Hank's Balanced Salt Solution Supplemented With L‐Ascorbic Acid Modulates Periodontal Ligament Cell Inflammation and Regenerative Responses: An In Vitro and Ex Vivo Study
Li Fuan Poh, Dalia Abdullah, Sook Luan Ng, Eason Soo, Amy Kia Cheen LiewABSTRACT
Background
Successful management of an avulsed tooth depends on preserving periodontal ligament viability during the extraoral period. Although Hank's Balanced Salt Solution (HBSS) is recommended for storage of avulsed teeth, it lacks bioactive components that actively support tissue repair. This study evaluated whether supplementation with L‐ascorbic acid (L‐AA) could enhance biological responses associated with periodontal healing.
Material and Methods
Complementary in vitro and ex vivo models were used. Cultured human periodontal ligament cells were exposed to HBSS, HBSS supplemented with L‐AA, or Dulbecco's Modified Eagle Medium containing fetal bovine serum. Wound closure, inflammatory cytokine expression, matrix metalloproteinase expression, and collagen type I alpha 1 chain gene and protein expression were evaluated. In the ex vivo model, 18 freshly extracted premolars were stored in the respective media for 24 h at 4°C before assessment of periodontal ligament tissue viability and gene expression. Statistical significance was set at p < 0.05.
Results
The optimized concentration of L‐AA was 175 μg/mL. Supplementation of HBSS with L‐AA resulted in significantly greater wound closure than HBSS alone. In both in vitro and ex vivo models, L‐AA supplementation significantly reduced the expression of pro‐inflammatory cytokines and matrix metalloproteinases while enhancing the expression of collagen type I alpha 1 chain. Although periodontal ligament tissue viability was numerically higher following supplementation, differences were not statistically significant.
Conclusions
Supplementation of HBSS with L‐AA favorably modulated inflammatory and extracellular matrix‐related responses in cultured and ex vivo periodontal ligament tissues. Although no significant improvement in tissue viability was observed, the findings suggest potential biological benefits relevant to periodontal repair. Further studies are required to validate these findings in animal replantation models and determine their clinical relevance.