title: Sphere-shaped nano-hydroxyapatite/chitosan/gelatin 3D porous scaffolds increase proliferation and osteogenic differentiation of human induced pluripotent stem cells from gingival fibroblasts.
abstract: Hydroxyapatite (HA) is an important component of human bone and bone tissue engineering scaffolds. A plethora of bone tissue engineering scaffolds have been synthesized so far, including nano-HA/chitosan/gelatin (nHA/CG) scaffolds; and for seeding cells, stem cells, especially induced pluripotent stem cells (iPSCs), have been a promising cell source for bone tissue engineering recently. However, the influence of different HA nano-particle morphologies on the osteogenic differentiation of human iPSCs (hiPSCs) from human gingival fibroblasts (hGFs) is unknown. The purpose of this study was to investigate the osteogenic differentiation of hiPSCs from hGFs seeded on nHA/CG scaffolds with 2 shapes (rod and sphere) of nHA particles. Firstly, hGFs isolated from discarded normal gingival tissues were reprogrammed into hiPSCs. Secondly, hiPSCs were seeded on rod-like nHA/CG (rod-nHA/CG) and sphere-shaped nHA/CG (sphere-nHA/CG) scaffolds respectively and then cell/scaffold complexes were cultured in vitro. Scanning electron microscope, hematoxyline and eosin (HE) staining, Masson's staining, and quantitative real-time polymerase chain reaction techniques were used to examine hiPSC morphology, proliferation, and differentiation on rod-nHA/CG and sphere-nHA/CG scaffolds. Finally, hiPSCs composited with 2 kinds of nHA/CG were transplanted in vivo in a subcutaneous implantation model for 12 weeks; pure scaffolds were also transplanted as a blank control. HE, Masson's, and immunohistochemistry staining were applied to detect new bone regeneration ability. The results showed that sphere-nHA/CG significantly increased hiPSCs from hGF proliferation and osteogenic differentiation in vitro. hiPSCs and sphere-nHA/CG composities generated large bone, whereas hiPSCs and rod-nHA/CG composities produced tiny bone in vivo. Moreover, pure scaffolds without cells almost produced no bone. In conclusion, our work provided a potential innovative bone tissue engineering approach using clinically discarded gingival tissues and sphere-nHA/CG scaffolds.
authors.affiliation: Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing 210008, People's Republic of China. Nanjing Key Laboratory, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing 210008, People's Republic of China.
authors.forename: Jun
authors.initials: J
authors.lastname: Ji
authors.forename: Xin
authors.initials: X
authors.lastname: Tong
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Huang
authors.forename: Tiancong
authors.initials: T
authors.lastname: Wang
authors.forename: Zitong
authors.initials: Z
authors.lastname: Lin
authors.forename: Yazhou
authors.initials: Y
authors.lastname: Cao
authors.forename: Junfeng
authors.initials: J
authors.lastname: Zhang
authors.forename: Lei
authors.initials: L
authors.lastname: Dong
authors.forename: Haiyan
authors.initials: H
authors.lastname: Qin
authors.forename: Qingang
authors.initials: Q
authors.lastname: Hu
mesh_terms: D018786:Bone Substitutes / Q000138:chemical synthesis; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry; D017886:Durapatite / Q000737:chemistry*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology; D005780:Gelatin / Q000737:chemistry; D005881:Gingiva / Q000166:cytology / Q000502:physiology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D008422:Materials Testing; D054118:Nanospheres / Q000737:chemistry* / Q000648:ultrastructure; D010012:Osteogenesis / Q000502:physiology*; D010316:Particle Size; D016062:Porosity; D054457:Tissue Scaffolds
chemical_list: D018786:Bone Substitutes; D005780:Gelatin; D048271:Chitosan; D017886:Durapatite