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Background Although several studies on the Mg-Zn-Ca system have focused on alloy compositions that are restricted to solid solutions, the influence of the solid solution component of Ca on Mg-Zn alloys is unknown. Therefore, to broaden its utility in orthopedic applications, studies on the influence of the addition of Ca on the microstructural, mechanical, and corrosion properties of Mg-Zn alloys should be conducted. In this study, an in-depth investigation of the effect of Ca on the mechanical and bio-corrosion characteristics of the Mg-Zn alloy was performed for the optimization of a clinically approved Mg alloy system comprising Ca and Zn. Methods The Mg alloy was fabricated by gravitational melting of high purity Mg, Ca, and Zn metal grains under an Ar gas environment. The surface and cross-section were observed using scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to analyze their crystallographic structures. Electrochemical and immersion tests in Hank’s balanced salt solution were used to analyze their corrosion resistance. Tensile testing was performed with universal testing equipment to investigate the impact of Ca addition. The examination of cytotoxicity for biometric determination was in line with the ISO10993 standard. Results In this study, the 0.1% Ca alloy had significantly retarded grain growth due to the formation of the tiny and well-dispersed Ca2Mg6Zn3 phase. In addition, the yield strength and elongation of the 0.1% Ca alloy were more than 50% greater than the 2% Zn alloy. The limited cell viability of the 0.3% Ca alloy could be attributed to its high corrosion rate, whereas the 0.1% Ca alloy demonstrated cell viability of greater than 80% during the entire experimental period. Conclusion The effect of the addition of Ca on the microstructure, mechanical, and corrosion characteristics of Mg-Zn alloys was analyzed in this work. The findings imply that the Mg-Zn alloy system could be optimized by adding a small amount of Ca, improving mechanical properties while maintaining corrosion rate, thus opening the door to a wide range of applications in orthopedic surgery.

A paramagnetic NiTi substrate was coated with diamagnetic carbon materials, i.e., graphene, graphene oxide (GO), and carbon nanotubes (CNTs), in order to reduce magnetic resonance (MR) image artifacts of NiTi implants. The present study focused on the effect of magnetic susceptibility variations in NiTi caused by the carbon coating on MR image artifacts. In the case of the graphene and GO coatings, the reduction of the magnetic susceptibility was greater along the perpendicular direction than the parallel direction. In contrast, the CNT coating exhibited a larger reduction along the parallel direction. The reduction of magnetic susceptibility measured in CNT-coated NiTi (CNT/NiTi) was smaller than the theoretical prediction especially when measured along the parallel direction, because CNTs on the NiTi surface were randomly arranged, rather than in a single direction. MR image artifacts were substantially reduced in all carbon-coated NiTi specimens, which is due to the reduction of magnetic susceptibility in NiTi by the carbon coating. This method can also be applied to other paramagnetic bio-metallic materials such as Co-Cr.

Bone graft materials are commonly used to regenerate various bone defects, but their application is often limited because of the complex defect shape in various clinical conditions. Hence, customized bone grafts using three-dimensional (3D) printing techniques have been developed. However, conventional simple bone defect models are limited for evaluating the benefits and manufacturing accuracy of 3D-printed customized bone grafts. Thus, the aim of the present study was to develop a complex-shaped bone defect model. We designed an 8-shaped bony defect that consists of two simple circles attached to the rabbit calvarium. To determine the critical-sized defect (CSD) of the 8-shaped defects, 5.6- and 7-mm-diameter trephine burs were tested, and the 7-mm-diameter bur could successfully create a CSD, which was easily reproducible on the rabbit calvarium. The rate of new bone formation was 28.65% ± 8.63% at 16 weeks following creation of the defect. To confirm its efficacy for clinical use, the 8-shaped defect was created on a rabbit calvarium and 3D computed tomography (CT) was performed. A stereolithography file was produced using the CT data, and a 3D-printed polycaprolactone graft was fabricated. Using our 8-shaped defect model, we were able to modify the tolerances of the bone graft and calvarial defect to fabricate a more precise bone graft. Customized characteristics of the bone graft were then used to improve the accuracy of the bone graft. In addition, we confirmed the fitting ability of the 3D-printed graft during implantation of the graft. Our 8-shaped defect model on the rabbit calvarium using a 7.0-mm trephine bur may be a useful CSD model for evaluating 3D-printed graft materials.

Bioengineered skin substitute offers new opportunities for treating various skin ailments. To compensate the structural integrity problems of scaffolds prepared from natural components, we mechanically developed highly modified electrospun nanofibrous membranes, incorporating poly(l-lactide-co-e-caprolactone) (PLCL) into gelatin [poly(l-lactide-co-e-caprolactone)/ gelatin membrane, (P/G (3/7)]. Subsequent to our previous in vitro study, our goal was to evaluate the in vivo performance of PLCL, gelatin, and P/G (3/7) membranes, and investigate the feasibility of the newly developed P/G (3/7) membrane for wound healing. Histological analysis using the mathematical model of wound healing and contraction, revealed the association between stiffness of skin substitute with cytokeratin production and wound contraction rate, and the defect site covered with the stiffer membrane showed lower cytokeratin production, and inversely, higher wound contraction rate. Overall, the P/G (3/7) membrane induced a satisfactory wound healing outcome. However, lower cytokeratin production rate with the mechanically modified P/G membrane involves the importance of the conditional blending of PLCL. Conversely, the condition of PLCL showed some incompatibility and hindrance of skin regeneration, consistent with previous in vitro results. With proper mechanical strength and cell viability, the P/G (3/7) membrane could successfully be used as a suitable skin substitute scaffold.

We report here the effect of micro-environmental changes from biodegradable magnesium alloys on the activities of cells - osteoblasts, osteoclasts and macrophages - which play critical roles in each phase of the bone-regeneration process. Despite positive bone formation effects from several in vivo studies, minimal progress has been made in identifying underlying mechanisms through in vitro studies, which are currently concentrated on osteoblastic activities. The observed in vitro and in vivo results indicated that alkaline pH and released magnesium and zinc ions derived from Mg-5 wt% Ca-1 wt% Zn alloy biodegradation promote the progress of bone formation. In contrast, alkaline pH and magnesium ions remarkably suppressed osteoclastic activities and pro-inflammatory cytokine production, closely related to osteolysis and prosthesis failure. Findings from the present study conclude that the degradation of Mg-5 wt% Ca-1 wt% Zn alloys can promote new bone formation by simultaneously affecting the complex combination of variable cellular activities and phases. Copyright © 2016 John Wiley & Sons, Ltd.

Biodegradable metallic materials represent a potential step‐change technology that may revolutionize the treatment of broken bones. Implants made with biodegradable metals are significantly stronger than their polymer counterparts and fully biodegradable in vivo, removing the need for secondary surgery or long‐term complications. Here, it is shown how clinically approved Mg alloy promotes improved bone repair using an integrated state of the art fetal mouse metatarsal assay coupled with in vivo preclinical studies, second harmonic generation, secretome array analysis, perfusion bioreactor, and high‐resolution 3D confocal imaging of vasculature within skeletal tissue, to reveal a vascular‐mediated pro‐osteogenic mechanism controlling enhanced tissue regeneration. The optimized mechanical properties and corrosion rate of the Mg alloy lead to a controlled release of metallic Mg, Ca, and Zn ions at a rate that facilitates both angiogenesis and coupled osteogenesis for better bone healing, without causing adverse effects at the implantation site. The findings from this study support ongoing development and refinement of biodegradable metal systems to act as crucial portal technologies with significant potential to improve many clinical applications., An integrated state‐of‐the‐art in vivo and in vitro approach reveals clinically approved Mg5Ca1Zn samples stimulate accelerated bone healing by releasing anabolic metallic ions into the surrounding tissues to enhance the growth of type H blood vessels, which localize at active sites of bone remodeling and actively recruit Osterix‐positive osteoprogenitors to the degrading implant site.

In the present study, the osseointegration of additively manufactured screws with a micro-rough surface structure and conventionally machined Ti6Al4V ELI screws with a smooth surface structure is compared. Screws were implanted in rabbit femurs or tibiae for two weeks. The additively manufactured screws exhibited a micro-rough surface with attached semi-molten powder with a size of 50–100 μm and blunt threads, whereas the conventionally machined screws showed a smooth surface and sharp threads. The bonding strength of the additively manufactured screws was significantly higher than that of the conventionally machined screws. The additively manufactured screws showed higher bone-to-implant contact than the conventionally machined screws. Considerable amount of bone tissue on the surface of the additively manufactured screws remained after the push-out tests, and the rabbit tibiae with the additively manufactured screws were significantly damaged, indicating a higher osseointegration rate. Active osteogenesis was observed around the semi-molten powder of the additively manufactured screws, and new bone was well developed along the micro-rough surface. Overall, this study shows that the micro-rough surface improved the bone ingrowth and suggests that additively manufactured screws may be an alternative to conventionally machined screws.

In clinical conditions, the reconstructions performed in the complex and three-dimensional bone defects in the craniomaxillofacial (CMF) area are often limited in facial esthetics and jaw function. Furthermore, to regenerate a bone defect in the CMF area, the used scaffold should have unique features such as different mechanical strength or physical property suitable for complex shape and function of the CMF bones. Therefore, a three-dimensional synthetic scaffold with a patient-customized structure and mechanical properties is more suitable for the regeneration. In this study, the customized kagome-structure scaffold with complex morphology was assessed in vivo. The customized 3D kagome-structure model for the defect region was designed according to data using 3D computed tomography. The kagome-structure scaffold and the conventional grid-structure scaffold (as a control group) were fabricated using a 3D printer with a precision extruding deposition head using poly(e-caprolactone) (PCL). The two types of 3D printed scaffolds were implanted in the 8-shaped defect model on the rabbit calvarium. To evaluate the osteoconductivity of the implanted scaffolds, new bone formation, hematoxylin and eosin staining, immunohistochemistry, and Masson's trichrome staining were evaluated for 16 weeks after implantation of the scaffolds. To assess the mechanical robustness and stability of the kagome-structure scaffold, numerical analysis considering the ‘elastic-perfectly plastic’ material properties and deformation under self-contact condition was performed by finite element analysis. As a result, the kagome-structure scaffold fabricated using 3D printing technology showed excellent mechanical robustness and enhanced osteoconductivity than the control group. Therefore, the 3D printed kagome-structure scaffold can be a better option for bone regeneration in complex and large defects than the conventional grid-type 3D printed scaffold.

There has been a tremendous amount of research in the past decade to optimize the mechanical properties and degradation behavior of the biodegradable Mg alloy for orthopedic implant. Despite the feasibility of degrading implant, the lack of fundamental understanding about biocompatibility and underlying bone formation mechanism is currently limiting the use in clinical applications. Herein, we report the result of long-term clinical study and systematic investigation of bone formation mechanism of the biodegradable Mg-5wt%Ca-1wt%Zn alloy implant through simultaneous observation of changes in element composition and crystallinity within degrading interface at hierarchical levels. Controlled degradation of Mg-5wt%Ca-1wt%Zn alloy results in the formation of biomimicking calcification matrix at the degrading interface to initiate the bone formation process. This process facilitates early bone healing and allows the complete replacement of biodegradable Mg implant by the new bone within 1 y of implantation, as demonstrated in 53 cases of successful long-term clinical study.

Feasibility studies were performed to determine the suitability of a novel synthesis technique for fabricating multifunctional composite materials for orthopedic implants. By blending paramagnetic Ti powder with diamagnetic graphite and consolidating the resulting mixtures, Ti-C composites that cannot be feasibly obtained via conventional alloying techniques or ingot metallurgy were synthesized. The synthesized composite material exhibited extremely low magnetic susceptibility (χ=67.6×10(-6)), and, as a result, exhibited fewer artifacts during magnetic resonance imaging. The strength of the composite material (σ=770MPa) was such that it could support external loads to which the human body is subjected, but its Young's modulus was low (E=81.9 GPa) such that it could mitigate the stress-shielding effect. The material was also free from toxic elements such as Al and V and, thus, can be considered less harmful.

Magnesium metal and its alloys have been proposed as a novel class of bone implant biomaterials because of their biodegradability and mechanical properties. The purpose of this study was to determine whether magnesium ions, which are released abundantly from alloys, affect proliferation and differentiation of human bone marrow-derived stromal cells (hBMSCs). High levels of magnesium ions did not induce cytotoxicity in hBMSCs, but treatment with 2.5-10 mm magnesium ions for 48-72 h significantly increased hBMSC proliferation. The expression of integrins α2 and α3, but not β1, was upregulated compared with the control and shifted from α3 to α2 in hBMSCs treated with magnesium ions. Knockdown of integrins α2 and/or α3 significantly reduced magnesium-induced proliferation of hBMSCs. Magnesium exposure profoundly enhanced alkaline phosphatase (ALP) gene expression and activity even at a relatively low magnesium concentration (2.5 mm). Exposure to magnesium ions facilitated hBMSC proliferation via integrin α2 and α3 expression and partly promoted differentiation into osteoblasts via the alteration of ALP expression and activity. Accordingly, magnesium could be a useful biomaterial for orthopaedic applications such as bone implant biomaterials for repair and regeneration of bone defects in orthopaedic and dental fields. Copyright © 2014 John Wiley & Sons, Ltd.

Ni–Cu–Ce 0.8 Sm 0.2 O 1.9 anode-supported single cells were developed for the direct utilization of methane. An yttria-doped zirconia and Ce 0.8 Sm 0.2 O 1.9 bi-layer electrolyte and a La 0.6 Sr 0.4 Co 0.2 Fe 0.8 O 3 − δ cathode layer were fabricated by slurry spin-coating. Cu was added to the anode by impregnation with a nitrate solution. The effects of Cu on the electrochemical performance of the anode were investigated in dry methane with respect to times of impregnation. Impregnation with Cu twice was determined to be optimal. Incorporating Cu into the anode improved electrochemical performance of the cells, reducing ohmic resistance and suppressing carbon deposition. At 700 °C, the single cell exhibited a maximum power density of 406 mW/cm 2 in dry methane. At a current density of 500 mA/cm 2 , the cell maintained 98.6% of its initial voltage after operation for 900 min.

Objective This study was undertaken to generate and characterize human induced pluripotent stem cells (PSCs) from patients with osteoarthritis (OA) and to examine whether these cells can be developed into disease-relevant cell types for use in disease modeling and drug discovery. Methods Human synovial cells isolated from two 71-year-old women with advanced OA were characterized and reprogrammed into induced PSCs by ectopic expression of 4 transcription factors (Oct-4, SOX2, Klf4, and c-Myc). The pluripotency status of each induced PSC line was validated by comparison with human embryonic stem cells (ESCs). Results We found that OA patient–derived human synovial cells had human mesenchymal stem cell (MSC)–like characteristics, as indicated by the expression of specific markers, including CD14−, CD19−, CD34−, CD45−, CD44+, CD51+, CD90+, CD105+, and CD147+. Microarray analysis of human MSCs and human synovial cells further determined their unique and overlapping gene expression patterns. The pluripotency of established human induced PSCs was confirmed by their human ESC–like morphology, expression of pluripotency markers, gene expression profiles, epigenetic status, normal karyotype, and in vitro and in vivo differentiation potential. The potential of human induced PSCs to differentiate into distinct mesenchymal cell lineages, such as osteoblasts, adipocytes, and chondrocytes, was further confirmed by positive expression of markers for respective cell types and positive staining with alizarin red S (osteoblasts), oil red O (adipocytes), or Alcian blue (chondrocytes). Functional chondrocyte differentiation of induced PSCs in pellet culture and 3-dimensional polycaprolactone scaffold culture was assessed by chondrocyte self-assembly and histology. Conclusion Our findings indicate that patient-derived synovial cells are an attractive source of MSCs as well as induced PSCs and have the potential to advance cartilage tissue engineering and cell-based models of cartilage defects.

Ni-YSZ (Yttria Stabilized Zirconia) composite powders were fabricated by glycine nitrate process. The prepared powders were sintered at for 4 h in air and reduced at for 2 h in a nitrogen and hydrogen atmosphere. The microstructure, electrical conductivity, thermal expansion and mechanical properties of the Ni-YSZ cermets have been investigated with respect to the volume contents of Ni. A porous microstructure consisting of homogeneously distributed Ni and YSZ phases together with well-connected grains was observed. It was found that the open porosity, electrical conductivity, thermal expansion and bending strength of the cermets are sensitive to the volume content of Ni. The Ni-YSZ cermet containing 40 vol% Ni was ascertained to be the optimum composition. This composition offers sufficient open porosity of more than 30 %, superior electrical conductivities of 917.4 S/cm at and a moderate average thermal expansion coefficient of between room temperature and .

When viewed using a magnetic resonance imaging (MRI) system, invasive materials inside the human body, in many cases, severely distort the MR image of human tissues. The degree of the MR image distortion increases in proportion not only to the difference in the susceptibility between the invasive material and the human tissue, but also to the intensity of the magnetic field induced by the MRI system. In this study, by blending paramagnetic Ti particles with diamagnetic graphite, we synthesized Ti100-xCx composites that can reduce the artifact in the MR image under the high-strength magnetic field. Of the developed composites, Ti70C30 showed the magnetic susceptibility of χ = 67.6 × 10, which corresponds to 30% of those of commercially available Ti alloys, the lowest reported in the literature. The level of the MR image distortion in the vicinity of the Ti70C30 composite insert was nearly negligible even under the high magnetic field of 4.7 T. In this paper, we reported on a methodology of designing new structural materials for bio-applications, their synthesis, experimental confirmation and measurement of MR images. (Received December 1, 2011)

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.

A edible cuttlefish(Zoological name : Sepia esculenta) bone has a porous structure with all pores interconnected The purpose of this research is to develop porous hydroxyapatite prepared by hydrothermal treatment from cuttlefish bone and evaluate the biocompatibility using undecalcified materials through the in-vivo test of rabbits. In this study, the phase and substructure of a porous hydroxyapatite, prepared by hydrothermal treatment using edible cuttlefish bone as a calcium source, has been confirmed by X-ray diffractometer and scanning electronic microscope. After preparing the specimens with 5mm diameter and 7mm length, the specimens were implanted into the femoral condyles of rabbits. Each rabbits were sacrificed at each time period of 1, 2, 3, 4 weeks after operation, respectively and the stained section was examined by a transmission light microscope. The X-ray diffraction patterns of the edible cuttlefish bone was confirm for aragonite phase and of the sample after hydrothermal treatment showed mostly into hydroxyapatite phase. There was more bone density increase in porous HA rod around implant site than natural edible cuttlefish bone. Because the edible cuttlefish bone is a very pure and good calcium source, porous hydroxyapatite developed from this study is expected to be a biomaterial having a good biocompatibility to be used as a suitable bone substitute.

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.

We report here the effect of micro-environmental changes from biodegradable magnesium alloys on the activities of cells - osteoblasts, osteoclasts and macrophages - which play critical roles in each phase of the bone-regeneration process. Despite positive bone formation effects from several in vivo studies, minimal progress has been made in identifying underlying mechanisms through in vitro studies, which are currently concentrated on osteoblastic activities. The observed in vitro and in vivo results indicated that alkaline pH and released magnesium and zinc ions derived from Mg-5 wt% Ca-1 wt% Zn alloy biodegradation promote the progress of bone formation. In contrast, alkaline pH and magnesium ions remarkably suppressed osteoclastic activities and pro-inflammatory cytokine production, closely related to osteolysis and prosthesis failure. Findings from the present study conclude that the degradation of Mg-5 wt% Ca-1 wt% Zn alloys can promote new bone formation by simultaneously affecting the complex combination of variable cellular activities and phases. Copyright © 2016 John WileySons, Ltd.

Titanium alloy(Ti-6Al-4V, Samsung techwin, Korea) rods with diameter of 2.5mm were used as the implant materials. Polymethyl methacrylate(PMMA) bone cements(CMW, USA) were used as bone cement and -tricalcium phosphate(TCP)-based bone filler powder was used as osteoconductive additives. Hydroxyapatite(HA) was the desired end product after hydrolysis of the -TCP-based bone filler powder mixed with the blood. In animal study using rabbits, we divided the group into A, B, C and D. Rabbits were sacrificed at 1, 3, 9 weeks after implantation and the affinity index and bone density were calculated. X-ray diffraction patterns of HA formed by hydrolysis of -TCP-based bone filler powder showed higher and higher intensity in HA peak with increase of time period. There was more bone density increase in the group B and D containing - TCP-based bone filler powder around implant site than in the group A and C(p

In the present study we experimented on a multimodal therapeutic approach, such as combining chemotherapy agent (Bee venom) with cellular (NK-92MI) immunotherapy. Previously bee venom has been found to show anti-cancer effect in various cancer cell lines. In lung cancer cells bee venom showed an IC(50) value of 3 μg/ml in both cell lines. The co-culture of NK-92MI cell lines with lung cancer cells also show a decrease in viability upto 50 % at 48 h time point. Hence we used bee venom treated NK-92MI cells to co-culture with NSCLC cells and found that there is a further decrease in cell viability upto 70 and 75 % in A549 and NCI-H460 cell lines respectively. We further investigated the expression of various apoptotic and anti-apoptotic proteins and found that Bax, cleaved caspase-3 and -8 were increasing where as Bcl-2 and cIAP-2 was decreasing. The expression of various death receptor proteins like DR3, DR6 and Fas was also increasing. Concomitantly the expression of various death receptor ligands (TNFalpha, Apo3L and FasL) was also increasing of NK-92MI cells after co-culture. Further the DNA binding activity and luciferase activity of NF-κB was also inhibited after co-culture with bee venom treated NK-92MI cell lines. The knock down of death receptors with si-RNA has reversed the decrease in cell viability and NF-κB activity after co-culture with bee venom treated NK-92MI cells. Thus this new approach can enhance the anti-cancer effect of bee venom at a much lower concentration.

Magnesium metal and its alloys have been proposed as a novel class of bone implant biomaterials because of their biodegradability and mechanical properties. The purpose of this study was to determine whether magnesium ions, which are released abundantly from alloys, affect proliferation and differentiation of human bone marrow-derived stromal cells (hBMSCs). High levels of magnesium ions did not induce cytotoxicity in hBMSCs, but treatment with 2.5-10 mm magnesium ions for 48-72 h significantly increased hBMSC proliferation. The expression of integrins α2 and α3, but not β1, was upregulated compared with the control and shifted from α3 to α2 in hBMSCs treated with magnesium ions. Knockdown of integrins α2 and/or α3 significantly reduced magnesium-induced proliferation of hBMSCs. Magnesium exposure profoundly enhanced alkaline phosphatase (ALP) gene expression and activity even at a relatively low magnesium concentration (2.5 mm). Exposure to magnesium ions facilitated hBMSC proliferation via integrin α2 and α3 expression and partly promoted differentiation into osteoblasts via the alteration of ALP expression and activity. Accordingly, magnesium could be a useful biomaterial for orthopaedic applications such as bone implant biomaterials for repair and regeneration of bone defects in orthopaedic and dental fields. Copyright © 2014 John WileySons, Ltd.

This study was undertaken to generate and characterize human induced pluripotent stem cells (PSCs) from patients with osteoarthritis (OA) and to examine whether these cells can be developed into disease-relevant cell types for use in disease modeling and drug discovery.Human synovial cells isolated from two 71-year-old women with advanced OA were characterized and reprogrammed into induced PSCs by ectopic expression of 4 transcription factors (Oct-4, SOX2, Klf4, and c-Myc). The pluripotency status of each induced PSC line was validated by comparison with human embryonic stem cells (ESCs).We found that OA patient-derived human synovial cells had human mesenchymal stem cell (MSC)-like characteristics, as indicated by the expression of specific markers, including CD14-, CD19-, CD34-, CD45-, CD44+, CD51+, CD90+, CD105+, and CD147+. Microarray analysis of human MSCs and human synovial cells further determined their unique and overlapping gene expression patterns. The pluripotency of established human induced PSCs was confirmed by their human ESC-like morphology, expression of pluripotency markers, gene expression profiles, epigenetic status, normal karyotype, and in vitro and in vivo differentiation potential. The potential of human induced PSCs to differentiate into distinct mesenchymal cell lineages, such as osteoblasts, adipocytes, and chondrocytes, was further confirmed by positive expression of markers for respective cell types and positive staining with alizarin red S (osteoblasts), oil red O (adipocytes), or Alcian blue (chondrocytes). Functional chondrocyte differentiation of induced PSCs in pellet culture and 3-dimensional polycaprolactone scaffold culture was assessed by chondrocyte self-assembly and histology.Our findings indicate that patient-derived synovial cells are an attractive source of MSCs as well as induced PSCs and have the potential to advance cartilage tissue engineering and cell-based models of cartilage defects.

Pernicious anemia has recently been recognized as one of the risk factors for osteoporosis and bone fractures, but the underlying pathophysiologic mechanism is still unknown. To determine whether vitamin B12 has any direct effect on osteoblasts, we studied the effects of vitamin B12 on the proliferation and alkaline phosphatase activity in human bone marrow stromal osteoprogenitor cells (hBMSC) and UMR106 osteoblastic cells. Vitamin B12 at concentrations as low as 10(-12) mol/L significantly stimulated [3H]-thymidine incorporation in both types of cells, but concentrations higher than 10(-12) mol/L did not produce a greater effect. Vitamin B12 in the concentration range from 10(-12) to 10(-8) mol/L concentration-dependently increased alkaline phosphatase activity in both hBMSC and UMR106 cells. Based on these results, we suggest that a suppressed activity of osteoblasts may contribute to osteoporosis and fractures in patients with vitamin B12 deficiency.

Objectives: The purpose of this study is to identify whether the change of pH affects the proliferation and the differentiation of human bone marrow stem cells (hBMSCs) and what mechanism is underlied. Methods: To achieve objective of this study, hBMSCs were cultivated in the conditioned media adjusted to potential of hydrogen (pH) ranging from 6.4 to 8.0 using addition of hydrochloric acid (HCl) and sodium hydroxide (NaOH). The ratio of proliferation of hBMSCs according to the change of pH was measured for 24 h, 48 h, and 72 h using water-soluble tetrazolium salt (WST)-8 method. To elucidate the mechanism involved, hBMSCs was subjected to blocking extracellular signal-regulated kinases (ERK) and calcium sensing receptor (CaSR) activation. The Osteogenicrelated genes and alkaline phosphatase (ALP) activity were tested under the conditioned media. Results: The proliferation of hBMSCs was promoted under extracellular alkali conditions (pH 7.6~8.0) via CaSR/ ERK pathway. On the other hand, the differentiation was inhibited/delayed via decreased ALP activity besides gene expression at pH 8.0. Conclusion: Extracellular alkali or acidic surrounding according to pH alteration can play a crucial role in hBMSC behavior including the proliferation and the differentiation. [Korean Journal of Bone Metabolism, 19(1): 35-46, 2012]

Purpose: To elucidate the relation between fracture healing and angiogenesis, we checked expression of Hypoxia-inducible factor (HIF) and Vascular endothelial growth factor (VEGF) in hypoxic cell cultures and the callus from a rat femur fracture model. Materials and Methods: Human osteoblasts, chondrocytes, and rat ST2 cells were cultured in DME/F12 media with 10% FBS. Hypoxic DME/F12 media (PO2

Objective: To investigate the influence of associated medical diseases and complications on functional improvement after in-patient through stroke rehabilitation. Method: We performed a retrospective analysis on medical records of 183 stroke patients who had admitted to the department of rehabilitation medicine. Functional Independence Measure (FIM), Modified Barthel Index (MBI) at admission and discharge were used to assess the functional status. We investigated medical diseases, such as hypertension, diabetes, myocardial infarct, atrial fibrillation osteoarthritis, rheumatoid arthritis, previous history of stroke and complications such as dementia, post-stroke depression, central post-stroke pain, complex regional pain syndrome, neglect and aphasia. Results: Post-stroke patients with myocardial infarct, atrial fibrillation, osteoarthritis, dementia, aphasia and neglect significantly showed lower gain of FIM and MBI, lower FIM and MBI efficacy during inpatient rehabilitation compared to without those (p 0.05). Total numbers of associated medical diseases and complications negatively affect on FIM and MBI efficacy (p

Ganglioneuroblastoma is a rare tumor originating from the sympathetic chain. It is intermediate between the highly malignant neuroblastoma and the benign ganglioneuroma. The predilection sites of ganglioneur- oblastoma are the retroperitoneum and mediastinum but involvement of the structures around the spinal cord is infrequent. A 4 year-old girl who had progressive weakness in both lower extremities had a tiptoe gait for 2 months. A magnetic resonance imaging (MRI) study revealed a heterogeneous enhancing mass in the right mediastinum and paraspinal areas. A biopsy of the mass confirmed a ganglioneuroblastoma with many neuroblasts and nodules of ganglion cells. This is an unusual case of a child with spastic paraplegia caused by ganglioneuroblastoma of the thoracic cavity. (Korean J Hematol 2007;42:414-418.)