BACKGROUNDS: Type I diabetes mellitus (T1DM), an autoimmune disease, is associated with insulin deficiency due to the death of beta-cells. Bone marrow-derived mesenchymal stem cells (BM-MSCs) are capable of tissue repair and thus are a promising source of beta-cell surrogates. METHODS: In this study, the therapeutic potential of BM-MSCs as beta-cell replacements was analyzed both in vitro and in vivo. First, we used retinoic acid (RA) to induce rat BM-MSCs to differentiate into cells of endodermal/pancreatic lineage. Then, differentiated rat BM-MSCs were syngeneically injected under the renal capsule of rats. RESULTS: Analysis of gene expression revealed that rat BM-MSCs showed signs of early pancreatic development, and differentiated cells were qualitatively and quantitatively confirmed to produce insulin in vitro. In vivo study was performed for short-term (3 weeks) and long-term (8 weeks) period of time. Rats that were injected with differentiated MSCs exhibited a reduction in blood glucose levels throughout 8 weeks, and grafted cells survived in vivo for at least 3 weeks. CONCLUSIONS: These findings show that RA can induce differentiation of MSCs into the beta-cell lineage and demonstrate the potential of BM-MSCs to serve as therapeutic tools for T1DM.
Animals ; Autoimmune Diseases ; Blood Glucose ; Diabetes Mellitus ; Diabetes Mellitus, Type 1 ; Gene Expression ; Insulin ; Insulin-Secreting Cells ; Mesenchymal Stromal Cells* ; Rats* ; Transplants ; Tretinoin

BACKGROUND: AEB071, an orally available PKC inhibitor, prevents organ rejection after transplantation in rodents and man. Furthermore, pro-inflammatory cytokines and inflammatory processes are important mediators of transplanted organ rejection. We therefore examined whether single or combination therapies of AEB071 and/or tacrolimus affect cytokine profiles in a rat cardiac allograft model. METHODS: AEB071 (60 mg/kg twice a day) and tacrolimus (0.6 or 1.2 mg/kg once a day) were orally administered daily after cardiac transplantation. Interferon (IFN)-gamma, interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-10, and tumor necrosis factor (TNF)-alpha levels in serum were subsequently measured 5 days after cardiac transplantation using a multiplex protein assay system. RESULTS: All cytokine levels were significantly depressed in cardiac transplanted rats treated with AEB071, whereas tacrolimus only reduced IFN-gamma, IL-2, IL-4, IL-6, and IL-10 levels. When administered in combination, AEB071 and low- or high-dose tacrolimus had additive effects on IFN-gamma, IL-4, IL-6, and TNF-alpha. CONCLUSIONS: These results suggest that AEB071 inhibits T cell activation by blocking the production of proinflammatory cytokines, and that tacrolimus combined with AEB071 can effectively regulate inflammatory cytokines in the transplantation setting.
Animals ; Cytokines ; Heart Transplantation ; Immunosuppression ; Interferons ; Interleukin-10 ; Interleukin-2 ; Interleukin-4 ; Interleukin-6 ; Interleukins ; Pyrroles ; Quinazolines ; Rats ; Rejection (Psychology) ; Rodentia ; Tacrolimus ; Transplantation, Homologous ; Transplants ; Tumor Necrosis Factor-alpha

A giant cell tumor of tendon sheath is a slow growing benign soft tissue tumor that is known by a variety of names including fibrous histiocytoma of tendon sheath and fibrous xanthoma of the synovium. Clinically, it presents as a 1~3 cm firm, non-mobile, painless, nontender mass, and mostly occurs at interphalangeal joints of fingers. It shows female predominance and can occur at any age, but it is most common between the third and fifth decades and is rare in children. We now report the case of a 10-year-old girl with a giant cell tumor of tendon sheath on the toe.
Child ; Female ; Fingers ; Giant Cell Tumors ; Giant Cells ; Histiocytoma, Benign Fibrous ; Humans ; Joints ; Synovial Membrane ; Tendons ; Toes ; Xanthomatosis

Diabetic foot ulcer is a serious clinical problem with significant medical and economic effects on health systems worldwide. Some patients undergo amputation and others experience disability for an extended period of time. Treatment of diabetic foot ulcer is complex and difficult. Even with proper management, the wounds may not heal as well as expected. To promote wound healing, many advanced topical dressing materials have been developed. Among them, bi-layered collagen membrane, which is composed of collagen and hyaluronic acid, is believed to enhance wound healing. Herein we report two cases of diabetic foot ulcer which were successfully treated using bi-layered collagen membranes.
Amputation ; Bandages ; Collagen ; Diabetic Foot ; Humans ; Hyaluronic Acid ; Membranes ; Ulcer ; Wound Healing

Diabetic foot ulcer is a serious clinical problem with significant medical and economic effects on health systems worldwide. Some patients undergo amputation and others experience disability for an extended period of time. Treatment of diabetic foot ulcer is complex and difficult. Even with proper management, the wounds may not heal as well as expected. To promote wound healing, many advanced topical dressing materials have been developed. Among them, bi-layered collagen membrane, which is composed of collagen and hyaluronic acid, is believed to enhance wound healing. Herein we report two cases of diabetic foot ulcer which were successfully treated using bi-layered collagen membranes.
Amputation ; Bandages ; Collagen ; Diabetic Foot ; Humans ; Hyaluronic Acid ; Membranes ; Ulcer ; Wound Healing

STUDY DESIGN: In vitro experimental study. OBJECTIVES: To examine the cellular proliferation, synthetic activity and phenotypical expression of intervertebral disc (IVD) cells seeded on types I and II atelocollagen scaffolds, with the stimulation of TGF-beta1 and BMP-2. SUMMARY OF LITERATURE REVIEW: Recently, tissue engineering is regarded as a new experimental technique for the biological treatment of degenerative IVD diseases, and has been highlighted as a promising technique for the regeneration of tissues and organs in the human body. Research on cell transplantation in artificial scaffolds has provided that the conditions for tissue engineering have to be equilibrated, including the cell viability and proliferation, maintenance of characteristic phenotype, suitable scaffolds in organisms and biologically stimulated growth factor. MATERIAL AND METHOD: Lumbar IVD cells were harvested from 10 New Zealand white rabbits, with the nucleus pulposus cells isolated by sequential enzymatic digestion. Each of 1% types I and II atelocollagen dispersions were poured into a 96-well plate (diameter 5 mm), frozen at -70 degrees C, and then lyophilized at -50 degrees C. Fabricated porous collagen matrices were made using the cross-linking method. Cell suspensions were imbibed by surface tension into a scaffold consisting of atelocollagen. The cell cultured scaffolds were then treated with TGF-beta1 (10 ng/ml) or BMP-2 (100 ng/ml) or both. After 1 and 2 week culture periods, the DNA synthesis was measured by [3H] thymidine incorporation, and newly synthesized proteoglycan by incorporation of [35S] sulphate. Reverse transcription-polymerase chain reactions for the mRNA expressions of type I and II collagen, aggrecan and osteocalcin were performed. The inner morphology of the scaffolds was determined by scanning electron microscopy (SEM). RESULTS: The IVD cultures in collagen type II with TGF-beta1 demonstrated an increase in proteoglycan synthesis and up regulation of aggrecan and types I and II collagen mRNA expressions compared to the control. IVD cultures in the type I atelocollagen scaffold with growth factors exhibited an increase in DNA synthesis and up regulation of the type II atelocollagen mRNA expression. With all combinations of growth factor, the IVD cultures in types I and II atelocollagen scaffolds showed no up regulation of the osteocalcin mRNA expression. Furthermore, there was no synergistic effect of TGF-beta1 and BMP-2 in the matrix synthesis or for the mRNA expression of the matrix components. CONCLUSIONS: Nucleus pulposus cells from rabbit were viable in atelocollagen types I and II atelocollagen scaffolds. The type I atelocollagen scaffold was suitable for cell proliferation, but the type II atelocollagen scaffold was more suitable for extracellular matrix synthesis. The IVD cells in both scaffolds were biologically responsive to growth factors. Taken together, nucleus pulposus cells in atelocollagen scaffolds, with anabolic growth factors, provide a mechanism for tissue engineering of IVD cells.
Aggrecans ; Cell Proliferation ; Cell Survival ; Cell Transplantation ; Collagen ; Collagen Type II ; Digestion ; DNA ; Extracellular Matrix ; Human Body ; Intercellular Signaling Peptides and Proteins* ; Intervertebral Disc* ; Microscopy, Electron, Scanning ; Osteocalcin ; Phenotype ; Proteoglycans ; Rabbits ; Regeneration ; RNA, Messenger ; Surface Tension ; Suspensions ; Thymidine ; Tissue Engineering ; Transforming Growth Factor beta1 ; Transplants ; Up-Regulation

STUDY DESIGN: In vitro experimental study. OBJECTIVES: To examine the cellular proliferation, synthetic activity and phenotypical expression of intervertebral disc (IVD) cells seeded on types I and II atelocollagen scaffolds, with the stimulation of TGF-beta1 and BMP-2. SUMMARY OF LITERATURE REVIEW: Recently, tissue engineering is regarded as a new experimental technique for the biological treatment of degenerative IVD diseases, and has been highlighted as a promising technique for the regeneration of tissues and organs in the human body. Research on cell transplantation in artificial scaffolds has provided that the conditions for tissue engineering have to be equilibrated, including the cell viability and proliferation, maintenance of characteristic phenotype, suitable scaffolds in organisms and biologically stimulated growth factor. MATERIAL AND METHOD: Lumbar IVD cells were harvested from 10 New Zealand white rabbits, with the nucleus pulposus cells isolated by sequential enzymatic digestion. Each of 1% types I and II atelocollagen dispersions were poured into a 96-well plate (diameter 5 mm), frozen at -70 degrees C, and then lyophilized at -50 degrees C. Fabricated porous collagen matrices were made using the cross-linking method. Cell suspensions were imbibed by surface tension into a scaffold consisting of atelocollagen. The cell cultured scaffolds were then treated with TGF-beta1 (10 ng/ml) or BMP-2 (100 ng/ml) or both. After 1 and 2 week culture periods, the DNA synthesis was measured by [3H] thymidine incorporation, and newly synthesized proteoglycan by incorporation of [35S] sulphate. Reverse transcription-polymerase chain reactions for the mRNA expressions of type I and II collagen, aggrecan and osteocalcin were performed. The inner morphology of the scaffolds was determined by scanning electron microscopy (SEM). RESULTS: The IVD cultures in collagen type II with TGF-beta1 demonstrated an increase in proteoglycan synthesis and up regulation of aggrecan and types I and II collagen mRNA expressions compared to the control. IVD cultures in the type I atelocollagen scaffold with growth factors exhibited an increase in DNA synthesis and up regulation of the type II atelocollagen mRNA expression. With all combinations of growth factor, the IVD cultures in types I and II atelocollagen scaffolds showed no up regulation of the osteocalcin mRNA expression. Furthermore, there was no synergistic effect of TGF-beta1 and BMP-2 in the matrix synthesis or for the mRNA expression of the matrix components. CONCLUSIONS: Nucleus pulposus cells from rabbit were viable in atelocollagen types I and II atelocollagen scaffolds. The type I atelocollagen scaffold was suitable for cell proliferation, but the type II atelocollagen scaffold was more suitable for extracellular matrix synthesis. The IVD cells in both scaffolds were biologically responsive to growth factors. Taken together, nucleus pulposus cells in atelocollagen scaffolds, with anabolic growth factors, provide a mechanism for tissue engineering of IVD cells.
Aggrecans ; Cell Proliferation ; Cell Survival ; Cell Transplantation ; Collagen ; Collagen Type II ; Digestion ; DNA ; Extracellular Matrix ; Human Body ; Intercellular Signaling Peptides and Proteins* ; Intervertebral Disc* ; Microscopy, Electron, Scanning ; Osteocalcin ; Phenotype ; Proteoglycans ; Rabbits ; Regeneration ; RNA, Messenger ; Surface Tension ; Suspensions ; Thymidine ; Tissue Engineering ; Transforming Growth Factor beta1 ; Transplants ; Up-Regulation

A family of proteins, the bone morphogenetic proteins (BMPs), which promote osteoblast differentiation and bone mineralization, have recently been identified. One, BMP-7, has shown the ability to induce cartilage and bone formation processes. In this report, the possibility that other cell lines, to CHO cells, may also be available as host cells for the expression of hBMP-7 was validated. Recombinant human BMP (rhBMP) -7 was produced in COS-7 cells, as a processed mature disulfide-linked homodimer, with an apparent molecular weight of 36, 000. Examination of the expressions of the markers characteristic of osteoblast phenotypes showed that the rhBMP-7 specifically stimulated the inductions of alkaline phosphatase (ALP) (5-fold increase at 100 ng of rhBMP-7/ml), parathyroid hormone (PTH) -mediated intracellular cAMP production (4-fold increase at 100 ng of rhBMP-7/ml) and osteocalcin synthesis (5-fold increase at 100 ng of rhBMP-7/ml). In summary, the in vitro mineralization assay results provide evidence that the rhBMP-7 peptide, produced by COS-7 expression system, possesses intact biological activity. A similar pattern of biological activity was observed for the BMP-7 in COS-7 cells compared to the corresponding CHO cell expression system. Thus, these findings can be experimentally utilized for the production of rhBMPs for in vitro or in vivo studies.
Animals ; Animals, Newborn ; Bone Morphogenetic Proteins/*pharmacology ; COS Cells ; Cell Differentiation/drug effects ; Cell Division/drug effects ; Cells, Cultured ; Human ; Osteoblasts/*cytology ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins/pharmacology ; Skull/*cytology ; Support, Non-U.S. Gov't

Autograft is frequently used to restore anatomic morphology and functional properties in bone defects. Disadvantages of the autograft are related to donor site morbidity and include the risk of wound infection, increased blood loss and additional postoperative discomfort. Allograft and xenograft, which are currently employed as the most common alternative to autografts, encounter the complications such as fracture, resorption and nonunion secondary to immunologic rejection. These volumetric and immunologic concerns with biologic implants have stimulated interest in the potential for synthetic, bioinert materials as bone graft substitutes. Hydroxyapatite (HA), a calcium phosphate ceramic, is a well known biocompatible artificial bone substitite without induction of systemic toxic and foreign body reactions. Bone conduction usually occurrs by the implanted HA but biodegradation of HA is poor and the bone formation around HA is slow. Carbonate apatite has been known as that the physicochemical properties are similar of the natural bony apatite and demonstrates no toxic reactions with better biodegradation. Carbonate apatite collagen composite was reported to show more bone formation and biodegradation than hydroxyap atite. In this study, the composite (CA-C) consisted of carbonate apatite and type I collagen was implanted in rabbit tibiae to evaluate the possibility as an artificial bone substitute. Forty HA (HA group) and forty CA-C (CA-C group) were applied in 80 dissected rabbit tibiae and fixed by external fixators. For biomechanical study, the rabbits were sacrificed and the specimens were obtained in 2, 4, 6 and 8 weeks after implantation. Tensile load was applied to the prepared tibiae in Instron and biomechanical properties were investigated. The fracture at the callus occurred as transverse or short oblique fracture in the vertical direction to the axis of applied tensile load. In each group, the tensile strength at breaking point increased significantly with time and at 4 weeks more prominent increase in break strength was observed (p<0.05). HA group showed higher mean strength before 6 weeks and at 8 weeks CA-C group higher mean strength but statistical significance could not be found. There was no significant difference in extension length at breaking point between both groups and time intervals. In summary, carbonate apatite collagen composite revealed similar biomechanical properties as hydroxyapatite, suggesting its clinical usefulness as a bone substitute, but it will be necessary to improve biodegradational property, stiffness of carbonate apatite collagen composite.
Allografts ; Autografts ; Axis, Cervical Vertebra ; Bone Conduction ; Bone Substitutes ; Bony Callus ; Calcium ; Carbon* ; Ceramics ; Collagen Type I ; Collagen* ; Durapatite* ; External Fixators ; Foreign Bodies ; Heterografts ; Humans ; Osteogenesis ; Rabbits ; Tensile Strength ; Tibia* ; Tissue Donors ; Transplants ; Wound Infection

Autograft is frequently used to restore anatomic morphology and functional properties in bone defects. Disadvantages of the autograft are related to donor site morbidity and include the risk of wound infection, increased blood loss, and additional postoperative discomfort. Allograft and xenograft, which are currently employed as the most common alternative to autografts, encounter the complications. such as fracture, resorption, and nonunion secondary to immunologic rejection. Hydroxyapatite(HAp), a calcium phosphate ceramic, is a well known biocompatible artificial bone substitute without induction of systemic toxic and foreign body reactions. Bone conduction is usually occurred by the implanted HAp. Biodegradation of HAp is poor and the bone formation around HAp is slow and about more than 70% of the natural apatite in bone is composed of carbonate apatite that has carbonic radical(CO3) instead of hydroxyl radical(OH-) or Phosphate radical(PO4 ) of HAp. Carbonate apatite ahs been known as that the physicochemical properties are similar of the natural bony apatite. In vivo experiment, carbonate apatite demonstrates non-toxic reactions. In this study, the compostite consisted of carbonate apatite and type I collagen, which is the bony structural protein, was implanted in rabbit tibiae to evaluate possibility as an artificial bone substitute. Biodegradation rate of the composite, the rate and qiantity of the regenerative cortical and cancellous bone formation were investigated radiologically and histopathologically. HAp and the carbonate apatite coilagen composite(Ap-C) were applied in the dissected rabbit tibiae(7-8mm), and fixed by external fixators. Postoperative roentgenograms were taken 2, 4, 6, and 8 weeks after implantations. For histological ohservations, the rabbits were sacrificed and the specimens were obtained 2, 4, 6, and 8 weeks after operation. Radiological Lane scores of Ap-C were 1.16±0.92, 2.16±0.98, 3.33± 0.82, and 3.67±0.51 in 2. 4, 6 and weeks after operation, while the Lane score of HAp were 0.53±0.84, 0.83±0.75, 1.67±0.81, and 2.83±0.98 respectively. Ap-C demonstrated the faster degradation and more bone formation then HAp. Though physicochemical properties of carbonate apatite is similar to those of HAp, the carbonate apatite and type I collagen composite(Ap-C) revealed rather faster degradation and bone formation than HAp in both radiological and histological investigations.
Allografts ; Autografts ; Bone Conduction ; Bone Substitutes ; Calcium ; Carbon ; Ceramics ; Collagen Type I ; Collagen ; Durapatite ; External Fixators ; Foreign Bodies ; Heterografts ; Humans ; Osteogenesis ; Rabbits ; Tibia ; Tissue Donors ; Wound Infection