Zappa U, Boretti G, Graf H, Case D
Abteilung für Parodontologie, Universität Bern, Schweiz.
J Periodontal Res. 1992 Jul;27(4 Pt 1):274-84. doi: 10.1111/j.1600-0765.1992.tb01678.x.
The purpose of the present study was to develop an intracrevicular lavage technique, and to use it for assessment of the total number and % of vital leukocytes from untreated periodontitis pockets. A lavage device was developed, consisting of specially crafted cannulas, a vacuum pump and a charge amplifier. In vivo evaluations showed that there was a linear relationship between lavage time and lavage volume; 2-6 lavage samples were obtained from each of 20 patients with untreated advanced periodontitis, and the lavage time was measured in a subsample of 16 pockets. The lavage fluid was centrifuged, and the supernatant was separated from the cellular components. The cells were vital-stained using two methods, trypanblue exclusion method (TB) and ethidium-bromide fluorescein-diacetate stain (EB-FDA). Numbers of vital and non-vital leukocytes per sample were assessed using a Neubauer chamber. The number of erythrocytes per sample was evaluated using the same counting method. The results included 95 samples obtained from the 20 patients: 76% of all samples ranged in pocket depth between 4 and 8 mm. The lavage technique provided an overall mean lavage volume of 282.37 microliters in an average time of 16.44 seconds. The mean number of leukocytes per sample was 22.20 x 10(3) (TB) and 24.48 x 10(3) (EB-FDA). Percent vital leukocytes were 72.27 (TB) and 72.63 (EB-FDA). EB-FDA had a lower counting error than TB. The low erythrocyte counts per sample suggested that subgingival bleeding during sampling was negligible. Spearman correlation coefficients showed weak associations between pocket depth and leukocyte counts, % of vital leukocytes and erythrocytes. Due to the short sampling time this new intracrevicular sampling technique permits sampling of pockets before the tissue responds to the stimulus of the lavage device, and provides subgingival washings with high numbers of leukocytes.
本研究的目的是开发一种龈沟内灌洗技术，并将其用于评估未经治疗的牙周炎牙周袋内活白细胞的总数及百分比。开发了一种灌洗装置，其由特制的套管、真空泵和电荷放大器组成。体内评估显示灌洗时间与灌洗量之间存在线性关系；从20例未经治疗的重度牙周炎患者中的每例获取2 - 6份灌洗样本，并在16个牙周袋的子样本中测量灌洗时间。将灌洗液离心，使上清液与细胞成分分离。使用锥虫蓝排除法（TB）和溴化乙锭 - 荧光素二乙酸酯染色法（EB - FDA）这两种方法对细胞进行活细胞染色。使用血细胞计数板评估每个样本中活白细胞和非活白细胞的数量。使用相同的计数方法评估每个样本中的红细胞数量。结果包括从20例患者获取的95份样本：所有样本的76%牙周袋深度在4至8毫米之间。灌洗技术在平均16.44秒的时间内提供的灌洗总量平均为282.37微升。每个样本白细胞的平均数为22.20×10³（TB法）和24.48×10³（EB - FDA法）。活白细胞百分比分别为72.27（TB法）和72.63（EB - FDA法）。EB - FDA法的计数误差低于TB法。每个样本中红细胞计数较低表明采样期间龈下出血可忽略不计。Spearman相关系数显示牙周袋深度与白细胞计数、活白细胞百分比和红细胞之间的关联较弱。由于采样时间短，这种新的龈沟内采样技术允许在组织对灌洗装置的刺激产生反应之前对牙周袋进行采样，并提供含有大量白细胞的龈下冲洗液。
