Supramolecular hydrogel composed of aromatic short peptide gelator was an attractive biomaterial owing to its simple and convenient synthetic route, nano-fibrillar microstructure resembling natural collagen fibers and intelligent response to external stimulus. Herein, stimuli-responsive biphenyl-tripeptide supramolecular hydrogels was prepared to simulate extracellular matrix scaffolds by temperature switch, ion induction and pH switch. The amino acid arrangement substantially affected gelation behavior, only BPAA-βAFF and BPAA-FFβA could form nanostructured supramolecular hydrogels with 8-10 nm nanotubes or nanofibers by potential intermolecular hydrogen bond interactions and π-π stacking. The minimum gelation concentration (MGC) and maximum storage modulus were 0.4 mM (0.023 wt%) and around 8.2 KPa. The two supramolecular hydrogels could support adhesion and proliferation of L929 cells. Moreover, the BPAA-βAFF hydrogel promoted proliferation and ECM secretion of chondrocytes in vitro, and facilitate the phenotype maintenance of hyaline cartilage. All the results demonstrated that BPAA-βAFF hydrogel hold great potential application prospects in cartilage tissue engineering. Statement of SignificanceDiphenylalanine was served as a core segment conjugating with 4-biphenylacetic acid (BPAA) to produce biphenyl-tripeptide compounds with transforming amino sequence, and multiple external stimuli was applied to study the gelation properties of the aromatic short peptide gelators. “FF” brick (phenylalanine-phenylalanine) was crucial for formation of fibrous supramolecular hydrogels. Meanwhile, the sequence of amino acids arrangement also had an essential effect on the gelation behavior. Optimal BPAA-βAFF with ultra-low minimum gelation concentration (0.4 mM, about 0.023 wt%) and similar microstructure to extracellular matrix (ECM) of nature cartilage tissue could promote the proliferation and ECM secretion of chondrocytes in vitro, and facilitate the formation of hyaline cartilage.