Histochemical characteristics of calcium binding S100 proteins and bone morphogenetic proteins in chondro-osseous tumors.

Immunohistochemical distribution of the Ca2+ binding proteins S100A1, S100A2, S100A4, S100A6, S100B, and bone morphogenetic protein (BMP) in chondro-osseous tumors and lesions, both benign and malignant, was investigated using specific anti S100 protein and BMP antibodies. Chondrogenic tumor cells of chondro-osseous lesions were characterized by the presence of marked staining for S100B and BMP, while they were only faintly reactive for S100A1, S100A2, S100A4 and S100A6. Dense fibrous connective tissue in osseous tumor and ossifying fibroma showed moderate immunoreactivity for S100A1, S100A4 and BMP. Immunoreactivity of S100A2, prominent in epidermal basal cells and associated or homologous cells of epidermal tumors or skin appendages tumors, was not found in cartilage and bone forming cells. Biological roles of S100B in chondroid cells may involve Ca2+-signaling in precalcified tissue, cytoskeletal protein organization and matrix formation since glycosaminoglycan synthesis is mediated by calcium ions. S100B positive cells in chondro-osseous structures also strongly expressed BMP. The present study allowed us to conclude that among the S100 proteins, the S100B in particular and S100A1, S100A4 and S100A6 but not S100A2 may be involved in the process of tumorigenesis of chondro-osseous tumors and BMP may have an important role in the chondroid and osseous differentiation. The detailed biological role of S100 proteins in chondro-osseous tumors is under investigation.

Tenascin: growth and adhesion modulation--extracellular matrix degrading function: an in vitro study.

Tenascin (TN), a recently characterised extracellular matrix protein, largely confined to the process with the development of embryo in areas of epithelial-mesenchymal interactions and in areas where there are morphogenetic movements and tissue patterning, has a highly restricted expression in adult tissues. The expression of TN is enhanced in a variety of human neoplastic lesions. However, function(s) and molecular mechanisms of enhanced expression in neoplastic lesions remain unclear. We employed human tongue carcinoma cells (SCCKN), human salivary gland adenocarcinoma cells (SGT-1), normal mouse embryonic fibroblasts (NIH3T3-3) and K-ras-2 transformed fibroblasts (Cle-H3) in an in vitro study to elucidate the biological roles of TN. In in vitro studies, all the cell lines examined had enhanced secretion of TN in the presence of transforming growth factor-beta in a dose-dependent manner and TN itself was found to possess a growth-enhancing activity. Moreover, studies on adhesion of the cell lines on coated substrates of fibronectin (FN), laminin (LN), tenascin (TN), TN/FN and TN/LN showed that all the cells adhere and spread well on FN and LN. However, on TN they attach poorly and remain rounded. The relative concentrations of TN and FN affected the cellular adhesion and morphology. In SCCKN and SGT-1, but not in NIH3T3 and Cle-He3 fibroblasts, a higher concentration of TN inhibited cellular adhesion on fibronectin, suggesting that cells attach poorly on TN, it may interfere with the action of fibronectin, and the relative concentrations of TN, FN or LN may affect cellular adhesion and morphology which may differ in different cell types. When TN was added in the growth medium of exponentially growing cells, the cells lost their cell to cell contact and were seen to be separating. The presence of these extracellular matrix proteins were further tested to determine whether they could modulate the secretion of proteolytic enzymes responsible for extracellular matrix degradation by tumour cells, when the neoplastic cells but not the non-neoplastic cells grown on FN/TN substrate showed positive immunofluorescence for collagenase. FN, LN or TN alone did not induce collagenase in the tumour cells. If the same is true in vivo, although a number of factors and interactions may implicate the ultimate outcome, the enhanced expression of TN in neoplastic lesions may have potential implications for tumour growth, differentiation, cellular adhesion, invasion and metastasis.

Immunohistochemical evaluation of bone morphogenetic protein (BMP) in mixed tumor of skin.

Immunohistochemical reaction of bone morphogenetic protein (BMP) was assessed in 19 cases of skin mixed tumor and 5 cases of skin appendage tumors by using monoclonal antibody raised against BMP. All cases of skin mixed tumor showed positive staining for BMP in modified myoepithelial cells located at the periphery of tubulo-ductal or solid structures, and in plasmacytoid or tumor cells in hyalinous structures. Chondroidally changed cells also showed a strong BMP immunoreactivity. Tumors originating from sweat glands were devoid of BMP immunoreactivity. It is concluded that BMP is synthesized and produced in modified or transformed myoepithelial cells in skin mixed tumor and participates in the process of chondroid changes in the tumor.

Bone formation under the influence of bone morphogenetic protein/self-setting apatite cement composite as a delivery system.

Self-setting apatite cement (apatite cement) with a phase of hydroxyapatite (HAP) was employed as a delivery system for bone morphogenetic protein (BMP). A composite of BMP and apatite cement (BMP/HAP composite) was implanted both in thigh muscle and surgically created defect of a critical size of 5 mm, which is a size that does not heal spontaneously in the femur of mice, to evaluate its osteogenetic potential as an augmentation and reconstructive material for clinical usage. The histological and immunohistochemical assessment of proteoglycans reiterated osteogenesis in the muscle tissue. On day 14 postimplantation of BMP/HAP composite, chondroid tissue was formed in the muscle, and HAP particles were seen in newly formed chondroid tissue. On the 21st day, endo-chondral ossification had occurred, however, small HAP particles remained in the newly formed bone, i.e., HAP particles and newly formed osseous tissues coexisted in a central area. BMP/HAP composite was incorporated by newly formed osseous tissue in the experimented animals. HAP particle found in BMP/HAP composite implanted into the bone defect was resorbed and replaced by osseous tissue. The apatite cement was proved to have advantages for its shaping as well as collapsing properties, and thus, apatite cement containing BMP is suggested as a favorable augmentation material in clinical usage for delivery system of BMP.

Mandibular reconstruction using electrically stimulated periosteum.

Electrically stimulated periosteum (ESP) grafting to a 12-year-old female patient who had had a segmental mandibulectomy in the anterior region following a diagnosis of Ewing's sarcoma was tried for the mandibular reconstruction. One year after the ESP transplantation, examination of the patient showed that fine, radio-opaque bone formation had occurred. The new technique consists of 1) electrical stimulation of the tibial periosteum with 20 microamperes for five weeks, and then 2) ESP transplantation into a titanium mesh tray placed in the mandibulectomized region. In the present report, these new procedures are detailed, and the process of calcification in the transplanted ESP is discussed.

Pathologic and enzyme histochemical studies on bone formation induced by bone morphogenetic protein in mouse muscle tissue.

Bone morphogenetic protein (BMP) irreversibly induced the differentiation of mesenchymal-type cells into osteoprogenitor cells for endochondral ossification. During the process of BMP-induced differentiation in mice, 4 cell type (chondroblasts, osteoblasts, chondroclasts, and osteoclasts) were examined for phosphatase and succinate dehydrogenase using a wide range of buffers (4.0 less than or equal to pH less than or equal to 9.2). During the chondroid tissue-forming stage (1 week), chondroblast-like or osteoblast-like cells expressed phosphatase activity at 6.8 less than or equal to pH less than or equal to 9.2; chondroclast-like or osteoclast-like cells expressed phosphatase activity at 4.0 less than or equal to pH less than or equal to 5.8. However, mature chondrocytes found in hyaline cartilage expressed phosphatase activity between 6.6 less than or equal to pH less than or equal to 7.6 (2 weeks). During the process of endochondral ossification, alkaline phosphatase activity decreased in osteoblast-like cells with traces of acid phosphatase activity still detectable. Chondroclastic and osteoclastic giant cells were characterized by intense succinate dehydrogenase activity.

[The clinical use of Hotz-type orthopedic plate].

In the present study a Hotz type orthopedic plate was used to improve feeding and physiological growth. In 2 patients with cleft lip it was used to improve feeding and swallowing. In 4 patients with cleft lip and palate it was used to normalize functions and aid physiological growth. The effects of the Hotz orthopedic plate were determined by recording body weight growth, milk volume per day, feeding time and comparison of the study model between before and after using Hotz orthopedic plate. Good physiological growth and improved feeding, were obtoincdinalmostall the patients who wore the Hotz type orthopedic plate correctly.