Viability and Reproducibility of Periodontal Ligament Cells on Avulsed Teeth Stored in Ham's F-10 Solution

2018 ◽  
Vol 42 (3) ◽  
pp. 203-207 ◽  
Author(s):  
Maryam Talebi ◽  
Iman Parisay ◽  
Jalil Tavakol afshari ◽  
Arezoo Shajiei ◽  
Mostafa Sofiani Ghadim

Objectives: The purpose of the present study was to evaluate the efficacy of Ham's F-10 in maintaining the viability and reproducibility of PDL cells on avulsed teeth. Study design: Sixty mature, healthy extracted premolars were used. The experimental media used were Ham's F-10, Hank's balanced salt solution (HBSS), skim milk, and tap water (n = 15 specimens each). Cell viability was tested after 1, 3, 6, and 24 h storage in medium. Cell reproducibility was assessed by methyl-thiazol-tetrazolium (MTT) assay after1, 3, and 6 h storage in Ham's F-10, HBSS, and tap water. Results: The viability of PDL cells stored in Ham's F-10 and HBSS was significantly greater than that of samples stored in milk and tap water at all-time points (P<0.001). A significant difference in cell viability between samples stored in Ham's F-10 and HBSS (favoring the former) was observed only at 6h (P=0.04). MTT assay results were significantly better for samples stored in Ham's F-10 and HBSS than for those stored in tap water (P<0.001), with a significant difference between Ham's F-10 and HBSS observed only at 3h (P<0.001). Conclusions: Ham's F-10 is capable of preserving PDL cells viable and reproducible better than milk and tap water and similar to HBSS.

2008 ◽  
Vol 9 (6) ◽  
pp. 17-24 ◽  
Author(s):  
Fatih Ozan ◽  
Zübeyde Akin Polat ◽  
Bektaş Tepe ◽  
Kürşat Er

Aim The purpose of this study was to determine the ability of Salvia officinalis (S. officinalis) extracts to serve as a storage medium for the maintenance of periodontal ligament (PDL) cell viability of avulsed teeth. Methods and Materials PDL cells were obtained from healthy third molars and cultured in Dulbecco's Modified Eagle's Medium (DMEM). Cultures were subjected to 4, 2.5, 1.5, and 0.5% S. officinalis solutions, Hank's balanced salt solution (HBSS), phosphate buffered saline (PBS), and tap water. Tissue culture plates were incubated with experimental media at 37°C for 1, 3, 6, 12 or 24 hours. PDL cell viability was assessed by trypan blue exclusion. Statistical analysis of the data was performed by one-way analysis of variance (ANOVA) complemented by the Tukey's test. The level of significance was 5% (p< 0.05). Results The results showed 2.5% S. officinalis was a more effective storage medium than the other experimental solutions (p<0.05). Only at 1 hour and 3 hours was there found similar effect between 2.5% S. officinalis and HBSS. At 24 hours, 2.5% S. officinalis was found to be significantly better than the other solutions tested. Conclusion S. officinalis can be recommended as a suitable transport medium for avulsed teeth. Clinical Significance The findings of this study support the use of S. officinalis as another option for clinicians to use to store and transport avulsed teeth until reimplantation procedures can be done. Citation Özan F, Polat ZA, Tepe B, Er K. Influence of Storage Media Containing Salvia officinalis on Survival of Periodontal Ligament Cells. J Contemp Dent Pract 2008 September; (9)6:017-024.


2016 ◽  
Vol 35 (9) ◽  
pp. 983-990 ◽  
Author(s):  
Xin Ge ◽  
Ying-Feng Liu ◽  
Yong Wong ◽  
Li-Zheng Wu ◽  
Ling Tan ◽  
...  

Periodontitis is a common infectious disease associated with destruction of periodontal ligaments and alveolar bones. CD4+ T cell-mediated immune response is involved in the progression of periodontitis. Tobacco consumption increases the risk of periodontal disease. However, the impact of nicotine on the interaction between human periodontal ligament (PDL) cells and CD4+ T cells remains unrevealed. Our study aims to investigate the effect of nicotine on PDL cells and the cocultured CD4+ T cells. The PDL cell cultures were established by explants from healthy individuals, exposed to nicotine or α-bungarotoxin (α-BTX), and incubated solely or in combination with CD4+ T cells. Afterwards, cell viability, secreted cytokines, and matrix metalloproteinases (MMPs) were evaluated. In monoculture of PDL cells, nicotine dramatically repressed cell viability and increased apoptosis. Meanwhile, α-BTX largely reversed the nicotine-induced apoptosis and increased viability of PDL cells. Compared with the monoculture, MMP-1, MMP-3, interleukin (IL)-1β, IL-6, IL-17, and IL-21 in supernatant of cocultures were markedly elevated after treatment with nicotine. Moreover, α-BTX significantly attenuated nicotine-triggered production of these components either in mono- or co-cultures. In addition, PDL cell-derived CXCL12 following nicotine treatment recruited CD4+ T cells. Above all, nicotine deteriorated periodontitis partially by promoting PDL cell–CD4+ T cell-mediated inflammatory response and matrix degradation.


2016 ◽  
Vol 27 (4) ◽  
pp. 404-407 ◽  
Author(s):  
Beatriz Dulcineia Mendes de Souza ◽  
Ana Maria Hecke Alves ◽  
Luciane Geanini Pena dos Santos ◽  
Claudia Maria de Oliveira Simões ◽  
Wilson Tadeu Felippe ◽  
...  

Abstract The objective of this study was to evaluate the effectiveness of various storage media at 20 °C in maintaining the viability of human periodontal ligament fibroblasts (HPLF) over time. HPLF were maintained at 20 °C in skim milk (SM), whole milk (WM), freshly prepared Hank's balanced salt solution (HBSS), Save-A-Tooth(r), natural coconut water (NCW), coconut water industrialized (ICW) and tap water (negative control) for 3, 6, 24, 48, 72, 96 and 120 h. Cells maintained in Minimal Essential Medium (MEM-37) at 37 °C served as a positive control. Cell viability was determined by MTT assay. Statistical analysis was performed by Kruskal-Wallis test and Scheffe test (α = 5%). From 24 h, NCW was significantly better in maintaining cell viability than all other tested storage media (p<0.05). SM and WM were significantly better than HBSS for up to 72 h. Save-A-Tooth(r) and ICW were the worst conservation storage media. In conclusion, the effectiveness of the tested storage media to maintain the viability of the periodontal ligament cells was as follows, in a descending order: NCW > MEM-37> SM and IM> HBSS> ICW > Save-A-Tooth(r)> tap water.


2016 ◽  
Vol 40 (1) ◽  
pp. 49-52 ◽  
Author(s):  
Punit Fulzele ◽  
Sudhindra Baliga ◽  
Nilima Thosar ◽  
Debaprya Pradhan

Objective: To investigate the effectiveness of aloevera gel as a new storage medium in maintaining the viability of periodontal ligament cells. Study design: Premolars extracted for orthodontic reason were obtained. Confluent monolayers of fibroblasts were grown by cell culture method from the PDL cells isolated from the extracted teeth. One ml of this cell suspension was transferred to wells of culture plates, incubated for 24 hrs, followed by exposure to the three experimental media, Hank's balanced salt solution (HBSS), aloevera gel, and packaged drinking water. These plates were then assessed for viable cells using trypan blue dye exclusion test with haemocytometer after 15, 30, 60, 90 and 120 mins. The results obtained were statistically analysed using one-way analysis of variance (ANOVA). Results: At 15 min, HBSS presented maximum mean percentage of viable PDL cells (89%), followed by aloevera at 81% and packaged drinking water at 10%. Aloevera demonstrated 71%, 59%, 57% viable cells at 30, 60, 90 mins respectively. At 120 min, HBSS presented 57% viable cells followed by aloevera gel (45%) and packaged drinking water (3%). No statistical significant difference was observed between HBSS and aloevera gel. Conclusions: Within the parameters of this study, both aloevera gel and HBSS were effective in maintaining the viability of PDL cells. Hence, aloevera gel could be used as a storage media for avulsed tooth in situations where availability of HBSS is in question.


2008 ◽  
Vol 9 (6) ◽  
pp. 25-32 ◽  
Author(s):  
Abbas Ali Khademi ◽  
Saeed Saei ◽  
Mohammad Reza Mohajeri ◽  
Nooshin Mirkheshti ◽  
Fatima Ghassami ◽  
...  

Aim The purpose of this study is to determine the efficacy of egg white in maintaining the viability of human periodontal ligament (PDL) cells on avulsed teeth. Methods and Materials The experimental media were: egg white, milk, Hanks’ Balanced Salt Solution (HBSS) as the positive control, and tap water as the negative control. The storage times were 1, 2, 4, 8, and 12 hours. Extracted premolar teeth of healthy individuals were rinsed in the media. After trypsinization and subsequent treatment in collagenase, cell viability was determined using trypan blue staining. The two-way analysis of variance (ANOVA) statistical test was used to compare the results among different media. Results There was no difference in the cell viability between egg white and HBSS media, but there was a statistically significant difference between the viability of PDL cells in egg white medium in comparison with milk (P<0.05) and water (P<0.05). Conclusion Egg white could be suggested as a suitable storage medium. Its principle advantage is its availability. Citation Khademi AA, Saei S, Mohajeri MR, Mirkheshti N, Ghassami F, Torabi nia N, Alavi SA. A New Storage Medium for an Avulsed Tooth. J Contemp Dent Pract 2008 September; (9)6:025-032.


2016 ◽  
Vol 27 (4) ◽  
pp. 408-411 ◽  
Author(s):  
Meenakshi Sharma

Abstract The aim of this study was to evaluate the viability of periodontal ligament cells of avulsed teeth in three different storage media. Forty-five mature premolars extracted for orthodontic therapeutic purposes were randomly and equally divided into three groups according to the storage medium: milk (control), rice water and egg white. After placing extracted teeth for 30 min in storage media, the scrapings of the periodontal ligament (PDL) were collected in Falcon tubes containing collagenase in 2.5 mL of phosphate buffer saline and were incubated for 30 min and centrifuged for 5 min at 800 rpm. Cell viability was analyzed by Trypan blue exclusion. Rice water had a significantly higher number of viable cells compared to egg white and milk. There was no statistically significant difference between egg white and milk. Rice water may be able to maintain PDL cell viability of avulsed teeth better than egg white or milk.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Lorena Bortolini Navarro ◽  
Fabiane Barchiki ◽  
Wilson Navarro Junior ◽  
Everdan Carneiro ◽  
Ulisses Xavier da Silva Neto ◽  
...  

AbstractThis study analyzed the efficacy of autologous platelet-rich fibrin (PRF) in maintaining and recovering cell viability of the periodontal ligament (PDL). The PDL cells were isolated from 45 extracted teeth randomly distributed among 6 groups: 5 min, 1 h, 2 h, PRF 30 min, PRF 1 h and PRF 2 h. In the groups 5 min, 1 h and 2 h (n = 5), the teeth were kept dry in extra-alveolar times of 5 min, 1 h and 2 h respectively. The teeth of the groups PRF 30 min, PRF 1 h and PRF 2 h (n = 10) were kept dry at extra-alveolar times of 30 min, 1 and 2 h followed by immersion in PRF for 45 min. PDL cells were isolated by enzymatic digestion with type II collagenase and dispase, counted and analyzed for viability with Trypan blue vital dye in Neubauer chamber. The variables total number of cells and cell viability demonstrated that in the 5 min, 1 h and 2 h groups there was a decrease after the extra-alveolar dry times of 1 and 2 h. In comparison with the total number of cells, group 1 h, considered immediate reimplantation, did not present statistical difference when compared to the groups PRF 30 min, PRF 1 h and 2 h, a result that demonstrates that PRF assists in cell maintenance and recovery. PRF provided increased cell viability in relation to the different dry extra-alveolar times analyzed (p < 0.001). Autologous PRF presented effectiveness in maintaining and recovering PDL cells from extracted teeth and kept dry for up to 2 h.


2016 ◽  
Vol 10 (03) ◽  
pp. 408-412 ◽  
Author(s):  
Fahimeh Adeli ◽  
Ebrahim Zabihi ◽  
Zeinab Abedian ◽  
Samane Gharekhani ◽  
Mahdi Pouramir ◽  
...  

ABSTRACT Objective: Green tea extract (GTE) was shown to be effective in preserving periodontal ligament fibroblasts (PDLFs) of avulsed teeth. This study aimed at determining the potential of GTE in preserving the viability of PDLFs comparing with different storage media. Materials and Methods: Periodontal ligament cells were obtained from freshly extracted healthy impacted third molars and cultured in Dulbecco's Modified Eagle Medium (DMEM). Cell viability was determined by storing the cells in seven media; DMEM, tap water, Hank's balanced salt solution (HBSS), whole milk, hypotonic sucrose solution, GTE, and GTE + sucrose for 1, 2, 4, and 24 h at 37°C using tetrazolium salt-based colorimetric (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide) assay. Statistical analysis was performed by one-way analysis of variance and post hoc tests. Results: GTE showed significantly higher protective effect than HBSS at 2, 4, and 24 h (P = 0.009, P = 0.02, P = 0.016), DMED at 2 h (P = 0.003), and milk at 4 h (P = 0.039). Conclusion: Although with undesirable osmolality and pH, GTE had a good ability in preserving the PDLFs comparing with other studied media.


2006 ◽  
Vol 85 (8) ◽  
pp. 757-760 ◽  
Author(s):  
L. Liu ◽  
K. Igarashi ◽  
H. Kanzaki ◽  
M. Chiba ◽  
H. Shinoda ◽  
...  

Periodontal ligament (PDL) cells play an essential role in orthodontic tooth movement. We recently reported that clodronate, a non-N-containing bisphosphonate, strongly inhibited tooth movement in rats, and thus could be a useful adjunct for orthodontic treatment. However, it is not clear how clodronate affects the responses of PDL cells to orthodontic force. In this study, we hypothesized that clodronate prevents the mechanical stress-induced production of prostaglandin E2 (PGE2), interleukin-1β (IL-1β), and nitric oxide (NO) in human PDL cells. A compressive stimulus caused a striking increase in PGE2 production, while the responses of IL-1β and NO were less marked. Clodronate concentration-dependently inhibited the stress-induced production of PGE2. Clodronate also strongly inhibited stress-induced gene expression for COX-2 and RANKL. These results suggest that the inhibitory effects of clodronate on tooth movement and osteoclasts may be due, at least in part, to the inhibition of COX-2-dependent PGE2 production and RANKL expression in PDL cells.


2007 ◽  
Vol 86 (12) ◽  
pp. 1212-1216 ◽  
Author(s):  
D.C. Wescott ◽  
M.N. Pinkerton ◽  
B.J. Gaffey ◽  
K.T. Beggs ◽  
T.J. Milne ◽  
...  

The forces that orthodontic appliances apply to the teeth are transmitted through the periodontal ligament (PDL) to the supporting alveolar bone, leading to the deposition or resorption of bone, depending upon whether the tissues are exposed to a tensile or compressive mechanical strain. To evaluate the osteogenic potential of PDL cells, we applied a 12% uni-axial cyclic tensile strain to cultured human PDL cells and analyzed the differential expression of 78 genes implicated in osteoblast differentiation and bone metabolism by real-time RT-PCR array technology. Sixteen genes showed statistically significant changes in expression in response to alterations in their mechanical environment, including cell adhesion molecules and collagen fiber types. Genes linked to the osteoblast phenotype that were up-regulated included BMP2, BMP6, ALP, SOX9, MSX1, and VEGFA; those down-regulated included BMP4 and EGF. This study has expanded our knowledge of the transcriptional profile of PDL cells and identified several new mechanoresponsive genes.


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