Optimization of conditions for in vitro modeling of subgingival normobiosis and dysbiosis.
Optimization of conditions for in vitro modeling of subgingival normobiosis and dysbiosis.
复制标题
DOI：
10.3389/fmicb.2022.1031029
复制
发表时间：
2022
影响因子：
5.2
通讯作者：
Al-Hebshi, Nezar N. N.
中科院分区：
文献类型：
--
作者：
Baraniya, Divyashri;Do, Thuy;Chen, Tsute;Albandar, Jasim M. M.;Chialastri, Susan M. M.;Devine, Deirdre A. A.;Marsh, Philip D. D.;Al-Hebshi, Nezar N. N.
对健康和疾病中的牙龈下微生物组进行建模是识别生物失调的驱动因素和研究微生物组调控的关键。在这里，我们优化了我们前面描述的体外龈下微生物组模型的生长条件。以健康受试者和牙周炎受试者的龈下菌斑标本为接种物，在MBEC试板上培养正菌和反菌微生物。唾液中添加1%、2%、3.5%或5%(v/v)的热灭活人血清，在摇动或非摇动条件下作为生长介质。分别于生长第4、7、10、13 天采集微生物群，进行16SRRNA基因测序分析。随着血清浓度和孵化时间的延长，生物量显著增加。与生长条件无关，健康和牙周炎来源的微生物群分别与各自的接种物聚集在一起。物种丰富度/多样性随时间略有增加，但受到较高的血清浓度的不利影响，特别是在牙周炎来源的微生物群中。随着时间的延长和血清浓度的增加，微生物的微生态失调程度增加。以链球菌、梭杆菌和普雷沃特氏菌为代价，卟啉单胞菌和别雷沃氏菌的血清浓度大大增加。随着时间的推移，最显著的变化是卟啉单胞菌、类杆菌和嗜热杆菌的增加，而普雷沃泰氏菌、卡通氏菌和双胞菌的减少。摇晃只有很小的影响。总体而言，健康来源的微生物在1%的血清中生长4 天，牙周炎来源的微生物在3.5%-5%的血清中生长7 天与各自的接种组最相似。综上所述，在含血清的唾液中可以重复生长正常生物和非生物的龈下微生物群，但对于健康和牙周炎来源的微生物群，需要不同的时间和血清浓度进行调整，以最大限度地提高与体内接种的相似性。优化后的模型可用于识别生物失调的驱动因素，并评估微生物组调节剂等干预措施。
Modeling subgingival microbiome in health and disease is key to identifying the drivers of dysbiosis and to studying microbiome modulation. Here, we optimize growth conditions of our previously described in vitro subgingival microbiome model. Subgingival plaque samples from healthy and periodontitis subjects were used as inocula to grow normobiotic and dysbiotic microbiomes in MBEC assay plates. Saliva supplemented with 1%, 2%, 3.5%, or 5% (v/v) heat-inactivated human serum was used as a growth medium under shaking or non-shaking conditions. The microbiomes were harvested at 4, 7, 10 or 13 days of growth (384 microbiomes in total) and analyzed by 16S rRNA gene sequencing. Biomass significantly increased as a function of serum concentration and incubation period. Independent of growth conditions, the health- and periodontitis-derived microbiomes clustered separately with their respective inocula. Species richness/diversity slightly increased with time but was adversely affected by higher serum concentrations especially in the periodontitis-derived microbiomes. Microbial dysbiosis increased with time and serum concentration. Porphyromonas and Alloprevotella were substantially enriched in higher serum concentrations at the expense of Streptococcus, Fusobacterium and Prevotella. An increase in Porphyromonas, Bacteroides and Mogibacterium accompanied by a decrease in Prevotella, Catonella, and Gemella were the most prominent changes over time. Shaking had only minor effects. Overall, the health-derived microbiomes grown for 4 days in 1% serum, and periodontitis-derived microbiomes grown for 7 days in 3.5%–5% serum were the most similar to the respective inocula. In conclusion, normobiotic and dysbiostic subgingival microbiomes can be grown reproducibly in saliva supplemented with serum, but time and serum concentration need to be adjusted differently for the health and periodontitis-derived microbiomes to maximize similarity to in vivo inocula. The optimized model could be used to identify drivers of dysbiosis, and to evaluate interventions such as microbiome modulators.
登录
查看更多内容
影响因子：
--
作者：
Walker, C.;Sedlacek, M. J.
通讯作者：
Sedlacek, M. J.
影响因子：
4.5
作者：
Al-Hebshi NN;Nasher AT;Idris AM;Chen T
通讯作者：
Chen T
影响因子：
15.5
作者：
Edlund A;Yang Y;Hall AP;Guo L;Lux R;He X;Nelson KE;Nealson KH;Yooseph S;Shi W;McLean JS
通讯作者：
McLean JS
影响因子：
4.3
作者：
Mallick H;Rahnavard A;McIver LJ;Ma S;Zhang Y;Nguyen LH;Tickle TL;Weingart G;Ren B;Schwager EH;Chatterjee S;Thompson KN;Wilkinson JE;Subramanian A;Lu Y;Waldron L;Paulson JN;Franzosa EA;Bravo HC;Huttenhower C
通讯作者：
Huttenhower C
影响因子：
4.2
作者：
Kistler JO;Pesaro M;Wade WG
通讯作者：
Wade WG