Optimization of platelet isolation and extraction of autogenous TGF-beta in cartilage tissue engineering.
Platelets are enriched with Transforming Growth Factor-beta (TGF-beta). However, information is limited concerning TGF-beta's effects at the molecular level. Nevertheless, it has been demonstrated that TGF-beta activates cell proliferation and its positive influence on cartilage formation has been proven within the field of Tissue Engineering (TE). As Platelet Rich Plasma (PRP) contains TGF-beta, it was the purpose of this study to optimize PRP-isolation for further TGF-beta 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-beta levels were determined using ELISA. The technique of platelet-/ TGF-beta-extraction described here proves to be more effective than other methods, is easily repeatable and not time-consuming, which predisposes it for TE requirements.
Staudenmaier
Rainer
R
Department of Otorhinolaryngology, Head and Neck Surgery, Hospital Rechts der Isar - Munich, Technical University Munich, Munich, Germany. r.staudenmaier@lrz.tum.de
Froelich
K
K
Birner
M
M
Kindermann
J
J
The Hoang
Nguyen
N
Pueschel
R C
RC
Mandlik
V
V
D000818:Animals; D001792:Blood Platelets / Q000166:cytology*; D002356:Cartilage / Q000166:cytology; D002469:Cell Separation / Q000379:methods*; D002498:Centrifugation; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D010976:Platelet Count; D023822:Tissue Engineering / Q000379:methods*; D016212:Transforming Growth Factor beta / Q000032:analysis / Q000302:isolation & purification*
D016212:Transforming Growth Factor beta