No report of lipids analysis of human gingiva in different degrees of chronic marginal periodontitis has so far appeared except OTA et al. of our department earlier reported. It was suggested that the content of cholesterol ester in human gingiva with periodontal disease revealed a tendency decrease.
In the present study, the author investigated the measurement of content of free cholesterol, cholesterol ester and total cholesterol in the gingiva, that occured in association with the progress of chronic marginal periodontitis. At the same time, a qualitative analysis was also made to identify main fatty acid composition of cholesterol ester.
A study material consisted of 19 males and 11 females, ranging from 18 to 50 in age, who were diagnosed as periodontitis in the department of periodontology, Nihon University Dental Hospital, and were divided into three clinical groups of P₁, P₂ and P₃ by OKADA's classification. As controls, P₀ group who were diagnosed as healthy gingiva consisted of 6 males and 4 females, ranging from 20 to 40 in age.
The obtained gingival specimen was extracted for total lipids by FOLCH et al's method. An aliquot of total lipids was spotted on a Silicagel G glass plate. After developing, free cholesterol and cholesterol ester fractions were identified. Free cholesterol and cholesterol ester in appropriate areas of the individual fractions were then scraped off and re-extracted. Gas liquid chromatography was carried out for qualitative and quantitative analysis.
Results were obtained as follows:
1. The study on content of free cholesterol and total cholesterol in different degrees of chronic marginal periodontitis, there was no statistical significance among each clinical group.
2. But the content of cholesterol ester revealed a tendency of decrease according to the severity of the disease, and it showed significantly different to compare P₁ group with P₂, P₃ group.
3. The main fatty acid composition of cholesterol ester was C_14:0, C_16:0, C_16:1, C_18:1 and C_18:2. However, there was no change in different degrees of chronic marginal periodontitis.

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The mouthrinses be carried out for prevention and inhibition of periodontal diseases and dental caries. Clinical studies have shown that mouthrinses with 0.2 per cent solution of Chlorhexidine prevent the formation of dental plaque (Löe and Schiött). This clear preventive effect of dental plaque formation considered that Chlorhexidine had strong bactericidal action and antibacterial durability.
The purpose of the present study was to compare to what contents the sialic acid of human mixing saliva were influencened by Chlorhexidine and each mouth-washs be on the market (Acrinol, Oradol and Lavoris).
The author carried out the experiments as follows: In vitro, variations of sialic acid contents were compared to influence in cases that 0.2 per cent solution of Chlorhexidine and other mouth-washs be on the market which were annexed to human mixing saliva and then collected from the subjects.
In vivo, it observed that the variations of sialic acid contents at interval of an hour from 8:30a.m. to 5:30p.m. by mouthrinses with 0.2 per cent solution of Chlorhexidine after taking regular meal.
Forty males dental student, 19 to 24 years of age included in these investigations.
Quantitative analysis of sialic acid contents was colourimetric determination by Aminoff's method.
The following conclusions are to be drawn.
1. In vitro, sialic acid decreased in saliva after 0.2 per cent of Chlorhexidine used that was more than other mouth-washs be on the market.
2. In vivo, sialic acid contents indicated considerable decrease in which mouthrinses with 0.2 per cont solution of Chlorhexidine after taking regular meal, but this value retrieved about one hour.
3. From the above, Chlorhexidine used for salivary mucin reduction, is due to effectiveness in inhibiting of dental plaque formation. The continuous usage was advised because the effective duration is limited.

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The color of gingiva plays an important role in the treatment of the periodontal diseases in that furnish valuable guides concering the diagnosis, observation of curative processes and prognostic judgement. It is said to be greatly influenced by such factors as the conditions of blood circulation, blood vessels, nature of the blood, the thickness of gingival epithelium, the degree of keratinization and deposition of certain pigments.
Little work has been done to study on the correlation between the gingival color and those factors. The author compared the gingival color with histopathological findings in the purpose of investigating with the correlation between the gingival color and those factors.
A study material consisted of 18 males and 19 females (except the menstrual cycles), ranging from 19 to 27 in age, who were diagnosed as clinically normal gingiva, and 10 males and 5 females, ranging from 19 to 58 in age, who were diagnosed as necessity of gingivectomy in the labial gingiva of both upper and lower anterior teeth in the department of periodontology, Nihon University Dental Hospital.
With each subjects, measurements were made of the gingival color in the interdental papilla and the marginal gingiva, by using new type color meter (CM-5A).
In inflamed gingiva, immediately after the measurements, the same portions were photographed by the capillary-microscope and then were provided for biopsy.
The obtained gingival specimen were divided into three groups by Munsell hue. Each group was observed capillary-microscopically and histpathologically.
The results were as follows:
1. The measured values of the clinically normal gingiva.
In the interdental papilla
4.6 R-5.0 YR 3.9-4.9/1.4-3.1
In the marginal gingiva
5.3 R-6.5 YR 4.0-4.9/1.4-4.0
Those measured values agreed approximately with the report of KANEKO.
2. The measured values of the inflamed gingiva.
In the interdental papilla
2.1 R-4.2 YR 3.3-4.6/1.6-4.0
In the marginal gingiva
3.0 R-4.5 YR 3.8-5.2/1.5-3.5
One third of the specimen was not in the range of the color in the clinically normal gingiva.
3. Histopathologically, in the inflamed gingiva, when the type of the keratinization and the thickness of the keratinization and the gingival epithelium were in certain range, they had few effects on the hue. Dut the correlation btween the grade of dilatation of blood vessels in gingival papilla and the hue was observed. In particular, the hue was seemed to be influenced by the width and the height of the papilla, and the thickness of the epithelium at the apex of the papilla.
4. Capillary-microscopically in the inflamed gingiva, the correlation between the hue and both the mean diameters of gingival capillaries, and the mean number of gingival capillary loops was. not remarkable as well as the report of HIRATSUKA.
As were confined by the results above, this apparatus is handy clinically, can be operated in a short time, and that the measured values on this study is the same as the experimental data in the past. These values suggest to be more available for the observation of the progressing than the diagnosis of the periodontal diseases.

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A comparison between macroscopic evaluation and cytological findings in the labial gingiva of maxillae and mandible. Histopathological findings were also examined, including the therapeutic course was made on periodontal diseases of 81 patients selected randomly. Macroscopic evaluation was made on four items, reddening, swelling, bleeding, pus discharge by a predetermined standard. The overall results were determined as the inflammation grade (IG). In classifying exfoliative cytology, exofoliative epithelium were divided in terms of positional morphology into 4 types, deep layer type (Dt), intermediate layer type (It), nucleated superficial layer type (S₁t), non-nucleated superficial layer type (S₂t). According to the staining attitude, they were divided into blue type (Bt), red-blue mixed type (RBt), red type (Rt), yellow type (Yt).
Additionally, an all type (At), blue type seen in the intermediate layer type (ItBt) were added. As a result of the examination, the following results were obtained.
(1) As the inflammation grade increases, Dt or Bt increases, whereas S₂t or Yt decreases. It, irrespective of the inflammation grade, occupied the majority of the exfoliative epithelium.
(2) Changes by inflammation grade is obscure excepting that turbidity, poor staining tendecy, indistinctness of the border, small vacuole and large vacuole become conspicuous at inflammation grade 8 as changes in the cytoplasm.
Nuclear changes show a tendency towards hyperchromatosis or hypochromasia, and karyosome increases as inflammation grade increases.
(3) In all inflammation grade, no change was seen on cytoplasm-nucleus ratio (long axis).
(4) As inflammation grade increase, appearance of leukocytes and bacteria tends to increase.
(5) The observation of the course using various local treatments showed a decrease both Dt and Bt from the second week and a tendency towards recovery was seen. Bacteria attachment decreased in the first week in periodontal curettage, gingivectomy and flap operation. Subsequently the same condition was maintained.
(6) The appearance of cells in each type according to positional morphology and staining attitude corresponding to inflammation grade, and changes of epithelial layer were confirmed histologically.
The above results demonstrate that the exfoliative cytological examination can express morphological changes of the pocket epithelium in the periodontal diseases to a considerable degree. Consequently, it is of use to accurately grasp the fluctuation in the disease process.

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The author attempted to measure the tissue respiration in gingiva by minimal blood with accurate analytic apparatus instead of using the Warburg's manometric technic.
So that, the pH value, pO₂, pCO₂, HCO₃^-, totalCO₂, base excess (B. E.), buffer base (B. B.) and standard bicardonate (S. B.) of the venous blood and the arterial blood were evaluated to the basic data with the Corning Evans pH/Blood Gas 165.
The data of the result were compared with the reports of the other authors.
Then, the author measured the arterial blood of the periodontal patients in order to check out its correlation with the degree of the inflammation.
The subjects were divided into two groups. The clinical normal group consisted of 18 students of Nihon University School of Dentistry from 18 to 25 in age, observed no oral lesion and other systemic disease.
The other group included 100 periodontal patients from 18 to 65 in age that were diagnosed by Periodontal Department of Nihon University Dental Hospital.
After the patient were sat in resting position, the venous blood was taken from the left medial cubital vein for about 6.0ml and the arteral blood was taken from the earlobe for about 0.125ml.
The following results were obtained.
1. The basic data of the tissue respiration in gingiva were obtained from the minimal venous and arterial blood with the above apparatus that were accurate and consistend with the value of the references.
2. The value of pO₂ determined by the arterial blood taken from the earlobe in the clinical is noted.
3. The value of pO₂ and base excess of the arterial blood were much higher in males than in females that were consistent with the other author's reports. But the value of HCO₃^-, total CO₂ and buffer base were not compared with the others.
4. The measured value between the clinical normal and the periodontal patients didn't have any obvious difference but in the metabolic activity the latter had a slight decreasing tendency than the former. Because the pH value of the blood and partial gas pressure were influenced by many local and systemic facter.
It is difficult, to state at this time that the cause of the deviation is the exact character of the periodontal disease.
5. The result that the value of pO₂ in arterial blood many decrease when the physiological age increase were consistent with the other author's reports.

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Germfree JCL: ICR strain mice were inoculated intraorally with 1.0×10⁸ colony forming units (CFU) of a plaque-forming streptococcus strain, Streptococcus mutans PK1, bred in the plastic isolator up to 10 weeks and tested at appropriate intervals for the establishment of the organism in the mouth and digestive tract. All specimens isolated from the tooth surface, stomach, duodenum, ilum and colon gave, in every mice tested, a positive reaction for qualitative cultural examinations using thioglycollate culture medium.
Growth of the organism was the greatest in the colon among the mouth and digestive tract and followed by the growth in the ileum. Measurement of viable numbers of the organism resulted in an average of 2.5×10⁹CFU/ml for the specimens from the colon. Number of the organism isolated from the digestive tract other than colon varried between less than 2.0×10⁵ to 10⁹CFU/ml. The organism isolated from the specimens of tooth surface were in the level of less than 1.4×10⁴ to 2.7×10⁶CFU/g and 1.8×10⁵ to 9.9×10⁶CFU/g at 2 and 6 weeks after the inoculation, respectively. Addition of sucrose in the drinking water gave no statistically significant increase viable cell numbers in any specimens at least within 4 weeks after inoculation.
In conclusion, it was found that Str. mutans PK1 could be established in the mouth and digestive tract of JCL: ICR strain germfree mice by a single inoculation of the organism.

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The clinical influence of exposed cementum is a matter of interest for the periodontist. It may often be speculated that the exposed cementum left in situ during the periodontal flap surgery would affect the healing resulting in an incomplete attachment of the flap, and bring about some unfavourable biological events. However, no controlled experiment has been found out in this respect.
In this experiment, the alveolar bone of Rhesus monkeys were chiseled away, and the bone defect with standardized shape and size were created. The copper strip covered by a silk cloth was placed into the bone defect, so that the cementum had been soarked in the artificially produced periodontal pocket for 4 weeks. At the flap surgery with reverse bevel incision, the exposed cementum was curetted away completely in the control group, but it was left unattached and covered by the gingival flap in the experimental group.
Sixteen weeks after the surgery, two groups were compared both by the extent of alveolar bone regeneration and by the degree of the new attachment. To evaluate the bone regeneration rate objectively, planometric measurement were made.
As the results of the experiment, obvious bone regeneration was observed in the control group, on the other hand, the marked bone destruction was noticed in the experimental group which difference was statistically significant (P<0.05). On the histologic specimens stained by H. E. and Azan-Mallory, no great difference was found between two groups. However, the functional orientation of periodontal fiber was insufficient in the experimental group.
The vital and the non-vital tooth (which root canal treatment had been completed) was compared by the same way, but no difference was found histologically and planometrically.
It could be concluded that the complete curettement of the exposed cementum is essential f or the more predictable periodontal flap surgery.

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The method of facilitating accumulation of dental plaque by preparating the cavities on the mesiolinguales at the cervical portions of the upper molars of rat was devised.
Following the intramuscular injection with 1ml bismuth/kg/day for experimental period, the changes with lapse of the time after bismuth injection in gingival tissue were studied macroscopically, histopathologieally, and by means of electron probe microanalysis (EPMA).
In the conttrol group, preparatting no cavities on the teeth, injected with bismuth.
1. In the control group, no bismuth deposition was observed in the gingival tissue macroscopically and histopathologically.
2. Bismuth line formation in the gingival tissue.
1) Light microscopic observation.
a: After one day, no bismuth deposition was seen in the gingiva.
b: After 3 days, bismuth granules were deposited around the capillaries in the lamina propria of gingiva beneath the bottom of the sulcus epithelium.
c: After 7 days, a large amount of bismuth depositions was seen along the sulcus epithelium.
d: After 14-35 days, a large amount of bismuth depositions was seen over the whole lamina propria of the gingiva.
2) Electron microscopic observation.
a: After one day, fine bismuth granules were found around the capillaries in the lamina propria of gingiva beneath the bottom of the sulcus epithelium and basement membrane of the epithelium.
b.: After 3 days, these fine bismuth granules were observed in the lysosome-like vesicles of the fibroblast and histiocyte-like cells.
c: After 7 days, these fine granules gradually grew in size to form larger precipitation in the lysosome-like vesicle, and f ibroblast and histiocyte-like cells which had taken up a large amount of fine bismuth granules exhibited some degenerations which were at first apparent in the membrane system such as rough-surfaced endoplasmic reticulum and Golgi apparatus.
d: After 14-35 days, some degrees of degenerations were found in the cells in the lamina propria of gingiva and bismuth depositions were composed of degenerated nuclei, intracellular organelles of the cells and many aggregation of bismuth granules.
e: Throughout the entire experimental period, no fine bismuth granules were found within the epithelial cells of the gingiva and intercellular space.
3. The dark brown depositions within gingival tissue were indicated to be the sulfide bismuth by means of electron probe microanalysis.

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This study was performed to examine the effect of modern dentifrices on the abrasion of acrylic resin and human teeth.
Those specimens were brushed with a tooth brushing machine for two hours, corresponding to 3000 back and forth strokes. The surface irregularities of the specimen were analyzed by a surface analyzer, and observed by scanning electron microscopy before and after the brushing procedure. The weight loss of the test specimens of acrylic resin materials was determinded, weighing them before and after brushing by a microbalance.
The effect of hardness of toothbrush on the abrasion was tested, using three different toothbrushes.
The abrasiveness by the dentifrices showed a wide range.
All dentifrices except one dentifrice which did not contained abrasive materials had considerable abrasive effects on both acrylic resin and human teeth. There was a high correlation (r=0.96) between the weight loss and the surface irregularities in the case of acrylic resin.
Without using dentifrices, abrasion was not observed on both acrylic resin and human teeth.

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We made examination of braided silk, chromic catgut, polyglycolic acid and nylon in human periodontal tissues. Operative procedures were performed on 57 male and female patients. They had sutures placed 3, 7, or 14 days preoperatively in the tissue at the prospective operative site and at the time of surgery tissue reaction to sutures was graded clinically and studied histologically. The results were as follows.
1) Clinically tissue reactions increased respectively from nylon, PGA, braided silk to catgut.
2) Histologically neutrophiles around catgut and braided silk infiltrated more than nylon and PGA, so the former experienced more severe reaction than the latter.
3) The fact that the sutures were multifilament or monofilament had no relation to the physical reactions.

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How to obtain good graft tissue in free mucosal grafts is key point in successful grafts.
We have devised a mucotome (I T O type) that can obtain graft tissue from palatal mucosa (donor site).
It has been clinically applied in periodontal surgical procedures and it has proved to be satisfactory. The structure of this instrument consists of a frame, a handle and a blade holder, and it is designed as simply as possible in consideration of the operative field in the oral cavity.
As the blade (edge of a stainless steel razor) is disposable, a sharp blade can be used every time, and the curve of the blade can be adapted to the individual palate by adjusting the screws of the blade handle, thereby causing the blade within a range of 20 to 30mm from end to end of the bent blade.
This article describes formation and features of our devised mucotome and a technique for obtaining partial thickness palatal mucosal graft.

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