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.     

Potential of the propolis as storage medium to preserve the viability of cultured human periodontal ligament cells: an in vitro study

Abstract – Aim: In vitro experiments were carried out to evaluate the potential of propolis, a natural resin known for its wide therapeutic window, as storage medium to preserve the viability of cultured human periodontal ligament (PDL) cells. Materials and Methods: Primary cultures of human PDL cells were subjected to either independent exposure of propolis (2.5%, 5.0%, 10.0%, and 20.0%), Hank’s balanced salt solution (HBSS), milk (0.5%), artificial saliva, Dulbecco’s modified Eagle’s medium (DMEM) or combination of propolis 10% + DMEM, propolis 20% + DMEM for 30 min to 24 h at 37°C. Cell viability was assessed using standard endpoints i.e., tetrazolium bromide salt (MTT), neutral red uptake, and trypan blue dye exclusion assay. Results: In general, combinations of propolis 10% + DMEM, propolis 20% + DMEM, and DMEM alone were found to be better than other media used in this study. The difference in the potentials of these media to maintain the cell viability reached to the statistically significant levels by 24 h, when compared with other media used viz., propolis 2.5% (P < 0.01), propolis 5.0% (P < 0.05), propolis 10.0% (P < 0.05), propolis 20.0% (P < 0.001), HBSS (P < 0.001), and milk (P < 0.01). Trypan blue dye exclusion assay could be recorded the most sensitive among all the assays selected to study the cell viability of PDL cells. Conclusions: Study indicates that combinations of propolis 10% + DMEM, propolis 20% + DMEM, and DMEM alone are equally good as storage media of choice to keep PDL cells viable during extra‐alveolar period up to 24 h. Other more readily available medium such as milk may serve as appropriate alternative storage medium for shorter time periods i.e., up to 12 h.     

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.     

Use of soymilk as a storage medium for avulsed teeth

Abstract

Objective. Tooth avulsion is one of the most severe forms of dental trauma. In these cases, immediate reimplantation is ideal; however, it almost never happens. The purpose of this study was to evaluate the viability of cells stored in soymilk and compare with other several storage media. Materials and methods. The media tested were: long-shelf-life coconut water, long-shelf-life whole milk, long-shelf-life soymilk, Gatorade, egg white, and Hank's Balanced Salt Solution. Cells cultured in DMEM and distilled water served as positive and negative controls, respectively. Plates containing confluent 3T3 fibroblast were soaked in the various media for 2, 12 and 24 h. After incubation at 37°C, viability of the cells was determined using the MTS assay. Data were analyzed by using one-way ANOVA and complemented by Tukey test with a significance level of 5%. Results. Statistical analysis showed that DMEM, whole milk, HBSS and soymilk were the most effective media for maintaining cell viability at all tested times (p < 0.05), followed by coconut water, egg white and Gatorade. The least amount of viable cells was observed in the distilled water group. Conclusions. The present study shows that the efficacy of soymilk in maintaining the viability of 3T3 fibroblasts is similar to that of HBSS and milk. Therefore, it can be concluded that soymilk could be a suitable alternative storage medium for avulsed teeth.     

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。     

Effect of chlorophyllin on normothermic storage of human periodontal ligament cells

The purpose of the present study was to evaluate whether chlorophyllin could serve as an effective constituent of a storage medium to enhance the human periodontal ligament (PDL) cell viability.Freshly isolated PDL cells from premolars extracted from healthy people were stored at 37 degrees C for 6 h in various solutions: F-medium and Hank's balanced salt solution (HBSS), supplemented with chlorophyllin. From MTT viability assays, the highest cell viability was found in the PDL cells stored in HBSS supplemented with 500 nM chlorophyllin, and the chlorophyllin-treated cells showed a dose-dependent response to concentration. Additionally, the results from flow cytometry showed that 77 to 80% of the PDL cells were in the G0/G1 phases of the cell cycle, which suggested that most were in a stable stage.These result showed that HBSS, supplemented with chlorophyllin, may be a useful solution for preserving the viability of PDL cells.     

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.     

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.     

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.     

不同离体时间和保存液对犬离体牙牙周膜细胞活力影响的实验研究

目的:探讨完全脱位牙不同离体时间和保存液对牙周膜细胞活力的影响。方法:麻醉拔除犬牙35个,首先将20个牙随机分为5组,分别为室温干燥放置0、30、60、120、240 min组,另15个牙室温干燥放置30 min后,随机分为3组,分别放入牛奶、HBSS液,100 g/L蜂胶液中浸泡2 h。各组处理完成后,采用全牙消化法获得牙周膜细胞,并通过4 g/L台盼蓝染色法检测各组牙周膜活细胞数和存活率。结果:室温干燥放置30、60、120、240 min后,牙周膜细胞存活率依次为33.6%、23.6%、18.5%、0.8%,而0 min的牙周膜细胞存活率可达95.5%。拔后30 min,经牛奶、HBSS液和100 g/L蜂胶液中保存2 h后,牙周膜细胞均有活力,其细胞存活率大小依次为100 g/L蜂胶液、HBSS液和牛奶,其中100 g/L蜂胶液与HBSS液相比无统计学差异(P>0.05),但与牛奶组相比,均有统计学差异(P<0.05)。结论:随着离体时间延长,完全脱位牙根面牙周膜细胞活力明显下降。100 g/L蜂胶液和HBSS液保存犬牙牙周膜细胞活力优于牛奶液。     

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.     

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.     

Human periodontal ligament cell viability in milk and milk substitutes

The purpose of this study was to determine the efficacy of several milk substitutes compared to whole milk in maintaining the viability of human periodontal ligament (PDL) cells on avulsed teeth. PDL cells were obtained from freshly extracted, healthy third molars and cultured in Eagle's minimal essential media (EMEM). The cells were plated onto 24-well culture plates and allowed to attach for 24 h. EMEM was replaced with refrigerated whole milk (positive control), reconstituted powdered milk, evaporated milk, or one of two baby formulas (Similac or Enfamil). Tap water served as the negative control. Tissue culture plates were incubated with the experimental media at 37 degrees C for 1, 2, 4, or 8 h. Cell viability was determined by a cell proliferation assay (CellTiter 96 AQ Assay), with absorbance read at 450 nM. A two-way ANOVA (p < 0.001) indicated that at 1 h there was no difference in the effect on PDL cell viability between any of the materials and whole milk. At 2 h, Enfamil and Similac performed significantly better than whole milk, whereas evaporated milk performed worse. At 4 h, Enfamil performed better than whole milk, whereas all other milk substitutes performed worse. At 8 h, all substitutes performed worse than whole milk. These results suggest that Enfamil, which is supplied in powder form that does not require special storage and has a shelf life of 18 months, is a more effective storage medium for avulsed teeth than pasteurized milk for at least 4 h.     

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.     

Survival of human periodontal ligament cells in media proposed for transport of avulsed teeth

Abstract –  Many solutions have been examined as possible storage media for avulsed teeth. In this report, human periodontal ligament (PDL) cells were exposed for 1 h to culture medium, milk, Hanks Balanced Salt Solution (HBSS), Soft Wear, Opti Free, and Solo Care contact lens solutions, Gatorade^®, and tap water, at room temperature and on ice. The number of viable cells was counted using the trypan blue exclusion technique, immediately after exposure (0 h) and at 24 and 48 h, to test the proliferative capacity of the cells after treatment. The results indicated that a significantly higher number of cells survived and proliferated when the exposures were performed at 0°C. Water had a detrimental effect on the cells, whereas culture medium and HBSS preserved significantly more viable cells than the other experimental solutions. Within the parameters of this study, it appears that HBSS is the optimal storage medium for avulsed teeth. Low-fat milk could serve as an alternative if ice is available. Contact lens solutions or Gatorade^® on ice could serve as short-term (1 h) storage media if the other solutions are not readily available.     

(-)-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.     

Storage media enhance osteoclastogenic potential of human periodontal ligament cells via RANKL‐independent signaling

Abstract – Background: Hank’s balanced salt solution (HBSS) and milk have gained wide acceptance as storage media for avulsed tooth. However, the effect of the media and storage time on the periodontal ligament (PDL) cells involvement in the development of root resorption is still unclear. The purpose of this study was to evaluate whether precultured PDL cells in HBSS, milk, or modified Eagle’s medium alpha (α‐MEM) would affect osteoclastogenesis. Materials and methods: PDL cells were precultured in HBSS, milk, or α‐MEM for 1 h or 6 h before being co‐cultured with RAW 264.7 cells for an additional 3 days for mRNA analysis and 11 days for osteoclastogenesis assay. Results: Cyclooxygenase‐2 (COX‐2) mRNA was detected immediately in PDL cells precultured in the three storage media. The expression was up‐regulated markedly in all co‐cultures when compared with RAW cells alone. As a result of the co‐culture, interleukin‐1β (IL‐1β) expression was detectable in both PDL and RAW cells. TRAP+ multinucleated, osteoclast‐like cells developed in all co‐cultures; the number of TRAP+ cells was highest (P < 0.05) in the co‐cultures that PDL cells precultured in milk for 6 h. The mRNA level of receptor activator of nuclear factor‐kappa B ligand (RANKL) was not detected in PDL cells. Osteoprotegerin (OPG) mRNA expression reduced with increased preculture time, but the difference was not significant (P > 0.05). Conclusions: PDL cells kept in the three storage media led to TRAP+ multinucleated, osteoclast‐like cells formation via RANKL‐independent signaling. The ability to induce osteoclastogenesis may be considered as one of the factors to evaluate the ability of storage medium to maintain PDL viability after tooth avulsion.     

Effect of HBSS storage time on human periodontal ligament fibroblast viability

Abstract – Hank’s balanced salt solution (HBSS) is recommended for the storage of avulsed teeth. The objective of this study was to evaluate if the HBSS storage time influences its ability to maintain the viability of human periodontal ligament fibroblasts (PDLF) by the analysis of cell metabolic function using MTT assay. PDLF were kept at 20°C for 3, 6, 24, 48, 72, 96 and 120 h in recently prepared HBSS (HBSS), HBSS stored for 6 months (HBSS 6 M), HBSS stored for 12 months (HBSS 12 M), and in Save‐A‐Tooth system’s HBSS (Save). Minimum essential medium (MEM) at 37°C and tap water at 20°C served as positive and negative controls, respectively. Cell viability was determined by the tetrazolium salt‐based colorimetric (MTT) assay. Data were statistically analyzed by the Kruskal–Wallis and Scheffé tests (α = 5%). Starting with the 6 h time‐point, HBSS was significantly more effective than HBSS 6 M, HBSS 12 M and Save in maintaining cell viability. HBSS 6 M effectiveness was similar to that of HBSS 12 M for up to 48 h, becoming higher at 72 h. In conclusion, the storage time of HBSS had a negative influence on its ability to maintain PDLF viability.     

The use of green tea extract as a storage medium for the avulsed tooth

Introduction

Green tea extract (GTE) has been reported to have remarkable anti-inflammatory, antioxidant, and anticarcinogenic effects and to prolong allograft survivals. The purpose of the present study is to investigate in vitro the efficacy of GTE as a storage medium for avulsed teeth. We estimated the possibility for storage medium by maintaining the viability of human periodontal ligament (PDL) cells.

Methods

Human PDL cells were cultured and stored in the following media: (1) Hank’s balanced salt solution (HBSS), (2) tap water, (3) milk, (4) GTE, and (5) commercial green tea. After 1, 3, 6, 12, and 24 hours, cells in different media were examined under the optical microscope, and their viabilities were analyzed by using a nucleocounter and 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethonyphenol)-2-(4-sulfophenyl)-2H-tetrazolium assay. The data were statistically analyzed by analysis of variance tests with post analysis using the Duncan method (P < .05).

Results

The result indicates that there was no difference in cell viability between GTE and HBSS media, whereas GTE showed higher cell viability than other media (P < .05).

Conclusions

Our study shows that the efficacy of GTE in maintaining the viability of human PDL cells is similar to that of HBSS and higher than that of milk. Therefore, we conclude that GTE could be a suitable, alternative storage medium for avulsed teeth.     

A Quantitative Analysis of a Probiotic Storage Media for Avulsed Teeth

Aim

The aim of the present in vitro study was to investigate the potential of a storage medium, probiotic yogurt (Bifidibacterium animalis DN 173010) in comparison with Hank''s balanced salt solution (HBSS), saline and milk in maintaining viable periodontal ligament (PDL) cells on simulated avulsed teeth.

Materials and methods

Thirty-six freshly extracted single-rooted human teeth with closed apices were divided into six experimental groups (N=6). The teeth were extracted as atraumatically as possible and washed in sterile saline solution to eliminate residual blood. Following extractions, the coronal 3 mm of PDL tissues were scraped with a #15 scalpel to remove cells that may have been damaged. The positive and negative controls corresponded to 0 minutes and an 8-hour dry time, respectively. After extraction, the positive control teeth were immediately treated with dispase and collagenase. The negative control teeth were bench-dried for 8 h, with no follow-up storage solution time, and then placed in the dispase and collagenase. The number of viable protective least significant difference PDL cells were counted under a light microscope with a hemocytometer at 20× magnification and analyzed. Statistical analysis of the data was accomplished using Nonparametric ANOVA complemented by Kruskal-Wallis Test and Dunn''s Multiple Comparisons Test.

Results

Positive control was found to be significantly better than the others, there were statistically significant differences between positive control and other test groups (p=0.000). The teeth stored in positive control demonstrated the highest number of viable PDL cells followed in order by probiotic yogurt, HBSS, saline and milk.

Conclusion

Bifidibacterium animalis DN 173010 seems to be an alternative for the temporary storage of avulsed teeth, due to high number of viable PDL cells. Probiotics may be suitable transport media for avulsed teeth, but further research is warranted using the commercially available products.Key words: dental trauma, probiotic, storage media