Hydrogen peroxide (H2O2) is originally an endogenous small molecule which is reduced into water in cells. In order to know the H2O2-induced oxidative stress in RAW 264.7 cells, first of all, the optimum concentration of exogenous H2O2 which show reactive cellular responses was determined as 40 μM by MTT assay, and followed by 40 μM H2O2 application in RAW 264.7 cells for 30 min, 1, or 2 hours. The expressional changes of essential proteins for cellular proliferation, epigenetic modification, inflammation, apoptosis, survival, and protection were assessed by immunoprecipitation high performance liquid chromatography (IP-HPLC) using 51 antisera. 40 μM H2O2 treatment down-regulated proliferation-related proteins, Ki-67, PCNA, CDK4, cyclin D2, cMyc, and PLK4, induced histone methylation/ deacetylation and DNA methylation by increasing levels of HDAC10 and DMAP1 and by decreasing levels of DNMT1 and KDM4D, activated inflammatory reaction by increasing levels of MCP-1, COX-2, CD68, LTA4H, CXCR4, and lysozyme, and dramatically up-regulated cellular apoptosis-, survival-, and protection-related proteins, AIF, PARP-1, caspase 9, c-caspase 9, pAKT1/2/3, SOD-1, HO-1, NF-kB, NRF2, and GSTO1 in RAW 264.7 cells. These observations suggest exogenous 40 μM H2O2-induced oxidative stresses which resulted global cellular responses including not only antioxidant, inflammation, and apoptosis but also proliferation and epigenetic modification. Particularly, 40 μM H2O2-induced apoptosis was mainly derived from PARP-1/AIF signaling leading parthanatos, and 40 μM H2O2-induced suppression of cMyc/MAX/MAD network was relevant to reduction of RAW 264.7 cell proliferation. Accordingly, H2O2 appears to affect RAW 264.7 macrophages in several ways eliciting not only oxidative stresses but also genome-wide DNA damage.

Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin's lymphoma, and usually showed painless neck swelling, fever, sweat, and weight loss. Although about 5% of all lymphomas appeared in the oral area, the primary maxillofacial lymphomas were rare and sometimes clinically tended to be misdiagnosed such as chronic periodontitis, osteomyelitits, etc. This study demonstrated three cases of primary DLBCL mimicking localized osteomyelitis at mandible or maxilla. A series of histological and immunohistochemical examination using different biomarkers of lymphoreticular cells were performed to characterize the neoplastic cells of DLBCL. The first case occurred in a 45 years old male exhibiting mandibular osteomyelitis and neck swelling. The second case simply showed a gingival swelling at left upper premolar area in a 55 years old male. And the third case is from an 84 years old female who felt numbness at left lower lip and had severe periodontitis involving regional alveolar bone resorption. All of three cases had experienced no systemic manifestation of lymphoreticular malignancy before the diagnosis of oral lymphoma. Immunostainings of CD3, CD20, TNFα, BCL-2, Ki-67, PCNA, and c-Myc were strongly positive in these tumor cells, while those of p53 and CD31 were slightly positive, and CD56 immunoreaction was negative. These three cases were diagnosed as DLBCL and referred to the hemato-oncology unit for treatment. Therefore, every chronic granulomatous periodontal lesion hardly cured by simple medical treatment should be carefully explored through pathological examination, and it was presumed that DLBCL is closely related to the chronic inflammatory periodontal lesions recruiting mucosa-associated lymphoid cells in older patients. It was also suggested that DLBCL be differentially diagnosed from T-cell lymphoma, Burkitt’s lymphoma, and Hodgkin’s disease, etc. with immunohistochemical determination of tumor cell subtypes as soon as possible in order to be treated with appropriate therapy.

A 31 years old female had been suffered from a bony swelling in right premolar region of the mandible for 12 years, recently grown rapidly. A fistula tract developed on the right anterior mandibular border, but the lesion was relatively asymptomatic. In the radiological examination, the tumor mass was irregularly mixed with radiolucent and radiopaque areas, forming multiple cystic spaces. Under the diagnosis of calcifying odontogenic cyst, the mandibular mass was resected and examined pathologically. After decalcification, the dissected tumor mass showed multiple small cystic spaces and calcifying fibrous tissue, mimicking calcifying odontogenic cyst or ameloblastoma. Histological observation showed many calcifying cementoid materials and ossifying trabeculae. The cystic spaces were turned out to be dilated vascular channels lined by endothelial cells, containing plasma fluid. However, the main lesion was diagnosed as cemento-ossifying fibroma (COF), and the atypical vascular channels were greatly dilated and gradually expanded the whole tumor mass. The present COF was examined through immunohistochemical (IHC) array, and investigated for tumor cell characteristics, exhibiting abnormal ossification and aneurysmal cystic changes. IHC array disclosed that the tumor cells grew progressively in the lack of apoptosis, and that they showed lower expression of RUNX2 than BMP-2, RANKL, and OPG, and increases of protein expression in HIF-1α, VEGF-A, and CMG2. These data suggested that the reduced expression of RUNX2, osteoblast differentiation factor, be relevant to abnormal ossification of COF, and that the consistent expressions of angiogenesis factors be relevant to de novo angiogenesis in COF, subsequently resulted in aneurysmal cystic changes.

5-fluorouracil (5-FU) is a pyrimidine analog which can work as antineoplastic antimetabolite by blocking thymidylate synthetase conversion of deoxyuridylic acid to thymidylic acid in DNA synthesis. This study is aimed to know the anticancer effect of 5-FU on the expressions of important signaling proteins in KB cells through immunoprecipitation high performance liquid chromatography (IP-HPLC). KB cells were treated with 5 μM 5-FU and cultured for 12, 24, 48, 72, and 96 hours, and followed by IP-HPLC analysis using 32 antisera. 5-FU suppressed the proliferation of KB cells by decreases in the expressions of proliferation-related proteins, Ki-67, PCNA, CDK4, and MPM2 to 82.6%, 92.4%, 95.2%, and 95.9%, respectively, but increases of antiproliferation-related proteins, p16 and p21 to 106.7% and 125.5%, respectively, during 96 hours of experiment. This proliferation reduction was also negatively regulated by cMyc/MAX/MAD network signaling. The cellular protection and survival were consistently arrested by 5-FU treatment in KB cells. The expressions of NFkB, MDR, p-mTOR, and TNFα were decreased to 95.1%, 92.8%, 93.4%, and 90.3% in 48-72 hours, respectively, while cellular stress was increased by upregulation of p38 to 111.3% in 48 hours. And the expressions of pAKT1/2/3, hTERT, and AMPK were also decreased to 93.3%, 97.4%, and 89.3% in 24-48 hours, respectively, while the cellular transformation might be undergone by upregulation of TGF-β1 to 117% until 96 hours. Particularly, 5-FU treatment greatly induced the cellular apoptosis in KB cells by increased expressions of PARP, cPARP, caspase 9, c-caspase 9, caspase 8, and caspase 3 in the lack of p53/BAX and FASL/FAS signaling. The expressions of PARP and c-PARP were increased maximum to 119.2% in 24 hours, and followed by increases of caspase 9, c-caspase 9, caspase 8, and caspase 3 to 111.2%, 125.9%, 108.6%, and 116.3% in 72-96 hours. Therefore, it is presumed that 5-FU induced cellular apoptosis in KB cells may be derived from the overexpression of PARP due to the increased DNA defect caused by 5-FU, which can lead to ATP depletion and subsequent cellular apoptosis.

With the multiple practices of bone graft using different artificial bone regenerative substitutes, the bone graft procedures have been widely performed to increase the bony stabilization of dental implant. Xenogenic bone graft materials have been well developed because of their good biocompatibility and abundant source of bone materials. The present study demonstrated the histological findings from excellent bony remodeling in xenogenic bone graft biopsies compared to those findings in autogenous bone graft. For the graft bone biopsies which were usually done in 5-9 months after graft bone insertion, five types of histological grades including excellent, favorable, partial, degenerative, and poor bony remodeling could be assessed to give prognostic information for dental implant. However, recently the xenograft bone materials have been much improved and produced strong osteogenic effect. Among 239 cases of trephine bur-supported core bone biopsy the excellent bony remodeling was found in 20 cases (13.1%) out of 153 xenogenic bone grafts and in 13 cases (43.3%) out of 30 autogenous bone grafts. They produced abundant new bones on the surface of the graft bones in 5–9 months, and the graft bones were partly resorbed and also surrounded by the repetitive deposition of new bone. The osteophytic new bones showed strong birefringence under polarizing microscope, and were gradually elongated and anastomosed with each other to form trabecular bony networks which became proper stress-baring structures for dental implant. Their marrow stromal tissues were composed of loose connective tissue which was well vascularized but rarely infiltrated with inflammatory cells. The present study compared the histological features of excellent bony remodeling between xenogenic and autogenous bone grafts. Although the ratio of excellent bony remodeling in xenogenic bone graft was still low, 13.1%, the recent advance of xenogeic bone products was remarkable in biological aspect and almost comparable to the autogenous bones. Therefore, it was suggested that the xenogenic bone graft will be applicable to the bone regeneration procedures for dental implant with beneficial output in the near future.

The native human saliva obtained through the centrifugation of whole saliva showed characteristic salivary protein complex (SPC) peaks in gel filtration high performance liquid chromatography (HPLC) using Superose 12 column1,2). In the previous study the SPC peaks in chromatography were explored to know their composition and functions by different detection methods, but still the nature of SPCs was not clearly elucidated so far. In this study the SPC peaks were examined by direct antibody interaction in order to target different antimicrobial and protective proteins distributed in the SPCs via gel filtration HPLC. As the SPC peak shape and migration speed can be changed by antibody binding to specific proteins of SPC, it was found that mucin1 is evenly distribution in all SPCs, while PRPs are more abundant in the late dominant SPC than the early dominant SPC and also in the intermediated SPCs. Most of antimicrobial proteins including lysozyme, LL-37, lactoferrin, β-defensin-1, -2, -3, IgA, mucocidin, and α1-antitrypsin were more abundant in the late dominant SPC than the early dominant SPC, while histatin showed relatively even distribution in all SPCs. Therefore, it was presumed that the late dominant SPC containing abundant antimicrobial and protective proteins could be applied as a biomarker to measure the defensive potential of whole saliva in oral diseases.

A nineteen years old male patient showed a cystic lesion in left maxillary canine to premolar area (#23-#25). This lesion was asymptomatic, and found during his routine radiological check in local clinic. In the radiological observation the cystic lesion showed round radiolucent image containing many calcified bodies which were usually small but irregular in shape, expanding tumorously and resulted in the displacement of canine and second premolar in the absence of first premolar. The lesion was surgically enucleated, and a cystic fibrous tissue containing abnormal teeth was removed and examined pathologically. With the histological observation of tumorous odontogenic epithelium including many ghost cells, which were closely associated with abortive teeth, the lesion was finally diagnosed as CCOT associated with complex odontoma. The ghost cells of CCOT was strongly positive for β-catenin, GADD45, and LC3, and slightly positive for MMP-9, while they were rarely positive for BCL2, Wnt1, HSP-70, and p38. Therefore, it was presumed that the ghost cells of CCOT might undergo dormant cell state through altered cytodifferentiation stimulated by severe growth arrest, DNA damage signaling, and abundant autophage formation.

Aneurysmal bone cyst (ABC) in maxilla is a rare and benign lesion but shows extensive bony destruction, occasionally accompanied with secondary osseous lesions, i.e., central giant cell granuloma, ossifying fibroma, fibrous dysplasia, etc. As the pathogenesis of ABC has not been clearly defined, ABC is diagnostically challenged due to its variable histological features. A 17-year-old boy showed a huge radiolucent lesion at right anterior maxilla, which was accidentally found in routine dental-radiological examination for orthodontic treatment. He had no medical history of systemic disease, and did not remember any traumatic experience on his right anterior maxilla. The radiolucent lesion involved periapical area from right central incisor to right first premolar, and was clinically diagnosed as odontogenic keratocyst. During surgical operation a cyst-like sac was enucleated with severe hemorrhage. In the histological observation the thick fibrous sac showed no lining epithelium, and its luminal side disclosed multiple aneurysmal spaces which were shrunken and almost obliterated. The fibrous sac itself was hyperplastic with abundant vascular channels, and produced fibromatous thickening associated with ossifying trabecular bones. This fibro-osseous tissue was hamartomatous, which was not directly connected and organized with marrow bone of maxilla. Finally, the present case was diagnosed as secondary type ABC differentially from traumatic bone cyst (TBC), odontogenic cyst, and central reparative granuloma. And it was presumed that the hamartomatous proliferation of fibro-osseous tissue in the cystic sac of ABC could produce the swelling pressure effect in the bone marrow similar to the overgrowth of central giant cell granuloma, ossifying fibroma, fibrous dysplasia, etc., in the secondary type ABC.

In order to know the characteristic roles of salivary protein complex (SPC) the gel-filtration chromatography was performed using the unstimulated and the stimulated whole saliva separately. The first and second dominant SPC peaks were fractionated and analyzed by immunoprecipitation HPLC (IP-HPLC) using antibodies against the essential salivary proteins including α-amylase, mucin-1, proline rich proteins (PRPs), histatin, cystatin, LL-37, lysozyme, lactoferrin, -defensin-1, -2, -3, IgA, transglutaminase 4 (TGase 4), mucocidin, α1-antitrypsin, cathepsin G. In the gel-filtration chromatography the stimulated whole saliva showed much reduced amount of SPCs than the unstimulated whole saliva, but the proportional patterns of both whole saliva were almost similar each other. Through IP-HPLC analysis both of the first and second dominant SPCs were variably positive for the essential salivary proteins, however, α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G were predominant in the first dominant SPC, while cystatin, lactoferrin, β-defensin-1, -2,-3, IgA, mucocidin, TGase 4, and α1-antitrypsin were predominant in the second dominant SPC. And more, the α1-antitrypsin and cathepsin G which were mostly derived from gingival crevicular fluid were also consistently found in the SPCs. These data may suggest that the first dominant SPC, rich in α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G, may play a role in food digestion, protein degradation, and mucosa lubrication, while the second dominant SPC, rich in cystatin, lactoferrin, β-defensin-1, -2, -3, mucocidin, IgA, TGase 4, and α1-antitrypsin, may play a role in the mucosa protection and antimicrobial defense.

Immunoprecipitation-based high performance liquid chromatography (IP-HPLC) is a type of modified enzyme-linked immunosorbent assay (ELISA) that uses protein A/G (or antibody)-conjugated beads instead of the antibody-conjugated wells used in ELISA. In order to determine the fidelity of IP-HPLC, the author used 83 antisera to identify protein expression changes caused by cisplatin treatment in KB human oral cancer cells. KB cells were cultured for 12 or 24 hours with 10 ug/mL cisplatin. The results obtained by IP-HPLC were comparable with published cisplatin data, although ELISA was not conducted in the present study. Cisplatin dominantly reduced the levels of proteins associated with cell proliferation, transcription factors, growth factors, cytoskeletal proteins, and cellular differentiating factors, but on the other hand, apoptosis-related factors, oncogenes, and protective proteins were usually up-regulated, presumably to address cisplatin-induced DNA damage. In particular, cisplatin directly inactivated genomic DNA by down-regulating histone H1 and demethylase and by up-regulating deacetylase. Cisplatin also rapidly induced p53 overexpression and mitochondria-mediated endogenous apoptosis occurred after 12 hours of cisplatin treatment, although this was almost completely replaced by FASL/FAS-mediated exogenous apoptosis after 24 hours. This preliminary study was conducted to investigate the anticancer effect of cisplatin on the KB human oral cancer cells and to determine the fidelity of IP-HPLC data. It was concluded that IP-HPLC is useful for identifying profile changes of genome wide essential proteins and signaling changes of major molecular pathways.

Caliber persistent artery (CPA) is a vascular anomaly presenting as a bluish and pulsatile artery in the subepithelial tissue. Although the incidence of CPA was debated, many CPAs occurred in the perioral and facial tissues at which the embryonal strapedial artery networks were distributed. The present study demonstrated a case of CPA occurred in the retromolar buccal mucosa in a 37 years old male. The lesion showed many pinkish granular spots, but was asymptomatic except biting irritation during mastication. It had slowly increased in size up to 20 × 25 mm for 3 years, and recently became hemorrhagic due to the biting injury between left upper and lower second molars. With the fear of oral cancer an incisional biopsy was performed, and followed by histological and immunohistochemical study. Histologically the lesion showed many tortuous artery localized at the submucosa area, and the arterial wall was thick and its lumen was narrowed and shrunken. In the immunochemistry α-SMA was positive for thick smooth muscle layer of artery and arterioles, TGase 2 was weakly positive for the luminal surface of arterial intima, and bFGF was consistently positive for the perivascular fibrous tissue. But PCNA, VEGF, CD31, CMG2, TGF-β1, HSP-70, and 14-3-3 were almost negative for the vascular tissue. Therefore, it was presumed that the lesion was not actively proliferative nor degenerative but still retained its cellular stability and slow growing potential. It was finally diagnosed as CPA differentially from arterio-venous malformation, hemangioma, lymphangioma, and squamous cell carcinoma. The retromolar buccal mucosa CPA is first reported in this study and may present usual clinical findings depending on its size and location. This asymptomatic lesion could be severely hemorrhagic by minor biting injury, therefore, precise differential diagnosis should be made through biopsy, and careful therapy be followed.

A 57 years old female received xenogenic bone graft for the extraction socket augmentation of right maxillary molars and for the sinus floor elevation six months ago. The bone graft sites were healed uneventfully and showed marked radiopacity in the postoperative X-ray view. Before dental implant insertion the bone biopsy was made using trephine bur and examined pathologically. The graft bones showed minimum new bone deposition with dysplastic epithelium. The epithelium was proliferative on the surface of graft bones forming epithelial strands and nests, similar to the odontogenic epithelium. The immunohistochemical study was performed using different antisera of odontogenic markers, growth factors, oncogenes, etc. The epithelial cells were strongly positive for pan-keratins, EGF, pAKT, and HSP-70, consistently positive for PCNA, p53, EGFR, 14-3-3, and survivin, slightly positive for ameloblastin, but rarely positive for amelogenin. Particularly the matrix of graft bone was slightly positive for EGF. Taken together, it is presumed that the abnormal epithelium on the graft bones was derived from odontogenic epithelial elements, Malassez epithelial rests, distributed at the periodontal tissue of maxillary molars, and that they might undergo dysplastic proliferation affected by the release of growth factors and osteogenic proteins from the graft bones. It is also suggested that the graft bone substitutes inserted for the dental implant possibly have a potential to induce the proliferation of odontogenic epithelial rests leading to the pathogenesis of odontogenic cysts and tumors.

Salivary proteins include numerous functional proteins which play important roles not only for the food-intake but also for the protective and defensive mechanisms. In the present study the compositions of salivary proteins were analyzed by different methods, including electrophoresis and high performance liquid chromatography (HPLC). In hydrophobic protein HPLC analysis the parotid saliva gradually produced macromolecular complexes when agitated in refrigerator until 30 minutes. These salivary protein complexes were digested by neuraminidase, and then migrated more rapidly in native tris glycine gel than the control. Therefore, it was assumed that the glycosylated proteins of parotid saliva became gradually aggregated to form salivary protein complexes similar to those of whole saliva. The salivary protein complexes were easily degenerated in different experimental buffers, i.e., SDS buffer, tris glycine buffer, methanol, etc., and resulted non-specific patterns in electrophoresis and HPLC. Therefore, it was presumed that the salivary protein complexes was made by the hydrophobic interaction as well as electrostatic attraction between salivary proteins. These data indicated that to know the real pattern of salivary protein complexes in vivo the whole saliva should be analyzed by HPLC using non-adhering column with isoelectric buffer. Consequently, the whole saliva was analyzed by HPLC using reverse phase SuperoseTM column with 20 mM potassium phosphate buffer, and two prominent peaks of salivary protein complexes were consistently found in every people. These salivary protein complex peaks were relatively stable up to 6 hours after saliva collection when the whole saliva was kept in refrigerator during experiment. Therefore, it is suggested that the salivary protein complex patterns are characteristic macromolecular structures of whole saliva, which are also applicable as a diagnostic point in different saliva-associated diseases

An 18 years old female patient suffered from cerebrovascular occlusive disease, moyamoya disease, showed a huge cyst in her left mandibular body in the radiological observation. The lesion was asymptomatic and found during routine dental check. She had no experience of traumatic injury on her jaw. The cystic lesion was ovoid with irregular scalloping margin and multilobular image, and occupied the whole marrow space of mandibular body with slight expansion of buccal cortical bone. During operation the lesion showed an empty space covered with grayish white fibrous tissue. The luminal fibrous tissue and underlying bony tissue were curettaged and examined pathologically. In the histological observation the lesion was a pseudocyst lined by thick fibrous tissue. Some large vessels underwent atherosclerotic change, exhibiting thickened vessel walls which were partly distorted with hemorrhage and thrombi, and some small capillaries were extremely dilated with hemorrhage and subsequently resulted in perivascular ischemic change with chronic vasculitis. This mandibular cystic lesion was finally diagnosed as simple bone cyst (SBC) associated with moyamoya disease differentially from aneurysmal bone cyst (ABC), traumatic bone cyst (TBC), periapical odontogenic keratocyst, and central giant cell granuloma. Therefore, it was presumed that the thromboembolic and atherosclerotic vessels of moyamoya disease might increase the hemodynamic pressure of mandibular bone marrow tissue and subsequently was able to induce SBC.

A 67 years old female showed diffuse erosive ulceration at left buccal mucosa. She had received tegretol to treat the patient’s pain and anxiety of trigeminal neuralgia for 18 months. Otherwise her medical history was nonspecific. Under the clinical diagnosis of lichen planus she received anti-inflammatory therapies using antibiotics and steroid ointment, which were not effective. Consequently her oral ulceration was gradually expanded and aggravated. In the biopsy examination mucosa epithelium was irregularly keratinized and focally detached from underlying connective tissue by thin cleft spaces, accompanied with inflammatory cell infiltration into the subepithelial area. The epithelium was generally acanthomatous with short rete ridges. Many spots of acantholysis were found in the basal and suprabasal layers of epithelium, into which melanocytes were migrated. Particularly, many keratinocytes not only in the spinous layer but also in the suprabasal layer contained atypical keratohyalin granules in their cytoplasms. In the immunohistochemistry the epithelium was rarely positive for PCNA and IgK, but strongly positive for HSP-70, and many keratinocytes showed strong positive reaction of lysozyme in their cytoplasms. Taken together, with the characteristic cytotoxic changes of keratinocytes, which are usually found in the oral epithelium damaged by certain drug abuse, the present case of pemphigus-like oral lesion was diagnosed as drug-induced pemphigus caused by long time intake of tegretol, carbamazepine derivative. The acute oral drug-induced pemphigus should be differentially diagnosed from oral lichen planus, recurrent aphthous ulceration, oral leukoplakia, candidiasis, autoimmune pemphigus, etc., in order to treat properly in the absence of biohazards of systemic therapeutic drugs

The molecular mechanisms of the carcinogenesis of oral squamous cell carcinomas (OSCCs) are highly variable and result in different features of tumor progression, i.e., local tissue destruction and metastasis to regional lymph nodes. A case of OSCC arising from proliferative verrucous leukoplakia (PVL) was analyzed for its protein expression profile by immunoprecipitation (IP) – high performance liquid chromatography (IP-HPLC) by using 72 antisera and comparing results with those of KB cells. OSCC arising from PVL showed stronger expressions of proteins associated with cell proliferation (MPM2, PCNA, eiF5A, DHS, DOHH), cell survival (pAKT, MDM2, survivin), matrix proteolysis (elaffin), tumor suppression (p16, p21, PTCH1), the WNT/β-catenin pathway (SHH, WNT1, APC, β-catenin, snail), proinflammation (TNFα), angiogenesis (HIF, CMG2, vWF), and cellular protection (HSP-70, FAK, caveolin) and of oncoproteins (STAT3, 14-3-3, K-RAS, PUMA, PIM1) and growth factors (EGFR, bFGF) than KB cells. On the other hand, KB cells showed stronger expressions of proteins associated with apoptosis (caspase-3, caspase-8, caspase-9, PARP, FAS, FASL, TGase-1, BCL2, BAD, BID, BAK, FLIP), matrix proteolysis (MMP-2, MMP-9), transcription signaling (NFkB, p38, E2F-1, HO-1), and tumor suppression (p53, RB1, PTEN) and of oncoproteins (DMBT1, CEA) and growth factor (TGF-β1, c-erbB2, VEGF) than OSCC arising from PVL. These data indicate the cells of OSCC arising from PVL are more resistant and more robust than KB cells. Furthermore, they suggest the oncogenic signalings of OSCC arising from PVL play important roles in the aggressive growth and rapid tumor metastasis to regional lymph nodes

Exostosis is a phenomenon of exophytic growth of compact bone, and the oral exostosis, so called a torus is usually found in the lingual or labial areas of mandible and maxilla, and the hard palatal area. It is not a pathological nor tumoral formation but a localized bony protuberance relevant to developmental and environmental origins. However, the pathogenesis of exostosis has not been clearly elucidated so far. In the present study total 51 cases of oral exostosis were examined by radiological, histological, and immunohistochemical methods to observe the osteogenetic potential existed in the sclerosed bony tissue of exostosis. Particularly, the unilateral mandibular exostoses occurred in the vicinity of mandibular primary growth centers which were more prominent than the contralateral ones in the radiological observation. Histologically the peripheral area of exostosis was composed of lamellated bone and covered with periosteal tissue which showed sparse osteoblastic activity, while the central area of exostosis was composed of thickened and anastomosed trabecular bones with small amount of marrow connective tissue. The latter stained blue in Masson trichrome method, while the former stained red. The immunohistochemical reactions of BMP-2, bFGF, CMG2, and TGF-β1 were clearly positive in the central trabecular bones, while almost negative in the peripheral cortical bones of exostosis. These findings may indicate that the central area of exostosis is less mineralized than the peripheral area of exostosis, and the former expresses different osteogenetic proteins to produce bony tissue contrary to the latter. Therefore, it is suggested that the strong osteogenetic potential in the central area of exostosis be relevant to the growth potential of mandibular and maxillary primary growth centers and play an important role for the latent expansile growth of exostosis in adult life.

A 72 years old male complained of gingival ulceration and whitish discoloration on the marginal and attached gingival epithelium of left mandibular premolar and molar area, where a porcelain-fused metal (PFM) crown and an ill-fitting gold crown were applied for 10 years, respectively. Recently he had a cancer phobia due to this whitish lesion unhealed even after intensive anti-inflammatory and antibiotic treatment. In the pathological examination the epithelium was hyperkeratotic and acanthomatous with severe inflammatory reaction, and subsequently its basement membrane was distorted and the intercellular spaces between keratinocytes were widened. Particularly, the nuclei of keratinocytes were elongated in the same direction towards the electric current between the dissimilar metallic crowns of PFM gold. In the immunohistochemistry KL1, β-catenin, and S-100 were strongly positive in the epithelium, but consistently weak for TNFα, HSP-70, and β-defensin-1, -2, -3. On the other hand, PCNA, p53, E-cadherin, and pAKT were rarely positive for the epithelium. Interestingly, the hyperkeratinized and inflamed epithelium was strongly positive for a calcium binding protein (S-100), while it showed almost reduced expression of protective molecules (HSP-70, β-defensin-1, -2, -3, and pAKT). Therefore, it was presumed that this localized lichenoid gingivitis was caused by the galvanic current phenomenon with lower cellular and immunological responses contrary to the ordinary oral lichen planus and leukoplakia

Oral erythroleukoplakia is characterized by severe dyskeratosis intermingled with multifocal erosive spots on the buccal mucosa, dorsal tongue, and lower lip, etc. A case of oral erythroleukoplakia was diagnosed among 83 cases of common oral leukoplakia since 1997. The pathological examination showed the typical features of leukoplakia with severe epithelial dysplasia, exhibiting dyskeratosis, acanthosis, and basal hyperplasia. The oral erythroleukoplakia was explored in comparison with a representative common oral leukoplakia by the immunohistochemical method using PCNA, β-catenin, EGFR, p53, TNFα, pAKT, and STAT3. Oral erythroleukoplakia showed strong positive reaction of PCNA, p53, EGFR, TNFα, pAKT1 and STAT3 in its spinous layer cells and these reactions were reduced in its basal layer cells, while common oral leukoplakia showed diffusely weak reaction of those proteins. Particularly, β-catenin was positive in the nuclei of some basal and spinous layer cells of oral erythroleukoplakia contrast to the common oral leukoplakia. These findings indicated that the present oral erythroleukoplakia was proliferative with the activation of β-catenin pathway, revealed the dysplastic changes of epithelium by the overexpression of EGFR, p53, and pAKT, and also produced inflammatory reaction through the activation of cytokine-dependent signalings of TNFα and STAT3. These data indicated that the present oral erythroleukoplakia might undergo the early stage of multi-step carcinogenesis via the overexpression of different oncoproteins, especially β-catenine, p53, pAKT, and STAT3.

Pemphigus is an autoimmune blistering disease characterized by autoantibodies against epidermal adhesion molecules, desmogleins. Pemphigus vulgaris is most common and shows intraepidermal vesicles caused by the breaking apart of epidermal cells, acantholysis. A 65 years old male patient complained of severe mucosa ulceration on his right mandibular retromolar pad area where traumatic injuries occurred during mastication. He also had multifocal round skin ulcerations, less than 7~8mm in diameter and showed habitual onset and disappeared soon. At this time he was anxious about his oral ulceration with a cancer phobia, thereby a biopsy was made to rule out any malignancy in the ulceration. The histology examination showed multifocal suprabasal splits forming vesicles and erosion. The suprabasal splits were linear and parallel to the basal cell layer. The immunostain of IgK was strongly positive in the vesicular fluid as well as the cell membranes of dissociating keratinocytes, and also positive in many plasma cells infiltrated into the subepithelial zone. TNFα, IL‐1, ‐8, ‐28 for the pro‐inflammatory reaction were weakly expressed, while IL‐6 was strongly positive in the acantholytic keratinocytes of vesicle forming area. β‐defensin‐1, ‐2, ‐3 for the innate immunity were diffusely positive in the involved epithelium. The cell survival proteins, pAKT and HSP‐70 were diffusely positive in the epithelium, while the apoptosis protein, PARP was consistently positive in some acantholytic keratinocytes. These findings indicated that the vesicle formation occurred by autoantibody reaction without the activation of pro‐inflammatory and cell‐mediated immune reactions. The lesion was diagnosed pemphigus vulgaris with abrupt onset of epithelial vesicles at the predisposing areas of traumatic injuries by type II hypersenstitive immune reaction.