Optimization of Platelet Isolation and Extraction of Autogenous TGF-ß in Cartilage Tissue Engineering

Abstract

Platelets are enriched with Transforming Growth Factor-ß (TGF-ß). However, information is limited concerning TGF-ß's effects at the molecular level. Nevertheless, it has been demonstrated that TGF-ß activates cell proliferation and its postitive influence on cartilage formation has been proven within the field of Tissue Engineering (TE). As Platelet Rich Plasma (PRP) contains TGF-ß, it was the purpose of this study to optimize PRP-isolation for further TGF-ß extraction. Red blood cell count (RBC) was separated from whole blood by centrifugation. From the supernatant PRP and platelet poor plasma (PPP) layer, the latter supernatant was re-centrifuged to extract PRP. Various experimental series were run to investigate influences concerning anticoagulating alternatives, different amounts of buffer, various centrifugal forces, or substituting centrifugation for sedimentation. TGF-ß levels were determined using ELISA. The technique of platelet-/ TGF-ß-extraction described here proves to be more effective than other methods, is easily repeatable and nottime-consuming, which predisposes it for TE requirements.

Keywords::

• autologous platelets
• centrifugation
• TGF-ß
• cartilage
• tissue engineering

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.