALP
by whole-blood stimulation assay. Of a total of 105 gene expression increasing significantly after 7 days.
different serum peptide mediators, 23 were detected in Further in vitro analyses showed correspondingly that an
medium control, 24 in PLA control, 31 in LPS control, increasing BG fraction did not lead to an improved OD of
and 23 in BG20 group. Figure 9 shows the relative the MSC, measured by calcium deposition. Interestingly,
expression levels of the mediators depending on the kind the BG fraction significantly inhibited prototypic
of stimulation applied to whole-blood samples. Table 1 inflammatory processes in MSC at the mRNA level. IL-6

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