不同处理液对脱位牙牙周膜修复作用的实验研究

目的观察比较Hanks'平衡盐溶液(简称HBSS)、牛奶及2%Na F这几种不同处理液对干燥放置的脱位牙牙周膜作用效果的差异,为临床选择适宜的处理液提供实验依据。方法收集因正畸治疗需要而拔除的前磨牙277颗模拟外伤脱位牙,在不同干燥放置时间点分别用HBSS、牛奶及2%Na F进行浸泡处理后,采用台盼蓝染色计数法及HE染色技术,观察比较各种条件下根面牙周膜细胞的存活状况及组织学表现。结果干燥时间为0.5 h时,HBSS组牙周膜细胞存活率(94.60±0.72)%,大于2%Na F组(84.37±3.98)%,差异有显著性(P<0.05);干燥时间为2、4 h时,HBSS组牙周膜细胞存活率(63.03±3.05)%、(13.38±1.04)%大于牛奶组(48.9±3.47)%、(5.73±1.29)%及2%Na F组(51.93±1.72)%、(8.60±0.74)%,有统计学意义(P<0.05)。结论不同处理液对于离体干燥保存的脱位牙的牙周膜的修复作用各有不同,HBSS的效果相对较好。     

不同处理条件对脱位牙牙周膜形态影响的扫描电镜观察

目的 观察不同方式处理的脱位牙牙周膜形态特点。方法 取临床拔除的牙根已发育完成的前磨牙,分别在空气中自然干燥放置1、2、4 h后,再以生理盐水或Hank′s平衡盐溶液(HBSS)浸泡处理0.5 h,制成扫描电镜标本,在扫描电镜下观察和比较牙颈部牙根表面牙周膜的形态特点。结果 脱位牙牙周膜表面形态随着干燥放置时间的增加其受损程度逐渐加重,经过保存液的处理,其牙周膜表面细胞和纤维的形态有了一定程度的恢复。HBSS处理组比生理盐水处理组牙周膜的形态恢复略好。结论 干燥放置的脱位牙在HBSS中浸泡,其牙周膜表面的细胞形态、胶原纤维的排列都优于生理盐水,这两种保存液都对干燥放置的脱位牙牙周膜的形态具有一定的恢复作用。?     

两种保存液对脱位牙再植术效果影响的实验研究

目的：比较常温下脱脂牛奶与汉克斯平衡盐溶液（Hank′s balanced salt solution,HBSS）保存脱位牙对再植牙牙根愈合过程的影响。方法：48只6周龄 SPF级 Wistar雄性大鼠,随机抽取3只为空白对照,直接处死取材拍摄根尖 X线片,标本切片组织学观察;余45只随机分为3组,将上颌第一磨牙脱位后分别采用 HBSS、脱脂牛奶浸泡或干燥室温保存,30 min 后植回牙槽窝,术后1、3、7、14、21 d处死大鼠,拍摄根尖 X线片进行图像分析,标本切片进行组织学观察。结果：再植后各时间点牛奶组与 HBSS 组近中根阴影面积无统计学差异,皆小于干燥组（P＜0．01）;牛奶组根尖组织炎症反应较轻,根吸收少,根周修复类型与HBSS 基本相同。结论：脱脂牛奶与HBSS 在常温下均为良好的脱位牙体外保存液,可在普通人群中作为再植牙保存液推广使用。     

改良HBSS液对体外培养人牙周膜细胞存活和克隆能力的影响

目的比较室温下改良HBSS液对体外培养人牙周膜细胞（PDLC）存活和克隆能力的影响。方法采用台盼蓝染色法和平板克隆形成试验检测体外培养的第4代人PDLC。在HBSS中添加青霉素、链霉素各100U／m1、地塞米松8mg／LTZ1、3、5mmol/L的还原型谷胱甘肽（GSH）制成改良HBSS液,并与HBSS液比较,观察PDLC在上述溶液中保存不同时间后的细胞活性及克隆能力的变化。结果3mmol/L和5mmol／LGSH组保存的细胞存活率在2。24h显著高于HBSS液组,且5mmol／LGSH组保存的PDLC克隆能力在1-12h显著高于HBSS液组。结论含5mmol／LGSH改良HBSS液保存人PDLC活性的效果在12h内明显好于HBSS液。     

全脱位牙牙周膜细胞活性在牛奶中的保存及其影响因素

全脱位牙回植可在外伤后短时间内重建咬合功能,恢复患者的容貌.牙回植成功与否关键在于保存患牙牙周膜细胞的活性,而保存时间和保存介质在其中起着十分重要的作用.牛奶廉价易得,可及时获取,有利于全脱位牙牙周膜细胞活性的及时保存,但不同种类、成分和温度的牛奶,其保存牙周膜细胞活性的能力和有效保存时间也不尽相同.此外,牛奶作为一种短期的全脱位牙牙周膜细胞活性保存介质,定期更新牛奶并没有太大的临床意义.保存在牛奶中的全脱位牙回植前需在汉克平衡盐溶液中浸泡30 min,以补充牙周膜细胞所需的营养物质,提高其增殖分化能力.     

人牙周膜成纤维细胞来源外泌体对大鼠延期牙再植术后牙根吸收的调控作用

目的: 探讨健康的人牙周膜成纤维细胞（hPDLFs）来源外泌体对大鼠延期牙再植术后牙根吸收的作用及其可能机制。方法: 分离提取hPDLFs来源的外泌体并鉴定。选择30只6周龄雄性SD大鼠,随机分为对照组和外泌体组,建立上颌第一磨牙延期牙再植术模型,牙脱位30 min后植回牙槽窝。对照组牙周膜局部注射40 μL汉克斯平衡盐溶液（HBSS）,实验组牙周膜局部注射40 μL含外泌体的HBSS。术后1、2、4周收样,苏木精-伊红（H-E）染色观察牙根表面吸收陷窝,抗酒石酸酸性磷酸酶（TRAP）染色观察破骨细胞数量,免疫组织化学染色观察牙周膜内骨保护素（OPG）的表达。采用SPSS 17.0软件包对数据进行统计学处理。结果: 鉴定表明提取的细胞外囊泡为外泌体。与对照组相比,hPDLFs来源的外泌体减少了延期牙再植术后牙根吸收陷窝的数量,降低TRAP阳性破骨细胞的表达（P<0.05）,促进牙周膜内OPG的表达（P<0.05）。结论: 延期牙再植术后,hPDLFs来源的外泌体减少了破骨细胞的数量,促进牙周膜内OPG表达,减少了再植术后的牙根吸收。     

外伤脱位离体牙再植23例成功

作者二十多年来对所遇到的 9例患者 ,2 3颗上前牙外伤性脱位离体 ,及时予以再植术 ,并取得满意远期效果。1　临床材料9例患者 ,男 7例 ,女 2例 ,年龄 9～ 33岁 ,均为牙体完整 ,无断根的上前牙 ,脱落原因主要是跌摔伤 ,拳击伤 ,离体时间 1 :30～ 1 5小时 (指牙脱位离体距就诊时间 )。2　手术方法2 .1　牙体处理情况 :对根尖发育尚未完成 ,牙根表面有轻度污染 ,牙周膜完好的脱落牙 ,就诊在 8小时之内者 ,将脱位离体牙置入 0 .9%生理盐水中浸泡6min后取出 ,清除牙根表面污物 ,尽力保存残余牙周膜纤维 ,然后将处理完毕的离体牙继续浸泡 0 .9%盐…     

六种保存液对牙周膜成纤维细胞活性的影响

目的评价6种不同保存液对牙周膜成纤维细胞活性的影响。方法培养因正畸拔除的前磨牙分离出的牙周膜成纤维细胞,采用CCK-8法检测细胞在自来水、Hank平衡盐溶液、豆浆、生理盐水、牛奶和DMEM高糖培养基6种保存液中1h、2h、4h、8h和24h五个时间点时的细胞生物学活性。结果在五个时间点,DMEM组细胞残存率大于生理盐水组,两组差异有统计学意义(P<0.01),豆浆、HBSS和DMEM三组之间细胞残存率无统计学差异(P>0.05),在1h、4h和24h 3个时间点,牛奶组细胞残存率大于DMEM组和豆浆组,其两组差异均有统计学意义(P<0.05)。结论牛奶、豆浆、HBSS和DMEM高糖培养基4种溶液是有效的牙周膜成纤维细胞临时保存液,而生理盐水和自来水保存效果差,不适合作为保存液。     

牙齿保存液的研究进展

牙脱位在儿童和青少年中的发生率呈逐渐上升趋势,牙齿离体后应尽快保存于合适储存介质中以备再植。理想的保存液应能保持牙周膜细胞的形态与活性,减少牙再植后牙根发生置换性吸收,提高牙再植成功率。笔者对牙齿保存液的研究进展作一综述。     

外伤后脱位牙保存方法的研究进展

颌面部的意外损伤将会导致不同类型的牙外伤,牙脱位是牙外伤中严重而常见的一种形式。解决这一问题的首选方法是脱位牙再植术,而牙再植的预后与脱位牙的急救措施息息相关。正确的急救方法可以提高脱位牙牙周膜细胞的存活率,这也是牙再植成功的关键。因此,国内外众多学者依据牙周膜细胞的生存环境,从不同的角度来寻求脱位牙的最佳保存条件和保存介质等,从而提高再植牙的成活率,减轻患者的痛苦。因此,本文就近期保存脱位牙的研究成果作一综述。     

A quantitative analysis of Propolis: a promising new storage media following avulsion

Abstract –  Both length of extra-alveolar time and type of storage media are significant factors that can affect the long-term prognosis of replanted teeth. Numerous studies have examined various media in an attempt to determine the ideal material for storage of the avulsed tooth. The purpose of this study was to use a Collagenase–Dispase assay to investigate the potential of a new storage media, Propolis, in maintaining viable periodontal ligament (PDL) cells on simulated avulsed teeth. Seventy freshly extracted human teeth were divided into five experimental groups and two control groups. The positive and negative controls corresponded to 0-min and an 8-h dry time, respectively. The experimental teeth were stored dry for 30 min and then immersed in one of the five media (Hank's balanced salt solution (HBSS), milk, saline, Propolis 50%, and Propolis 100% for 45 min). The teeth were then treated with dispase grade II and collagenase for 30 min. The number of viable PDL cells were counted with a hemocytometer and analyzed. Statistical analysis demonstrated that both Propolis groups kept significantly more PDL cells viable compared to either milk, saline, or HBSS. Within the parameters of this study, it appears that Propolis may be a better alternative to HBSS, milk, or saline in terms of maintaining PDL cell viability after avulsion and storage.     

Assessment of post-traumatic PDL cells viability by a novel collagenase assay

Abstract – Both length of extra-alveolar time and type of storage media are significant factors that can affect the long-term prognosis for replanted teeth. Numerous studies have examined various media in an attempt to determine the ideal material for storage of the avulsed tooth. The purpose of this study was to compare the number of viable periodontium ligament (PDL) cells in different storage media using a collagenase assay. Thirty-three freshly extracted human teeth were divided into four experimental and two control groups. The positive and negative controls corresponded to 0 min and an 8-h dry time, respectively. The experimental teeth were stored dry for 30 min and then immersed in one of four media (Hank's balanced salt solution (HBSS), milk, saline, water) for 45 min. The teeth were then treated with dispase grade II and collagenase for 30 min. The number of viable and nonviable PDL cells was counted with a hemocytometer and analyzed. An anova demonstrated no statistically significant differences in the viability of PDL cells among saline, HBSS and milk. Within the parameters of this study, it appears that milk or saline is an equally viable alternative to HBSS for storage of avulsed teeth.     

Viability of fibroblasts in a novel probiotic storage media

Abstract – A number of storage media have been investigated as to their ability to maintain the viability of the periodontal ligament (PDL) cells and thus to permit longer extra‐alveolar periods prior to replantation of avulsed teeth. The aim of the present in vitro study was to evaluate the number of viable PDL cells of avulsed teeth treated by Hank’s Balanced Salt Solutions (HBSS), saline, a novel probiotic solution and milk. Thirty‐six freshly extracted single‐rooted human teeth with closed apices were divided into one of the four experimental groups and two control groups (N = 6 each). The positive and negative controls corresponded to 0 min and an 8‐h dry time respectively. Following extraction, the coronal 3 mm of PDL tissue was scraped with a #15 scalpel to remove cells that might have been damaged. The experimental teeth were dried for 30 min followed by a 45 min immersion in one of the four experimental media. Each experimental tooth, after drying and soaking, was incubated for 30 min with a 2.5 ml solution of 0.2 mg ml^?1 of collagenase CLS II and a 2.4 mg ml^?1 solution of dispase grade II in phosphate buffer saline (PBS). The cells were then labelled with 0.4% Trypan blue for determination of viability. The teeth stored in positive control demonstrated the highest number of viable PDL cells followed in rank order by HBSS, saline, Lactobacillus reuteri solution and milk. There was no significant difference in the number of viable PDL cells between HBSS, milk, L. reuteri solution and saline. Within the parameters of this study, it appears that probiotic may be able to maintain PDL cell viability as HBSS, milk, or saline.     

Study of the effectiveness of propolis extract as a storage medium for avulsed teeth

Abstract – The purpose of the present study was to evaluate the efficacy of propolis extract in maintaining the viability of human periodontal ligament (PDL) cells, and to radiographically analyze tooth replantation and the adjacent periodontium in dogs after storage in this extract. Human PDL cells were incubated with the experimental media propolis, milk, saliva, Hank’s balanced salt solution (HBSS), and Dulbecco’s modified Eagles medium (DMEM, positive controls), and distilled water (negative control). Cell viability was determined 0, 1, 3, 6, 12, and 24 h later by colorimetric MTT assay. Thirty incisors from dogs were divided into two storage time blocks (1 and 3 h) and were maintained in the experimental media. HBSS served as a positive control, and dry teeth (on gauze) as a negative control. The replanted teeth were radiographed once per month for 6 months. The radiographic images were standardized by the shortening/lengthening factor, and were both qualitatively and quantitatively analyzed. The in vitro results showed that the efficacy of propolis in maintaining functional viability of PDL cells was similar to that of milk. Propolis and milk were significantly better than controls from the 6‐h time period. The in vivo results showed that teeth maintained in propolis medium exhibited replacement resorption with significant reduction in tooth length, similar to teeth maintained in saliva and dried teeth. This resorption was less intense with the 3‐h storage time than the 1‐h storage time. Conditions close to normal were found in teeth maintained in milk, similar to the HBSS control. Therefore, although propolis was effective in maintaining the viability of human PDL cells, resorption of the tooth replantation in dogs occurred under these experimental conditions.     

(-)-Epigallocatechin-3-gallate: a novel storage medium for avulsed teeth

The purpose of the present study was to evaluate the efficacy of (-)-epigallocatechin-3-gallate (EGCG) in maintaining the vitality of human periodontal ligament (PDL) cells when used as a storage medium for avulsed teeth prior to replantation. Thirty freshly extracted single-rooted human teeth with closed apices were randomly assigned to three experimental groups with 10 samples per group and immersed in one of the storage media: EGCG, Hank's balanced salt solution (HBSS), or milk for 2 h. The PDL cells were dissociated by an enzyme treatment with collagenase and trypsin. The cells were then labeled with 0.4% Trypan blue for the determination of viability. The result showed that EGCG group had the highest percentage of cell viability, followed by HBSS and milk group, in descending order.     

Effect of soaking in Hank's balanced salt solution or milk on PDL cell viability of dry stored human teeth

Abstract— This study was designed to evaluate the effect of soaking in either Hank's balanced salt solution (HBSS) or milk on peri⁻odontal ligament (PDL) cell viability in avulsed teeth. Dry storage times of 30, 60, and 90 min were evaluated. PDL cell viability was determined after removal of the cells from the root surfaces of extracted teeth using a modification of the procedure described by Nakashima (Arch Oral Biol 1991;36:655–63). After trypsinization and subsequent treatment in collagenase, the cells were stained with trypan blue, and viable and non-viable cells were counted using a hemocytometer and converted to percentages for statistical comparison. The results of this study demonstrated no significant difference in the number of viable cells with or without soaking in HBSS or milk at any of the dry storage times. In addition, there was no significant difference in PDL cell viability between the 30-and the 60-min dry periods. Although the soaking procedure had no obvious negative consequence, no simcant improvement in PDL cell viability by the addition of this step was demonstrated under the conditions of this study.     

Pedialyte Promotes Periodontal Ligament Cell Survival and Motility

IntroductionThe search still continues to find the best storage media for avulsed teeth. Unfortunately, some of the recommended storage solutions are not commonly found in households or do not preserve the periodontal ligament (PDL) cells long-term. The purpose of the present study was to determine whether Pedialyte is a viable alternative storage solution for avulsed teeth by assessing its ability to preserve human PDL cell viability.MethodsHuman PDL cells were exposed to 6 different storage solutions (minimal essential medium [MEMα], Hank's balanced salt solution [HBSS], non-fat milk, coconut water, Pedialyte, or tap water) for 2, 6, 24, or 48 hours at 4°C or 25°C. Cell viability was quantified immediately or 1 week after exposure. The effects of these storage solutions on PDL cell motility and bacterial proliferation were also examined. The results were statistically analyzed by analysis of variance.ResultsPedialyte at 4°C and 25°C showed significantly (P < .001) higher cell survival compared with water after all time intervals. No significant difference was noted between control (MEMα), HBSS, coconut water, and Pedialyte at 4°C after 2 hours. Cells stored in Pedialyte for 24 hours at 25°C and assayed 1 week later showed significantly higher cell survivability compared with milk. Pedialyte supported significantly less bacterial growth compared with non-fat milk and coconut water. No difference in cell motility was observed for cells stored for 24 hours in Pedialyte, MEMα, HBSS, milk, or coconut water.ConclusionsPedialyte is a viable alternative as a storage solution for avulsed teeth.     

In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in six different media

Abstract— The choice of storage medium for preserving traumatic-ally avulsed teeth is important for the success of future replantation. The objectives of this study were to evaluate the effectiveness of six different media: culture medium, α minimal essential medium (αMEM), milk, Hank's balanced salt solution (HBSS), ViaSpan and conditioned medium (CM) to preserve cultured periodontal ligament fibroblasts (PDLF). Periodontal ligament fibroblasts were obtained from explants of human healthy extracted teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at 4°C. A control group was incubated with culture medium at 37°C. After incubation, cell viability was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Storage of PDLF up to 24 h decreased their vitality by only 2%-14%. Vitality of the PDLF after 2, 8 and 24 h was highest when stored in milk or HBSS (91%-97%) and lowest when stored in ViaSpan or CM (82%-93%). PDLF stored for 2–8 h in various media had a mitogenic capacity comparable to the control. However, increasing the storage period to 24 h decreased the mitogenicity of the cells by 3%-39%. The highest mitogenicity was found in PDLF stored in milk or HBSS and the lowest in CM or ViaSpan. The clonogenic capacity of the cells dropped by 38%-71% after 24 h and was the best indicator of the deteriorating effect of long storage. Milk and HBSS were the most effective in preserving the clonogenic capacity. Nevertheless, reduction in the viability, mitogenicity or clonogenic capacity was statistically significant in nearly all the tested media only after 24 h of incubation. In conclusion, HBSS and milk were the most effective media for preserving the viability, mitogenicity and clonogenic capacity after storage for up to 24 h at 4°C.     

Comparison of coconut water, propolis, HBSS, and milk on PDL cell survival

Coconut water is biologically pure and sterile, with a rich presence of amino acids, proteins, vitamins, and minerals. The purpose of this study was to use a collagenase-dispase assay to investigate the potential of a new storage medium, coconut water, in comparison with propolis, Hank's balanced salt solution (HBSS), and milk in maintaining viable periodontal ligament (PDL) cells on simulated avulsed teeth. Seventy freshly extracted human teeth were divided into 4 experimental groups and 2 control groups. The positive and negative controls corresponded to 0-minute and 8-hour dry times, respectively. The experimental teeth were stored dry for 30 minutes and then immersed in 1 of the 4 media (coconut water, propolis, HBSS, and milk). The teeth were then treated with dispase grade II and collagenase for 30 minutes. The number of viable PDL cells was counted with a hemocytometer and analyzed. Statistical analysis showed that coconut water kept significantly more PDL cells viable compared with propolis, HBSS, or milk. Coconut water can be used as a superior transport medium for avulsed teeth.     

Expression of proliferating cell nuclear antigen in pulp cells of extracted immature teeth preserved in two different storage media

Abstract – A specially composed medium for storing avulsed teeth has been developed. In experimental and clinical studies it could be shown that PDL cells could be kept viable during storage in the medium for up to 53 h. In the present study the medium was tested on pulp cells. A total of 40 immature unerupted third molars with open apices were removed surgically and the teeth were stored in a special cell culture medium (SCCM) or in Hank's balanced salt solution (HBSS) at room temperature for 6, 12, 18 or 24 h. Five teeth were assigned to each group. A total of seven consecutive pulp cross‐sections per tooth were examined, resulting in a total of 280 specimens. Viable cells were marked using proliferating cell nuclear antigen (PCNA). The pulp was divided in three regions: apical region (0–0.5 mm), middle region (>0.5–1.5 mm) and coronal region (>1.5 mm). The labelling index (LI) was calculated for the whole cut (regions 1, 2 and 3) and for each region separately. The statistical evaluation was made using the One‐way anova and Mann–Whitney Test. Pulp cells of all teeth expressed PCNA. About 110 of 140 specimens in the SCCM and 101 of 140 specimens in the HBSS group showed PCNA‐positive cells. The highest LI was observed within the apical region and decreased with increased distance from the medium. No marked cells were observed at a distance of more than 1.5 mm. The LI for both media showed a significant increase with storage intervals (P < 0.05). The pulp cells of teeth stored in SCCM showed a LI nearly twice as high compared to pulp cells of teeth stored in HBSS for the apical and middle region (time interval 6, 18 and 24 h: P < 0.05). The LI for the apical region was found to be 8.43% for the SCCM and 4.50% for the HBSS after 24 h. For the middle region the LI was found to be 2.02% for the SCCM and 0.81% for the HBSS after 24 h. Within the parameters of this study, it appears that the SCCM is able to maintain pulp cell viability better than HBSS. The use of special cell culture media in case of tooth avulsion may be beneficial.