Human Periodontal Ligament Cell Viability in Milk and Milk Substitutes

Abstract

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 a Probiotic Storage Media for Avulsed Teeth

“…Svaki zub je nakon sušenja i namakanja 30 minuta bio pohranjen u 15-mililitarskim epruvetama Falcon koje su sadržavale 2,5ml 0,2 mg ml −1 otopine kolagenaze CLS II (Biochrom, Berlin, Njemačka) i 2,4 mg ml −1 otopine dispaze II (Sigma-Aldrich, St.Louis, MO, SAD) u fiziološkoj physiologic saline, cell culture media, probiotic media, propolis, coconut water as storage media (3,(5)(6)(7)(8)(9)(10) Host bacteria are a natural surrounding for avulsed teeth. Probiotic bacteria are live microbial food supplements that may benefit the host by influencing the balance between the many species of the commensal flora both in oral cavity and the intestinal tract (11)(12).…”

Medios de almacenamiento y transporte para dientes avulsionados

“…• Agua: La osmolalidad del agua varía de 3 -16 mOsm/Kg y su pH es de 7,41 4,20 es el medio de transporte menos adecuado pues al ser hipotónica desencadena la lisis celular, adicionalmente e l a l m a c e n a m i e n t o e n a g u a más de veinte minutos provoca grandes reabsorciones radiculares 14,20,23,25,27,29 • Saliva: Por tener una osmolalidad de 60 -80 mOsm/Kg y pH de 6,76 -7,35 [12][13][14][15][16]19,20 como por contener bacterias no es muy idónea. No obstante, los estudios demuestran que si el diente se pone en el vestíbulo bucal, los fibroblastos pueden mantenerse vitales unas dos horas.…”

“…26,29 • L e c h e : E n 1 9 8 1 A n d r e a s e n (Dinamarca) y Blomlof, Lindskog y Hammarstrom (Suecia) fueron los primeros en reportar sobre la viabilidad de la leche fresca descremada pasteurizada como medio para conservar un diente avulsionado. La leche si no fuera por el contenido de lípidos 3,20,23,25 sería un excepcional medio; no obstante, en las condiciones en que se produce un traumatismo dentario es el mejor medio de transporte porque es fácil de conseguir y porque es un líquido estéril debido a la pasteurización…”

“…17 Sin embargo, sólo previene la muerte celular, pero no restituye la forma ni restablece la capacidad mitótica de las células. 2,21,25 Estudios recientes indican que a nivel celular el almacenamiento en leche es similar a la solución de Hank's, siempre que el período en seco no exceda de treinta minutos. Por tanto, la leche es un muy buen medio de almacenamiento a corto plazo, si se coloca el diente en ella como máximo media hora después del trauma.…”

Avulsed Tooth - A Storage Medium Dilemma an Update

“…Considering the unphysiologic media, tap water although is the most convenient medium, it causes rapid cell lysis similar to dry storage due to its hypotonicity, unphysiological pH and osmolality and microbial contamination [8,13,14] and therefore Ozan et al [15] in their study concluded that water was worse in comparison to saline [15]. Likewise saliva emphasizes similar properties as water and therefore inefficient to maintain the cellular integrity [16][17][18][19].…”