Treatment of delayed bone healing and non‐unions after fractures, osteotomies or arthrodesis still is a relevant clinical challenge. Artificially applied growth factors can increase bone healing and progressively gain importance in clinical routine. The aim of this study was to determine the effects of rhPDGF‐BB, rhVEGF‐165, and rhBMP‐2 in fibrin matrix on bone healing in a delayed‐union rat model. Thirty‐seven rats underwent a first operation where a standardized femoral critical size defect was created. A silicone spacer was implanted to impair vascularization within the defect. At 4 weeks the spacer was removed in a second operation and rhPDGF‐BB, rhVEGF‐165, or rhBMP‐2 were applied in a fibrin clot. Animals in a fourth group received a fibrin clot without growth factors. At 8 weeks fibrin bound rhBMP‐2 treated animals showed a significantly increased union rate and bone volume within the defect compared to the other groups. Single application of fibrin bound rhPDGF‐BB and rhVEGF‐165 failed to increase bone healing in our atrophic non‐union model. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:1563–1569, 2012