The effects of lipophilic substances on the shape of erythrocytes demonstrated by a new in vitro-method
Författare
Summary, in English
Abstract
Low aqueous solubility of lipophilic agents, such as free fatty acids, hampers proper in vitro demonstration of biological effects, yielding an ambiguous in vitro-in vivo correlation. We have therefore developed a method for evaluating the acute effects of lipophilic substances on the shape of erythrocytes and estimated EC50 and Hill coefficient according to the sigmoidal Emax model.
The test substance dissolved in medium-chain triglyceride is coated on a polycarbonate slide which serves as a cover sheet of a Bürker chamber. Freshly collected finger-tip blood is diluted with autologous EDTA-plasma and introduced into the chamber. After ten min at 37 C, the cells are photographed under microscope and the fractions of normal and defect cells are evaluated. No staining is needed and the cells are kept viable during the test period.
With increasing chain length, fatty acids, aliphatic amines and alcohols all increased the fraction of defect erythrocytes in a concentration-dependent manner. The results indicate that several fatty acids are very potent in their acute actions on erythrocytes, and that this effect is due to chain length rather than conformation.
Conclusion: The technique offers a screening method for testing the harmful effects of small amounts of lipophilic substances on erythrocytes.
Low aqueous solubility of lipophilic agents, such as free fatty acids, hampers proper in vitro demonstration of biological effects, yielding an ambiguous in vitro-in vivo correlation. We have therefore developed a method for evaluating the acute effects of lipophilic substances on the shape of erythrocytes and estimated EC50 and Hill coefficient according to the sigmoidal Emax model.
The test substance dissolved in medium-chain triglyceride is coated on a polycarbonate slide which serves as a cover sheet of a Bürker chamber. Freshly collected finger-tip blood is diluted with autologous EDTA-plasma and introduced into the chamber. After ten min at 37 C, the cells are photographed under microscope and the fractions of normal and defect cells are evaluated. No staining is needed and the cells are kept viable during the test period.
With increasing chain length, fatty acids, aliphatic amines and alcohols all increased the fraction of defect erythrocytes in a concentration-dependent manner. The results indicate that several fatty acids are very potent in their acute actions on erythrocytes, and that this effect is due to chain length rather than conformation.
Conclusion: The technique offers a screening method for testing the harmful effects of small amounts of lipophilic substances on erythrocytes.
Avdelning/ar
- Department of Food Technology, Engineering and Nutrition
- Avdelningen för klinisk kemi och farmakologi
Publiceringsår
2009
Språk
Engelska
Sidor
458-464
Publikation/Tidskrift/Serie
European Journal of Pharmaceutical Sciences
Volym
36
Issue
4-5
Dokumenttyp
Artikel i tidskrift
Förlag
Elsevier
Ämne
- Pharmacology and Toxicology
- Medicinal Chemistry
Status
Published
ISBN/ISSN/Övrigt
- ISSN: 1879-0720