Hyaluronic acid (HA) fillers are the most popular filler agents for skin rejuvenation. Although 1,4-butanediol diglycidyl ether (BDDE) is regarded as a relatively safe crosslinker, it still exhibits certain cytotoxicity. We presented here an amino acid crosslinked HA (ACHA) which was obtained by the amidation reaction with lysine and HA. This study aimed to investigate ACHA's efficacy and safety for skin augmentation and rejuvenation. Rheology, compressive tests, and swelling experiments were conducted to investigate ACHA's mechanical and viscoelastic properties. The effect of ACHA on the human keratinocytes (HaCaT) cells, and the human dermal fibroblast (HDF) was investigated by Transwell and wound healing assays. Its impact on the epithelial thickness and collagen synthesis was further examined in a mouse experimental model. We recruited 50 patients with moderate to severe nasolabial folds. The patients were randomly allocated to receive ACHA or Restylane injections. The retention rate of HA, Wrinkle Severity Rating Scale (WSRS) and Global Aesthetic Improvement Scale (GAIS) were evaluated and compared. ACHA exhibited good viscoelasticity. It can not only promote migration and proliferation of HaCat and HDF and secretion of various growth factors but also increase skin thickness and promote the generation of collagen. Patients who received ACHA had more residual volume 12 months after treatment. And ACHA exhibited a promising augmentation effect in NLF correction with few adverse reactions. ACHA has shown promise as a biomaterial with excellent biocompatibility and viscoelastic characteristics both in research and the clinic.