Neoplastic odontogenic epithelial cells express bone sialoprotein

Bone sialoprotein (BSP) is synthesized and secreted by bone-, dentine- and cementum-forming cells and has been implicated in de novo bone formation and mineralization. In this study, we used histological sections of odontogenic neoplasms and performed immunohi stochemical and in situ hybridization analyses. In ameloblastoma, BSP mRNA signals were seen in the neoplastic epithelial cells forming nests, strips and islands. BSP deposition was also seen in the stellate reticulum of the tumour masses revealed by immunohistochemistry using human BSP antibodies. In calcifying epithelial odontogenic tumour, the calcified masses demonstrated positive immunoreactivity to the human BSP antibodies, and the hybridization signals for BSP were located in the cells near the calcified particles. In the calcifying odontogenic cyst, strong BSP signals were seen in cells surrounding the characteristic nests of ghost cells, which often calcify subsequently. BSP protein was also found in these cells by immunohistochemistry. The active expression of BSP in the epithelial elements of the odontogenic tumours of adult patients suggests the activation of this matrix protein gene in the neoplastic process, and that BSP may play an important role in tumour formation and differentiation with respect to pathological calcification. © Chapman & Hall     

Botryoid odontogenic cyst. Exploration of proliferative activity,apoptosis and expression of TP53 and BCL2 compared to the histologically identical lateral periodontal and gingival cysts

The botryoid odontogenic cyst (BOC) is a rare, locally more aggressive variant of the usually indolent lateral periodontal cyst (LPC) and gingival cyst (GC). A recent case of BOC provided an opportunity for an exploratory study on the causes of its more aggressive behavior. The limited objective was to see if the BOC was sufficiently different from the other cysts to warrant an investment in a large study. Sections of neutral buffered formalin fixed, paraffin-embedded tissues from the BOC and archival specimens of four GCs, four LPCs and three odontogenic keratocysts (OKCs) were stained using immunohistochemistry for Ki-67, a marker of proliferating cells, caspase-3, a marker of cells undergoing apoptosis, tumor suppressor p53, and the apoptosis inhibitor BCL2. The mean labeling index (LI) of immunoreactive cyst epithelial cells was computed for each antibody and type of cyst. Compared to the LPCs and GCs, the BOC exhibited a moderately larger Ki-67/caspase-3 LI difference, which indicates that the BOC had a net higher rate of growth. We found a much higher level of LI, therefore likely dysregulation of p53. We also found a much higher LI of BCL2. The LIs of p53 and BCL2 in the BOC were similar to and more than twice that of the OKCs, respectively. Although meaningful statistical analysis was precluded by our use of only one case of BOC and a small number of the other cysts, the high p53 and very high BCL2 labeling indices of the BOC offer a potential explanation for its reportedly more aggressive behavior that clearly is worthy of further investigation.     

Peripheral calcifying cystic odontogenic tumour of the maxillary gingiva

ABSTRACT: BACKGROUND: Odontogenic tumors are lesions that are derived from remnants of the components of the developing tooth germ. The calcifying cystic odontogenic tumor or calcifying odontogenic cyst is a benign cystic neoplasm of odontogenic origin that is characterized by an ameloblastoma-like epithelium. Calcifying cystic odontogenic tumor may be centrally or peripherally located, and its ghost cells may exhibit calcification, as first described by Gorlin in 1962. Most peripheral calcifying cystic odontogenic tumors are located in the anterior gingiva of the mandible or maxilla. CASE PRESENTATION: Authors report a rare case of a peripheral calcifying cystic odontogenic tumor of the maxillary gingiva. A 39-year-old male patient presented with a fibrous mass on the attached buccal gingiva of the upper left cuspid teeth. It was 0.7-cm-diameter, painless and it was clinically diagnosed as a peripheral ossifying fibroma. After an excisional biopsy, the diagnosis was peripheric calcifying cystic odontogenic tumor. The patient was monitored for five years following the excision, and no recurrence was detected. CONCLUSIONS: All biopsy material must be sent for histological examination. If the histological examination of gingival lesions with innocuous appearance is not performed, the frequency of peripheral calcifying cystic odontogenic tumor and other peripheral odontogenic tumors may be underestimated.     

A mathematical model of the dynamics of odontogenic cyst growth

OBJECTIVE: To formulate a mathematical model of odontogenic cyst growth and establish the dynamics of cyst enlargement and role of osmotic pressure forces throughout its growth. STUDY DESIGN: The model assumed a spherical cyst with a semipermeable lining of living cells and a core consisting of degraded cellular material, including generic osmotic material, fed by the continuous death of epithelial cells in the lining. The lining cells were assumed to have both elastic and viscous properties, reflecting the action of physical stresses by the surrounding cyst capsule, composed of fibroblasts and collagen fibers. The model couples the cyst radius and osmotic pressure differences resulting in a system of 2 nonlinear ordinary differential equations. RESULTS: The model predicts that in all parameter regimens the long-time behavior of the cyst is the same and that linear radial expansion results. CONCLUSION: In the early and intermediate stages of cystic growth, osmotic pressure differences play an important role; however, in very large cysts, this role becomes negligible, and cell birth in the lining dominates growth.     

Ghost cells in calcifying odontogenic cyst express enamel-related proteins

The so-called ghost cell is a unique cell type occurring in a variety of odontogenic and non-odontogenic lesions. However, the true nature of ghost cells has not been determined. In the present study, we examined the immunoreactivity of ghost cells in calcifying odontogenic cysts and dermal calcifying epitheliomas, with antibodies against amelogenin, enamelin, sheath protein (sheathlin) and enamelysin, in an attempt to clarify the nature of this unique cell. The cytoplasm of ghost cells in calcifying odontogenic cysts demonstrated distinct immunolocalization of the enamel-related proteins, while similar in the calcifying epitheliomas of the skin showed a negative reaction. The results indicate that the ghost cells in calcifying odontogenic cysts, as opposed to ghost cells in dermal calcifying epitheliomas, contain enamel-related proteins in their cytoplasm accumulated during the process of pathological transformation.     

Calcifying odontogenic cyst--Gorlin's cyst--report of two cases

The authors present two cases of calcifying odontogenic cysts, which were confirmed by histological examination. In the first case the radiographic findings and clinical status did not indicate the presence of a calcifying odontogenic cyst. In the second case, differential diagnosis included COC. The histopathological findings showed that what appeared to be simple cases of bone translucencies was in fact an unusual odontogenic lesion. The two cases point out the possibility of incorrect assessment of deceptively banal cases in daily specialist practice.     

Submicroscopic localization of acid phosphatase in the dental cyst epithelium

Summary Distribution of acid phosphatase was studied in the dental cyst epithelium. Enzyme activity was found to be localized in primary and secondary lysosomes and vacuoles of the Golgi complex. Longer incubation revealed positive reaction in the Golgi complex vesicles and cisternae, in the endoplasmic reticulum, and perinuclear' space. Acid phosphatase reaction was bound also to membranous structures occurring in the intercellular spaces. These structures originated probably from disintegrated cells. Localization of acid phosphatase reaction in the dental cyst epithelium was found to be essentially the same as in the oral cavity epithelium. Smaller differences between both types of epithelium may be conditioned by different type and arrangement of the organelles in the corresponding epithelial layers.     

Localization of activated caspase-3-positive and apoptotic cells in the developing tooth germ of the mouse lower first molar

This study examined the immunohistochemical detection of activated caspase-3, and its association with apoptosis, during tooth morphogenesis of the mouse lower first molar. The distribution of cells positive for caspase-3 closely corresponded with the localization of the terminal deoxynucleotidyl transferase-mediated deoxyuridine-5-triphosphate-biotin nick end labelling (TUNEL)-positive apoptotic cells through the developmental course of tooth germs from embryo day 12 (E12) to E19, thus showing that the apoptosis occurring in the developing odontogenic tissue was induced by the activation of the caspase family. The specific distribution pattern of apoptotic cells in the developing odontogenic epithelial tissue from the initiation (E12) of tooth germ to the completion of tooth crown morphology (E19) also suggests that apoptotic events are related not only to a deletion of functionally suspended cells, but also participate in initiation and the completion of tooth morphogenesis. Electron microscopic examination revealed that apoptotic cells were present in the primary enamel knot, and these apoptotic cells were phagocytized by neighbouring odontogenic epithelial cells, thus indicating the prompt disposal of any dead cells by epithelial cells.     

Microstructural Reconstruction for the Testicular Cysts of Wolf Spider Using 3D Volume Rendering Technique

In insects, the alignment of neighboring spermatid in the late stages is nearly perfect, so that a transverse section of a cyst containing late spermatids transects all the spermatids at approximately the same level. However, the testicular cysts of spiders are spherical, most cysts are arranged in order of increasing maturity from the periphery to the center of the testis. For this reason, it is difficult to observe the whole spermatids within a single microscopic slide and count them. Therefore, we demonstrate microstructural reconstruction technique enabling to count exact number of sperm cells per cyst with aid of 3D volume rendering. For image processing and reconstruction, serially sectioned histologic specimens were scanned with microscopy and 3D images were reconstructed using Amira 5.3.2 software from the image stacks of the germ cells and surrounding testicular cysts subsequentially. With the information gathered by 3D reconstruction, it has finally been counted that exactly 32 (2⁵) cells of the secondary spermatocytes per cyst. This means that most cysts in P. laura contain exactly 64 (2⁶) spermatids or spermatozoa, which presumably arose from four synchronous mitotic and two meiotic divisions. In addition, the number of divisions occurring in a cyst appears to be constant for this spider because it has been known that the number of spermatids per cyst is characteristic for each species.     

Ependymal cyst of the subarachnoid space. Cytologic diagnosis and developmental considerations

Fluid aspirated from a subarachnoid cystic lesion that covered and compressed part of the left frontal lobe was examined cytologically and compared with histologic sections of the cyst wall. The fluid contained epithelial and histiocytelike cell populations. The epithelial cells were tall columnar, occurring singly or in clusters or sheets. Many cells were ciliated and their cytoplasm showed characteristic refractile granules. The differential diagnosis of this rare type of subarachnoid cyst and the mechanism of the development are discussed. Cytologic evaluation of the fluid of the subarachnoid cysts is potentially a more accurate method of classification of these lesions than is random biopsy of the cyst wall. It is of particular importance in cases with a history of growth, in which the progressive expansion results in attenuation of the diagnostic epithelial lining of the cyst.     

Fine needle aspiration cytology of bursal cyst

OBJECTIVE: To evaluate the cytomorphology of bursal cyst and assess the efficacy of aspiration cytology in its diagnosis and treatment. STUDY DESIGN: Nineteen cases of bursal cyst seen over four years were studied. Material was obtained by fine needle aspiration. The smears were stained with May-Grünwald-Giemsa stain and hematoxylin and eosin. RESULTS: Eight cysts were in the popliteal fossa, 4 on the elbow, 3 on the knee, 2 on the shoulder and 2 in the calf. Gelatinous material was aspirated in all the cases. In some cases the cyst collapsed after aspiration. The key diagnostic features were hypocellular smears in a mucoid background. Histiocytelike (synovial) cells were seen lying in all cases and as pseudopapillary structures in two. CONCLUSION: The presence of a cyst at a classic location with aspiration of gelatinous material and the presence of singly occurring histiocytelike cells in a mucoid background in smears is diagnostic of bursal cyst. The procedure is therapeutic in some cases.     

The orosomucoid 1 protein (alpha1 acid glycoprotein) is overexpressed in odontogenic myxoma

ABSTRACT: BACKGROUND: Odontogenic myxoma (OM) is a benign, but locally invasive, neoplasm occurring in the jaws. However, the molecules implicated in its development are unknown. OM as well as Dental Follicle (DF), an odontogenic tissue surrounding the enamel organ, is derived from ectomesenchymal/mesencyhmal elements. To identify some protein that could participate in the development of this neoplasm, total proteins from OM were separated by two-dimensional electrophoresis and the profiles were compared with those obtained from DF, used as a control. RESULTS: We identified eight proteins with differential expression; two of them were downregulated and six upregulated in OM. A spot consistently overexpressed in odontogenic myxoma, with a molecular weight of 44-kDa and a pI of 3.5 was identified as the orosomucoid 1 protein. Western blot experiments confirmed the overexpression of this protein in odontogenic myxoma and immunohistochemical assays showed that this protein was mainly located in the cytoplasm of stellate and spindle-shaped cells of this neoplasm. CONCLUSION: Orosomucoid 1, which belongs to a group of acute-phase proteins, may play a role in the modulation of the immune system and possibly it influences the development of OM.     

Intercellular bridges and factors determining their patterns in the grasshopper testis

Intercellular bridges joining cells contained in cysts of Chortophaga viridifasciata testes were studied with light and electron microscopy. Preparations consisted of expressed whole cells (living, or fixed and stained) as well as sections. The secondary spermatogonia of each cyst are joined centrally by persisting fused interzonal bodies (fusomes) of incompletely cleaved cells. Shifts in cell orientation during anaphase are apparently responsible for central as opposed to chain linkage of cells. In the primary spermatocytes, the central fusome is replaced by a chain linkage, apparently resulting from the breakdown of the fusome into its original interzonal body components. Intercellular bridges are also present in spermatids, but there is no evidence to indicate the time of their formation (in the immediately preceding meiotic divisions or in the secondary spermatogonial divisions). The function of the compact centrally situated fusome in the secondary spermatogonial cyst is discussed as it relates to synchrony, number of cell divisions, spermatodesm formation, and fertility.     

Immunocytochemical study of the epithelial lining of naturally occurring cysts in the rat intermediate lobe.

An immunocytochemical study of the epithelial lining of naturally occurring cysts in the rat intermediate lobe (IL) has been carried out. Paraffin-embedded sections, in which cysts were identifiable, were treated either with anti-serotonin or anti-S-100 protein sera. S-100-positive cells were intermingled with glandular cells surrounding the cyst lumen. These S-100-positive cells sent slender cytoplasmic processes as if to cover the apical surface of neighbouring cells. Rarely were 5-HT-immunopositive cells seen in the cyst epithelial lining. Most cells of the marginal layer of the IL were found reactive either to an S-100 or a-5-HT serum. The presence of an epithelial lining positive to S-100 protein sera is in keeping with the notion that cysts in the IL might form as evaginations of the epithelial lining of the pituitary cleft. The lack of correspondence between 5-HT-positive cells in the marginal layer and the cyst lining is controversial. A peculiar spatial relationship of 5-HT cells with the vascular network of the IL is suggested.     

The type, origin and function of the odontogenic cells of continuously growing guinea-pig molars

Little is known about which cell types act as the origin of odontogenic stem cells and how these stem cells differentiate to become functional. In the present study 3-day-old guinea-pigs were injected with tritiated thymidine, killed at intervals and their molars studied. The odontogenic cells were found to originate from the stem cells which by autoradiography were shown to be slow cycling and lightly labelled. As they differentiated they became heavily labelled and were designated transit 'dividing cells'. With further differentiation the cells were unlabelled and were designated 'simple transit cells'. It was concluded from the present study that outer enamel epithelium functioned as the source of all odontogenic epithelium; that primitive mesenchyme around the cervical loop functioned as the source of all odontogenic mesenchyme and that all of the transit cells which differentiated to become functional migrated coronally.     

Newt RAD51: Cloning of cDNA and analysis of gene expression during spermatogenesis

A cDNA encoding a newt homolog of Escherichia coli RecA and yeast RAD51 from a testis cDNA library was isolated. The newt RAD51 (nRAD51) cDNA predicted a 337 amino acid protein with a 95-96% amino acid identity to Xenopus and mammalian RAD51. Northern blot analysis showed that nRAD51 mRNA, 1.7 kb in length, was expressed strongly in the testis and ovary, but weakly in the liver, kidney and brain. In situ hybridization revealed that expression of nRAD51 mRNA was barely observed in primary spermatogonia (one cell in a cyst) and early secondary spermatogonia (two to four cells in a cyst), but increased in late secondary spermatogonia (> or =eight cells in a cyst), reaching a maximum level in leptotene-zygotene spermatocytes, and thereafter declined. These results suggest that nRAD51 is involved in mitotic recombination in spermatogonia as well as in meiotic recombination in spermatocytes.     

Daughterless coordinates somatic cell proliferation,differentiation and germline cyst survival during follicle formation in Drosophila

During Drosophila oogenesis two distinct stem cell populations produce either germline cysts or the somatic cells that surround each cyst and separate each formed follicle. From analyzing daughterless (da) loss-of-function, overexpression and genetic interaction phenotypes, we have identified several specific requirements for da(+) in somatic cells during follicle formation. First, da is a critical regulator of somatic cell proliferation. Also, da is required for the complete differentiation of polar and stalk cells, and elevated da levels can even drive the convergence and extension that is characteristic of interfollicular stalks. Finally, da is a genetic regulator of an early checkpoint for germline cyst progression: Loss of da function inhibits normally occurring apoptosis of germline cysts at the region 2a/2b boundary of the germarium, while da overexpression leads to postmitotic cyst degradation. Collectively, these da functions govern the abundance and diversity of somatic cells as they coordinate with germline cysts to form functional follicles.     

三维联合培养牙髓细胞和血管内皮祖细胞促进成牙本质向/成骨向分化

目的:探讨三维联合培养牙髓细胞(dental pulp cells, DPCs)和血管内皮祖细胞(endothelial progenitor cells, EPCs)对成牙本质向/成骨向分化的影响。方法:取单独培养DPCs及联合培养的DPCs和EPCs进行三维培养后成牙本质向/成骨向诱导,使用茜素红染色及半定量分析、RT-PCR和细胞免疫荧光检测成牙本质向/成骨向分化能力。采用SPSS 23.0统计软件对数据进行统计学分析。结果:茜素红染色显示联合培养组和单独培养组之间未见显著差异。RT-PCR和细胞免疫荧光显示成牙本质向/成骨向相关基因m RNAs和蛋白表达水平联合培养组显著高于单独培养组。结论:三维联合培养的DPCs和EPCs促进成牙本质向/成骨向分化,为牙髓再生提供可能实验依据。     

The orosomucoid 1 protein (α1 acid glycoprotein) is overexpressed in odontogenic myxoma

Background

Odontogenic myxoma (OM) is a benign, but locally invasive, neoplasm occurring in the jaws. However, the molecules implicated in its development are unknown. OM as well as Dental Follicle (DF), an odontogenic tissue surrounding the enamel organ, is derived from ectomesenchymal/mesencyhmal elements. To identify some protein that could participate in the development of this neoplasm, total proteins from OM were separated by two-dimensional electrophoresis and the profiles were compared with those obtained from DF, used as a control.

Results

We identified eight proteins with differential expression; two of them were downregulated and six upregulated in OM. A spot consistently overexpressed in odontogenic myxoma, with a molecular weight of 44-kDa and a pI of 3.5 was identified as the orosomucoid 1 protein. Western blot experiments confirmed the overexpression of this protein in odontogenic myxoma and immunohistochemical assays showed that this protein was mainly located in the cytoplasm of stellate and spindle-shaped cells of this neoplasm.

Conclusion

Orosomucoid 1, which belongs to a group of acute-phase proteins, may play a role in the modulation of the immune system and possibly it influences the development of OM.