RESUMEN
The purpose of this study was to evaluate the influence of fibrin glue and aprotinin on the growth of adult human skin keratinocytes in defined serum-free conditions. The keratinocytes were cultured on cell culture plastics and on a fibrin matrix prepared from fibrin glue. The cell growth was measured by MTT assay, while the growth of clonogenic keratinocytes was evaluated by colony assay and expressed as colony-forming efficiency (CFE). The clonogenic potential of keratinocytes released from subconfluent and confluent cultures grown on fibrin glue was also studied by the colony assay. In comparison to a plastic culture surface the fibrin glue had significantly (P<0.05) increased the clonogenic potential of keratinocytes, as well as enhanced their growth. Keratinocytes released from subconfluent cultures grown on fibrin glue attained a significantly (P<0.05) higher percentage of clonogenic cells than their confluent parallels. At 75, 150, 300 and 450 KIU/ml aprotinin did not influence the growth of keratinocytes (P>0.2). A fibrin-based skin substitute produced in the defined keratinocyte medium could be safely used to treat a number of skin defects.
Asunto(s)
Evaluación Preclínica de Medicamentos/métodos , Adhesivo de Tejido de Fibrina/síntesis química , Queratinocitos/química , Piel Artificial , Aprotinina/farmacología , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Adhesivo de Tejido de Fibrina/metabolismo , Humanos , Queratinocitos/citología , Queratinocitos/efectos de los fármacos , Sales de TetrazolioRESUMEN
Autologous fibrin glue was used in 20 patients undergoing lung resection to reduce pulmonary air leaks and improve hemostasis. The fibrinogen in the glue was prepared by ethanol precipitation of plasma separated from 88 ml of the patient's blood. The mean volume of fibrinogen concentrate +/- SD was 4.9 +/- 0.5 ml with a fibrinogen concentration of 28 +/- 5 mg/ml. The yield obtained by the separation was 81% +/- 9%. One part of fibrinogen concentrate was converted to solid fibrin by means of 0.3 parts of thrombin solution. The outcome was 6.4 ml of two-component fibrin glue. The preparation was performed in a closed system to ensure sterility, and was completed within 90 min. Pulmonary air leak decreased following sealing of the resection lines with autologous fibrin glue and the hemostasis was effective. No adverse effects were observed, and all cultures from the glue were negative. Autologous fibrin glue has the obvious advantages of safety from transmission of viral diseases and from immunological reactions. In summary, we report a new technique for preparing autologous fibrin glue with a high concentration of fibrinogen making it a safe and effective sealant of pulmonary air leak and hemostatic agent in thoracic surgery.