Bone regeneration of induced pluripotent stem cells derived from peripheral blood cells in collagen sponge scaffolds

Stem cell-based regeneration therapy offers new therapeutic options for patients with bone defects because of significant advances in stem cell research. Although bone marrow mesenchymal stem cells are the ideal material for bone regeneration therapy using stem cell, they are difficult to obtain. Induced pluripotent stem cells (iPSCs) are now considered an attractive tool in bone tissue engineering. Recently, the efficiency of establishing iPSCs has been improved by the use of the Sendai virus vector, and it has become easier to establish iPSCs from several type of somatic cells. In our previous study, we reported a method to purify osteogenic cells from iPSCs. Objective: This study aimed to evaluate the osteogenic ability of iPSCs derived from peripheral blood cells. Methodology: Mononuclear cells (MNCs) were obtained from human peripheral blood. Subsequently, T cells were selectively obtained from these MNCs and iPSCs were established using Sendai virus vectors. Established iPSCs were evaluated by the expression of undifferentiated markers and teratoma formation assays. Osteoblasts were induced from these iPSCs and evaluated by the expression of osteoblast markers. Additionally, the induced osteoblasts were transplanted into rat critical size calvaria bone defect models with collagen sponge scaffolds. Samples were evaluated by radiographical and histological assessments. Results: Induced osteoblasts expressed several osteoblast-specific markers. The results of radiographical and histological assessments revealed that the cell transplant group had bone formations superior to those of the control group. Conclusions: This study suggests that peripheral blood MNCs have the potential to differentiate into osteoblasts. Although there are some hurdles in iPSC transplantation, osteoblasts obtained from MNC-iPSCs could be applied to bone regeneration therapy in the future

Targeted reversion of induced pluripotent stem cells from patients with human cleidocranial dysplasia improves bone regeneration in a rat calvarial bone defect model

BackgroundRunt-related transcription factor 2 (RUNX2) haploinsufficiency causes cleidocranial dysplasia (CCD) which is characterized by supernumerary teeth, short stature, clavicular dysplasia, and osteoporosis. At present, as a therapeutic strategy for osteoporosis, mesenchymal stem cell (MSC) transplantation therapy is performed in addition to drug therapy. However, MSC-based therapy for osteoporosis in CCD patients is difficult due to a reduction in the ability of MSCs to differentiate into osteoblasts resulting from impaired RUNX2 function. Here, we investigated whether induced pluripotent stem cells (iPSCs) properly differentiate into osteoblasts after repairing the RUNX2 mutation in iPSCs derived from CCD patients to establish normal iPSCs, and whether engraftment of osteoblasts derived from properly reverted iPSCs results in better regeneration in immunodeficient rat calvarial bone defect models.MethodsTwo cases of CCD patient-derived induced pluripotent stem cells (CCD-iPSCs) were generated using retroviral vectors (OCT3/4, SOX2, KLF4, and c-MYC) or a Sendai virus SeVdp vector (KOSM302L). Reverted iPSCs were established using programmable nucleases, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas-derived RNA-guided endonucleases, to correct mutations in CCD-iPSCs. The mRNA expressions of osteoblast-specific markers were analyzed using quantitative reverse-transcriptase polymerase chain reaction. iPSCs-derived osteoblasts were transplanted into rat calvarial bone defects, and bone regeneration was evaluated using microcomputed tomography analysis and histological analysis.ResultsMutation analysis showed that both contained nonsense mutations: one at the very beginning of exon 1 and the other at the initial position of the nuclear matrix-targeting signal. The osteoblasts derived from CCD-iPSCs (CCD-OBs) expressed low levels of several osteoblast differentiation markers, and transplantation of these osteoblasts into calvarial bone defects created in rats with severe combined immunodeficiency showed poor regeneration. However, reverted iPSCs improved the abnormal osteoblast differentiation which resulted in much better engraftment into the rat calvarial bone defect.ConclusionsTaken together, these results demonstrate that patient-specific iPSC technology can not only provide a useful disease model to elucidate the role of RUNX2 in osteoblastic differentiation but also raises the tantalizing prospect that reverted iPSCs might provide a practical medical treatment for CCD

Experimental investigation of performance differences between Coherent Ising Machines and a quantum annealer

Physical annealing systems provide heuristic approaches to solving NP-hard Ising optimization problems. Here, we study the performance of two types of annealing machines--a commercially available quantum annealer built by D-Wave Systems, and measurement-feedback coherent Ising machines (CIMs) based on optical parametric oscillator networks--on two classes of problems, the Sherrington-Kirkpatrick (SK) model and MAX-CUT. The D-Wave quantum annealer outperforms the CIMs on MAX-CUT on regular graphs of degree 3. On denser problems, however, we observe an exponential penalty for the quantum annealer ($\exp(-\alpha_\textrm{DW} N^2)$) relative to CIMs ($\exp(-\alpha_\textrm{CIM} N)$) for fixed anneal times, on both the SK model and on 50%-edge-density MAX-CUT, where the coefficients $\alpha_\textrm{CIM}$ and $\alpha_\textrm{DW}$ are problem-class-dependent. On instances with over $50$ vertices, a several-orders-of-magnitude time-to-solution difference exists between CIMs and the D-Wave annealer. An optimal-annealing-time analysis is also consistent with a significant projected performance difference. The difference in performance between the sparsely connected D-Wave machine and the measurement-feedback facilitated all-to-all connectivity of the CIMs provides strong experimental support for efforts to increase the connectivity of quantum annealers.Comment: 12 pages, 5 figures, 1 table (main text); 14 pages, 12 figures, 2 tables (supplementary

Scaling advantages of all-to-all connectivity in physical annealers: the Coherent Ising Machine vs. D-Wave 2000Q

Physical annealing systems provide a heuristic approach to solve NP-hard Ising optimization problems. It is believed that the connectivity between spins in such annealers significantly impacts the machine's computational effectiveness. In this paper we study the performance of two types of annealing machines that have very different connectivity -- a commercially available quantum annealer built by D-wave Systems, which has sparse connectivity, and coherent Ising machines based on optical parametric oscillator networks, which have all-to-all connectivity. We demonstrate an exponential (e^(−O(N^2))) penalty in performance for the D-wave quantum annealer relative to coherent Ising machines when solving Ising problems on dense graphs, which is attributable to the differences in internal connectivity between the machines. This leads to a several-orders-of-magnitude time-to-solution difference between coherent Ising machines and the D-wave system for problems with over 50 vertices. Our results provide strong experimental support to efforts to increase the connectivity of physical annealers

Experimental investigation of performance differences between coherent Ising machines and a quantum annealer

Physical annealing systems provide heuristic approaches to solving combinatorial optimization problems. Here, we benchmark two types of annealing machines—a quantum annealer built by D-Wave Systems and measurement-feedback coherent Ising machines (CIMs) based on optical parametric oscillators—on two problem classes, the Sherrington-Kirkpatrick (SK) model and MAX-CUT. The D-Wave quantum annealer outperforms the CIMs on MAX-CUT on cubic graphs. On denser problems, however, we observe an exponential penalty for the quantum annealer [exp(–α_(DW)N^2)] relative to CIMs [exp(–α_(CIM)N)] for fixed anneal times, both on the SK model and on 50% edge density MAX-CUT. This leads to a several orders of magnitude time-to-solution difference for instances with over 50 vertices. An optimal–annealing time analysis is also consistent with a substantial projected performance difference. The difference in performance between the sparsely connected D-Wave machine and the fully-connected CIMs provides strong experimental support for efforts to increase the connectivity of quantum annealers

Hedgehog-Related Mutation Causes Bone Malformations with or without Hereditary Gene Mutations

The hedgehog (Hh) family consists of numerous signaling mediators that play important roles at various stages of development. Thus, the Hh pathway is essential for bone tissue development and tumorigenesis. Gorlin syndrome is a skeletal and tumorigenic disorder caused by gain-of-function mutations in Hh signaling. In this review, we first present the phenotype of Gorlin syndrome and the relationship between genotype and phenotype in bone and craniofacial tissues, including the causative gene as well as other Hh-related genes. Next, the importance of new diagnostic methods using next-generation sequencing and multiple gene panels will be discussed. We summarize Hh-related genetic disorders, including cilia disease, and the genetics of Hh-related bone diseases

Interleukin-22 Exerts Detrimental Effects on Salivary Gland Integrity and Function

Interleukin-22 (IL-22) affects epithelial tissue function and integrity in a context-dependent manner. IL-22 levels are elevated in salivary glands of Sjögren’s syndrome (SS) patients, but its role in the pathogenesis of this disease remains unclear. The objective of this study is to elucidate the impact of IL-22 on salivary gland tissue integrity and function in murine models. We showed that IL-22 levels in sera and salivary glands increased progressively in female non-obese diabetic (NOD) mice, accompanying the development of SS. Administration of IL-22 to the submandibular glands of NOD mice prior to the disease onset reduced salivary secretion and induced caspase-3 activation in salivary gland tissues, which were accompanied by alterations in multiple genes controlling tissue integrity and inflammation. Similarly, IL-22 administration to submandibular glands of C57BL/6 mice also induced hyposalivation and caspase-3 activation, whereas blockade of endogenous IL-22 in C57BL/6 mice treated with anti-CD3 antibody mitigated hyposalivation and caspase-3 activation. Finally, IL-22 treatment reduced the number of viable C57BL/6 mouse submandibular gland epithelial cells cultured in vitro, indicating a direct impact of this cytokine on these cells. We conclude that IL-22 exerts a detrimental impact on salivary gland tissues