J.H. Kim

Generation and characterization of integration-free induced pluripotent stem cells from patients with autoimmune disease

Autoimmune diseases (AIDs), a heterogeneous group of immune-mediated disorders, are a major and growing health problem. Although AIDs are currently treated primarily with anti-inflammatory and immunosuppressive drugs, the use of stem cell transplantation in patients with AIDs is becoming increasingly common. However, stem cell transplantation therapy has limitations, including a shortage of available stem cells and immune rejection of cells from nonautologous sources. Induced pluripotent stem cell (iPSC) technology, which allows the generation of patient-specific pluripotent stem cells, could offer an alternative source for clinical applications of stem cell therapies in AID patients. We used nonintegrating oriP/EBNA-1-based episomal vectors to reprogram dermal fibroblasts from patients with AIDs such as ankylosing spondylitis (AS), Sjögren’s syndrome (SS) and systemic lupus erythematosus (SLE). The pluripotency and multilineage differentiation capacity of each patient-specific iPSC line was validated. The safety of these iPSCs for use in stem cell transplantation is indicated by the fact that all AID-specific iPSCs are integrated transgene free. Finally, all AID-specific iPSCs derived in this study could be differentiated into cells of hematopoietic and mesenchymal lineages in vitro as shown by flow cytometric analysis and induction of terminal differentiation potential. Our results demonstrate the successful generation of integration-free iPSCs from patients with AS, SS and SLE. These findings support the possibility of using iPSC technology in autologous and allogeneic cell replacement therapy for various AIDs, including AS, SS and SLE.

Resorption Mechanism of Hydroxyapatite and β-Tricalcium Phosphate Coating Layer

Beta-tricalcium phosphate(β-TCP) coating layer is known to be resorbed much faster than hydroxyapatite(HA), however, there has been no report to explain the exact reason of these results. Eighty titanium discs, coated with HA(n=40) or β-TCP(n=40) by dip and spin coating method, were divided into 2 subgroups respectively; Dissolution(D, n=20) and osteoclast culture(C, n=20). The coated discs in D group were immersed in the cell culture media for 5 days, whereas, in C group, osteoclasts were seeded on the specimens and cultured for 5 days. After simple dissolution test, β-TCP coating layer showed much more cracks and denudation as compared to HA. In osteoclast culture group, mean area fraction of resorption pits in HA-C group was 11.62%, which was significantly higher than that of 0.73% in β-TCP-C group(p=0.001). In conclusion, the resorption mechanisms of HA and β-TCP coating layers were different each other in vitro study. The coated β-TCP was degraded mainly by dissolution and separation from implant, on the other hand, the HA coating layer was resorbed by osteoclastic activity.

The Influence of Bioactive Glass Particles on Osteolysis

We observed the cytotoxicity of human bone marrow stromal cells(hBMSCs) by microparticles of bioactive glass with four particle groups(same chemical composition-45S5 but produced by two different manufacturer and two different size groups). In vivo test using rat calvaria were also carried out. The apoptosis rates of all small particle groups(10-20 ㎛) were increased than large(500-700 ㎛ or 200-900 ㎛) particle groups in any culture time and any amount of particles with statistical significance. In vivo study we observed pathologic signs such as macrophages and foreign-body giant cells in rat calvaria by micro-particles of bioglass. Small(10- 20 ㎛) sized particles induced foreign body reaction and bone resorption. There was proliferation of macrophages and cells in large number. But in large particle groups, only fibroblasts were surrounding the particles. The micro-particles of bioglass induced apoptosis of hBMSC and foreign body reaction in calvaria of rat, therefore micro-particles of bioglass may cause osteolysis if used in replacement arthroplasty.