The aim of this study was to functionalize synthetic poly-(L-lactic) (PLLA) nanofibers by direct incorporation of cRGD, in order to promote adhesion, growth and osteogenic differentiation of human mesenchymal stem cells (hMSC) in vitro. The cRGD was incorporated into PLLA nanofibers either by emulsion [PLLA-cRGD (d)] or suspension [PLLA-cRGD (s)]. Matrices were seeded with hMSC and cultivated over a period of 28 days under growth conditions and analyzed during the course. Although the mode of incorporation resulted in different distributions of the RGD peptide, it had no impact on the fiber characteristics when compared to corresponding unblended PLLA control fibers. However, hMSC showed better adherence on PLLA-cRGD (d). Nevertheless, this advantage was not reflected during the course of cultivation. Furthermore, the PLLA-cRGD (s) fibers mediated the osteogenic potential of collagen (determined as the expression and deposition of collagen and osteocalcin) to some extent. Further studies are needed in order to optimize the RGD distribution and concentration.