Proliferation and Morphological fine assessment of human periodontal ligament fibroblast cultures towards bovine pericardial membranes

Regenerative dental medicine increased the development of performing scaffold materials. These scaffolds,which induce cell adhesion, support, and guide the tissues’ growth, include membranes which are eitherresorbable or not. This study investigated the proliferation abilities and attachment of human periodontalligament fibroblasts (HPLFs) over two bovine pericardial membranes with different thicknesses, namely0.2mm and 0.4mm respectively. The HPLFs were cultured at the standard condition and exposed to the testedmaterials. XTT (a colorimetric assay for non-radioactive quantification) was performed to assess cellproliferation, while light microscopy (LM) and scanning electron microscopy (SEM) observation assessedfibroblast morphology at different times (T1-T2-T3). Proliferations assays reported a statistically significantdifference in growth at T1 (p<0.05) in the cells cultured with a thicker membrane than the thinner one. LMobservations showed healthy fibroblasts in contact with the membranes, appearing larger and with apolygonal shape. In addition, the SEM images displayed a thickening of the fibroblasts, which continued toadhere to the membrane’s surfaces, with an enlarged polygonal shape and developed filopodia andlamellipodia. These results showed a similar cell behaviour over the two bovine pericardial membranes.Furthermore, they demonstrated a cellular migration along and within the membrane layers, binding withmembrane fibres using filopodial extensions. The findings on the effects of the collagen membranes oncellular behaviour will help clinicians to choose the best type of scaffolds according to the clinical need.