RABBIT PYROGEN TEST STUDY OF INFUSIBLE PLATELET MEMBRANE AS A PLATELET SUBSTITUTE FOR BLOOD TRANSFUSION

Abstract

Blood transfusion centers are under considerable pressure to produce platelet concentrates with a shelf life limited to 3-5 days. Many approaches have been investigated experimentally to produce new hemostatically active platelet products that are capable of long term storage. In this article infusible platelet membrane (IPM) as a platelet substitute was investigated with regard to rabbit pyrogen test as a requirement for parenteral drugs according to the EU Pharmacopoeia monograph to demonstrate its safety. Lyophilized IPM was prepared from fresh and outdated platelet concentrates. Platelet concentrates were pooled, disrupted by freeze-thaw procedure and pasteurized for 20 hours to inactivate possible viral or bacterial contaminants. Sterility test, size particle test and rabbit pyrogen test were then performed. Administration of IPM at various dosage forms were carried out on a group of three rabbits with a total of five groups. The summed response of body temperature of each group was calculated. At dosage forms of 1.0, 2.0, 4.0, 6.0 and 9.0 mg per kg of body weight, the summed responses of each group were 0.1, 0.2 , 0.5, 0.7 and 1.0 degree centigrade, respectively. As might be expected, there was shown a strong direct correlation between dose and response with r=0.996. Preparations being examined passed the test according to the EU Pharmacopoeia monograph. It was concluded that IPM as a platelet substitute is sterile and safe without endotoxins and non-endotoxin pyrogens that may be originated from bacteria, intracellular and extracellular proinflamatory cytokines and other biologic response modifiers in conventional platelet concentrates. Keywords: Infusible platelet membrane, rabbit pyrogen test, platelet substitute