A　novel　electrochemiluminescence　(ECL)　biosensor　for　hyaluronidase　(HAase)　based　on　the　adjustable　electrostatic　interaction　between　the　surface-charge-controllable　nanoparticles　and　negatively　charged　electrode　has　been　devised.　Hyaluronic　acid　(HA)-coated　amino-modified　ruthenium　bipyridine-doped　silica　nanoparticles　(Ru@SiO2-NH2@HA　NPs)　have　been　synthesized　and　act　as　ECL　indicators,　and　the　surface　of　this　particle　is　negatively　charged　because　HA　contains　a　large　amount　of　OH-　and　COO-.　The　strong　electrostatic　repulsion　between　the　Ru@　SiO2-NH2@HA　NPs　and　negatively　charged　indium　tin　oxide　(ITO)　electrode　surface　leads　to　the　detection　of　a　low-intensity　ECL　signal.　In　the　presence　of　HAase,　the　HA　on　the　surface　of　the　Ru@SiO2-NH2@HA　NPs　can　be　decomposed,　and　the　particles　can　be　transformed　into　positively　charged　amino-modified　ruthenium　bipyridine-doped　silica　nanoparticles　(Ru@SiO2-NH2　NPs),　which　can　be　concentrated　near　the　surface　of　the　ITO　electrode　through　electrostatic　attraction,　and　result　in　the　detection　of　an　enhanced　ECL　signal.　The　ECL　of　the　system　has　a　good　linear　relationship　with　HAase　concentration　in　the　range　of　2.0-60　U/mL,　and　the　limit　of　detection　was　0.37　U/mL.　The　designed　biosensor　had　been　applied　to　detect　the　target　in　real　samples　with　satisfied　results.