골조직의 신속한 진단을 위한 탈회방법의 비교 연구

김성철 ( Sung Chul Kim ),백운철 ( Oun Chul Back ),김태전 ( Tai Jeon Kim ),배형준 ( Hyung Joon Bae ),강희규 ( Hee Gyoo Kang ) 대한임상검사과학회 2005 대한임상검사과학회지(KJCLS) Vol.37 No.1

These studies were done to know decalcification methods to reduce the time of decalcification for quick bone tissue diagnosis. When bone tissue was decalcified with 10 % formic acid at room temperature, decalcification and hematoxylin & eosin (H&E) stains were complete and satisfactory after 12 hours, but some of the tissue sections fell off during staining. In this way, decalcification, H&E stains were complete and satisfactory after 24 hours, 36 hours and 48 hours, tissue sections didn``t fall off during staining. When bone tissue was decalcified with 10 % formic acid in a 60 ℃ paraffin oven, decalcification and H&E stains were complete and satisfactory after 6 hours, but some tissue sections fell off during staining. In this way, decalcification and tissue sections were complete, with no falling off during staining after 8 hours, 10 hours, 12 hours, 14 hours, 24 hours, or H&E stains were satisfactory from 8 hours to 12 hours, but H&E stains appeared to reddish nucleus after 14 hours and 24 hours. Bone tissue was decalcified with 10 % formic acid for 6 hours, 12 hours and 24 hours at DECAL machine frequencies of 15 Hz and 45 Hz, and for 6 hours, 12 hours and 24 hours at a DECAL machine frequency of 90 Hz. Decalcification and H&E stains were complete and satisfactory after 6 hours at the 15 Hz and 45 Hz DECAL settings. Some of the tissue sections fell off during staining at the 15 Hz DECAL machine setting. At the 90 Hz setting, decalcification, H&E stains, and tissue sections were complete and satisfactory with no falling off during staining after 4 hours. In this way, decalcification, H&E stains, and tissue section were complete and satisfactory with no falling off during staining after 12 hours, 24 hours at all machine settings. Bone tissue was decalcified with 10 % formic acid for 6 hours, 12 hours and 24 hours at 37 ℃ 3 hours, 6 hours and 12 hours at 45 ℃ and 1 hours, 5 hours and 10 hours at 60 ℃ with the RHS-1 machine setting at 60Hz. At the temperatures of 37 ℃, 45 ℃, and 60 ℃ decalcification, H&E stains, and tissue sections were complete and satisfactory, with no falling off during staining except for after 6 hours at 37 ℃. 3 hours, 1 hours, or decalcification, H&E stains, and tissue sections were complete and satisfactory with no falling off during staining after 12 hours and 24 hours at 37 ℃, 6 hours and 12 hours at 45 ℃, and 5 hours at 60 ℃. But H&E stains appeared to reddish nucleus after 10 hours at 60 ℃. From the above reults, the authors were able to deduce that decalcification is accelerated by heat and frequency. We therefore think that it is necessary for machines which are similar to the RHS-1 machine to be maintained at the temperature evenly with agitation effect for quick decalcification.

An Experimental Study for Minimum Level of Decalcification to Detect the Osteolytic Bone Metastasis of Long Bone on Plain Radiography

백준호,박일형,서성화 대한골대사학회 2016 대한골대사학회지 Vol.23 No.3

Background: In 1951, Ardran reported that metastatic bone lesions could be detectable on plain radiography with 30% to 50% of decalcification. Authors performed experimental study for minimum level of decalcification to detect the osteolytic bone metastasis of long bone with recent technique of radiographs. Methods: One pair of fibula and humerus from two cadavers was cut into specimen 1 inch in length. Distal half of specimen was dipped into hydrochloride (HCl) with 15 min interval. All 16 specimens were checked by film-type radiography (FR), computed radiography (CR), digital radiography (DR). To exclude inter-observer’s variance, 3 radiologists evaluated images. Calcium amount before and after decalcification was measured and expressed in percentage of decalcification. Results: Osteolytic changes were detectable with 11% to 16% of decalcification for fibula and 3% to 8% for humerus on plain radiography with FR, CR, and DR. Conclusions: Our study showed that minimum of 3% and maximum of 16% of decalcification is necessary when osteolytic metastatic bone lesions of long bone could be detected on plain radiography.

한국인 고정식 교정 환자의 치태, 치은염 및 탈회의 초기 변화에 관한 연구

강국진,손병화 대한치과교정학회 1999 대한치과교정학회지 Vol.29 No.3

구강내 고정식 교정장치의 장착으로 인해 치은염 및 치주염, 법랑질 탈회 및 치아 우식증, 치근 흡수, 치수변화 등 일시적 혹은 영구적 손상이 야기될 수 있다. 이러한 부작용발생의 원인으로 치태의 증가, 세균수의 증가와 조성의 변화 등을 들 수 있고 이러한 변화는 치은의 염증과 탈회를 유발한다. 이에 본 연구는 한국인 고정식 교정장치 장착환자에서 장치장착 전후의 치태, 치은염 그리고 탈회의 변화를 시간에 따른 변화, 남녀간의 차이 그리고 좌.우 소구치 부위간의 차이를 통해 알아보고자 전신질환이 없고, 여자의 경우 초경이 지난 사람을 대상으로 대조군은 연세대학교 치과대학생48명(남자 26명, 여자 22명)과 실험군으로 고정식 교정장치로 치료할 환자 73명(남자 36명, 여자 37명)을 모두 잇솔질교육(TBI)을 실시한 후, 치태치수, 치은염지수 그리고 탈회지수에 대하여 대조군은 3주 간격으로 2회를, 실험군은 최초측정을 하고 공정식 교정장치를 부착한 뒤 3주, 6주, 9주에 걸쳐 총 4회 측정을 실시하였다. 이상의 자료를 분석한 결과 다음과 같은 결론을 얻었다. 1. 치태지수(PI)는 고정식 교정장치 장착후 3주의 측정 이후 서서이 증가하였다. 2. 치은염지수(GI)는 고정식 교정장치 장착후 3주의 측정 이후 치태 지수의 증가보다 좀더 바른 속도로 증가하였다. 3. 탈회는 3주와 6주 사이에서 발생하기 시작하며, 탈회지수(DI)는 처음과 비교하여 6주 측정시부터 증가하기 시작하였으나 통계적으로 유의한 차이를 발견할 수는 없었다. 4. 좌.우 소구치 부위의 비교에서는 실험군의 치태지수와 치은염지수에서 우측에서 높은 값을 보였다. Intraoral fixed type of orthodontic appliance can cause reversible or irreversible damages such as gingivitis, periodontitis enamel decalcification, dental caries, root resorption, and pulpal changes. Such adverse effects are brought by increase in dental plaque as well as oral flora. Such an increase cause gingival inflammation and enamel decalcification. The purpose of this study is to get knowledge on initial changes in dental plaque, gingivitis, and enamel decalcification after bonding fixed orthodontic appliances according to time flow, gender, and sides(right/left) of premolar region. For control group, 48 students of dental college, Yonsei university(26 males, 22 females) were chosen; for experimental group, 73 orthodontic patients(36males, 37 females) who will be treated with fixed appliances were chosen. All the subjects had no systemic disease, juvenile periodontitis and all the females had passed their menarche. Tooth brushing instruction was given to all the subjects prior to the experiment. for control group, plaque index, gingival index, and decalcification index were measured twice at 3 weeks interval ; for experimental group, the same was done prior to, 3, 6, 9 weeks after bonding fixed appliances. The following results were obtained: 1. In plaque index 3 weeks after placement of appliances, and it showed gradual increase afterwards. 2. In gingival index 3 weeks after placement of appliances, and afterwards it showed increase at a faster rate than plaque index. 3. Enamel decalcification began to show between 3 and 6 weeks after bonding fixed appliances. Decalcification index began to increase 6 weeks appliance placement, but there was no statistical significance. 4. When the comparison was made between two sides of premolar region, the right side showed greater index in plaque and gingival index of experimental group.

Microbially induced calcium precipitation based anaerobic biosynthetic crystals for removal of F− and Ca2+ in groundwater: Performance optimization, kinetics, and reactor operation

Zhenyu Zhai,Amjad Ali,Junfeng Su,Zhenle Hao,Jiaran Liu,Zhao Wang 한국화학공학회 2022 Korean Journal of Chemical Engineering Vol.39 No.11

Anaerobic biosynthetic crystals (ANBC) were prepared based on microbially induced calcium precipitation (MICP) and their potential explored for groundwater defluoridation and decalcification. The preparation conditions of ANBC were optimized and the influence of key factors (initial fluoride ions (F−) concentration, pH, and initial calcium ions (Ca2+) concentration) on the crystals was investigated. During the operation of the reactor, at pH of 7.0, the hydraulic retention time (HRT) of 6 h, and Ca2+ concentration of 180 mg L−1, a maximum removal efficiency reached 93.31%, while 66.20% of Ca2+ could be removed. The adsorption dynamics study showed that the adsorption of ANBC was most in line with the pseudo-second-order model. The stability of ANBC operation was studied and failure reaction showed that the crystals maintained a stable removal ability after 35 times of repeated use. Further studies found that this was attributed to the continuous growth and synthesis of the crystals. The defluoridation and decalcification mechanism was further explored by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), and X-ray diffraction spectra (XRD). This study innovatively proposes a method for biosynthesis of crystals under anaerobic conditions based on MICP, which can efficiently and stably remove F− and Ca2+ in groundwater, providing a valuable strategy for groundwater contaminant remediation and energy saving.

불소바니쉬가 법랑질 탈회에 미치는 영향

윤명옥,이난영,이상호 大韓小兒齒科學會 2008 大韓小兒齒科學會誌 Vol.35 No.3

이 연구의 목적은 법랑질 탈회부위에 불소 바니쉬 적용의 효과를 평가하는 것이다. 80개의 소의 법랑질 블록을 무작위로 4군으로 나누었다. 1군은 대조군으로 바니쉬 적용을 하지 않았다. 2군은 APF gel을 도포하고 4분후에 세척하였다. 3군과 4군은 Fluor Protector^(ⓡ)와 CavityShield™를 도포하고 1분 후에 세척하였다. 모든 표본을 인공우식용액 속에 넣어서 탈회시켰다. 가시광선 형광을 이용해 병소의 광밀도를 측정하고 병소의 깊이를 현미경으로 측정하고 통계적으로 비교하였다. 1. 48시간 후 광밀도 측정시 2군 광밀도는 대조군에 비해서 높았으나 바니쉬군보다 낮았고,두 가지 바니쉬 간에 유의한 차이는 없었다. 2. 72시간 후 광밀도 측정결과 3군에서만 약간의 광밀도 감소가 관찰되고,4군이 가장높았다. 3. 72시간의 탈회 후 병소의 깊이를 측정한 결과 대조군에서는 평균 205.36±42.85㎛, 2군에서 병소의 평균깊이는 21O.81±44.60㎛로 두 군간에 유의한 차이는 없었다. 4. 불소 바니쉬군의 병소 깊이는 3군이 80.03±21.66㎛, 4군이 77.46±27.72㎛로 대조군이나 2군에 비해 탈회가 억제되었음을 알 수 있었으나 두 가지 바니쉬 사이에 차이가 없었다. The aim of this study was to evaluate the effect of fluoride varnish application on enamel decalcification. Eighty bovine enamel blocks divided randomly into 4 groups. Group I is the control group. Group II was treated with the APF gel and washed after 4 minutes. Group III and IV was treated with Fluor Protector^(ⓡ) and CavityShield™ and washed after 1 minutes. Decalcification were created by placing all specimen into artificial acidic solution(pH4.0). Then the optical density of the lesions were measured by visible light fluorescence and the lesion depths were measured. The results were : 1. The optical density of group II was higher than group I but lower than group III, IV(p<0.05) and there was no difference between group III, IV(p>0.05) at 48 hours. 2. The optical density of group IV was highest at 72 hours(p<0.05). 3. Mean lesion depths were 205.36±42.85㎛ and 210.81±44.60㎛ in group I, II but no significant difference between two groups(p>0.05). 4. Mean lesion depths were 80.03±21.66㎛ and 77.46±27.72㎛ in group III, IV but no significant difference between two groups(p>0.05). Fluoride varnish treatment resulted in a significant reduction in lesion depth compared with APF gel. Fluor Protector^(?) and CavityShield™ provided the similar effect.

골수검사시 탈회용액의 조건에 관한 연구

임형선 ( H S Lim ) 대한임상검사과학회 1991 대한임상검사과학회지(KJCLS) Vol.23 No.1

Examination of marrow helps diagnosis of blood disease, inflammatory disease, transfer of malignant tumor to the marrow, and determination of their curing methods, judgement of convalescence and post curing course. It is very important that decalcification process for making the marrow soft by removal of calcification due to calcium salt in the marrow does not affect other intracellular materials in the examination of marrow. And therefore, in this study, experiments were made on stability and proper condition of decalcification solution during decalcification processes with marrow of rabgbit using four types of decalcification solution such as formic acid solution, acid modify solution, Zenker``s solution, and rapid solution during different time at different pH, respectively, in order to solve problems in production of specimen of marrow by determination of stability and proper condition of decalcification solution in decalcification process and furthermore to seek methods to help the examination of marrow. The results are as follows though ther are some differences depending upon the type of decalcification solution ; 1. 5% Formic acid solution Seven (7) hours decalcificaiton at pH 4. 0 was proper for cutting. Intracellular iron was detected to be positive in seven hours decalcification at pH 3.0-4.0 2. Acid modify solution Cutting and/ or dyeing of iron was good in case of five(5) to seven(7) hours decalcifcation at pH 3.0-4.0 3. Zenker``s solution At least ten (10) hours or more decalcification was good for cutting at pH 4. 0, and iron was detected to be positive in fifteen (15) to twenty (20) hoUI~s decalcification at pH 2. 0- 3. 0 or that at pH 3.0-4.0. 4. Rapid decalcifcaiton solution More than thirty (30) minutes process is good for cutting and for detection of iron, at least twenty minutes or more is required. Thus, it is thought that Zenker``s solution which stabilizes intracellular material shows less fluctuation according to time change and relatively much iron detecting property according to time and variation of pH, for examination of marrow in rabbit.

청소년 교정환자들의 치은염 및 치아탈회 조절을 위해 사용한 겔형 불화주석(SnF_2 gel)의 장기간 평가

Boyd, Robert L.,전윤식 대한치과교정학회 1995 대한치과교정학회지 Vol.25 No.3

이 논문의 목적은 청소년 교정환자들의 치은염과 치아탈회를 조절하기위해 사용된 수종의 화학요법들에 관해 최근에 보고된 2개의 연구를 재검토하기 위함이다. 첫번째 연구(치은염 연구)는 고정성 교정장치를 장착하고 있는 경우, 기존의 칫솔법에 매일 2회 유효 주석이온이 90%이상 함유된 0.4%의 겔형 불화주석(SnF_2 gel)을 함께 사용하는 경우와 기존의 칫솔법만을 사용하는 것 중 치내침착과 치은염을 조절하는데 있어서 어느 것이 더 효과적인지를 결정하기 위하여 시행 되었다. 두번째 연구(치아탈회 연구)는 교정환자들을 대상으로 1100 ppm의 불소가 함유된 치약만을 사용할 때와 이와 똑 같은 치약과 0.05% 불화나트륨 양치액(NaF rinse)을 병용하여 양치하거나 이 치약과 함께 0.4% 겔형의 불화주석을 사용할 때 치아탈회의 조절효과를 비교하기 위하여 시행 되었다. 치은염 연구에서는 모든 치아를 고정성 장치로 계속 치료를 받고 있는 청소년 교정환자 65명을 연령과 성별에 따라 두 군으로 지정하였다. 마찬가지로 치아탈회 연구에서도 30명을 추가(총 95명)하여 제 3군으로 지정하였다. 제1군(대조군, 35명)은 단지 표준화 불소(1100 ppm 불소) 치약만 사용하였다. 제 2군은 1 군과 같은 치약에 겔형의 0.4% 불화주석이 함유된 치약을(겔형 불화주석군, 표본수=30) 매일 2회씩 18개월 동안 사용하였다. 제 3군은(단지 치아탈회군만으로 이용) 같은 종류의 치약과 0.05% 불화나트륨 양치액을 사용하였다(불화나트륨 양치구, 표본수=30). 치태침착의 임상평가는 Plaque Index로, 치은의 염증은 Gingival Index로, 치관의 착색은 단일맹검(single-blind)으로, 고정성 교정장치를 장착하기 전과 장착한 수 1, 3, 6, 9, 12, 18 개월마다 실시하였다. 치아탈회의 임상평가는 맹출한 모든치아의 순측면에 고정성 교정장치를 장착하기 전과 장치 제거 3개월 후에 단일맹검으로 실시하였다. 치은염 연구에서 겔형 불화주석군(SnF_2 gel group)이 대조군에 비해 교정치료 기간동안 시행한 모든 검사에서 Plaque Index(p<0.01)와 Gingival Index(p<0.001)가 상당히 낮은것으로 나타났다. 겔형 불화주석군에서 한 증례는 미미한 치관착색을, 두 증례는 보통정도의 치관착색을 보였다. 치아탈회 연구에서는 겔형 불화주석군과 불화나트퓸 양치액군이 치료후 치아탈회값에서 치료전 치아탈회값을 뺀 치아탈회값이 대조군에 비해 구강전체및 제1대구치에서 현저하게 낮은 값(p<0.05)을 보였다. 비록 겔형 불화주석군이 불화나트륨 양치액군보다 일관되게 낮은 치아탈회값을 보였을지라도 통계적으로 그 차이는 단지 유의성을 보이는 정도였다. The purpose of this paper is to review two recently reported, long-term studies of several chemical methods to control gingivitis and decalcification in adolescent orthodontic patients. The first study(gingivitis study) was designed to determine whether conventional toothbrushing and twice daily use of a brush-on 0.4 per cent SnF_2 gel containing more than 90 per cent available Sn^2+ would be more effective for controlling plaque accumulation and gingivitis in the presence of orthodontic appliances than conventional toothbrushing alone. The second study(decalcification study) was designed to compare the effectiveness of controlling decalcification in orthondontic patients with either a 1100 ppm F tooth paste used alone, this same toothpaste and a 0.05 percent NaF rinse or this toothpaste and a 0.4 percent SnF_2 gel. In the gingivitis study, sixty-five consecutively treated adolescents who were to receive full-mouth fixed orthodontic appliances were assigned to two groups according to age and sex criteria. In the decalcification study and additional 30 subjects(95 total) were similarly assigned to a third group. The first group(control, n=35) used only toothbrushing with a standard fluoride(1100 ppm F) toothpaste. The second group used toothbrushing with a similar dentifrice supplemented with a 0.4 percent SnF_2 gel(SnF_2 gel group, n=30) used twice daily for the entire 18-month study period. The third group(in the decalcification study only) used a similar toothpaste and 0.05 percent NaF rinse(NaF rinse group, n=30). Clinical assessments of plaque accumulation using the Plaque Index, gingival inflammation using the Gingival Index, and coronal staining were completed single-blinded before appliances were placed and 1, 3, 6, 9, 12 and 18 months after appliances were placed. Decalcification was assessed single blind on all labial surfaces of all erupted teeth before appliances were placed and 3 months after appliances were removed. The results of the gingivitis study indicated that the SnF_2 gel gorup had significantly lower scores for the Plaque Index(p<0.01) and Gingival Index(p<0.001) at all examinations during orthodontic treatment than did the control group. In the SnF_2 gel group, one subject developed mild coronal staining and two subjects developed moderate staining. In the decalcification study, when pre-treatment levels of decalcification were subtracted from post-treatment values, significantly lower decalcification scores(p<0.05) were found for both whole mouth and first molars in the NaF rinse and gel groups as compared with the control group(toothpaste alone). Although the gel group consistently had less decalcification than the rinse group, this difference only approached statistical significance.

맥동성 단극 자기장에 의한 탈회 촉진 효과

이석근,정은영,김기진,송대범,김주호,지제근 대한병리학회 1993 Journal of Pathology and Translational Medicine Vol.27 No.2

Proposal of an Appropriate Decalcification Method of Bone Marrow Biopsy Specimens in the Era of Expanding Genetic Molecular Study

최성은,윤선옥,홍순원 대한병리학회 2015 Journal of Pathology and Translational Medicine Vol.49 No.3

Background: The conventional method for decalcification of bone specimens uses hydrochloric acid (HCl) and is notorious for damaging cellular RNA, DNA, and proteins, thus complicating molecular and immunohistochemical analyses. A method that can effectively decalcify while preserving genetic material is necessary. Methods: Pairs of bilateral bone marrow biopsies sampled from 53 patients were decalcified according to protocols of two comparison groups: EDTA versus HCl and RDO GOLD (RDO) versus HCl. Pairs of right and left bone marrow biopsy samples harvested from 28 cases were allocated into the EDTA versus HCl comparison group, and 25 cases to the RDO versus HCl comparison group. The decalcification protocols were compared with regards to histomorphology, immunohistochemistry, and molecular analysis. For molecular analysis, we randomly selected 5 cases from the EDTA versus HCl and RDO versus HCl groups. Results: The decalcification time for appropriate histomorphologic analysis was the longest in the EDTA method and the shortest in the RDO method. EDTA was superior to RDO or HCl in DNA yield and integrity, assessed via DNA extraction, polymerase chain reaction, and silver in situ hybridization using DNA probes. The EDTA method maintained intact nuclear protein staining on immunohistochemistry, while the HCl method produced poor quality images. Staining after the RDO method had equivocal results. RNA in situ hybridization using kappa and lambda RNA probes measured RNA integrity; the EDTA and RDO method had the best quality, followed by HCl. Conclusions: The EDTA protocol would be the best in preserving genetic material. RDO may be an acceptable alternative when rapid decalcification is necessary.

골수생검조직의 조직병리검사에서 탈회방법에 따른 결과 분석

박지영 ( Ji Young Park ),한경희 ( Kyung Hee Han ) 대한임상검사과학회 2016 대한임상검사과학회지(KJCLS) Vol.48 No.4

Decalcification is routinely performed to obtain a pathological diagnosis using bone marrow biopsy. During the decalcification process using a conventional acidic solution, such as HCl, the antigenicity of tissue is damaged. Especially DNA and RNA in the bone marrow are impaired. Hence, there is the need for a standardized decalcification protocol that preserves the antigenicity of tissue. To this end, we compared the effects of two commonly used decalcifiers: Commercial decalcifier (Calcl-Clear Rapid, HCl) and the EDTA (12.5%, pH 7.0). Bone marrow biopsies sampled from 71 patients were decalcified in accordance with the protocols of respective groups―HCI versus EDTA. The differences of decalcification protocols were analyzed with respect to Hematoxylin & Eosin staining, Gomori`sreticulum staining, and immunohistochemical staining and molecular analysis. Immunohistochemical staining used Ki-67, CD20 and CD138 as primary antibodies and molecular analysis was conducted through the DNA concentration analysis, in situ hybridization (ISH) and immunoglobulin heavy chain (IGH) gene rearrangement. On the routine histopathology analysis, there was no difference between HCl and EDTA. Moreover, in case of immunohistochemical staining, the cytoplasmic membrane or cytoplasmic CD markers was well preserved. However, nuclear proteins, such as Ki-67, were stained with low quality. Conversely, according to the molecular analysis, the EDTA protocol preserved the DNA and RNA compared with the HCI. The differences of DNA quantity and quality were statistically significant between protocols of HCl and EDTA. We used 38 cases in HCl and 12 cases in EDTA. Consequently, the EDTA protocol maintains the antigenicity of the protein on tissue and is acceptable for examination with molecular base analysis. Decalcification of bone marrow biopsy by EDTA is highly recommended for the examination of immunohistochemical staining and molecular analysis.