The current clinical treatment of cartilage defects involves autologous chondrocyte implantation into cartilage defect sites. However, one of the complications associated with this method is the lack of bonding between the implanted materials and natural tissue. Helical rosette nanotubes (HRNs) are novel biomimetic self-assembled supramolecular structures whose basic building blocks are DNA base-pairs. HRNs are similar in size to collagen in cartilage. Moreover, previous studies have shown that HRNs are biocompatible and increase the adhesion of numerous cells compared to other commonly used cartilage implant materials (like hydrogels and Ti). In addition, HRNs can solidify into a viscous gel at body temperatures under short periods of time. Thus, it is hoped that HRNs can serve as a novel in situ tissue implant to improve cartilage cell adhesion and functions. In this study, in order to heal cartilage rupture and regenerate cartilage during possible implantation, the mechanical properties of select hydrogel/HRN composites were tested. In addition, electro-spinning was used to generate three-dimensional, implantable, composite fibers encapsulated with chondrocytes and fibroblast-like type-B synoviocytes (SFB cells, a type of mesenchymal stem cell). Importantly, results showed that drug-delivered HRNs enhanced hydrogel adhesive strength and created a scaffold with nanometer-rough surface structures pertinent for cartilage regeneration. In this manner, this study provided an alternative cartilage regenerative material which relies on nanotechnology that can be injected as a liquid, solidify at body temperatures under short periods of time, have suitable mechanical properties to collagen, and promote cell functions.