Abstract：
BACKGROUND: When infants cannot consume breast milk, the most commonly available alternative milk formula is cow milk-based. Due to a rise in the prevalence of cow milk protein allergy (CMPA) among children, this study aimed to assess the biofilm formation and acidogenicity of cow milk-based formulas as well as milk formulas suggested for children with CMPA.
METHODS: Cow milk-based formulas with 0%, 10%, or 18% sucrose added, partially hydrolyzed formula (pHF), extensively hydrolyzed formula (eHF), amino acid-based formula (AAF), and soy-based formulas with 0%, or 11% sucrose added were evaluated. Streptococcus mutans was used as a representative microorganism associated with caries. The acidogenicity after 24-h incubation was assessed by the pH of the formed biofilm and lactic acid formation. Biofilm formation was quantified using crystal violet staining. Additionally, the biofilm characteristics were determined using confocal laser scanning microscopy (CLSM). Comparisons were made among formulas without added sucrose to observe protein-based differences. Furthermore, formulas with different sucrose percentages were compared to explore the impact of sucrose content.
RESULTS: When comparing the formulas without added sucrose, the biofilm formation in the cow milk-based formula and pHF were significantly greater than the soy-based formula, eHF, and AAF. In the presence of S. mutans, all formulas reduced the biofilm pH below the critical enamel pH. The cow milk-based formula and AAF showed a significantly lower biofilm pH than the pHF, soy-based, and eHF groups, while the lactic acid production was markedly higher in the cow milk-based formula, pHF and AAF, compared with the eHF and soy-based formula. Adding sucrose into the cow milk-based and soy-based formulas substantially increased biofilm mass. The biofilm pH of the cow milk-based formulas, with or without sucrose, was significantly lower than that of the soy-based formulas. The CLSM indicated distinct biofilm characteristics among the different protein-based formulas, with sucrose supplementation promoting S. mutans aggregation in cow milk-based formula biofilm and increased density and intact biofilm in the soy-based formula.
CONCLUSIONS: All assessed milk formulas had caries-inducing factors, including those without supplemental sucrose. Among them, the eHF demonstrated the least caries-inducing factors, attributed to its minimal biofilm formation and the highest biofilm pH.
METHODS: Cow milk-based formulas with 0%, 10%, or 18% sucrose added, partially hydrolyzed formula (pHF), extensively hydrolyzed formula (eHF), amino acid-based formula (AAF), and soy-based formulas with 0%, or 11% sucrose added were evaluated. Streptococcus mutans was used as a representative microorganism associated with caries. The acidogenicity after 24-h incubation was assessed by the pH of the formed biofilm and lactic acid formation. Biofilm formation was quantified using crystal violet staining. Additionally, the biofilm characteristics were determined using confocal laser scanning microscopy (CLSM). Comparisons were made among formulas without added sucrose to observe protein-based differences. Furthermore, formulas with different sucrose percentages were compared to explore the impact of sucrose content.
RESULTS: When comparing the formulas without added sucrose, the biofilm formation in the cow milk-based formula and pHF were significantly greater than the soy-based formula, eHF, and AAF. In the presence of S. mutans, all formulas reduced the biofilm pH below the critical enamel pH. The cow milk-based formula and AAF showed a significantly lower biofilm pH than the pHF, soy-based, and eHF groups, while the lactic acid production was markedly higher in the cow milk-based formula, pHF and AAF, compared with the eHF and soy-based formula. Adding sucrose into the cow milk-based and soy-based formulas substantially increased biofilm mass. The biofilm pH of the cow milk-based formulas, with or without sucrose, was significantly lower than that of the soy-based formulas. The CLSM indicated distinct biofilm characteristics among the different protein-based formulas, with sucrose supplementation promoting S. mutans aggregation in cow milk-based formula biofilm and increased density and intact biofilm in the soy-based formula.
CONCLUSIONS: All assessed milk formulas had caries-inducing factors, including those without supplemental sucrose. Among them, the eHF demonstrated the least caries-inducing factors, attributed to its minimal biofilm formation and the highest biofilm pH.
摘要：
背景：当婴儿不能食用母乳时，最常见的替代牛奶配方是以牛奶为基础的。由于儿童中牛奶蛋白过敏（CMPA）的患病率上升，这项研究旨在评估基于牛奶的配方以及建议用于CMPA儿童的牛奶配方的生物膜形成和产酸性。
方法：含0%的牛乳配方，10%,或添加18%蔗糖，部分水解配方(pHF)，广泛水解配方(eHF)，基于氨基酸的配方（AAF），以及0%的大豆配方，或添加11%蔗糖进行评价。变形链球菌被用作与龋齿相关的代表性微生物。通过形成的生物膜的pH和乳酸形成来评估24小时孵育后的产酸性。使用结晶紫染色定量生物膜形成。此外,使用共聚焦激光扫描显微镜（CLSM）确定生物膜特征。在不添加蔗糖的配方之间进行比较以观察基于蛋白质的差异。此外，比较不同蔗糖含量的配方，探讨蔗糖含量的影响。
结果：当比较不添加蔗糖的配方时，基于牛奶的配方和pHF的生物膜形成明显大于基于大豆的配方，eHF,AAF。在S.mutans面前，所有配方将生物膜pH降低到低于临界牙釉质pH。基于牛奶的配方和AAF显示出显著低于pHF的生物膜pH,以大豆为基础，和eHF组，虽然牛奶配方中的乳酸产量明显更高，pHF和AAF，与eHF和大豆配方相比。将蔗糖添加到基于牛奶和基于大豆的配方中显著增加了生物膜质量。基于牛奶的配方的生物膜pH值，有或没有蔗糖，明显低于大豆配方。CLSM在不同的基于蛋白质的配方中显示出不同的生物膜特征，蔗糖补充剂可促进基于牛奶的配方食品生物膜中的变形链球菌聚集，并增加基于大豆的配方食品中的密度和完整生物膜。
结论：所有评估的牛奶配方都有龋齿诱发因素，包括那些没有补充蔗糖的。其中,eHF表现出最少的龋齿诱发因素，归因于其最小的生物膜形成和最高的生物膜pH。
方法：含0%的牛乳配方，10%,或添加18%蔗糖，部分水解配方(pHF)，广泛水解配方(eHF)，基于氨基酸的配方（AAF），以及0%的大豆配方，或添加11%蔗糖进行评价。变形链球菌被用作与龋齿相关的代表性微生物。通过形成的生物膜的pH和乳酸形成来评估24小时孵育后的产酸性。使用结晶紫染色定量生物膜形成。此外,使用共聚焦激光扫描显微镜（CLSM）确定生物膜特征。在不添加蔗糖的配方之间进行比较以观察基于蛋白质的差异。此外，比较不同蔗糖含量的配方，探讨蔗糖含量的影响。
结果：当比较不添加蔗糖的配方时，基于牛奶的配方和pHF的生物膜形成明显大于基于大豆的配方，eHF,AAF。在S.mutans面前，所有配方将生物膜pH降低到低于临界牙釉质pH。基于牛奶的配方和AAF显示出显著低于pHF的生物膜pH,以大豆为基础，和eHF组，虽然牛奶配方中的乳酸产量明显更高，pHF和AAF，与eHF和大豆配方相比。将蔗糖添加到基于牛奶和基于大豆的配方中显著增加了生物膜质量。基于牛奶的配方的生物膜pH值，有或没有蔗糖，明显低于大豆配方。CLSM在不同的基于蛋白质的配方中显示出不同的生物膜特征，蔗糖补充剂可促进基于牛奶的配方食品生物膜中的变形链球菌聚集，并增加基于大豆的配方食品中的密度和完整生物膜。
结论：所有评估的牛奶配方都有龋齿诱发因素，包括那些没有补充蔗糖的。其中,eHF表现出最少的龋齿诱发因素，归因于其最小的生物膜形成和最高的生物膜pH。
