The purpose of this study was to determine the types of inflammatory cells and bacterial contamination on expanded polytetrafluoroethylene (ePTFE) membranes which might affect new tissue formed by guided tissue regeneration (GTR). Forty periodontal bony defects were treated by the flap procedure, which included the use of an ePTFE membrane. Twelve months after the second surgery, the defect sites were re-evaluated for changes in probing depth and clinical attachment level. The ePTFE membranes were retrieved after 4 to 6 weeks of healing and sectioned serially at 3 microm in a coronal-apical plane. The ePTFE membrane was divided into 3 portions: cervical, middle, and apical, each of which was subdivided into outer, central, and inner segments, providing a total of 9 fields. Cells and bacteria were analyzed by light microscopy for their types: mononuclear cell, erythrocyte, fibroblast, neutrophil, plasma cell, T lymphocyte, B lymphocyte, macrophage, and oral bacteria. Both cells and bacteria decreased in number towards the apical portion and were present even in the central part. Most cells were mononuclear cells. Erythrocytes, fibroblasts, neutrophils, and plasma cells were rarely encountered. Bacteria, most of which were Gram-positive, were observed in almost the same number in the outer and inner parts. The results indicate that numerous inflammatory cells adhered to and invaded the ePTFE membranes accompanied by bacterial contamination and that there was a tendency for a negative correlation between the increment number of bacteria and the gain of clinical attachment level.