An Ultrastructural Perspective on Cell Death

Najdawi, Zaid; Abu‐Asab, Mones

doi:10.35516/jmj.v56i1.232

Cited by 5 publications

(4 citation statements)

References 55 publications

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“…The outcome of the cytotoxic assays was further supported by the TEM analysis, where an increased amount of autophagic vesicles and cytoplasmic autolysosomes were detected in some cells on day 7 following exposure to MTA 48h and Uncaria gambir at 500 µg/mL. The presence of autophagic vesicles and autolysosomes is associated with deranged and degradation of cellular compartment including the proteins and organelles that maintains cell survival [24]. This explained the reduction in cell viability of M3CT3-E1 cells over time after being exposed to both materials.…”

Section: Discussionmentioning

confidence: 76%

Preliminary in vitro cytotoxic evaluation of Uncaria gambier (Hunt) Roxb extract as a potential herbal-based pulpotomy medicament

Tan

Mahyuddin

Sockalingam

et al. 2022

Preprint

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Background The downfall of formocresol as a pulpotomy medicament highlights the importance of cytotoxic evaluation and the establishment of a safe concentration of dental material prior to its usage in the oral cavity. Uncaria gambir is a herbal plant that possesses antimicrobial, anti-inflammatory and antioxidant properties, suggesting its potential as an alternative medicament for pulpotomy. However, a study on its cytotoxicity is lacking. Here, we investigated the concentration and time-dependent toxicity of Uncaria gambir extract towards the M3CT3-E1 cell line and compared it with the gold standard pulpotomy medicament: mineral trioxide aggregate (MTA). Methods Uncaria gambir extracts at concentrations ranging from 1000 to 7.8 µg/ml and MTA eluates at 4- and 48 hours setting times were prepared. 10% dimethyl sulfoxide (DMSO) and culture media were used as positive and negative control respectively. Cell viability on days 1, 2, 3 and 7 were analysed using Alamar Blue and Live and Dead Cell assay. Any morphological cellular changes were evaluated using transmission electron microscopes (TEM). Data were analysed using a two-way mixed Analysis of Variance (ANOVA). Results The interaction between the concentration and exposure time on the fluorescence intensity of Uncaria gambir extract and MTA 48 hours was found to be statistically significant (p < 0.001). No cytotoxic effects on the cells were exerted by both MTA 48 hours and Uncaria gambir extract at a concentration below 500 µg/mL. TEM analysis and Live and Dead Cell assay for both materials were comparable to the negative control. No significant differences in fluorescent intensity were observed between Uncaria gambir extract at 500 µg/mL and MTA 48 hours (p > 0.05). Conclusion Uncaria gambir extracts at a maximum concentration of 500 µg/mL are non-cytotoxic over time and are comparable to the MTA.

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Section: Discussionmentioning

confidence: 76%

Preliminary in vitro cytotoxic evaluation of Uncaria gambier (Hunt) Roxb extract as a potential herbal-based pulpotomy medicament

Tan

Mahyuddin

Sockalingam

et al. 2022

Preprint

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“…Further, at similar concentrations, TEM analysis of the cells showed intact cell and nucleolar membranes and preservation of the organelle structure comparable to the untreated negative control. In contrast, the presence of autophagic vacuolisation and organelle derangement, a feature that is associated with cell apoptosis [47] and autophagy [48], were detected in cells exposed to EEP at a 12.5% concentration. This explains and validates the gradual reduction in cell viability following exposure to EEP at a concentration higher than 6.25%, as shown by the Alamar Blue cytotoxic assay.…”

Section: Discussionmentioning

confidence: 84%

Cytotoxic Evaluation of Effective Ecoproduce (EEP) as a Potential Root Canal Irrigant: A Preliminary In Vitro Study

Hung,

Mahyuddin,

Sockalingam

et al. 2023

Applied Sciences

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Concerns have been raised about the usage of sodium hypochlorite (NaOCl) in endodontics following its toxic effects. Effective ecoproduce (EEP), an organic solution produced through the fermentation of fruit peels, exhibits antibacterial and antibiofilm action, suggesting its potential as an endodontic irrigant. However, studies on its cytotoxicity are limited. This in vitro study aimed to evaluate the cytotoxic effects of EEP at different concentrations and fermentation periods against the MC3T3-E1 cell. EEP derived from orange and pineapple peel waste and fermented for 3 and 6 months was prepared from 100% to 0.78% concentration. Briefly, 2.5% NaOCl was used as the comparison group. Cell viability was analysed using Alamar Blue and Live and Dead Cell assay. A transmission electron microscope (TEM) was used to evaluate ultrastructural changes to the cells. Data analysis was performed using a two-way mixed Analysis of Variance (ANOVA). EEP exhibited concentration-dependent cytotoxicity regardless of the fermentation period (p > 0.05). A concentration below 6.25% was non-cytotoxic and comparable to the negative control (p > 0.05). Live and Dead Cell assay and TEM analysis complement the findings. The mean cell viability of EEP at all concentrations for both fermentation periods was significantly higher than that of 2.5% NaOCl (p < 0.05). Conclusively, 6.25% EEP fermented for 3 and 6 months are non-cytotoxic and can serve as an alternative endodontic irrigants.

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“…The outcome of the cytotoxic assays was further supported by the TEM analysis, where an increased amount of autophagic vesicles and cytoplasmic autolysosomes were detected in some cells on day 7 following exposure to MTA 48h and Uncaria gambir at 500 µg/mL. The presence of autophagic vesicles and autolysosomes is associated with the deranged and degradation of cellular compartment including the proteins and organelles that maintains cell survival [ 28 ]. This explained the reduction in cell viability of M3CT3-E1 cells over time after being exposed to both materials.…”

Section: Discussionmentioning

confidence: 97%

Preliminary in vitro cytotoxic evaluation of Uncaria gambier (Hunt) Roxb extract as a potential herbal-based pulpotomy medicament

Tan,

Mahyuddin,

Sockalingam

et al. 2023

BMC Complement Med Ther

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Background The downfall of formocresol as a pulpotomy medicament highlights the importance of cytotoxic evaluation and the establishment of a safe concentration of dental material prior to its usage in the oral cavity. Uncaria gambir is an herbal plant that possesses antimicrobial, anti-inflammatory and antioxidant properties, suggesting its potential as an alternative medicament for pulpotomy. However, there are not many studies published on its cytotoxicity, with some using non-standardised techniques and reported variable outcomes. Here, we investigated the concentration and time-dependent toxicity of Uncaria gambir extract towards the M3CT3-E1 cell line and compared it with the gold standard pulpotomy medicament: mineral trioxide aggregate (MTA). Methods Uncaria gambir extracts at concentrations ranging from 1000 to 7.8 µg/ml and MTA eluates at 4- and 48 h setting times were prepared. 10% dimethyl sulfoxide (DMSO) and culture media were used as positive and negative controls respectively. Cell viability on days 1, 2, 3 and 7 was analysed using Alamar Blue and Live and Dead Cell assay. Any morphological cellular changes were evaluated using transmission electron microscopes (TEM). Data were analysed using a two-way mixed Analysis of Variance (ANOVA). Results The interaction between the concentration and exposure time on the fluorescence intensity of Uncaria gambir extract and MTA 48 h was found to be statistically significant (p < 0.001). No cytotoxic effects on the cells were exerted by both MTA 48 h and Uncaria gambir extract at a concentration below 500 µg/mL. TEM analysis and Live and Dead Cell assay for both materials were comparable to the negative control. No significant differences in fluorescent intensity were observed between Uncaria gambir extract at 500 µg/mL and MTA 48 h (p > 0.05). Conclusion Uncaria gambir extracts at a maximum concentration of 500 μg/mL are non-cytotoxic over time and are comparable to the MTA.

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An Ultrastructural Perspective on Cell Death

Cited by 5 publications

References 55 publications

Preliminary in vitro cytotoxic evaluation of Uncaria gambier (Hunt) Roxb extract as a potential herbal-based pulpotomy medicament

Preliminary in vitro cytotoxic evaluation of Uncaria gambier (Hunt) Roxb extract as a potential herbal-based pulpotomy medicament

Cytotoxic Evaluation of Effective Ecoproduce (EEP) as a Potential Root Canal Irrigant: A Preliminary In Vitro Study

Preliminary in vitro cytotoxic evaluation of Uncaria gambier (Hunt) Roxb extract as a potential herbal-based pulpotomy medicament

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