Adiponectin (APN) is the most abundant adipocyte-secreted adipokine; it regulates energy homeostasis and exerts well-characterized insulin-sensitizing properties. Previous studies have verified that globular adiponectin (gAPN) is also involved in bone metabolism, although observations have been controversial. The purpose of the current study is to use an APN-knockout (APN-KO) mouse model to evaluate the local delivery of gAPN to new bone formation. Using chitosan microspheres (CMs), we found that following an initial burst at 1 week, the release behavior of gAPN from the scaffold was sustained in a linear manner for the first 4 weeks, followed by a slower, more stable release from week 5 onwards. Interestingly, PLGA/β-TCP/CM-loaded gAPN scaffolds implanted in APN-KO mice increased bone formation and mineralization, and enhanced osteogenic marker expression 28 days post-implantation. gAPN also promoted preosteoblast (MC3T3-E1) cellular proliferation in vitro. In MC3T3-E1 cells, adaptor protein-containing pleckstrin homology domain, phosphotyrosine domain, leucine zipper motif (APPL1) and phosphoinositide 3-kinase (PI3K) expression was upregulated in a time-dependent manner upon gAPN treatment, while APPL1 small interfering RNA (siRNA) pre-treatment reversed this enhanced expression. In conclusion, modified bone graft substitutes loaded with gAPN increase bone formation and mineralization in part by promoting osteoblast proliferation via the APPL1/PI3K pathway.