[Gene mutation and expression of SH-3BP-2 in cherubism].

OBJECTIVE: To detect the mutation and expression of SH-3BP-2 in Chinese patients of cherubism and to investigate the possible relationship of gene mutation and multinucleated giant cells in lesions. METHODS: Genomic DNA was extracted from paraffin-imbedded tissues and peripheral blood samples of 10 cases of cherubism (6 familial cherubism and 4 sporadic cherubism). SH-3BP-2 mutations were detected by PCR-direct sequencing. The nature of multinucleated giant cells in lesions was detected by enzyme histochemical staining and immunohistochemical staining using paraffin-imbedded tissues sections. The SH-3BP-2 protein was detected by immunohistochemical staining. RESULTS: Three missense mutations (G1520A, G1505A, G1505C) in exon 9 of SH-3BP-2 were identified which led to 3 transitions (Gly420Glu, Arg415Gln, Arg415Pro). There were no abnormalities in exon 3 of SH-3BP-2 except 1 case which had not PCR products. The protein SH-3BP-2, the calcitonin receptor and the tartrate-resistant acid phosphatase were detected in the cytoplasm of all multinucleated giant cells and parts of monokaryon matrix cells in 8 paraffin-imbedded samples. CONCLUSIONS: The SH-3BP-2 mutation may participate in the differentiation and maturation of osteoclast-like cells in the lesion of cherubism.

[In vitro study of the effects of copper ion on osteoclastic resorption in various dental mineralized tissues].

OBJECTIVE: To investigate the effects of copper ion on osteoclastic resorption in various dental mineralized tissues. METHODS: Osteoclasts were separated from long-limb bones of neonatal rabbits, and cultured with de-activated human tooth slices and glass slices. The cells in the experiment group were treated with (1 x 10(-14))-1 x (10(-4)) mol/L copper + 10% (volume fraction) fetal calf serum (FCS) + alpha-MEM, while the cells in control group cells were grown in 10% FCS + alpha-MEM. Osteoclasts on glass slices were stained by TRAP staining. The absorption pits on tooth slices were observed by inverted phase contrast microscope. The resorbing activity was evaluated with the concentration of calcium in the supernatant liquid of osteoclasts. The ratio between the concentration of calcium in the experiment group and control group was termed as the resorption index. RESULTS: The isolated cells were multinuclear and TRAP positive in cytoplasma. Osteoclasts resorbed teeth slices first on the cementum and dentin. Compared with those on bone slices, the lacunae on the dental slices numbered less, with smaller volume and shallower in depth. Microscopy showed that the number and area of absorption pits formed on treated tissues were less than those on control tissues. The content of calcium in the supernatant liquid decreased in the concentrations of 1 x 10(-14) mol/L - 1 x 10(-4) mol/L copper, especially in the group of 1 x 10(-10) mol/L copper at 3rd day (P < 0.05) and 1 x 10(-4) mol/L, 1 x 10(-10) - 1 x 10(-12) mol/L copper at 7th day (P < 0.05). Their resorption index was lower than one. CONCLUSIONS: Extracellular copper ion can inhibit osteoclastic resorption on dental slices.

[Effect of extracellular zinc on osteoclastic resorption in dental mineralized tissues].

OBJECTIVE: To investigate the bone resorption caused by osteoclasts and modulating functions of zinc ion on dental slices. METHODS: Osteoclasts were separated from long-limb bones of neonatal rabbits, cultured with de-activated human tooth slices and glass slices. The cells in the experiment group were treated with 1x10(-14)mol/L-1x10(-4)mol/L zinc+10% (volume fraction) fetal calf serum (FCS)+alphaMEM, while those in the control group were grown in 10%FCS+alphaMEM. Osteoclasts on glass slices were stained by TRAP staining. The absorption pits on tooth slices were observed by inverted phase contrast microscope. The resorbing activity was evaluated with the concentration of calcium in the supernatant liquid of osteoclasts. The ratio between the concentration of calcium in the experiment group and that of the control group was termed the resorption index. RESULTS: The isolated cells were multinuclear and showed positive in cytoplasma by TRAP staining. Usually, osteoclasts resorbed tooth slices first on the cementum and dentin, which had lower content of mineralized tissue. Compared with those on bone slices, the lacunae on the dental slices appeared less in amount, less in area and shallower in depth. They often showed shallow pits in a large area. Microscopy showed that the number and area of absorption pits formed on treated tissues were less than those on the control tissues. The content of calcium in the supernatant liquid increased at the concentrations of 1x10(-4)-1x10(-14)mol/L zinc, especially in the group of 1x10(-8)mol/L, 1x10(-10) mol/L, 1x10(-14)mol/L zinc on the 3rd day (P<0.05). But they were reversed on the 7th day, except in the group of 1x10(-14)mol/L zinc. At the end of culture, the resorption indexes of 1x10(-4)-1x10(-7)mol/L, 1x10(-9)mol/L, 1x10(-12)mol/L and 1x10(-13)mol/L group were lower than 1, but those of 1x10(-8)mol/L, 1x10(-10)mol/L, 1x10(-11)mol/L and 1x10(-14)mol/L group were higher than 1. CONCLUSION: The effect of zinc ion on osteoclastic resorption in dental slices is associated with phase and dosage closely.

A novel mutation in the SH3BP2 gene causes cherubism: case report.

BACKGROUND: Cherubism is a rare hereditary multi-cystic disease of the jaws, characterized by its typical appearance in early childhood, and stabilization and remission after puberty. It is genetically transmitted in an autosomal dominant fashion and the gene coding for SH3-binding protein 2 (SH3BP2) may be involved. CASE PRESENTATION: We investigated a family consisting of 21 members with 3 female affected individuals with cherubism from Northern China. Of these 21 family members, 17 were recruited for the genetic analysis. We conducted the direct sequence analysis of the SH3BP2 gene among these 17 family members. A disease-causing mutation was identified in exon 9 of the gene. It was an A1517G base change, which leads to a D419G amino acid substitution. CONCLUSION: To our knowledge, the A1517G mutation has not been reported previously in cherubism. This finding is novel.

[Mutation detection in SH3BP2 gene in a cherubism family].

OBJECTIVE: To detect SH3BP2 gene mutation in a cherubism family. METHODS: Peripheral blood samples were obtained from the family of cherubism. Genomic DNA was extracted. Polymerase chain reaction and direct sequencing were performed to identify the mutation. RESULTS: A transition in exon 9 in SH3BP2 gene was detected in the family, which led to a missense mutation (Arg 415 Pro). CONCLUSIONS: Missense mutation in the SH3BP2 gene was responsible for the phenotypes of this Chinese cherubism family.

[Expression of macrophage inflammatory protein-1alpha, a disintegrin-like and metalloproteinase 8 and 12, and CD68 protein in giant cell lesions of jaw and giant cell tumors of long bone].

OBJECTIVE: To detect the expression of macrophage inflammatory protein-1alpha (MIP-1alpha), a disintegrin-like and metalloproteinase (ADAM) 8 and 12 and CD68 protein in giant cell lesions of jaw and giant cell tumors of long bone, and to study their effects on the histogenesis of giant cells in such lesions. METHODS: MIP-1alpha, ADAM8, ADAM12 and CD68 were detected by immunohistochemistry in 24 paraffin-embedded specimens of central giant cell lesions of jaw and giant cell tumors respectively. RESULTS: MIP-1alpha positive signal was located in blood vessels and bone. ADAM8, ADAM12 and CD68 positive signals were located in the cell membrane and cytoplasm of all multinucleated giant cells and some round mononuclear cells in the lesions. In addition, some spindle mononuclear stromal cells were positive for ADAM12 in both lesions. CONCLUSION: Multinucleated giant cells probably originate from CD68-postive round mononuclear cells, which are recruited from monocyte-macrophage system by chemokines, such as MIP-1alpha, followed by cell fusion mediated by ADAM8 and ADAM12.

[Comparison of two methods of isolation and culture of osteoclasts and the dynamic observation of bone resorption].

OBJECTIVE: To study the origin, morphological structure, and functional regulation of osteoclast (OC) for further investigation on the mechanism and regulation of bone resorption. METHODS: The OCs were isolated by two kinds of traditional method. Osteoclasts were isoclated from neonatal rat long bones. The cytochemistry was observed. The osteoclast-like cells (OLC) were derived from the mouse bone marrow cells in the presence of 1,25(OH)2VitD3 in vitro. RESULTS: Both morphological and functional studies showed that the isolated cells shared some of the typical characteristics of osteoclasts, that is A. multinuclearity; B. developing spreading and pseudopodial activity when cultured on glass; C. high tartrate-resistant acid phosphatase (TRAP); D. resorption lacunae could be found when the cells were cocultured with devitalized bone slices and the number was increased as the time followed. OLC had the same histological and structural traits as the OCs by the former method. The concentration of Ca(2+) and acid phosphatase (ACP) increased gradually. CONCLUSION: Different kinds of method fit different experiments. The OC obtained by the first method has more activity of bone resorption. The OLC by the second method has more in quantity and can be used in the study of cell differentiation.

Ameloblastic fibroma and related lesions: a clinicopathologic study with reference to their nature and interrelationship.

BACKGROUND: Ameloblastic fibroma (AF) and related lesions constitute a group of lesions, which range in biologic behavior from true neoplasms to hamartomas. The aim of this study was to elucidate the nature and interrelationship of this group of lesions. METHODS: Clinical and pathological studies were undertaken retrospectively on 13 cases of AF and seven cases of ameloblastic fibro-odontoma (AFO). Thirty-three complex odontomas and 33 compound odontomas were also included for comparative purpose. Relevant follow-up data were recorded and the literature was reviewed. RESULTS: The majority of patients with AF (nine cases, 69.2%) were over the age of 22 years with frequent involvement (76.9%) of the posterior mandible. Tumors recurred in four of 11 patients with follow-up information and two recurrent tumors showed malignant transformation. There was no case in this series that could be designated as the so-called ameloblastic fibrodentinoma, apart from one recurrent AF in which further maturation to form only tubular dentin materials was identified. AFO tended to occur at a younger age group with an average of 9.6 years. Recurrence was noted in two of five patients with follow-up data and both recurrent lesions showed limited growth potential and further maturation into a complex odontoma. Significant differences were noted in the age and site distribution between the complex and the compound odontomas. CONCLUSION: Whilst the majority, if not all, of AFs are true neoplasms with a potential to recur and/or of malignant transformation, some, especially those occurred during childhood, could represent the primitive stage of a developing odontoma. Our data also suggests that some AFOs are hamartomatous in nature, representing a stage preceding the complex odontoma.

[Expression of receptor activator of NF-kappa B ligand and osteoprotegerin protein in the giant cell lesions of jaw].

OBJECTIVE: To detect the expression of RANKL and OPG protein in the giant cell lesions of jaw and to study the mechanism of this lesion. METHODS: RANKL and OPG were detected by immunohistochemistry (SP) in 24 paraffin-embedded and 2 frozen specimens of central giant cell lesion of jaw. RESULTS: RANKL signals were strongly positive in the vascular epithelial cells. They also could be found in fibrous stroma, bone matrix, and stromal spindle cells, even in some cytomembrane of multinucleated giant cells. OPG was detected in multinucleated giant cells and a fraction of round mononuclear cells. CONCLUSIONS: Active vascular epithelial cells are contributed to the formation of multinucleated giant cells through regulating RANKL, and RANKL could play its role by paracrine and autocrine, which might be inhibited by OPG.

Characteristics of benign lymphoadenosis of oral mucosa.

AIM: To investigate the pathological characteristics and carcinogenesis mechanism of benign lymphoadenosis of oral mucosa (BLOM). METHODS: The expressions of Ki-67, CD34 and apoptosis were evaluated by immunohistochemical SP staining in 64 paraffin-embedded tissue samples. Of them, 9 were from BLOM with dysplasia, 15 from BLOM without dysplasia, 15 from oral squamous cell carcinoma (OSCC), 15 from oral precancerosis, and 10 from normal tissues. Cell proliferation, apoptosis and angiogenesis of tissue samples were also analyzed. RESULTS: The expression of Ki-67 in BLOM with dysplasia, oral precancerosis and OSCC was significantly higher than in BLOM without dysplasia and normal mucosa. The microvascular density (MVD) in BLOM with and without dysplasia, oral precancerosis, and OSCC was significantly higher than in normal mucosa. Apoptosis in BLOM and oral precancerosis was significantly higher than in OSCC and normal mucosa. CONCLUSION: Benign lymphoadenosis of oral mucosa has potentialities of cancerization.

[Effect of genistein on the proliferation, differentiation and apoptosis of the osteoblasts].

OBJECTIVE: To study the effect of genistein on the proliferation, differentiation and apoptosis of the osteoblasts in vitro. METHODS: The primary osteoblasts (OBs) were obtained from the rat calvaria and the cell line of osteosarcoma-UMR-106 served as control. The cells in the experiment group were grown in 10% (volume fraction) fetal calf serum (FCS) + alphaMEM + various concentrations of genistein. The control groups were grown in 10% FCS + alphaMEM. The growth of OBs was assessed by flow cytometry and MTT method. The differentiation of OBs was examined by the alkaline phosphatase (ALP) activity. RESULTS: Flow cytometry analysis and MTT showed that genistein could prompt primary OB from stage G(0) (G(1)) to stage S, G(2) or M. By contrast, genistein had no effect on the cell cycle of UMR-106, but could induce its apoptosis. Additionally, the results of ALP activity showed that genistein stimulated the differentiation of primary OB. CONCLUSIONS: Genistein can stimulate the proliferation and differentiation of the primary osteoblasts in some degree, and induce the apoptosis of osteosarcoma cells.

[Central giant cell lesions of the jaws: a clinicopathological study of 31 cases].

OBJECTIVE: To study the clinicopathologic features of central giant cell granuloma (CGCG) of the jaws and the relationship between the pathologic features and its clinical behavior. METHODS: Histologic, radiographic and follow-up information for 31 cases of central giant cell granuloma were reviewed. The histopathologic patterns were analyzed between nonrecurrent and recurrent cases for which the following-up information was available. RESULTS: The majority of the giant cell granuloma of the jaws occurred in patients under 30 with a predilection of females and mostly were involved in the mandible. The radiographic features of CGCG non-specific. The multinucleated giant cell scattered unevenly, the numbers of the nuclei were few and mostly 10-19. The marked fibrosis, the multiple area of hemorrhage, abundant hemosiderin and newly formed bone were always present in the lesions. No significant difference exited between the recurrence and nonrecurrence groups in the pathologic features. The patients with aggressive behavior showed more consistent with the recurrence. CONCLUSIONS: CGCG was a non-neoplastic lesion of the jaws which was different from the giant cell tumor. It was difficult to distinguish between the CGCG and giant cell tumor (GCT), and to predict its clinical behavior only by the histopathological patterns. It was helpful to combine the clinical presentation of CGCG with its treatment.

[Genistein inhibits the promotive effect of IL-1beta on osteoclastic bone resorption].

OBJECTIVE: To investigate the effect of genistein, a soybean-derived isoflavone, on the stimulating effect on bone resorption of IL-1beta. METHODS: The osteoclasts (OCs) were isolated with the methods of Yu Shifeng. The rat calvaria were cultured as an organ. The cells in the experiment were grown in four respectively: Control (without genistein or IL-1beta), 10(-6) mol/L genistein,10 microg/L IL-1beta and 10(-6)mol/L genisteine + 10 microg/L IL-1beta. The area of bone resorption, the concentration of Ca(2+) in the supernatant liquid of OCs cultures and mice calvaria were tested. The contents of acid phosphatase(ACP) were also examined by biochemistry method. The index of bone resorption was counted as the ratio of the experiment average and control ones, which indicated the increase in bone resorption when it was above 1.0. RESULTS: The area of bone resorption of 10(-6) mol/L genisteine + 10 microg//L IL-1beta increased compared with that of 10(-6) mol/L genistein, while the concentration of Ca(2+) in the supernatant liquid of OCs cultures decreased significantly. The index of bone resorption of 10(-6) mol/L genisteine + 10 microg//L IL-1beta lied between 10(-6) mol/L genistein and 10 mug/L IL-1beta. In the organ culture, there was no difference in the content of ACP among all the groups. The index of bone resorption of 10(-6) mol/L genisteine + 10 microg//L IL-1beta was below that of 10 mug/L IL-1beta, but both were above 1.0. The index of bone resorption was below 1.0 in the group of 10(-6) mol/L genistein. CONCLUSION: Genistein can suppress obviously the bone resorption simulated by single IL-1beta.

[Immunohistochemical study of benign lymphoadenosis of oral mucosa].

OBJECTIVE: To study the expression of Ki-67 and the changes of MVD and apoptosis in benign lymphoadenosis of oral mucosa (BLOM). METHODS: The expression of Ki-67, CD34 and apoptosis were evaluated by SP immunohistochemical staining in 15 BLOM, 9 BLOM with dysplasia, 15 oral squamous cell carcinoma (OSCC). RESULTS: The expression of Ki-67 in BOLM with dysplasia and OSCC was significantly higher than that of BLOM without dysplasia and normal oral mucosa (P < 0.05). The MVD in all BLOM and OSCC was significantly higher than that in normal mucosa (P < 0.05). Apoptosis in BLOM was higher than in normal mucosa and OSCC (P < 0.05). CONCLUSIONS: The expression of Ki-67 and MVD in BLOM with dysplasia were between normal oral mucosa and oral carcinoma. The occurrence of apoptosis in BLOM was significantly higher than in normal oral mucosa. The results suggest that BLOM had the potentiality of malignant transformation.

[Expression of a disintegrin-like and metalloproteinase protein 8 and 12 in the giant cell lesions of jaw].

OBJECTIVE: To detect the expression of a disintegrin-like and metalloproteinase (ADAM) 8 and 12 gene in the giant cell lesions of jaw and to study their effects on the histogenesis of cells in these lesions. METHODS: ADAM8 and ADAM12 was detected by immunohistochemistry (SP) in 40 paraffin-embedded specimens of central giant cell lesions of jaw, 10 peripheral giant cell lesions, 9 cherubisms, 6 aneurysmal bone cysts. RESULTS: ADAM8 and ADAM12 were positive in the cytomembrane and cytoplasm of all multinucleated giant cells and some round mononuclear cells of the lesions; ADAM12 was positive for some spindle mononuclear stromal cells in central and peripheral giant cell lesions. CONCLUSIONS: Multinucleated giant cells probably originated from the fusion of the round mononuclear cells, and ADAM8 and ADAM12 were involved in this process. In addition, ADAM12 might play a role in the maturation of spindle mononuclear stromal cells.

[Clinicopathologic study of a family cherubism].

OBJECTIVE: To investigate the clinicopathologic features of familial cherubism and its differentiation from other giant cell lesions in jaws and the results of surgical treatments with a long-term follow-up. METHODS: Four cases of familial cherubism were reviewed and their clinical and radiographic features, histopathologic appearance, biochemical markers and surgical treatments analysed. RESULTS: Clinically, cherubism was characterized by bilateral painless swelling of jaws, mandibular deformity was common. Radiographs showed multilocular radiolucencies with sclerotic thickening border. Histopathologically, numerous randomly distributed multinucleated giant cells and vascular spaces within a fibrous connective tissue stroma with or without eosinophilic collagen perivascular cuffing were shown. The lesion regressed without treatment in 1 cases. Curettage was performed in 3 cases with good results. CONCLUSIONS: Cherubism can be diagnosed according to its typical clinical and radiographical features with a positive family history. It might regress without treatment. But surgery intervention is suggested to improve physiological function and to solve the psychologic problem of the patients.

[The beneficial effect of genistein on mandible bone metabolism in ovariectomized rats].

OBJECTIVE: To detect the effect of genistein on mandible metabolism in ovariectomized rats. METHODS: Forty 12 week-old female SD rats were randomly assigned to four groups: (1) sham operated; (2) ovariectomized; (3) ovariectomized and treated with estradiol; (4) ovariectomized and received genistein, 45 mg/kg body weight per day. After 12 weeks, bone mineral density (BMD), serum level of alkaline phosphatase (ALP), acid phosphatase (ACP), osteocalcin, IL-1beta, TNF-alpha, IL-6 and calcitonin (CT) were evaluated. In addition, the serum estradiol and the weight of uteri were also examined to indicate the side effect of genistein to the uteri. RESULTS: Ovariectomized animals had a significant decrease in BMD, and increased serum level of ALP, ACP, IL-1beta and osteocalcin compared with sham rats. After treated with genistein, BMD and the serum level of ALP, ACP, osteocalcin increased significantly, while the serum level of IL-1beta and TNF-alpha decreased. Especially, the increase of ALP and osteocalcin was higher than that of estradiol-treated animal. Additionally, the uterus weight index and the serum estradiol in genistein-treated rats were lower significantly than those of estradiol-treated rats. CONCLUSIONS: Genistein can improve the mandible bone metabolism as well as its effect on femur through the promotion of bone formation and the prevention of bone resorption with slight side effect. Genistein provides an additional viable way to therapy for osteoporosis in the jaw bones.

Giant cell granuloma of the temporal bone: a case report with immunohistochemical, enzyme histochemical, and in vitro studies.

A case of giant cell granuloma (GCG) that occurred in the right temporal bone is reported. The lesion showed histologic features identical to GCG. The multinuclear giant cells (MGCs) in the lesion showed strong reactivity with CD68, but patchy staining for myeloid/histiocyte antigen, alpha-1-antitrypsin, alpha-1-antichymotrypsine, and lysozyme. Activity of tartrate-resistant acid phosphatase was also consistently detected in the MGCs. Some of the mononuclear cells of the lesion exhibited similar immunocytochemical and histochemical reactivity as the MGCs. Ki-67 staining, however, was only detected in the mononuclear cells. The MGCs isolated from the lesion presented characteristic morphology of osteoclasts and possessed the ability to excavate bone in vitro. Thus, the MGCs in GCG appeared to express both macrophage- and osteoclast-associated phenotypes. The mononuclear cells were the major proliferative elements in the lesion and a subpopulation of these cells may represent precursors of the MGCs.