A Subset of Paracrine Factors as Efficient Biomarkers for Predicting Vascular Regenerative Efficacy of Mesenchymal Stromal/Stem Cells.

Mesenchymal stromal/stem cells (MSCs) have been developed as a promising source for cell-based therapies of ischemic disease. However, there are some hurdles in their clinical application such as poor cell engraftment and inconsistent stem cell potency. In this study, we sought to find biomarkers for predicting potency of MSCs for proangiogenic therapy to improve their beneficial effects. Large variations were observed in proangiogenic factor secretion profiles of conditioned media derived from nine different donor-derived Wharton's jelly (WJ)-derived MSCs and 8 factors among 55 angiogenesis-related factors were secreted at considerable levels. Two distinct WJ-MSCs that had the lowest or the highest secretion of these eight factors showed corresponding proangiogenic activities in in vitro angiogenesis assays. When four additional different donor-derived WJ-MSCs were further examined, proangiogenic activities in migration and tube formation of endothelial cells and in in vivo Matrigel plug assay were highly consistent with secretion levels of four major factors (angiogenin, interleukin-8, monocyte chemoattractant protein-1, and vascular endothelial growth factor). Such correlation was also observed in vascular regenerative effect in a mouse hind limb ischemia model. Blocking of these four factors by neutralizing antibodies or knockdown of them by siRNA treatment resulted in significant inhibition of proangiogenic activities of not only WJ-MSCs, but also bone marrow-derived MSCs. These results suggest that these four factors may represent efficient biomarkers for predicting vascular regenerative efficacy of MSCs. Stem Cells 2019;37:77-88.

Enhancement of Healing of Long Tubular Bone Defects in Rabbits Using a Mixture of Atelocollagen Gel and Bone Marrow Aspirate Concentrate.

We evaluated the bone-forming potential of a mixture of atelocollagen and bone marrow aspirate concentrate which was transplanted into bone defects. Radial shaft defects of about 10 mm in size were created in 30 New Zealand white rabbits. Ten rabbits in the control group were not treated further, 10 rabbits in the first experimental group (E1) received an atelocollagen injection, and 10 rabbits in the second experimental group (E2) received an injection of a mixture of atelocollagen and bone marrow aspirate concentrate. The groups were compared radiologically at 8 weeks. Osteogenesis in group E2 progressed more rapidly than that in the other groups, and osteogenesis in group E1 progressed faster than that in the control group. Thus, the administration of a mixture of atelocollagen and bone marrow aspirate concentrate in bone defects was found to enhance bone defect healing.

Autologous collagen induced chondrogenesis (ACIC: Shetty-Kim technique) - A matrix based acellular single stage arthroscopic cartilage repair technique.

The defects of articular cartilage in the knee joint are a common degenerative disease and currently there are several established techniques to treat this problem, each with their own advantages and shortcomings. Autologous chondrocyte implantation is the current gold standard but the technique is expensive, time-consuming and most versions require two stage procedures and an arthrotomy. Autologous collagen induced chondrogenesis (ACIC) is a single-stage arthroscopic procedure and we developed. This method uses microfracture technique with atelocollagen mixed with fibrin gel to treat articular cartilage defects. We introduce this ACIC techniques and its scientific background.

Potential application of temozolomide in mesenchymal stem cell-based TRAIL gene therapy against malignant glioma.

Because the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) selectively kills tumor cells, it is one of the most promising candidates for cancer treatment. TRAIL-secreting human mesenchymal stem cells (MSC-TRAIL) provide targeted and prolonged delivery of TRAIL in glioma therapy. However, acquired resistance to TRAIL of glioma cells is a major problem to be overcome. We showed a potential therapy that used MSC-TRAIL combined with the chemotherapeutic agent temozolomide (TMZ). The antitumor effects of the combination with MSC-TRAIL and TMZ on human glioma cells were determined by using an in vitro coculture system and an in vivo experimental xenografted mouse model. Intracellular signaling events that are responsible for the TMZ-mediated sensitization to TRAIL-induced apoptosis were also evaluated. Treatment of either TRAIL-sensitive or -resistant human glioma cells with TMZ and MSC-TRAIL resulted in a significant enhancement of apoptosis compared with the administration of each agent alone. We demonstrated that TMZ effectively increased the sensitivity to TRAIL-induced apoptosis via extracellular signal-regulated kinase-mediated upregulation of the death receptor 5 and downregulation of antiapoptotic proteins, such as X-linked inhibitor of apoptosis protein and cellular FLICE-inhibitory protein. Subsequently, this combined treatment resulted in a substantial increase in caspase activation. Furthermore, in vivo survival experiments and bioluminescence imaging analyses showed that treatment using MSC-TRAIL combined with TMZ had greater therapeutic efficacy than did single-agent treatments. These results suggest that the combination of clinically relevant TMZ and MSC-TRAIL is a potential therapeutic strategy for improving the treatment of malignant gliomas.

Autologous collagen-induced chondrogenesis using fibrin and atelocollagen mixture.

For articular cartilage defect treatment, many treatment modalities have been developed. We evaluate the cartilage repair potential of an atelocollagen and fibrin mixture transplanted to cartilage defects. A circular, articular cartilage defect 4 mm in diameter was made in the trochlear region in each of 20 New Zealand white rabbits. The 10 rabbits in the control group were kept without treatment and the 10 rabbits in the experimental group underwent injection of atelocollagen mixed with fibrin. At week 12 following surgery the cartilage was observed and histologically compared in both groups. The surface of the newly generated cartilage was very smooth and even, and we also noted that the entire area was completely regenerated in the experimental group. The control group showed incomplete and irregular cartilage formation in the defect. Regarding the histological scoring, comparison of the two groups differed significantly (p < 0.001). Injection of a mixture of atelocollagen and fibrin used to treat articular cartilage defects of the knee appears to be an effective method for cartilage regeneration.

A multi-center, randomized, clinical study to compare the effect and safety of autologous cultured osteoblast(Ossron) injection to treat fractures.

BACKGROUND: We performed a multicenter, open, randomized, clinical study of autologous cultured osteoblast injection for long-bone fracture, to evaluate the fracture healing acceleration effect and the safety of autologous cultured osteoblasts. METHODS: Sixty-four patients with long-bone fractures were randomly divided into two groups, i.e. those who received autologous cultured osteoblast injection and those who received no treatment. The sum of the difference in the callus formation scores after four and eight weeks, was used as the first efficacy variable. RESULTS: The autologous cultured osteoblast injection group showed fracture healing acceleration of statistical significance, and there were no specific patient complications when using this treatment. CONCLUSION: Autologous cultured osteoblast injection should therefore be considered as a successful treatment option for treating long-bone fracture.

Treatment of osteonecrosis of the femoral head using autologous cultured osteoblasts: a case report.

INTRODUCTION: Osteonecrosis of the femoral head is a progressive disease that leads to femoral head collapse and osteoarthritis. Our goal in treating osteonecrosis is to preserve, not to replace, the femoral head. CASE PRESENTATION: We present the case of a patient with bilateral osteonecrosis of the femoral head treated with autologous cultured osteoblast injection. CONCLUSION: Although our experience is limited to one patient, autologous cultured osteoblast transplantation appears to be effective for treating the osteonecrosis of femoral head.

Treatment of long tubular bone defect of rabbit using autologous cultured osteoblasts mixed with fibrin.

The osteogenic potential of autologous cultured osteoblasts mixed with fibrin when transplanted to bone defects was evaluated. Radial shaft defects over 15 mm were made in 30 New Zealand white rabbits. A total of 15 rabbits in the control group underwent an iliac bone graft and 15 rabbits in the experimental group underwent an autologous cultured osteoblast injection mixed with fibrin. Both groups were compared radiologically and 5 rabbits in each group were sacrificed for histological evaluation using H-E and Masson's trichrome stain at 3, 6, and 9 weeks. Osteogenesis in the control group progressed more rapidly than in the experimental group. However, at 9 weeks, bone formation in both groups were similar and showed no significant difference in terms of the amount of bone formation and the quality of bone union. Autologous cultured osteoblast transplantation mixed with fibrin in bone defects was found to produce bone efficiently.