We report on the self-assembly based fabrication of fibrous polymers for tissue engineering applications. Directed self-assembly followed by polymerization of lysineappended diacetylenes generated a variety of polymers (P1. P5) with distinct chemical properties. The self-assembly along with the conjugated double and triple bonds and rigid geometry of diacetylene backbone imposed a nanofibrous morphology on the resulting polymers. Chemical properties including wettability of the polymers were tuned by using lysine (Lys) with orthogonal protecting groups (Boc and Fmoc). These Lys-appended polydiacetylene scaffolds were compared in terms of their efficiency toward human mesenchymal stem cells adhesion and spreading. Interestingly, polymer P4 containing Lys Nα-NH2and Lys Nε-Boc with balanced wettability supported cell adhesion better than the more hydrophobic polymer P2 with Nε-Boc and Nα-Fmoc or more hydrophilic polymer P5 containing free Nεand Nαamino groups. The molecular level control in the fabrication of nanofibrous polymers compared with other existing methods for the generation of fibrous polymers is the hallmark of this work.