It has been hypothesized that lymphokine-activated killer (LAK) cells may be effective against antitumor drug resistant (ATDR) squamous cell carcinoma. The objective of this study was to determine 1) the susceptibility of ATDR human tumor cell lines to interleukin-2 (IL-2)-induced LAK cell-mediated cytotoxicity, and to assess 2) the characteristics of the population of LAK cells as effector cells on the tumor cell lines tested and 3) the relationship between the susceptibility of ATDR cells and their expression of surface antigens. Parent strains of CEM (derived from leukemia), KB (derived from oral squamous cell carcinoma), and TCM (derived from metastasized lung cell carcinoma from oral squamous cell carcinoma) cells were used as drug-sensitive cells. The ATDR cell lines were established from the parent cells used; the ATDR cell lines were CEM (resistant to vinblastine, VLB; CEM/VLB0.1 and CEM/VLB1.0), KB (resistant to cisplatin, CDDP, and bleomycin, BLM; KB/CDDP1.0 and KB/BLM1.0, respectively) and TCM (resistant to CDDP; TCM/CDDP1.0). Peripheral blood lymphocytes were obtained from nine healthy volunteers and fractionated; the fractions consisted of unfractionated whole cells, CD 4 + cells, CD 8 + cells, and CD16+ cells. Each fraction was cultured with IL-2, and the induced LAK cells were used as effector cells. Whole-LAK cells were the most cytotoxic against all ATDR cell lines tested (as compared with the parent cells). However, the antitumor activity of the fractionated cells was limited (antitumor activity: CD16+-LAK, 60-80% on CEM/VLB0.1 and 1.0; CD 8+-LAK, about 30% on KB/CDDP1.0 and about 50% on TCM/CDDP1.0). The expressions of P-glycoprotein (Pgp), major histocompatibility complex (MHC) class I, intercellular adhesion molecule-1 (ICAM-1), and lymphocyte function-associated antigen-3 (LFA-3) were respectively high in CEM/VLB; KB/CDDP1.0, KB/BLM1.0 and TCM/CDDP1.0; KB/CDDP1.0 and KB/BLM1.0; and TCM/CDDP1.0. In contrast, the susceptibility of the ATDR cells to LAK killing was not affected by anti-P-gp, anti-MHC class I, anti-ICAM-1, or anti-LFA-3 monoclonal antibodies. These results indicate that the effective population of LAK cells varies according to the tumor cell line, and are consistent with the assumption that the susceptibility of drug resistant cells to LAK cells depends on the property of the parent tumor cells and the type of antitumor drug that induces ATDR cells.
The second position of codon 61 (A to T) in the H-ras gene is a specific mutational site of DMBA-induced hamster cheek pouch squamous cell carcinoma (SCC). We previously reported a high frequency of mutations in the H-ras gene at codon 61 in the late stage of this model of carcinogenesis. However, it is difficult to detect small numbers of genetic abnormalities in a heterogenous population. Mutant allele specific amplification (MASA) is a very sensitive method for detecting micrometastasis in “histologically negative” lymph nodes. We used this method to detect H-ras mutations in premalignant lesions as well as SCCs. Using PCR and direct sequencing, we detected mutations in the H-ras gene in only 30% of advanced cancers but not in early cancer. However, mutations were detected in 15% of dysplastic lesions, 58% of superficial cancer, and 76% of invasive cancers with the use of MASA. These results suggest that 1) MASA is useful for detecting small numbers of genetic abnormalities in a heterogenous population in this model of carcinogenesis and 2) mutations in the H-ras gene occur in the early stage of DMBA-induced hamster cheek pouch carcinogenesis.
The saliva of patients with oral cancer was subjected to reversed-phase HPLC and a peak (P14) eluted with a linear gradient of acetonitrile. In contrast, minimal P14 was detected in the saliva of healthy volunteers. P14 showed strong absorbance at a λ max of 278nm, and showed high fluorescence at λ em of 304nm and 348nm with a λ ex of 278nm. It seemed that P14 was a peptide containing tryptophan and tyrosine. The molecular weight of P14 was 3, 442 daltons on a mass spectrometry. Amino acid analysis by Edman degradation and homology studies showed that P14 was consistent with human α-defensin 1 (HNP-1), a peptide that belongs to the defensin family. HNP-1 was detected in the whole saliva of cancer patients, but only low levels were found in the serum, the parotid saliva, and the submandibular and sublingual saliva. The concentrations of HNP-1 in the whole saliva of the 8 preoperative and 13 postoperative carcinoma patients were 11.3±7.8 and 5.9±5.6 μg/ml (mean± S. D.), respectively. The concentration in the preoperative patients was higher than that in the postoperative patients and gradually decreased after operation. However, HNP-1 was found in the saliva of patients even a half year after operation. The origin of HNP-1 in the saliva of oral cancer patients was not clear, but it was suggested that HNP-1 might act as a defensive substance against carcinoma.
In oral and maxillofacial surgery, we often require wound dressing materials to cover bone or soft tissue defects and promote healing. We previously showed that materials made of chitin are suitable for this purpose. Chitin changes its physical and biological properties according to its deacetylation level. However, no report has described the influence of the deacetylation of chitin on biocompatibility. To evaluate this issue, we performed in-vitro experimental studies using materials containing various levels of deacetylated chitin. Human gingival fibloblast-like cells (HGF) and mouse osteoblast cells (MC 3 T3-E 1) were cultured on sheets made of non-deacetylated chitin and, 35%, 50%, and 70% deacetylated chitin (DAC-0, DAC-35, DAC-50 and DAC-70, respectively). The growth of HGF and MC 3 T 3-E 1, and alkaline phosphatase (ALPase) activity of MC 3 T 3-E 1 were measured. The growth of these cells on the materials was also studied by SEM. The biocompatibility of DAC-35 with these cells was high. Levels lower or higher than 35% deacetylated chitin were less biocompatible. Especially, DAC-50 and DAC-70 suppressed the growth and differentiation of these cells.
We studied osteoinduction by recombinant human bone morphogenetic protein-2 (rhBMP-2) with the use of porcine skin-derived atelopeptide type I collagen (CL) as a carrier. Ten μg of rhBMP-2 with 3 mg CL was implanted into the calf muscle of 10-week-old Wistar rats. In the control groups, 3 mg CL only was implanted. Ten days (BMP group I, CL group I; n=5) or four weeks (BMP group II, CL group II; n=5) after implantation, soft X-ray and light microscopic exminations were performed. Soft X-ray findings showed a radiopaque shadow in the implanted muscles in the BMP groups. In BMP group I, a radiopaque shadow was observed partially at the outermost edge of the implanted materials. In BMP group II, a radiopaque shadow was observed at all areas of the implant. In contrast, no such shadows were observed in the CL groups. Histological examination revealed new bone formation in the BMP groups. Woven bone appeared at the outermost edge of the implant material in BMP group I. Trabecular bone and osteoblast-like cells were observed around almost the entire circumference of the implant material in BMP group II. There was no evidence of bone or cartilage in the CL groups. From 10 days to 4 weeks after implantation, osteoinduction was continuously promoted by rhBMP-2 with CL, and the amount of CL decreased in the implant. These results suggest that rhBMP-2 may play an important role in reconstructive surgery for extensive bony defects and augmentation.
We evaluated the usefulness of operative arthroscopy with a Holmium: YAG laser for the treatment of internal derangement of the temporomandibular joint (TMJ) as compared with electric cautery. Thirty-four patients (43 joints) with deranged TMJ were treated with a Holmium: YAG laser and 55 (68 joints) were treated with electric cautery. After 9.5 months of follow-up, the surgical success rate with Holmium: YAG laser was 95.3%, which was similar to the success rate with electric cautery 9.7 months after operation. There was no significant difference between the two groups with respect to the size of the postoperative interincisal opening or arthralgia. Almost all patients recovered more rapidly and returned more promptly to normal daily activities by using a Holmium: YAG laser as well as by electric cautery. We conclude that TMJ arthroscopic surgery with a Holmium: YAG laser is useful for the treatment of internal derangement of the TMJ.
A bioactive bone cement was developed for the fixation of implants during femoral head replacement operations and the repair of bone defects. This new biomaterial has the potential to be used as a root canal retrofilling material, because it is sterile and has good biocompatibility. The canal sealing tests of root apexes of extracted teeth were performed using copper amalgam, which is widely used for retrofilling, as control. Bioactive bone cement showed less dye penetration than copper amalgam when 2% fuchsin dye solution was used. The results of this study suggest that this bioactive bone cement, which was easy to handle and promoted bony repair, may be used as a retrofilling material.
The dental and oral surgical treatment of medically compromised patients has been avoided or contraindicated because such patients are at high medical risk. However, the oral surgery of these patients is important for the relief of oral discomfort and may have a good influence on the course and outcome of their systemic disease. We analyzed the treatment items and times before, during, and after oral surgery in high risk patients, as compared with normal patients. The subjects comprised 96 patients (male, 58; female, 38) treated at 10 hospitals. Their systemic diseases were restricted to 6 kinds: cardiovascular disease, diabetes mellitus, hematologic disease, hemodialysis, infectiousdisease (viral hepatitis, HIV infection), and patients receiving steroids, anticoagulants, and antitumor drugs. The average number of treatment items before, during, and after oral surgery was 13, and 360.2 min were required for treatment per patient. Furthermore, for tooth extraction, the treatment items of the patients who were treated on an inpatient basis were 2-fold that of the patients treated on an outpatient basis, and the treatment items of medically compromised patients were 3-fold that of normal patients.
Bone formation in the autogenous periosteum of the tibia grafted to the floor of the mouth to bridge the mandible was studied by micro-CT to assess its efficacy in evaluating bone formation in rabbits. On soft radiographs, bone formation was observed from both ends of the periosteum on day 14. The bone increased in width and extended medially; contact was made in the center on day 28. The time course of the development of bone trabeculae was well demonstrated three-dimensionally on micro-CT. Indices of bone quality such as Tb-Th, Tb.N, and BV, which reflect the growth of trabeculae, increased gradually from days 14 to 21 and more rapidly from days 21 to 28, whereas Tb. S decreased gradually after grafting. The results suggest that micro-CT is useful in evaluating bone formation threedimensionally.
We performed secondary mandibular reconstruction with a mandibular reconstruction implant in a 58-year-old man after hemimandibulectomy for gingival carcinoma. The mandibular reconstruction implant was made by combining mandibular reconstruction plates with a cast chromium-cobalt alloy plate.The cast plate had to be accurately placed between the reconstruction plate and the mandibular condyle. We thesefored used a solid model based on three-dimensional computed tomography image data to manufacture the cast plate. The good fit of the cast plate demonstrated the high usefulness of the threedimensional solid model.
Basal cell adenocarcinoma represents only about 1% of all malignant salivary gland epithelial tumors. Most basal cell adenocarcinomas occur in the parotid gland. This report describes a rare case of this tumor arising in the minor salivary glands of the buccal mucosa in a 76-year-old woman. The tumor was resected and immunohistochemical analysis was performed. Reactivity for anti-smooth muscle actin was confirmed in luminal cells, anti-cytokeratin 14, and anti-S-100 protein in the cytoplasm of tumor cells as wellas in anti-vimentin in stromal elements of tumor cells and anti-ki-67 in peripheral tumor cells. We also performed image cytometric analysis of nuclear DNA content. The mean DNA index was 1.22, ranging from 0.89 to 2.67 and showing a diploid pattern, which suggested that this tumor had lowgrade malignancy.
Mucoepidermoid carcinoma is one of the most common malignant tumors arising in the salivary glands. The incidence in the major and minor salivary glands has been re-ported as 3 % to 11% and 10%, respectively. However, mucoepidermoid carcinoma in the lower lip is rare, and its incidence is about 1 % to 5 % among tumors in the minor salivary glands. This paper reports a case of mucoepidermoid carcinoma originating in the minor salivary glands of the lower lip. A 46-year-old man consulted our clinic because of a mass in the lower lip. The mass meas-ured 16×14mm and was movable on palpation. Total extirpation biopsy was performed under local anesthesia. Pathological examination showed high-grade mucoepidermoid carci-noma. Because tumor appeared to remain in the surrounding tissue, further resection of the lower lip was carried out under general anesthesia. No recurrence or metastasis has been found as of 8 years since the last resection.
A case of squamouse cell carcinoma arising in a pectoralis major myocutaneous flap used in the oral cavity is reported. A 64-year-old woman was treated for lower gingival squamouse cell carcinoma. After radiation therapy, radical neck dissection, mandibulectomy, total resection of oral floor, and resection of buccal mucosa and skin were performed. A PM-MC flap was used to reconstruct the defect. Six years later, recurrence in the gingiva of the maxilla was diagnosed, and the maxilla was resected partially. Eight years after the first operation, a new tumor showing papillary proliferation was noted in the center of the transplanted PM-MC flap. The tumor was simply resected and pathological examination showed that it was a well differenciated squamouse cell carcinoma in situ. In six months, a tumor was noted on the flap again. After complete resection of the tumor, there has been no recurrence for 3 years.
Tolosa-Hunt syndrome (THS) is characterized by unilateral ophthalmoplegia due to non-specific granuloma in the cavernous sinus. Steroid therapy is effective against THS. We report a case of malignant lymphoma showing symptoms similar to THS subsequent to paresthesia of the third division of the trigeminal nerve. The patient was a 56-year-old man who presented with paresthesia of the third division of the trigeminal nerve region. On initial examination, there were no radiographic or computed tomographic abnormalities of the head and neck region, including the cavernous sinus. After 2 weeks, highly enhanced images were observed in the cavernous sinus on magnetic resonance imaging (MRI), and the patient complained of ophthalmoplegia and neuralgia of the first and second division of the trigeminal nerve region. Daily administration of steroids improved symptoms. THS was suspected on the basis of neurological symptoms, MRI findings, and the response to steroids. Symptoms recurred after 2 months. Bone marrow puncture was performed, and acute lymphatic leukemia (ALL-L 3, Burkitt type) was finally diagnosed. Eighteen months after the first examination, the patient died. Autopsy was done, and a malignant lymphoma arising in the cavernous sinus was confirmed. We reported a case in which paresthesia of the third division of the trigeminal nerve region was an initial manifestation of the Tolosa-Hunt syndrome.
The patient was a 48-year-old man with an unusual papillary tumor of maxillary sinus origin extending from the left premolar to the right molar region of the maxilla. Clinical examination suggested that the tumor was malignant, because of repeated recurrence and extensive bone destruction. The tumor was removed en bloc after chemotherapy with pepleomycin, which was ineffective. Histopathological examination revealed an inverted papilloma with moderate atypism. Immunohistochemically, p53, Ki-67, and PCNA were positive in tumor cells. The labeling indices of these markers were between those of leukoplakia and squamous cell carcinoma. This is the first reported case of inverted papilloma of the maxillary sinus with extension into the oral cavity.
We report a case of a huge congenital hemangioma involving the oro-maxillofacial region, pharynx, neck, and mediastinum. A 28-year old man visited our clinic because of anorexia due to glossodynia and tongue swelling. He had a history hemangioma and had received irradiation, partial resection, and a skin graft between the ages of 1 to 5 years at other hospitals. The hemangioma extended from the head and neck to the mediastinum on initial examination. Angiographic and clinical findings suggested that this case was a purely venous malformation, and aggressive treatment was impossible. Nasal intubation under bronchofiberscopic guidance and needle cricothyrostomy for CO2 narcosis caused by the infected hemangioma were unsuccessful. The patient suffocated to death because of massive bleeding.
A case of pigmented ameloblastic fibrodentinoma and one of pigmented ameloblastic fibroodontoma, both arising in the mandible, are reported. Case 1. An 8-year-old girl visited our clinic for further evaluation of a radiolucent area in the left side of the mandible, found by a dentist. Radiographic examination demonstrated a circumscribed radiolucent mass at the root apex of the left second deciduous molar. The clinical diagnosis was a mandibular cyst. The lesion was removed with the patient under intravenous sediation. A postoperative pathological examination disclosed a pigmented ameloblastic fibrodentinama. Reexamination 7 years after the operation revealed no recurrence of the lesion. Case 2: A 10-year-old girl presented with delayed eruption of the right mandibular first molar. Radiographs revealed a multicystic radiolucent lesion, which contained some small radiopaque masses from the right second premolar to the acending ramus. The clinical diagnosis was a mandibular tumor. The tumor was extirpated with the patient under general anesthesia. The postoperative histopathological diagnosis was a pigmented ameloblastic fibroodontoma. There has been no evidence of recurrence as of 2 years 4 months postoperatively.
We report the case of an HIV infected patient who was discovered during dental treatment. The patient was a 25-year-old man in whom oral candidiasis was diagnosed by a dentist. The clinical diagnosis was oral candidiasis with immunodeficiency. The patient had a history of sexual relations with many partners. Laboratory tests revealed markedly decreased CD4 (69.9/μl) and a CD 4/CD 8 ratio of 0.09. HIV antibody was positive on both particle agglutination (PA) and Western blot (WB) analysis. Since HIV infected patients show numerous oral manifestations, dentists and oral and maxillofacial surgeons have an important role in the diagnosis and treatment of AIDS.
In this report, tooth extraction in a patient with congenital factor V deficiency is described. During transfusion of fresh frozen plasma (FFP), a wisdom tooth was extracted. The tooth socket was filled with thrombin powder and packed with periodontal dressing material and acrylic splints. On the next day, the patient again recived FFP. For extraction of an upper third molar, FFP was used as descrived above, and for local hemostatic management the extraction socket was applied an atelocollagen sponge soaked in liquid thrombin. A blood factor V level of 20% ensures adequate hemostasis.
We observed and treated mycosis affecting an extensive area of the paranasal sinuses. A 63-year-old woman was referred to our hospital because of left cheek swelling. Both maxillary sinuses were cloudy on X-ray examination. CT examination showed a low density area that included high density areas in both maxillary sinuses and the left ethmoid sinus. After these examinations, we diagnosed this case as bilateral maxillary and left ethmoid sinusitis dentalis with suspected mycosis. Surgery was performed under general anesthesia. Histological examination revealed fungi. Fungi were detected in culture specimens and identified as Aspergillus. At the same time, oral indigenous bacteria closely related to odontogenic infections, i.e., Streptococcus constellatus, Peptostreptococcus anaerobius, Prevotella intermedia, and Prevotella sp., were isolated for anaerobic culture. Clinically and microbiologically, these results suggest that this case was caused first by odontogenic infection, followed by the adherence and proliferation of Aspergillus.
We describe an intraoral foreign body that had not been noticed for a long time in a 9-year-old girl.When she was 1-year old, she mistakenly swallowed sewing needles.Some of them were removed with the use of an enema.However, further examination was not performed at that time.Subsequently, she had no symptoms for 8 years.Another needle-like radiopaque object was discovered by chance on an X-ray film.We confirmed that the object was situated in the right medial pterygoid muscle by computed tomographic and fluoroscopic examination.We removed it by an intraoral approach.