and biological properties, we hypothesize that the microphasesegregated DN concept could be applicable for developing injectable microsphere-based gel scaffolds with biological functionalities. It consists of two networks: a densely cross-linked biopolymer microspheres network (skeleton) and a sparsely cross-linked ductile biopolymer network. One can expected two advantages of this microsphere-based DN gel. First, microspheres incorporated at higher composition ratio can potentially increase strength of the DN gel due to their higher stiffness. Second, because GFs were encapsulated not in the ductile network but in microspheres, the GFs were expected to function better in this microsphere-based DN gel than in normal DN hydrogels. Importantly, unlike most microspheres that are chemically inert, our biopolymer microspheres would have residual functionalities that allow for further bioconjugation of a ductile network.Our strategy is to use heparin as the microspheres substrate and chitosan as the ductile network, respectively. In this system, heparin microspheres with higher composition ratio can potentially increase strength of the DN gel, while chitosan solution as a liquid phase can provide the injection. Heparin and chitosan are proteoglycans or glycosaminoglycans (GAGs), as highly charged polyelectrolytes, which could be additionally crosslinked by electrostatic interactions. Heparin is a negatively charged GAG, which is composed of repeated disaccharide units of alternating glucosamine and glucuronic residues heterogeneously modifi ed by carboxyl groups and N -or O -linked sulfate. Most importantly, the ability of heparin to sequester and stabilize GFs due to specifi c affi nity GF-immobilization has been exploited in the production of microspheres that can mediate cell proliferation and differentiation. [ 10 ] Chitosan is a positively charged polysaccharide composed of repeated glucosamine and N -acetylglucosamine, which also has been widely applied in drug delivery, gene therapy, and tissue engineering. [ 11 ] Mechanism for the formation of the microspheres, as well as for the formation of a 3D gel scaffold, is proposed in Scheme 1 . Our method for preparing heparin microspheres and gel scaffolds relies on in situ cross-linking of heparin with built-in fi brin-mimicking peptide of CTIGEGQQHHLGGAKQAGDV containing Lys and Gln functionalities. This peptide derived from the transglutaminase factor XIIIa (FXIIIa) crosslinking site of fi brin, with an additional N -terminal cysteine residue. [ 12a ] The peptides would couple and form covalent isopeptide bridges between Gln and Lys by enzymatic catalysis of the FXIIIa. [ 12 ] To do this, chemical modifi cations were employed to selectively introduce complementary reactive peptides to maleimide-functionalized heparin and chitosan via the Michael-type addition, respectively. Subsequently, peptide-functionalized heparin was formed microspheres via the inverse emulsion cross-linking Macromolecular microspheres have become an important structure in polymeric materials...