Laboratorijska analiza hemolitičkih svojstava materijala koji dolaze u kontakt sa krvlju - uporedna primena dve varijante metode ispitivanja prema standardu ASTM F756 u skladu sa ISO 10993-4

Abstract

U ovom radu prikazano je poređenje rezultata dve varijante in vitro metode ispitivanja prema standardu ASTM F756 u skladu sa ISO 10993-4 hemolitičkih svojstava šest polaznih materijala (Polipropilen Moplen EP 540 P, Polikarbonat bezbojan 164 R- 112, Polikarbonat braon 164 R-51918, Polietilen NG 3026 K, Polietilen NG - Purell GB 7250, Polietilen VG - Hiplex 5502) za proizvodnju medicinskih sredstava i jednog materijala (Polietilen NG granulat) za proizvodnju boca za infuzione rastvore. Jedna varijanta metode zasniva se na direktnom kontaktu polaznog materijala sa svinjskom krvlju, a druga na dobijanju ekstrakta polaznog materijala autoklaviranjem i kontaktu tako dobijenog ekstrakta sa svinjskom krvlju. Na osnovu dobijenih rezultata zaključeno je da svi materijali zadovoljavaju kriterijume prihvatljivosti i to u slučaju obe varijante metode ispitivanja, kao i da metoda direktnog kontakta ima veću osetljivost od metode u kojoj su korišćeni ekstrakti ovih materijala. .

The presence of hemolytic material in contact with blood may produce increased levels of blood cell lysis and increased levels of plasma hemoglobin. This may induce toxic effects or other effects which may stress the kidneys or other organs. In this paper two variants of in vitro method and obtained results' comparison were presented for testing of hemolytic properties of six raw materials (polipropylene Moplen EP 540 P, policarbonate colorless 164 R-112, policarbonate brown 164 R-51918, polietylene NG 3026 K, polietylene NG - Purell GB 7250, polietylene VG - Hiplex 5502) for medical device manufacturing and one raw material (polietylen NG granulate) used for infusion solutions plastic bottles manufacturing. One of the method variants relies on direct contact of the raw material with swine blood and the other on contact of extract of the material with swine blood. Both variants imply reading of the absorbance of the supernatant after tubes were incubated and centrifuged. According to the values obtained and using the standard curve free hemoglobin concentration is determined and based on this percentage hemolysis of raw material. Positive and negative controls were used in both variants where water for injection (WFI) was used as positive control in which partial or complete hemolysis of erythrocytes occurs due to osmotic shock and phosphate buffer saline was used as negative control with no hemolytic property. In this paper a comparison of results obtained by both method variants for testing of seven raw materials is presented, while these conclusions cannot be used neither for all materials, nor for all applications without preliminary testing using both variants and then choosing more sensitive and more reliable one. It was shown and stated in the paper as well that incubation time being 3, 15 or 24 h, had no impact on the variants with direct contact sensitivity. This comparative approach was used for drawing conclusions in terms of suitability for application of one or the other method variant, as well as for defining relevant incubation time and finally for choosing the more sensitive and more reliable variant for assessment of hemolytic properties of raw materials. The variant with direct contact was chosen from the aspect of less complexity regarding necessary laboratory equipment which makes it economically more favorable and fit for the purpose. .