Viability Of Periodontal Ligament Cells Research Articles

Storage Media for Dental Avulsion: A Literature Review

Tooth avulsion is a critical dental emergency where immediate and appropriate management is essential for the successful reimplantation of the tooth. The choice of storage medium during the extra-alveolar period significantly impacts the viability of periodontal ligament (PDL) cells, crucial for the long-term retention of the replanted tooth. This review examines various storage media proposed for avulsed teeth, assessing their pH, osmolality, and overall efficacy in maintaining PDL cell viability. A comprehensive literature search was conducted across databases including PubMed, Scopus, and Google Scholar, identifying media such as Hank’s Balanced Salt Solution (HBSS), milk, saline, propolis, coconut water, and more. Ideal storage media should exhibit antimicrobial properties, maintain physiological osmolality and pH, support PDL cell viability, and be readily available. HBSS is highlighted as the gold standard, with its ability to preserve PDL cells for up to 48 hours. Other effective media like milk, soy milk, and oral rehydration solutions are discussed for their accessibility and performance. Emerging media such as propolis and epigallocatechin-3-gallate show promise but require further research. This review emphasizes the importance of public awareness and accessibility to effective storage media to improve the prognosis of avulsed teeth. Keywords: Storage Media, Dental Avulsion, PDL viability.

A Comparative Evaluation of the Periodontal Ligament Cell Viability Using Placentrex, Propolis, and Hanks' Balanced Salt Solution: An In-Vitro Study.

Most of the dento-alveolar trauma that occurs frequently in childhood is often linked to avulsion injuries. Two considerable elements influencing the prognosis of tooth avulsion after replantation are extraoral dry duration and the characteristics of the storage media to support the viability of cells. The study aimed to compare and assess the effectiveness of Hanks' balanced salt solution (HBSS) (HiMedia Laboratories, Thane, India) and Placentrex (Albert David Limited, Kolkata, India) against propolis (5% and 10%) as storage media in preserving the vitality of periodontal ligament (PDL) cells. Four groups with 15 samples each were selected for the study. Sixty recently extracted premolars were left to incubate for 30 minutes in 15-ml falcon tubes containing 2.5 ml of collagenase 0.2 mg/ml in phosphate-buffered saline and 2.4 mg/ml of dispase in HBSS, Placentex, propolis 5%, and propolis 10%. After the addition of bovine serum, it was centrifuged for 4 minutes. Trypan blue 0.4% was utilized to recognize the cells, and a hemocytometer was employed for counting the live PDL cells under a light microscope. Propolis 5% and 10%, HBSS, and Placentrex all showed statistically significant differences in PDL cell viability; however, HBSS was significantly outperformed by Placentrex and propolis. Placentrex is a superior substitute storage medium in cases of tooth avulsion as opposed to alternatives such as propolis 5% and 10%.

In-vitro Evaluation of Wild Bitter Melon (Momordica charantia) Extracts against Periodontopathic Bacteria (Prevotella intermedia and Porphyromonas gingivalis) using Antibacterial, Anti-Inflammatory and Molecular Docking Analysis.

Wild bitter melon (Momordica charantia) extracts were prepared and their antibacterial and anti-biofilm assay were investigated against two different periodontopathic bacteria (Prevotella intermedia and Porphyromonas gingivalis) for the first time to the best of our knowledge based on the presence of different phytochemical compounds. Momordica charantia solvent extracts were prepared and phytochemical analysis was performed. Minimal inhibitory and bactericidal concentrations were determined. Antibacterial activity was evaluated using the standard well diffusion method. Anti-inflammatory studies on periodontal ligament (PDL) cell viability and lipopolysaccharide (LPS)-induced inflammation were performed. Molecular docking was investigated between the bioactive compound (Charantadiol A) of plant extract and biofilm-expressing genes in each test organism. Phytochemicals from ethanol extract showed promising results; alkaloids, flavonoids, phenols, and tannins were found present at considerable levels. The minimum inhibitory concentration was found to be 400 μg/mL for Prevotella intermedia and Porphyromonas gingivalis. Antibacterial activity expressed in terms of zone of inhibition showed 14 mm to 18 mm zones against the test organisms. The molecular docking report revealed the maximum binding energy of about -6.54 Kcal/Mol of binding energy between Charantadiol A and fimA of Porphyromonas gingivalis. Anti-biofilm study showed that the minimum biofilm eradication concentration (MBEC) of Momordica charantia expressed significantly good results against the test organisms. The PDL cell viability values expressed in percentage indicated the anti-inflammatory properties of Momordica charantia extracts at three different known concentrations. The findings concluded that Momordica charantia extracts have promising prospects as an anti-periodontopathic and anti-inflammatory agent.

Characterization of aTiO2 surfaces functionalized with CAP-p15 peptide.

Functionalization of Titanium implants using adequate organic molecules is a proposed method to accelerate the osteointegration process, which relates to topographical, chemical, mechanical, and physical features. This study aimed to assess thepotential of a peptide derived from cementum attachment protein (CAP-p15) adsorbed onto aTiO2 surfaces to promote the deposition of calcium phosphate (CaP) minerals and its impact on the adhesion and viability of human periodontal ligament cells (hPDLCs). aTiO2 surfaces were synthesized by magnetron sputtering technique. The CAP-p15 peptide was physically attached to aTiO2 surfaces and characterized by atomic force microscopy, fluorescence microscopy, and water contact angle measurement. We performed in vitro calcium phosphate nucleation assays using an artificial saliva solution (pH 7.4) to simulate the oral environment. morphological and chemical characterization of the deposits were evaluated by scanning electronic microscopy (SEM) and spectroscopy molecular techniques (Raman Spectroscopy, ATR-FTIR). The aTiO2 surfaces biofunctionalized with CAP-p15 were also analyzed for hPDLCs attachment, proliferation, and in vitro scratch-healing assay. The results let us see that the homogeneousamorphous titanium oxide coating was 70 nanometers thick. The CAP-p15 (1 μg/mL) displayed the ability to adsorb onto the aTiO2 surface, increasing the roughness and maintaining the hydrophilicity of the aTiO2 surfaces. The physical adsorption of CAP-p15 onto the aTiO2 surfaces promoted the precipitation of a uniform layer of crystals with a flake-like morphology and a Ca/P ratio of 1.79. According to spectroscopy molecular analysis, these crystalline deposits correspond to carbonated hydroxyapatite. Regarding cell behavior, the biofunctionalized aTiO2 surfaces improved the adhesion of hPDLCs after 24 h of cell culture, achieving 3.4-fold when compared to pristine surfaces. Moreover, there was an increase in cell proliferation and cell migration processes. Physical adsorption of CAP-p15 onto aTiO2 surfaces enhanced the formation of carbonate hydroxyapatite crystals and promoted the proliferation and migration of human periodontal ligament-derived cells in in vitro studies. This experimental model using the novel bioactive peptide CAP-p15 could be used as an alternative to increasing the osseointegration process of implants.

Efficacy of Natural Coconut Water, Pre-packaged Coconut Water, and Hank’s Balanced Salt Solution as Storage Media in Maintaining Periodontal Ligament Cell Viability: An In-vitro Study

Introduction: Avulsion of teeth is one of the most complex forms of dental injury, and the selection of an appropriate storage medium greatly influences the preservation of Periodontal Ligament (PDL) cell viability, which is crucial for the successful re-implantation of avulsed teeth. Therefore, identifying effective storage options such as natural coconut water and prepackaged coconut water holds significant promise in improving outcomes for this challenging dental injury. Aim: To evaluate the efficacy of natural coconut water, prepackaged coconut water, and Hank’s Balanced Salt Solution (HBSS) as storage media in maintaining PDL cell viability. Materials and Methods: A total of 32 non-carious freshly extracted human premolars were randomly divided into four study groups (n=8) and stored in the following storage media respectively: Group I-Natural coconut water group, Group IIPre-packaged coconut water group, Group III-Bench dry group, and Group IV-HBSS groups for 30 minutes. The PDL cells were collected and incubated in phosphate buffer saline for 30 minutes and then centrifuged at 800 rpm for five minutes. Following this, the cells were stained with trypan blue to observe their viability. The Analysis of Variance (ANOVA) with Tukey’s post-hoc test was used for analysing the data. Results: The mean percentage of viable cells in natural coconut water (80.6250) was higher than in HBSS (79.8750), prepackaged coconut water (79.2500), and the bench dry group (6.1250). Meanwhile, the mean percentage of non-viable cells was highest in the bench dry group (93.8750), followed by the pre-packaged coconut water (20.7500), HBSS (20.1250), and natural coconut water (19.3750). Conclusion: Natural coconut water and pre-packaged coconut water are equally effective in maintaining the viability of PDL cells. Therefore, pre-packaged coconut water can be used as a substitute for natural coconut water for tooth storage, depending upon availability.

Viability of periodontal ligament cells in selected transport media for avulsed teeth

To compare the viability of periodontal ligament (PDL) cells stored in Hanks Balanced Salt Solution (HBSS) with those in readily available transport media over a variable period of time. Methods: Periodontal ligament cells harvested from premolars freshly extracted for orthodontic reasons were cultured for exponential growth. The cells were exposed to egg white, evaporated milk, water and Hanks Balanced Salt Solution (HBSS) at room temperature. Their viability was evaluated after 30 minutes, 1 hour and 3 hours with the tetrazolium salt-based colorimetric assay (MTT assay). Statistical analysis was done using the IBM® SPSS version 23.0 software. Comparison between the Mean Optical Densities (MODs) of the cells stored in HBSS and other media at each time interval was done using the independent t test. Repeated measure ANOVA and Tukey post-hoc test were also carried out to compare the MOD of cells within each medium over time. The level of significance was set at p <0.05. Result: The PDL cells stored in egg white had higher MODs than those in HBSS at 30 minutes and 1 hour. Conversely, the MODs of the cells stored in milk and water were lower than those in HBSS at all the studied points. There was a significant difference in the viability of the cells stored in HBSS and water at all the time points (p<0.05). Conclusion: For up to an hour, egg white was found to perform better than HBSS in supporting the viability of PDL cells.

Novel Antibacterial and Biocompatible Nanostructured Gels Based on One-step Synthesis as a Potential Disinfectant for Endodontic Infection Control

AimThe objective of this study was to develop nanostructured gels as biocompatible intracanal disinfectants by one-step microwave radiation-assisted synthesis. MethodsPolyvinyl alcohol (PVA) and polyvinyl pyrrolidone (PVP) were used as a support network, and polyethylene glycol (PEG) was used as a reducing agent. The gels were characterized by measuring the swelling ratio (SR) and rheological properties and by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and atomic force microscopy (AFM). The antibacterial effects of each gel were evaluated against the endodontic clinical strain Enterococcus faecalis. Then, the viability of the 21-day mature multispecies bacterial biofilm was assessed using confocal microscopy in an ex vivo model, where the biofilm was exposed to the mix of nanogels. The cell proliferation, viability, and morphology of human periodontal ligament (HPDL) cells were quantified using a real-time IncuCyte® S3 Live-Cell System. Viability was measured by confocal microscopy using an ex vivo model exposing a 21-day mature multispecies bacterial biofilm to the mix of nanogels. ResultsThe antibacterial activity of the gels coincided with the superficial characterization and the solubility of the gel in the growth medium. Gels with higher viscosity (327.85-980.58 Pa s), higher dissolution (42-70%SR), and lower porosity (no porosity and 611.63 nm) showed excellent antibacterial activity against E. faecalis. Despite their physicochemical characteristics, CuNPs gels showed greater effectiveness against E. faecalis.These nanostructured gels with high PVA concentrations promote HPDL cells proliferation while still exerting antibacterial properties. Mix of nanogels showed an increase non-viable cells biomass from at of application. ConclusionsThe use of biocompatible polymers influences the physicochemical, bactericidal, and cytotoxic response, making these materials potential disinfectant agents against resistant bacteria with good biocompatibility and improved HPDL cells proliferation.

Effect of Polydeoxyribonucleotide on Human Periodontal Ligament Cells as a Storage Medium for Avulsed Tooth

Objective: This study aimed to evaluate the suitability of polydeoxyribonucleotides (PDRN) as a storage medium for avulsed teeth.Materials and Methods: The viability of human periodontal ligament (PDL) cells stored in Hank’s balanced salt solution and PDRN solutions (concentrations, 10, 25, 50, and 100 μg/mL) and tap water was measured using the Cell Counting Kit-8 and Live/Dead assays. In addition, Nitric oxide detection and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to evaluate the anti-inflammatory effect of PDRN.Results: The viability of PDL cells stored in a 100 μg/mL PDRN solution was significantly higher than that of cells stored in the other solutions (<i>p</i> < 0.01). Furthermore, cells stored in 100 μg/mL PDRN solution demonstrated a significantly reduced NO production (<i>p</i> < 0.0001), and cells stored in 50 and 100 μg/mL PDRN solutions expressed significantly lower levels of tumor necrosis factor α, interleukin (IL) -4, IL-6, and IL-10 (<i>p</i> < 0.01) compared to cells stored in HBSS.Conclusion: The PDRN solution exhibited cell-preserving and anti-inflammatory effects on the PDL cells. The findings of this study can serve as a basis for further experiments directed at the development of an effective storage medium for avulsed teeth.

The Comparative Analysis of the Effectiveness of Four Different Storage Media (Placentrex, Propolis 10%, Pomegranate Juice 5%, and Hank's Balanced Salt Solution) in Preserving the Viability of Periodontal Ligament Cells: An In Vitro Study.

Out of all traumas, dentoalveolar trauma occurs quite frequently and is commonly linked to avulsion injuries. Of all severe lesions to the permanent dentition, tooth avulsion accounts for 1%-16% of cases. The extraoral dry time and the storage medium in which the tooth is stored and its capacity to promote cell viability are two important variables influencing the prognosis of an avulsed tooth following replantation. The aim of this paper is to assess and contrast the effectiveness of placentrex, propolis 10%, pomegranate juice 5%, and Hank's balanced salt solution (HBSS) as a storage medium in preserving the periodontal ligament (PDL) cells' viability. Sixty freshly extracted premolars were put in four various media for 45 minutes each, including HBSS, placentrex, propolis 10%, and pomegranate juice 5%, and placed in an incubator for 30 minutes in 15 mL falcon tubes with 2.5 mL solutions of collagenase 0.2 mg/mL and dispase 2.4 mg/mL in phosphate-buffered saline. Bovine serum was added after incubation, and the mixture was centrifuged for four minutes. Trypan blue 0.4% wasused to identify the cells. Under a light microscope, a hemocytometerwasutilized to quantify the number of periodontal ligament cells that were still alive. Propolis, pomegranate, and placentrex are all significantly inferior to HBSS in the context of the sustainability of the cells. It was discovered that HBSS with placentrex and pomegranate juice 5% was statistically insignificant. The difference in periodontal ligament cell viability between propolis 10%, HBSS, placentrex, and pomegranate juice 5% was shown to be statistically significant. When compared to other media such as propolis 10% and pomegranate juice 5%, placentrex is a better alternative storage medium for avulsed teeth.

Comparative Evaluation of Maintenance of Cell Viability of an Experimental Transport Media "Ringer's Lactate" with Dextrose Normal Saline ORS Egg White and Infant Milk Formula for Transportation of an Avulsed Tooth.

The viability of the periodontal ligament (PDL) cells on the root surface of the avulsed tooth determines the prognosis of the replanted tooth, which in turn is determined by a suitable transport medium in which the tooth was stored. The aim of the present study is to evaluate and compare the effectiveness of Ringer's lactate (RL) as a storage medium for an avulsed tooth in maintaining the PDL cell viability with dextrose normal saline (DNS), oral rehydration salt (ORS), egg white (EW), and infant milk formula (IMF). A total of 85 freshly extracted human teeth were divided into five experimental groups and two control groups. The positive and negative controls corresponded to 0-minute and 8-hour dry time, respectively. The experimental teeth were stored dry for 30 minutes and then immersed in one of five experimental media (RL, DNS, ORS, EW, and IMF) for 45 minutes. The teeth were then treated with collagenase type III and trypsin for 10 minutes. The number of viable PDL cells was counted with a hemocytometer and analyzed. Statistical analysis showed that IMF, RL, and EW had no statistically significant differences among them in maintaining the viability of the PDL cells but were significantly better than DNS. No statistically significant difference between RL, EW, and ORS in the number of viable PDL cells. Infant milk formula (IMF), RL, and EW showed similar results within the parameters of the study; they can be used as alternative storage media for avulsed teeth. DNS showed poor results, and ORS could serve as short-term storage media if the other solutions are not readily available. The search for an appropriate storage media with favorable pH and osmolality along with easy availability is the basic thought behind this study. Kumar P, Kotumachagi SS, Fabi AJ, et al. Comparative Evaluation of Maintenance of Cell Viability of an Experimental Transport Media "Ringer's Lactate" with Dextrose Normal Saline ORS Egg White and Infant Milk Formula for Transportation of an Avulsed Tooth. Int J Clin Pediatr Dent 2023;16(3):453-458.

Advanced platelet rich fibrin demonstrates improved osteogenic induction potential in human periodontal ligament cells, growth factor production and mechanical properties as compared to leukocyte and platelet fibrin and injectable platelet rich fibrin.

This cross-sectional invitro research aimed to compare and contrast the macroscopic and microscopic, mechanical and biochemical features of leukocyte-rich platelet-rich fibrin, advanced platelet-rich fibrin, and injectable platelet-rich fibrin. In all, 150 samples were taken from males aged 18 to 25 with good systemic health (n = 50 each for i-PRF, A-PRF, and L-PRF). The samples were assessed for clot length, clot width, membrane length and width. Microscopic parameters assessed were the distribution of cells and fibrin structure. Mechanical tests were performed for tensile strength using a universal testing machine and growth factor analysis was performed for platelet derived growth factor (PDGF), vascular endothelial growth factor (VEGF), and transforming growth factor (TGF)- on Days 1, 3 and 7 using commercially available ELISA kits. The osteogenic potential was analyzed in a culture of human periodontal ligament cells for 21days using cell viability assay, alkaline phosphatase formation and alizarin red staining for mineralization. L-PRF demonstrates statistically superior clot length, width, weight, membrane length, width and weight in comparison to A-PRF (p < 0.05). L-PRF demonstrates a denser fibrin structure in comparison to A-PRF and i-PRF (p < 0.05). The cells in L-PRF are most commonly situated in the proximal of the clot where as they are distributed in the proximal and middle aspect for A-PRF(p < 0.05). A-PRF demonstrates the highest tensile strength followed by L-PRF (p < 0.05). When growth factor release was evaluated, A-PRF showed noticeably increased release of all growth factors, namely PDGF-BB, TGF-ß, and VEGF, in comparison to i-PRF and L-PRF (p < 0.05). On days 7 and 14, the cell viability of human periodontal ligament cells in co-culture with A-PRF was statistically substantially greater than that of L-PRF and i-PRF (p < 0.05). Alkaline phosphatase levels were statistically substantially higher in A-PRF, followed by i-PRF and L-PRF on days 14 and 21 (p < 0.05). After 21days of culture, A-PRF treated cultures had much more Alizarin Red staining than L-PRF and i-PRF cultures did (p < 0.05). It was determined that although L-PRF exhibits greater size and weight in comparison to A-PRF and i-PRF, A-PRF has superior mechanical properties, increased growth factor releases of TGF-b, PDGF-BB, and VEGF as well as superior cell viability, alkaline phosphatase production, and mineralization on human periodontal ligament cells. Based on these findings, A-PRF can be recommended for improved delivery of growth factors and osteogenesis whereas L-PRF is better-suited for applications relying on the size of membrane.

Evaluation of Viability of Periodontal Ligament Cells using Propolis, Coconut Water, Aleo Vera, and Soy Milk: An In vitro Comparative Study.

This research was done to assess the periodontal ligament cells viability using propolis, coconut water, aloe vera, and soy milk storage media. Periodontal ligament cells were taken from freshly extracted teeth and stored into four groups of storage media: Group A-propolis, Group B-coconut water, Group C-aloe vera, and Group D-soy milk. Later, the cell viability was assessed at 1, 12, and 24 hours. The periodontal ligament cell viability was significantly greater in propolis, followed by coconut water, aloe vera, and soy milk. We found that propolis, coconut water, aloe vera, and soy milk can be used as an effective storage media. These are naturally occurring and easily available storage medium.

An In Vitro Evaluation of Ice Apple as a Novel Storage Medium to Preserve the Viability of Human Periodontal Ligament Fibroblasts.

For a better prognosis of an avulsed tooth, it is important to store it in an appropriate storage medium prior to replantation. The purpose of this study was to evaluate the ability of ice apples to preserve the viability of periodontal ligament (PDL) fibroblasts. Periodontal ligament (PDL) fibroblasts were procured from the roots of healthy premolars and cultured in Dulbecco's Modified Eagle's Medium (DMEM). They were preserved with-ice apple water (IAW), 30 and 10% ice apple fruit pulp extract (IAFPE), DMEM, negative control without any agent, and positive control DMEM supplemented with fetal bovine serum (FBS). Culture plates were incubated with investigational media at 37°C for 1, 3, and 24 hours. Each experiment was repeated thrice. Assessment of cell viability was done using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium reduction (MTT) assay. After every test period, storage media was removed; 60 µL of MTT solution was added to each well and incubated for 3 hours at 37°C. The supernatant was aspirated and the formazan blue crystals formed were solubilized using dimethyl sulphoxide (DMSO) (100 µL). Optical density was measured at a wavelength of 490 nm. The effects of the test storage media at each time period were evaluated by a one-way analysis of variance (ANOVA) test, and this was followed by post hoc Tukey's multiple comparison tests (p < 0.05). A total of 10% IAFPE demonstrated the maximum capacity to maintain PDL cell viability in all three test periods (p = 0.001). Among the different forms of ice apple used in this study, IAFPE showed better results as compared to IAW (p = 0.001). Ice apple fruit pulp extract (IAFPE) at 10% concentration showed the maximum capacity to maintain PDL cell viability across all three test periods. Therefore, it can be considered a suitable alternative natural storage medium for avulsed teeth. However, further, more detailed studies are required in this field. Bijlani S, Shanbhog RS. An In Vitro Evaluation of Ice Apple as a Novel Storage Medium to Preserve the Viability of Human Periodontal Ligament Fibroblasts. Int J Clin Pediatr Dent 2022;15(6):699-703.

Evaluation of delphinidin as a storage medium for avulsed teeth

BackgroundDelphinidin (DP), an anthocyanidin found in blueberries, has antioxidant and anti-inflammatory effects. This study aimed to investigate the efficacy of DP as a storage medium for avulsed teeth.MethodsHuman periodontal ligament cells were cultured and exposed to DP solution (10, 50, and 100 μM), Dulbecco’s modified Eagle’s medium, Hank’s balanced salt solution and tap water. Cell counting kit-8 assays were performed after 0.5, 1, 6, and 24 h to measure the cell viability. Nitric oxide assays and gelatin zymography were performed to evaluate the anti-inflammatory effects of DP. Reverse transcription-polymerase chain reaction was used to determine the expression levels of inflammatory cytokines.ResultsThe viability of periodontal ligament cells was greatest at 100 μM DP. At 1 h, 100 μM DP decreased nitric oxide synthesis (p < .0167). Matrix metallopeptidase-9 activity was inhibited by DP in a dose-dependent manner (p < .0167). Moreover, treatment with 100 μM DP decreased the expression levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-8 in periodontal ligament cells (p < .0167).ConclusionsWithin the limits of this study, DP preserved the viability and suppressed the inflammatory response of periodontal ligament cells. These findings suggest that DP could be promising for preservation of avulsed teeth.

Effect on Cellular Vitality In Vitro of Novel APRF-Chlorhexidine Treated Membranes.

Chlorhexidine (CHX) has been used for some time in clinical practice as a local antiseptic agent with excellent efficacy. The combination of CHX with APRF (Advanced-platelet rich fibrin) membrane has the potential to stimulate tissue regeneration and to provide a bactericidal effect. We hypothesize that this may reduce the rate of infections development and protect cell viability. The aim of this study was two-fold-to create a stable APRF membrane treated with different concentrations of CHX (0.01% and 0.02%) and to monitor its effect on the viability of PDL cells in vitro. This benefits the introduction of a new protocol for APRF membrane production -CHX-PRF and enriches the available evidence on the effect of this antiseptic agent on PDL (Periodontal ligament) cells. APRF membranes were prepared by the addition of two concentrations (0.01% and 0.02%) of CHX. Membranes without the antiseptic were also prepared and used as control samples. PDL cells were cultivated on the membranes for 72 h. Cell number and vitality were examined by fluorescent cell viability assays. Our results demonstrated that a concentration of 0.01% CHX allowed the production of a stable APRF membrane. This concentration slightly reduced the viability of PDL cells to 96.7%, but significantly decreased the average number of cells attached to the membrane-149 ± 16.5 cells/field compared to controls -336 ± 26.9 cells/field. APRF-CHX 0.02% membranes were unstable, indicating a dose-dependent cytotoxic effect of CHX. The introduced novel protocol leads to the production of a new type of APRF membrane-CHX-PRF. The incorporation of an antiseptic into the APRF membrane can improve its bactericidal activity and might serve as an important step for the prevention of postoperative infections.

Impacts of long noncoding RNA MALAT1 on LPS-induced periodontitis via modulating miR-155/SIRT1 axis

Abstract Objectives Periodontitis, a dental disease that causes inflammation of gums is triggered by a bacterial infection. Long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (lncRNA MALAT1) has been reported to participate in inflammatory diseases. In the present study, we investigated the effects of lncRNA MALAT1 on periodontal ligament cells (PDLCs) with lipopolysaccharide (LPS) induction. Methods MALAT1, microRNA-155 (miR-155), Sirtuin 1 (SIRT1), Bax, and Bcl-2 RNA expressions were detected by using real-time quantitative polymerase chain reaction (RT-qPCR). PDLC viability and apoptosis were assessed by using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium salt (MTT) assay and flow cytometry. Enzyme-linked immunosorbent assay (ELISA) assessed secretions of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). Luciferase reporter test was applied for confirming the binding of miR-155 with MALAT1 and SIRT1, respectively. Results Overexpression of MALAT1 facilitated the LPS-induced periodontitis while MALAT1 suppression restrained LPS-induced injury. Also, miR-155 was expressed in LPS-induced PDLCs. Moreover, MALAT1 negatively regulated miR-155 followed by the up-regulation of SIRT1 which in turn reduced the inflammation in LPS-induced PDLCs. Conclusions We concluded that upregulated lncRNA MALAT1 could accelerate periodontitis through the regulation of miR-155/SIRT1. Our findings suggested a novel MALAT1/miR-155/SIRT1 pathway for the treatment of periodontitis.

Comparative evaluation of the efficacy of aloe vera juice, amla juice, and milk as storage media in maintaining the periodontal ligament cell viability

Background: Tooth avulsion implies total displacement of the tooth out of its socket. Many storage media such as milk, saliva, Hank’s Balanced Salt Solution (HBSS), propolis, Viaspan, have been studied as storage medias for preserving PDL cell viability. Amongst these, HBSS has been proposed as the storage medium of choice for treatment of avulsed teeth by the American Associations of Endodontists. However, the major disadvantage is that it is not easily available at places where these traumatic injuries occur. Aim: To evaluate and compare the efficacy of aloe vera juice, amla juice, milk and water in maintaining the periodontal ligament cell viability of avulsed teeth. Study Design: Forty premolars extracted for orthodontic therapeutic purposes were randomly and equally divided into four groups based on storage media used [Group I: aloe vera juice; Group II: amla juice; Group III: milk; Group IV: water. Following extractions, the teeth were dried for 30 mins and were placed in one of the four different storage media for 45 minutes, following which the scrapings of the PDL from these teeth were collected in Falcon tubes containing collagenase enzyme in 2.5 mL of phosphate buffered saline.

Comparative evaluation of periodontal ligament cell viability of permanent teeth in five different storage media followed by simulated avulsion injury

Aim: To evaluate and compare the viability of periodontal ligament cells of avulsed teeth in five different storage media followed by simulated avulsion injury. Settings and Design: Seventy-five premolars extracted for orthodontic therapeutic purposes were randomly and equally ivided into five groups based on storage media used [A: HBSS (control), B: Milk (experimental), C: Aloe Vera (experimental), D: Egg white (experimental), E: Coconut water (experimental)].Methods and Material: Following extractions, the teeth were placed in one of the five different storage media for 60 minutes, following which the scrapings of the PDL were collected in Falcon tubes which already contained collagenase enzyme in 2.5 mL of Phosphate buffered saline. The tubes were subsequently incubated and centrifuged. Then acquired PDL cells were stained with Trypan Blue dye and were counted under an optical microscope. Statistical Analysis Used: Results were subjected to statistical analysis using ANOVA test and Post Hoc Tukey test. p-value &lt; 0.05 is considered to be significant. Results: HBSS showed the highest percentage of viable cells (80.14%), followed by Egg white (74.01%) and Aloe Vera (73.68%). Milk and Coconut water showed the least percent of viable cells 63.20%, 63.58% respectively.

The different irradiation parameters of carbon dioxide laser effects on periodontal ligament cells

Background/purpose: Photobiostimulation (PBS) can affect cellular functions. The objective of the present study was to evaluate the cellular changes in periodontal ligament (PDL) cells that received different carbon dioxide (CO2) laser irradiation parameters under negative pressure culture. Materials and methodsThe negative pressure-cultured PDL cells on normal medium and differentiation medium were subjected to continuous irradiation with a CO2 laser at an energy density of 5 J/cm2 or 10 J/cm2. The irradiated PDL cells were harvested at Days 1, 5 and 7, and their viability was analyzed by the Presto Blue assay and the biologic markers alkaline phosphatase (ALP), bone sialopoietin (BSP), osteopontin (OPN), osteocalcin (OC), matrix metalloproteinase-3 (MMP-3) collagen I (Col I) and cyclooxygenase-2 (COX-2) expression by reverse transcription-polymerase chain reaction (RT–PCR). ResultsThe PDL cell viability showed that the differentiation medium groups were higher than the normal culture groups. The cell morphologies were all expressed as spindle type. The inflammatory markers in the laser-irradiated groups were higher on the first day and decreased on the seventh day (P < 0.05). Osteogenesis markers were highly expressed at different time periods (P < 0.05). The Col I and OPN genes were highly expressed on the first day, and the Col I high expression lasted until the seventh day. The OC gene was highly expressed on the seventh day. The effects of PDL cultured in differential medium and normal medium were the same in the present study. ConclusionA low-dose CO2 laser continuously irradiating cultured PDL cells can induce osteogenesis and reduce cell inflammatory expression.

Comparative Evaluation of Corneal Storage Medias Used as Tooth Avulsion Medias in Maintaining the Viability of Periodontal Ligament Cells Using the Cell Counting Kit-8 Assay.

PurposeThe prime factor in determining the success of reimplantation of an avulsed tooth is the maintenance of the viability of periodontal ligament fibroblast cells (PDFC). This study aims to evaluate and compare Mc Carey Kaufman media (MK), Cornisol, Dulbecco’s Modified Eagles Medium (DMEM), Hanks Balanced Salt Solution (HBSS) and distilled water in preserving the viability of the PDFC using the Cell Counting Kit-8 assay (CCK-8).MethodsCryopreserved PDFC were suspended in DMEM and incubated in CO2 incubator at 370C with 95% humidity and 5% CO2 for attachment. Once cells attained 80% confluence, they were trypsinised and passed into T-25 culture flasks to expand the culture population. Cells from passage 5 were pooled for experimentation. Trypan blue exclusion test was performed before each experiment to measure cell viability and batches showing more than 95% viability were used in the experiment. The viable PDFC with 1×105 were seeded in 96 well plates and incubated in CO2 incubator at 370C, 95% humidity and 5% CO2 for 24 hours to allow cell attachment. A 100µL of the experimental media were added in the wells and the cells were exposed for 1, 24 and 48 hours respectively. The viability was determined using the CCK-8. Experiment was performed in triplicates and data was subjected to statistical analysis.ResultsStatistical analysis was performed using repeated measure ANOVA, ANOVA, and post-hoc Bonferroni test with the significance level p<0.05. The values are as follows: MK (1.3146 ±0.0588, 1.9012±0.0511, 2.0723±0.1211) > Cornisol (1.2399±0.0548, 1.9596±0.0652, 1.9592±0.1361) >DMEM (1.1914±0.0691, 1.8479±0.0116, 2.0718±0.0795) > HBSS (0.3665±0.0814, 0.0184±0.0010, 0.0248±0.0042) >distilled water (0.0122±0.0033, 0.0225±0.0085, 0.0104±0.0008) at 1 hour, 24 hours and 48 hours respectively. MK >Cornisol>DMEM>HBSS>distilled water.ConclusionIt can be concluded that the corneal preservation solutions showed promising results in preserving periodontal ligament cell viability for extended time periods.