In-vitro cytocompatibility and growth factor content of GBR/GTR membranes

To assess the cytocompatibility of five commercially available xenogenic barrier membranes used for oral regenerative procedures and to determine the growth factor content of these membranes in-vitro.

Human mesenchymal stem cells (hMSCs) and immortalized periodontal ligament stem cells (PDL-hTERTs) were used to determine the cytocompatibility of xenogenic barrier membranes made of collagen (Biogide, BG, Geistlich Pharma AG, Switzerland; Biomend, BM, Zimmer Biomet, USA; Osseoguard OG, Zimmer Biomet, USA; OssixPlus, OX, Datum Dental, Israel) or extracellular matrix (ECM) (Dynamatrix, DM, Keystone Dental, USA) and of their eluates obtained by washing. Cells were cultured with previously washed and unwashed membranes (n=4) and in the medium used for washing (eluate). Cell proliferation at 3 days (eluates) and at 7 days (membranes) was assessed using the WST-1 cell proliferation kit. Growth factor content of the membranes was measured using multiplex ELISA.

The eluate of BG and BM significantly inhibited proliferation of hMSCs, whereas DM and OX showed stimulating effects. The highest impact was observed for DM, its eluate doubled the cell proliferation of adherent cells when compared to the control (p<0.001). The eluate of OG did not influence eluate cell cultures (p>0.05). The presence of membranes had different impact on hMSCs and PDLs. hMSCs seem to be more resistant to the inhibitory effects of BG, OG and BM. hMSCs are only affected by OX, which actually stimulates hMSCs when the specimens are not washed previously. PDLs however proliferate significantly less once they are placed into culture with BM and OG as well as BG-not washed. Once BG is washed no inhibitory effect on PDLs was observed, however overall the washing of membrane samples prior to the placement into the cell culture did hardly have any effect on the outcome. The strongest inhibition of proliferation was shown with the BM and OG membrane in PDL-hTERTs (p<0.001). Growth factor contents were quite similar quantitatively and qualitatively among the tested membranes with concentrations in the range of 50-500 pg/ml. Intriguingly DM contained considerably higher amounts of bFGF with up to 8000 pg/ml.

The collagen membranes cross-linked with aldehydes show poor outcomes in PDLs while the collagen membrane cross-linked with polysaccharides generally shows promising results similar to the ECM-membrane DM in both membrane and eluate tests. The findings may be due to various factors, especially differences observed in composition, processing and bFGF content.