title: Optimization of platelet isolation and extraction of autogenous TGF-beta in cartilage tissue engineering.
abstract: 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.
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, Hospital Rechts der Isar - Munich, Technical University Munich, Munich, Germany. r.staudenmaier@lrz.tum.de
authors.forename: Rainer
authors.initials: R
authors.lastname: Staudenmaier
authors.forename: K
authors.initials: K
authors.lastname: Froelich
authors.forename: M
authors.initials: M
authors.lastname: Birner
authors.forename: J
authors.initials: J
authors.lastname: Kindermann
authors.forename: Nguyen
authors.initials: N
authors.lastname: The Hoang
authors.forename: R C
authors.initials: RC
authors.lastname: Pueschel
authors.forename: V
authors.initials: V
authors.lastname: Mandlik
mesh_terms: 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*
chemical_list: D016212:Transforming Growth Factor beta