OBJECTIVEBiodegradable collagen membranes are widely used in guided bone regeneration. This study evaluated the effect of enamel matrix proteins (Emdogain®) combined with two collagen membranes (Bio-Gide® or Collprotect®) on osteoblastic cell cultures.METHODSHuman osteoblastic cells were cultured on Bio-Gide® or Collprotect® membranes coated with Emdogain® or left uncoated. The assessed parameters included amelogenin quantification at 1 h, 12 h, 1 day, 3 days, and 7 days; cell morphology at 1 day; cell proliferation at 1, 2, and 3 days; gene expression of COL1A1, IBSP, SPP1, and BGLAP at 1 day; Collagen type I quantification at 3 and 7 days; alkaline phosphatase (ALP) activity at 3 and 7 days; and mineralization at 14 days. Data were analyzed using two-way analysis of variance or the Kruskal-Wallis test.RESULTSHigher amelogenin levels were detected in Bio-Gide® cultures than in Collprotect® cultures after 3 and 7 days. Cells adhered and spread in all experimental groups. Cell proliferation was higher in Bio-Gide® cultures after 3 days (p<0.05). Gene expression of COL1A1, IBSP, and SPP1 was greater in Bio-Gide® cultures with Emdogain® after 1 day. There was higher collagen type I secretion by cultures grown on collagen membranes coated with Emdogain®, particularly on Bio-Gide® cultures at 7 days. ALP activity was also higher in Bio-Gide® cultures at 7 days (p<0.05). Greater mineralization was detected in cultures grown on Bio-Gide® with Emdogain® (p<0.05).CONCLUSIONCombining collagen membranes, particularly Bio-Gide®, with enamel matrix proteins promotes osteogenesis in vitro, potentially advancing bone tissue regeneration and preservation methods.