颌骨巨细胞瘤和巨细胞肉芽肿病变中多核巨细胞性质的探讨

目的 探讨颌骨巨细胞瘤(Giant cell tumor，GCT)和巨细胞肉芽肿(Giant cell granuloma，GCG)病变中多核巨细胞(Multinucleated giant cells，MGCs)的性质及组织来源。方法 对8例颌骨GET和8例GCG(中心性4例、外周性4例)进行酶组织化学和免疫组织化学观察，并与其它含MGICs的口腔病变(如异物反应、淋巴结结核等)进行比较。结果 颌骨GCT和GCG中的MGCs既可表达组织细胞／单核巨噬细胞相关抗原[如CD68、αl-抗胰蛋白酶(Alpha-1-antitrypsin，AAT)、α1-抗糜蛋白酶(Alpha-1-antichymotrypsin，AACT)、溶菌酶等]，又具有破骨细胞特异性酶-抗酒石酸酸性磷酸酶[Tartrate-resistant acid phosphatase(TRAP)]的活性，但不表达增殖细胞标记Ki-67抗原。结论 颌骨GCT和GCG中的MGCs可能是反应性终末分化细胞，由病变中处于不同分化阶段的前体破骨细胞融合而成．同时具有单核巨噬细胞和破骨细胞的某些表型特征。     

Multinucleated giant cells in various forms of giant cell containing lesions of the jaws express features of osteoclasts

BACKGROUND: The nature and the mechanism involved in the formation of the multinucleated giant cells (MGCs) in various giant cell-containing lesions of the jaws are not fully understood. The aim of this study is to clarify the osteoclastic features of the MGCs in central giant cell granuloma (CGCG), peripheral giant cell granuloma (PGCG), cherubism, and aneurysmal bone cyst (ABC), and the mechanism underlying the interrelations between cellular components in the formation of the MGCs. METHODS: Immunohistochemical study with a panel of antibodies including vacuolar H+-ATPase (V-ATPase), carbonic anhydrase II (CA II), Cathepsin K, matrix metalloproteinases-9 (MMP-9), CD68, and proliferating cell nuclear antigen (PCNA), and enzyme histochemical staining for tartarate-resistant acid phosphatase (TRAP) were applied on a total number of 53 cases of giant cell-containing lesions including CGCG (n = 34), PGCG (n = 6), cherubism (n = 7), and ABC (n = 6). In situ hybridization was also carried out to detect the mRNA expression of the receptor activator of NF-kappaB ligand (RANKL), a newly identified cytokine that is shown to be essential in the osteoclastogenesis, its receptor RANK (receptor activator of NF-kappaB ligand), and its decoy receptor OPG (osteoprotegerin) in these four types of lesions. RESULTS: Immunohistochemical and enzyme histochemical studies showed that both the MGCs and a fraction of mononuclear cells in these lesions were strongly positive for TRAP, V-ATPase, CA II, Cathepsin K, MMP-9, and CD68, while the spindle-shaped mononuclear cells were positive for PCNA. The results with in situ hybridization indicated that RANKL mRNA was mainly expressed in the spindle mononuclear cells while OPG was extensively distributed in both the MGCs and the mononuclear cells. RANK mRNA was expressed in the MGCs and some round mononuclear cells. CONCLUSIONS: These results suggest that MGCs in the four types of giant cell-containing lesions of the jaws show characteristics of the osteoclast phenotype. The mononuclear stromal cells, which show TRAP positively, may be the precursors of the MGCs. RANKL, OPG, and RANK expressed in these lesions may play important roles in the formation of the MGCs. The similar characteristics and mechanisms in the differentiation of MGCs in these lesions also suggest that there might be a similar kind of pathogenesis involved in the formation of the MGCs in these lesions     

PCNA and Ki-67 immunoreactivity in multinucleated cells of giant cell fibroma and peripheral giant cell granuloma

Immunohistochemical investigation of PCNA and Ki-67, two diverse nuclear proteins essential to the cell cycle, was undertaken in archival, formalin-fixed and paraffin-embedded specimens of giant cell fibroma (GCF) and peripheral giant cell granuloma (PGCG). GCF multinucleated cell nuclei were mostly PCNA⁺, although there was variability in staining intensity. This indicates heterogeneity in nuclear PCNA metabolism of GCF multinucleated cells, and it is possible that the most intensely stained nuclei have passed through the cell cycle more recently compared to the less immunoreactive nuclei. However, the absence of Ki-67 immunoreactivity in GCF multinucleated cells, and absence of mitoses in GCF multinucleated cells, suggests that cell cycling in the absence of cytokinesis is not involved in GCF multinucleated cell formation. Alternatively, GCF multinucleated cells possibly form by fusion of mononuclear cells previously identified as fibroblasts, although this theory cannot be confirmed by the data presented in this study, and the histogenesis of GCF multinucleated cells remains unclear. In contrast, absence of either PCNA or Ki-67 immunoreactivity in PGCG multinucleated cells is consistent with an osteoclast lineage and formation from differentiated mononuclear cells.     

A comparative immunohistochemical evaluation of CD68 and TRAP protein expression in central and peripheral giant cell granulomas of the jaws

J Oral Pathol Med (2011) 40 : 334–337 Background: Giant cell granulomas of the jaws are lesions that arise either peripherally in periodontal ligament and mucoperiosteum or centrally in the bone. The aim of this study was to evaluate expression of CD68 and tartrate‐resistant acid phosphatase (TRAP) proteins in multinucleated giant cells and mononuclear cells. Methods: Formalin‐fixed and paraffin‐embedded tissue section of 20 specimens of central giant cell granuloma and 20 cases of peripheral giant cell granuloma were immunohistochemically analyzed for CD68 and TRAP proteins expression rate using Biotin‐Streptavidin method. Result: In central giant cell granuloma, more than 99% of multinucleated giant cells were positive for TRAP antibody and about 90% were positive for CD68. In mononuclear cells of this lesion, 14% of cases were positive with TRAP antibody and 8% with CD68. In peripheral giant cell granuloma, TRAP antibody was positive in 99% of giant cells and in 13% of mononuclear cells. A proportion of 97% of giant cells and 6% of mononuclear cells reacted positively with CD68. Conclusion: Immunohistochemical evidence of this study shows that giant cells and a group of mononuclear cells of stroma in both peripheral and central giant cell granuloma express TRAP antibody severely that is specific for osteoclast. Also, these cells are positive reactive to CD68, which is the macrophage marker and therefore it can be mentioned that giant cells are osteoclast, although their origins are macrophagic/monocytic or their precursors, and maybe mononuclear cells in stroma have a role in formation of giant cells.     

Touton-like giant cells in periapical granulomas

Two types of multinucleated giant cells were observed in periapical granulomas--the foreign body type and the Touton type. In the Touton type, the nuclei were near the center of the cell, surrounded by foamy cytoplasm. Both types of giant cells reacted positively to lysozyme, alpha 1-antitrypsin and alpha 1-antichymotrypsin, indicating their histiocytic origin.     

CENTRAL GIANT CELL GRANULOMA OF THE JAWS AND GIANT CELL TUMOR OF LONG BONES - AN IMMUNOHISTOCHEMICAL COMPARATIVE STUDY

Objective:

This study investigated whether some components of the extracellular matrix and CD68 expression may drive the differences between the central giant cell granuloma (CGCG) of the jaws and giant cell tumor (GCT) of long bones, which present distinct evolution and clinical behavior.

Material and Methods:

Eight cases of CGCG and 7 cases of GCT were selected and immunohistochemically analyzed to verify the pattern of expression of CD68, tenascin (Tn) and fibronectin (Fn).

Results:

A large number of the mononuclear cells and multinucleated giant cells CD68+ was observed in both of the studied lesions, indicating histiocyte/macrophage origin. Seven cases of CGCG of the jaws showed intense staining of Fn, with uniform distribution predominantly. In all 7 cases of GCT of long bones the Fn displayed intense expression, with distribution pattern varying from uniform to reticulate/fibrillar. Six cases of CGCG were intensively stained by Tn, presenting focal expression in half of specimens, and reticulate/fibrillar pattern of expression in 4 cases. All cases of GCT of the long bones presented intense expression of Tn, uniform distribution, and reticulate/fibrillar pattern of expression in four cases.

Conclusions:

The immunoexpression of CD68 in mononuclear cells and multinucleated giant cells and staining patterns of Fn and Tn were similar in both entities. These findings indicate that these proteins could not be used to explain the differences between the CGCG of the jaws and GCT of the long bones.     

Expression of receptor activator of NF‐κB ligand and osteoprotegerin in peripheral giant cell granulomas of the jaws

J Oral Pathol Med (2010) 39 : 687–689 Background: Peripheral giant cell granuloma is a tumor of the jaw characterized by the presence of multinucleated giant cells and mononuclear cells within a fibrous stroma. These lesions are considered to be of a reactive nature rather than neoplastic. Although peripheral giant cell granulomas is a well‐described clinical entity, little is known on its pathogenesis. The aim of this study was to investigate the receptor activator of NF‐κB ligand (RANKL) and osteoprotegerin (OPG) expression and immunolocalization in giant cell granulomas. Methods: RANKL and OPG protein expression was evaluated in 22 peripheral giant cell granulomas samples, by means of immunohistochemistry. Staining was evaluated semi‐quantitatively, according to the extent and intensity of the stain. Results: RANKL was expressed in all cases with a cytoplasmic staining pattern, whereas OPG expression was detected in 21 of the 22 cases examined. Active multinucleated giant cells exhibited intense immunoreactivity for both proteins. Conclusion: RANKL and OPG are expressed in peripheral giant cell granulomas of the jaw in a manner supporting the osteoclastic nature of giant cells whereas the possible osteoclastic lineage of stromal monocytes remains ambiguous.     

梅罗综合征病理分型的免疫组织化学和电镜研究

目的：进一步实梅罗综合征的病理分型的并观察其肉芽肿瘤细胞的形态特征。方法：收集２０例,用抗溶菌酶抗体免疫组织化学的方法及透射电镜对其进行标记和观察。结果：肉芽肿型可见阳性细胞灶,电镜下有多核巨细胞、类上皮细胞等肉芽肿细胞,且有超微特征。而非肉牙肿型为阴性结果。结论：进一步证实了梅罗综合征有肉芽肿型和非肉芽肿型两型病变,并认为前者肉芽肿泖而小是其区别结节病的结核病等的形态特征之一。     

Expression of regulatory apoptotic proteins in peripheral giant cell granulomas and lesions containing osteoclast-like giant cells

Peripheral giant cell granuloma consists of monononuclear cells and osteoclast-like giant cells. The proliferative ability of peripheral giant cell granuloma is restricted to the mononuclear cell compartment, whereas multinucleated giant cells lack mitotic activity. Although the proliferative compartment of peripheral giant cell granuloma has been investigated in detail, the expression and distribution of proteins regulating apoptosis is unknown. The present study demonstrates strong expression of bak and bax in the majority of giant cells. In contrast, giant cells show only weak positivity for bcl-2 and moderate positivity for bcl-x. Mononuclear cells were negative to weakly positive for bcl-x. Only scattered mononuclear cells were positive for bak, bax and bcl-2. The frequency of apoptotic nuclei detected by TUNEL-staining compared to regular nuclei was 18 times higher in giant cells than in mononuclear cells. In summary, our findings support the presumption that giant cells of bone and soft tissue tumors are reactive cell forms and not of neoplastic origin.     

解整合素样-金属蛋白酶8和12在颌骨巨细胞病变组织中表达的研究

目的 检测解整合素样-金属蛋白酶(ADAM)8和12在颌骨巨细胞病变中的表达,探讨它们在颌骨巨细胞病变细胞发生中的作用。方法 采用免疫组织化学的方法检测40例颌骨中心性巨细胞病变,10例颌骨周围性巨细胞病变,9例巨颌症,6例颌骨动脉瘤性骨囊肿石蜡组织中的ADAM8、ADAM12的基因表达。结果 颌骨中心性巨细胞病变、颌骨周围性巨细胞肉芽肿、巨颌症、颌骨动脉瘤性骨囊肿中的几乎所有多核巨细胞和部分圆形单核基质细胞均为ADAM8、ADAM12阳性;颌骨中心性和周围性巨细胞病变中部分短梭形的细胞ADAM12阳性。结论 颌骨巨细胞病变中的多核破骨样细胞可能由圆形单核基质细胞融合而成,而ADAM8、ADAM12参与了此融合过程,同时,ADAM12可能还参与了病变中梭形单核基质细胞的成熟过程。     

The spectrum of giant cells in tumours of the salivary glands: an analysis of 11 cases

In view of the different terminology for salivary gland tumours with giant cells, eleven cases were analysed by histopathology and immunocytochemistry. Four cases (three pleomorphic adenomas, one carcinosarcoma in a pleomorphic adenoma) were classified as having a foreign-body giant cell reaction, and five cases (two mucoepidermoid carcinomas, one acinic cell carcinoma, two carcinomas in pleomorphic adenomas) as having a sarcomatoid osteoclast-like giant cell reaction. In two further cases a giant cell tumour and a giant cell granuloma were associated with carcinomas in pleomorphic adenomas. All giant cells showed characteristic expression of CD68 as a typical marker for histiocytes and macrophages with their origin in mononuclear haematopoetic stem cells. There was no evidence for an epithelial origin of the giant cells because all those examined had a negative reaction to cytokeratin. Foreign-body cells were characterized by cytoplasmic vacuoles and irregularly dispersed nuclei. They showed a focally circumscribed reaction mostly outside the connective tissue pseudocapsule of the tumours. The sarcomatoid osteoclast-like giant cell reactions in carcinomas were distinctly intermingled with the carcinomatous patterns. In contrast, the associated osteoclast-like giant cell tumour was distinctly separate from the salivary gland tumour tissue and was composed of numerous larger osteoclast-like giant cells with a greater number of nuclei (more than 20): these giant cells were uniformly distributed throughout the tumour tissue. The giant cell granuloma was also separate from the carcinoma and was composed of nests of smaller, more irregularly distributed giant cells.     

Characterisation of the inflammatory cell infiltrate in chronic hyperplastic candidosis of the oral mucosa

The inflammatory cell infiltrate in biopsy material of chronic hyperplastic candidosis (CHC) from the oral mucosa was characterised using immunocytochemical techniques. Nine specimens were stained for human kappa and lambda immunoglobulin light chains. CD68 antigen (macrophages), lysozyme (macrophages, granulocytes), CD3 antigen (T-lymphocytes), CD20 antigen (B-lymphocytes) and leucocyte common antigen (LCA). In addition, these and a further 13 specimens were also examined for immunoglobulin (Ig)-containing cells (IgA, IgG and IgM). The density of the infiltrate varied considerably between cases: T-lymphocytes were the dominant cell type (153.9%), with fewer B-lymphocytes (8.2%) and macrophages (14.2%). Many Ig-containing cells were seen, and although IgG-containing cells predominated, (60.8%, SD ±9.0) there was a high proportion of IgA-containing cells (36.7%, SD ± 9.1) with few IgM-containing cells (2.5%, SD ±3.0). Many neutrophils, together with smaller numbers of T-lymphocytes and macrophages were seen in the epithelium. It is suggested that mucosal defence to Candida infection involves a cell-mediated reaction in which there is recruitment of macrophages and local production of immunoglobulin with a prominent IgA component.     

FASN expression,angiogenesis and lymphangiogenesis in central and peripheral giant cell lesions

Central giant cell lesion (CGCL) and peripheral giant cell lesion (PGCL) are non-neoplastic proliferative processes of the jaws. PGCL is a reactive process induced by irritant local factors and CGCL is an intra-osseous lesion of unknown etiology. Both lesions exhibit similar histologic features showing abundant mononuclear cells, admixed with a large number of multinucleated giant cells and a rich vascularized stroma with extravasated erythrocytes, hemosiderin deposition, and blood-filled pools. Recent studies have linked fatty acid synthase (FASN) with angiogenesis.

Objective

To evaluate angiogenesis and lymphangiogenesis and their relationship with FASN expression in CGCL and PGCL.

Material and Methods

Thirteen CGCL and 14 PGCL of the jaws were selected for immunoexpression of FASN; CD34 and CD105 (to assess blood microvessel density [MVD] and microvessel area [MVA]); and D2-40 (to assess lymphatic MVD and MVA).

Results

Within PGCL and CGCL, MVD-CD34 was signifcantly higher than MVD-CD10S, followed by MVD-D2-40. Moreover, a signifcantly higher number of FASN-positive multinucleated giant cells than mononuclear cells were observed. Between PGCL and CGCL, only MVD-CD34 and all MVA were signifcantly higher in PGCL. Positive correlation between MVA-CD10S with FASNpositive mononuclear cells in both lesions was observed.

Conclusions

Our results show both lesions exhibiting similar levels of FASN expression and neoangiogenesis, suggesting constitutive processes that regulate tissue maintenance.     

破骨细胞分化因子和骨保护因子在颌骨中心性巨细胞病变组织中的表达

目的 检测破骨细胞核因子κB受体活化因子配体(RANKL，又称破骨细胞分化因子)和骨保护因子(OPG)蛋白在颌骨中心性巨细胞病变中的表达，探讨此类病变发生骨破坏的作用机制。方法采用免疫组织化学的方法检测26例颌骨中心性巨细胞病变中RANKL和OPG蛋白的表达。结果 RANKL在颌骨中心性巨细胞病变中的血管、基质中有强烈表达，某些病变中的部分短梭形单核基质细胞和多核巨细胞的胞膜也为RANKL阳性；病变中大部分的多核巨细胞和少部分圆形单核基质细胞的胞质OPG表达为阳性。结论 激活的血管内皮细胞通过调节RANKL的表达来促进颌骨中心性巨细胞病变中的多核破骨样巨细胞的形成；RANKL可以通过旁分泌和自分泌的方式发挥作用。同时在颌骨中心性巨细胞病变中存在由OPG介导的抑制多核破骨样巨细胞生成和骨吸收的负反馈机制。     

Myofibroblasts in peripheral giant cell granuloma. Light and electron microscopic study

Myofibroblasts are found in various pathologic conditions including lesions containing giant cells, such as central giant cell granuloma and the giant cell tumor of tendon sheath (nodular tendosynovitis). In the present study, myofibroblasts were found in peripheral giant cell granuloma (PGCG) using the TP-Levanol fast cyanine 5RN histochemical procedure and electron microscopy. A number of myofibroblasts displayed intracellular collagen fibrils. The presence of myofibroblasts supports the reactive nature of PGCG.     

Oral granular cell myoblastoma: an immunohistochemical study

Immunoperoxidase staining for a variety of antigens was performed on 15 cases of granula cell myoblastoma occurring within the oral tissues. All tumours gave intense intracytoplasmic granular staining for a CEA-related antigen with one rabbit antiserum to CEA (Dakopatts) whereas all were negative with a sheep antiserum to CEA (Seward). None of the tumours were positive for lysozyme, alpha 1 antitrypsin to cathepsin B whereas 12 lesions gave a granular reaction for cathepsin D. All granular cells showed a weak reaction for actin in contrast to the strong cytoplasmic staining found in smooth striated muscle and muscle derived tumours.     

Immunohistochemical and ultrastructural investigation of apoptotic cell death in granular cell ameloblastoma

Apoptotic cell death in granular cell ameloblastomas was examined by immunohistochemistry using anti-single-stranded DNA (ssDNA) antibody and transmission electron microscopy. Routinely prepared sections of granular cell ameloblastomas showed various quantities of granular cells with some apoptotic nuclear fragments. Immunoreactivity for ssDNA was higher in granular cells than in other neoplastic cells. Ultrastructural examination revealed abundant lysosomes in the cytoplasm of granular cells. Numerous apoptotic cell fragments with condensed nuclei in granular cell clusters were phagocytosed by adjacent granular cells. On immunohistochemical characterization of cellular differentiation, granular cells were positive for cytokeratin, CD68, lysozyme and alpha-1-antichymotrypsin, but negative for vimentin, desmin, S-100 protein, neuron-specific enolase and CD15, indicating epithelial origin and lysosomal aggregation. These features suggest that the cytoplasmic granularity in granular cell ameloblastomas might be caused by increased apoptotic cell death of neoplastic cells and associated phagocytosis by neighboring neoplastic cells.     

The Pathogenesis of Implant‐Related Reactive Lesions: A Clinical,Histologic and Polarized Light Microscopy Study

Background: Peri‐implant soft tissue reactive lesions (I‐RLs) may jeopardize implant success and survival. To the best of the authors’ knowledge, its pathogenesis is unknown. The objective of this study is to conduct a clinicopathologic and polarized light microscopy (PLM) analysis of 14 new I‐RLs and compare them with comparable tooth‐associated cases (T‐RLs) to better understand I‐RL pathogenesis. Methods: Fifty‐eight new cases of I‐RL and T‐RL were retrieved from the pathology department archives of Rambam Health Care Campus, Haifa, Israel. Retrospective analysis of histopathologic and clinical features was conducted, documented, and then compared for: 1) I‐RL (n = 14), 2) peri‐implant pyogenic granuloma (I‐PG) (n = 5), 3) peri‐implant peripheral giant cell granuloma (I‐PGCG) (n = 9), 4) T‐RL (n = 44), 5) tooth‐associated pyogenic granuloma (T‐PG) (n = 21), and 6) tooth‐associated peripheral giant cell granuloma (T‐PGCG) (n = 23). Presence of foreign bodies was assessed using PLM. Results: Foreign bodies were found more commonly in I‐RLs (n = 13/14; 93%) when compared with T‐RLs (n = 18/44; 41%), which was a statistically significant difference (P = 0.01) with an odds ratio of 7.9. Microscopically, I‐PGCG was associated with: 1) lower multinucleated giant cell count (P = 0.04); 2) lower density of mesenchymal cells (P = 0.05); and 3) more diffuse, non‐lobulated stromal morphology (P = 0.001). Clinically, I‐RLs were found in patients who were older, and all cases were located in the posterior region: mandible (n = 12/14; 86%) and maxilla (n = 2/14; 14%). Conclusions: In cases of implant failure, implantation of foreign bodies may play a role with subsequent development of I‐PG and I‐PGCG‐like lesions. Clinicians should be aware of this risk so they can implement measures to minimize adverse implant outcomes.     

原位杂交与免疫组化检测c-Src在家族性巨颌症的表达

目的:探讨c-Src基因在家族性巨颌症中的表达及意义。方法:应用原位杂交方法与免疫组织化学技术检测12例家族性巨颌症病变组织中c-Src蛋白与c-Src mRNA的表达。结果:12例家族性巨颌症病变组织中的多核巨细胞、10%~15%的单核圆形基质细胞胞浆表达c-Src蛋白与mRNA,二者阳性细胞表达率无显著性差别,c-Src mRNA表达结果与c-Src蛋白相一致,二者呈正相关关系。结论:c-Src表达可能与家族性巨颌症病变中多核巨细胞的形成和破骨特性有关。     

Peripheral giant cell granuloma. A report of five cases and review of the literature

Peripheral giant cell granuloma (PGCG) is a relatively frequent benign reactive lesion of the oral cavity, originating from the periosteum or periodontal membrane following local irritation or chronic trauma. PGCG manifests as a red-purple nodule located in the region of the gums or edentulous alveolar margins, fundamentally in the lower jaw. The lesion can develop at any age, though it is more common between the fifth and sixth decades of life, and shows a slight female predilection. PGCG is a soft tissue lesion that very rarely affects the underlying bone, though the latter may suffer superficial erosion. The present study reviews 5 cases of PGCG, involving 3 males and 2 females between 19-66 years of age, and with presentation in the upper jaw in three cases. Two patients showed radiological concave depression images corresponding to bone resorption. Treatment consisted of resection and biopsy, using a carbon dioxide laser in 2 cases and a cold scalpel in the remaining 3. There were no relapses during postoperative follow-up (range 10 months to 4 years). The differential diagnosis of PGCG includes lesions with very similar clinical and histological characteristics, such as central giant cell granuloma, which are located within the jaw itself and exhibit a more aggressive behavior. Only radiological evaluation can establish a distinction. The early and precise diagnosis of these lesions allows conservative management without risk to the adjacent teeth or bone.