Cell sheets technology is being available for fracture healing. This study was performed to clarify bone healing mechanism of undifferentiated (UCS) and osteogenic (OCS) differentiated mesenchymal stromal cell (MSC) sheets in the fracture model of dogs. UCS and OCS were harvested at 10 days of culture. Transverse fractures at the radius of six beagle dogs were assigned into three groups (n = 4 in each group) i.e. UCS, OCS and control. The fractures were fixed with a 2.7 mm locking plate and six screws. Cell sheets were wrapped around the fracture site. Bones were harvested 8 weeks after operation, then scanned by micro-computed tomography (micro-CT) and analyzed histopathologically. The micro-CT revealed different aspects of bone regeneration among the groups. The percentages of external callus volume out of total bone volume in control, UCS, and OCS groups were 42.1, 13.0 and 4.9% (p < 0.05) respectively. However, the percentages of limbs having connectivity of gaps were 25, 12.5 and 75% respectively. In histopathological assessments, OCS group showed well organized and mature woven bone with peripheral cartilage at the fracture site, whereas control group showed cartilage formation without bone maturation or ossification at the fracture site. Meanwhile, fracture site was only filled with fibrous connective tissue without endochondral ossification and bone formation in UCS group. It was suggested that the MSC sheets reduced the quantity of external callus, and OCS induced the primary bone healing.
Animals ; Bone Regeneration ; Bony Callus ; Cartilage ; Connective Tissue ; Dogs ; Extremities ; Fracture Healing ; Mesenchymal Stromal Cells ; Osteogenesis ; Radius

Mesenchymal stem cells (MSCs) are a heterogeneous population of cells that proliferate in vitro as plastic-adherent cells, have fibroblast-like morphology, form colonies in vitro and can differentiate into bone, cartilage and fat cells. Recent studies have shown that MSCs can be differentiated into nonmesordermal lineages. Although MSCs can be isolated from every type of connective tissues, the abundance, the easy and repeatable access to subcutaneous adipose tissue and the simple isolation procedures indicate that adipose tissue can be a preferable candidate in MSC isolation for clinical application. Therefore, in this review, the isolation, characterization, preclinical and clinical application, and the mechanisms and future roles of ADSC in cell therapy are discussed.
Adipocytes ; Adipose Tissue ; Cartilage ; Connective Tissue ; Mesenchymal Stromal Cells ; Subcutaneous Fat ; Tissue Therapy

According to the development of craniofacial surgery, various bone grafts were used for the contouring and reconstruction of craniofacial bone defects. Recently, autogenous bone graft has been used widely but there are several drawbacks associated to the autogenous bone grafting. In addition, using allogenic bone has a few problems. Alloplastic materials are expensive and sometimes infected. Therefore, our research is focused on the xenogenic bone graft which has advantage associated with no restriction in harvesting doner bone. The author undertook an experimental investigation as following: Skull and femur defects were made in size of 1x1cm each on 48 New Zealand white rabbits. The defects were grafted with human bone, and experimental group was divided into four groups depending on conditions of doner bone: Group 1 with deep frozen human bone, group 2 with autoclaving human bone, group 3 with deep frozen human bone and immunosuppression with FK-506, group 4 With autoclaving human bone and immunosuppression with FK-506. The specimens were obtained at 1, 2, 4, 8, 16, 24 weeks postoperatively. The author investigated new bone formation by gross appearance, microscopic and radiologic findings in grafted sites and calculated xenogenic antibody titer using ELISA in blood sample. The experimental results are as followings. Neovasculization was shown in connective tissue surrounding the grafted bone and new bone formation was started a week postoperatively and it is significantly increased at 4 weeks postoperatively. At 16 and 24 weeks, new bone formation was more remarkable in group 2 and 4, especially in group 2. Xenogenic antibody titer was higher in group 1 and 3 and immunosuppressed groups with FK-506 showed decreased xenogenic antibody titer. At 24 weeks postoperatively, callus formation and bone union were more remarkable in group 2, 4. According to this experimental studies, the author suggested that new bone formation was enhanced by osteoconductive properties of xenogenic bone and viability of bone was more injured in autoclaved bone than deep frozen bone. And immunologic reaction was suppressed satisfactorily by FK-506.
Bone Transplantation ; Bony Callus ; Connective Tissue ; Enzyme-Linked Immunosorbent Assay ; Femur ; Heterografts* ; Humans* ; Immunosuppression ; Osteogenesis ; Rabbits ; Skull ; Tacrolimus ; Transplants

According to recent advances in tissue engineering, several tissue types of mesodermal origin are successfully engineered In order to engineer connective tissues, a clinically practical and readily available source of cells must be identified Based on recent work relating to the existence of undifferentiated multipotential stem cells, they exist not only in bone marrow but also in many other connective tissue stromal components. We wondered if human fat tissue might also contain cells with developmental plasticity. More specifically, the purpose of this study was to induce cells isolated from the stromal fraction of human adipose tissue along the chondrogenic lineage in-vitro. Human adiopse tissue was harvested by conventional liposuction procedure. Collagenase type I was used for adipose tissue dissociation and stromal cell suspension was collected The cells were basically cultured in Opti-mem"". For chondrogenic digerentiation, a special culture condition, named micromass culture technique, was utilized and the media was supplemented with fetal bovine serum, L-ascorbic acid, ethanol, TGF-g and antibioticg antimycotic agents. The cells were cultured at 37C incubator with 5% COz for 3 weeks. For detection of chondroblast and/or cartilage matrix, alcian blue (pH 1.0) staining, alkaline phosphatase staining, indirect immunofluorescent labeling using ExtrAvidin'-FIZC conjugate were done. Scanning and transmission electron microscopic images were also taken for ultramicroscopic evaluation of the cells. Blue-colored positive reaction was detected in alcian blue staining, which means presence of sulfated mucin ('cartilage matrix). The cells expressed strong reaction of the tissue nonspecific isoform of the enzyme alkaline phosphatase. Bright fluorescence was detected in indirect immunofluorescent labeling representing the presence of collagen type II in the matrix, which is a specific component of cartilage. Based on our results, it can be concluded that human adipose tissue contains certain population of undigerentiated precursor cells, which could be differentiated into several connective tissue cell lineages. This population of cells may be used as an important future source for cellular building blocks for tissue engineering.
Adipose Tissue ; Alcian Blue ; Alkaline Phosphatase ; Ascorbic Acid ; Bone Marrow ; Cartilage ; Chondrocytes ; Chondrogenesis ; Collagen Type II ; Collagenases ; Connective Tissue ; Connective Tissue Cells ; Culture Techniques ; Ethanol ; Fluorescence ; Humans* ; Incubators ; Lipectomy ; Mesoderm ; Mucins ; Plastics ; Stem Cells ; Stromal Cells ; Tissue Engineering

Painful fat hemiation is characterized. by the soft, skin colored painful papules on the heels that appears only when weight is bome and disappears when weight. is taken off the heels. The papules were thought to be caused by hemiation of adipose tissue through connective tissue defect. To our knowledge, such a case or similar one has not been reported previously in Korea. The patient was 30 year-old housewife, who have had soft, skin colored painful papules on the medial aspect of the both heels when weight was placed on the foot. Routine laboratory findings were within normal limit. Skin biopsy revealed nodules of lobulated adipose tissue on the mid-dermis far above the original subcutaneous tissue and large venous channels with thick muscular walls. Hypertrophied peripheral nerve trunks were scattered adjacent to the above vascular structure. Diagnosis was confirmed by clinical and histopathological features. Literatun was briefly reviewed.
Adipose Tissue ; Adult ; Biopsy ; Connective Tissue ; Diagnosis ; Foot ; Heel ; Humans ; Korea ; Peripheral Nerves ; Skin ; Subcutaneous Tissue

BACKGROUND: Unlike bone, cartilage, or muscle, tendon-specific markers are not well established. The purpose of the study was to investigate expression pattern and level of 6 well-known tendon-specific markers, in various human musculoskeletal tissues, tenocytes, and mesenchymal stem cells (MSCs). METHODS: Musculoskeletal tissue samples of tendon, bone, cartilage, nerve, muscle, and fat were obtained from patients undergoing orthopedic surgery. Tenocytes, MSCs from bone marrow, adipose tissue, and umbilical cord were isolated from each tissue and cultured. Six tendon-specific markers, scleraxis (Scx), tenomodulin (TNMD), thrombospondin-4 (TSP-4), tenascin-C (TNC), type I collagen (Col I), and type III collagen (Col III) were investigated in tendon tissue, tenocytes, and MSCs. RESULTS: mRNA levels of 6 tendon-specific markers were significantly higher in tendon tissue that in other connective tissues levels of Scx, TNMD, TSP-4, and Col III immediately decreased after plating tenocytes in culture dishes whereas those of TNC and Col I did not. In comparison with tendon tissue, mRNA levels pattern of Scx, TNMD, and TSP-4 in tenocytes were significantly higher than that in MSCs, but lower than in tendon tissue whereas expression pattern of TNC, Col I and III showed different pattern with each other. CONCLUSION: This study demonstrated that 6 commonly used tendon-specific markers were mainly expressed in tendon tissue, but that expression level and pattern of the tendon-specific markers with respect to kinds of tissues, culture duration of tenocytes and sources of MSCs.
Adipose Tissue ; Biomarkers ; Bone Marrow ; Cartilage ; Collagen Type I ; Collagen Type III ; Connective Tissue ; Humans ; Mesenchymal Stromal Cells ; Orthopedics ; RNA, Messenger ; Tenascin ; Tendons ; Umbilical Cord

To examine the fate of cryopreserved meniscus used as orthotopic allograft on alternative to total removal of menisci, the authors have performed replacement of medial meniscus with cryopreserved one which had been preserved at −70℃ for 7 days in right side and with fresh autograft without cryopreservation in left side as a control group.Transplanted menisci were sampled at 4, 8, 12 ,16 weeks, postoperatively to observe their gross and histological findings. At 4 weeks, the allograft was healed to the perioheral capsular tissue and meniscal horn attachments of the host by fibrovascular scar tissue retained their normal appearance in gross findings. It showed an increased cellularity immediately adjacent to its peripheral attachment covered by the vascularized synovial tissues. At 8 weeks, it showed even distribution of cellularity in histological finding. At 12 weeks, gross findings of mild shrinkage and degeneration were seen and those were covered by both synovial and connective tissue with regenerated chondrocyte in histological findings. At 16 weeks, a little difference between allograft and normal meniscus was showed in gross and histological findings. There are regenerated chondrocytes in the deep area some degenerative changes were noted in the tibial and femoral cartilage not covered by meniscus. Above results showed cryopreserved meniscal allograft was able to regenerate and clinically suggest good treatment modality in case of unable to reshape to normal structure after total meniscectomy.
Allografts ; Animals ; Autografts ; Cartilage ; Chondrocytes ; Cicatrix ; Connective Tissue ; Cryopreservation ; Horns ; Knee ; Menisci, Tibial

The second case of this serial reports under the title of unusual cutaneous manifestations of connective tissue diseases is a woman with primary idiopathic dermatomyositis who developed cutaneous ulcerations on her arms and legs, which is thought to b a rare manifestation of adult dermatomyositis. These multiple ulcerations are about bean sized, polymorphous, and grouped on the poikilodermatous lesional skin. In this patient these ulcerative lesions became regressive and healed with disfiguring scars after the activity of the disease has controlled by prednisolone and methotrexate.
Adult ; Arm ; Cicatrix ; Connective Tissue Diseases* ; Connective Tissue* ; Dermatomyositis* ; Female ; Humans ; Leg ; Methotrexate ; Prednisolone ; Skin ; Ulcer*

Nevus lipomatosus cutaneous superficialis (NLCS) is an uncommon form of connective tissue nevus, which is manifested principally as the deposition of fatty tissue in the dermis. Lesions almost always arise unilaterally on the thigh or buttock. Although NLCS can affect any region of the skin, only five cases of NLCS on face have been reported. A 48-year-old female presented with a skin-colored, 1 cm sized, soft and edematous plaque on the right malar area. The histopathologic findings showed scattered ectopic fat lobules between collagen fibers in the mid to deep dermis. We herein reported a case of solitary and plaque-shaped NLCS on the malar area.
Adipose Tissue ; Buttocks ; Collagen ; Connective Tissue ; Dermis ; Female ; Humans ; Nevus ; Skin ; Thigh

Nevus lipomatosus superficialis is an uncommon form of connective tissue nevus and is manifest principally by the deposition of fatty tissue in the dermis. In this disease two clinical types are distinguished. The classic type consists of asymptomatic linear, soft, flesh-colored to yellowish papules or nodules that often coalesce into plaque. It is usually located in the pelvic girdle and lumbar area. The solitary form usually appears later in life and can occur at any site. Clinically, it may be confused with neurofibroma and acrochordon. The solitary form has been described as a pedunculated lipofibroma. There is no sexual predilection or hereditary trend. The authors experienced two cases of solitary nevus lipomatosus superficialis. The first case was a 57-year-old male who had a solitary, 1 cm-diametered, flesh-colored, polypoid nodule on the buttock for 4 years. The second case was a 32-year-old female who had a 1.5 cm-diametered, flesh-colored and light brown, polypoid nodule on the right flank for 10 years. Diagnosis was confirmed by clinical and histopathologic findings.
Adipose Tissue ; Adult ; Buttocks ; Connective Tissue ; Dermis ; Female ; Humans ; Light ; Male ; Middle Aged ; Neurofibroma ; Nevus