UKY DENTAL COBRE: POLYBACTERIAL PERIODONTITIS: N-3 PUFA AND ANTIOXIDANTS

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。最初的项目3侧重于用小鼠颅骨模型研究多微生物感染诱导炎症和骨质流失。该试验旨在检测牙龈假单胞菌、连翘假单胞菌和齿状假单胞菌的协同毒力效应。此外，目的是确定在免疫炎症和组织破坏后获得性免疫反应对感染和主动免疫的影响。在提交COBRE之后，在2004年9月获得资助之前，该研究获得了NIDCR的R01资助。因此，COBRE拨款为PI， Kesavalu博士，a ？过渡奖?以扩展他的研究，并协助提交第二份R01申请。该基金为一项初步研究提供了支持，该研究旨在记录牙龈卟啉单胞菌（Pg）、齿齿龈卟啉单胞菌（Td）和连翘单胞菌（Tf） （Pg/Td/Tf）感染大鼠口腔的能力。与此相比，将具核梭菌（桥接和聚集病原体）掺入这种混合物（Pg/Td/Tf/Fn）可能会提高感染方案的有效性。采用改良的感染程序，连续5天口服含有微生物混合物的2% CMC溶液。擦拭磨牙，从口腔样本中分离DNA，并使用PCR评估细菌的存在。结果证实了大鼠感染多微生物致病联合体的重要数据。对大鼠实施安乐死，收集牙龈组织并在液氮中快速冷冻。采集血和大鼠下颚进行血清IgG抗体检测和牙槽骨吸收测定。对多微生物感染成员的显著血清抗体反应被注意到，支持口腔组织感染和口腔感染的全身挑战。有趣的是，Pg/Td/Tf/Fn联合体的多微生物攻击（5天，12周内1次）似乎引发了更大的抗体。感染这些联合体的大鼠的牙槽骨丢失表现出两个主要结果。单一感染方案挑战Pg/Td/Tf引起最小的骨质流失，支持单一挑战方案不足以建立和维持疾病过程。然而，Pg/Td/Tf/Fn联合感染在12周内引起了明显的骨质流失，明显大于任何单一感染，在12周内接受多次感染。这些发现提供了额外的初步数据，以支持目前正在美国国立卫生研究院审查的R01申请。目的1：确定n-3 PUFA对病原菌P. gingivalis-T诱导的牙周炎症和牙槽骨丢失的调节作用。forsythia-T。老鼠的牙髓。具体目标2。确定膳食抗氧化补充剂的效果？-硫辛酸和维生素E (？-LA/VE)对大鼠牙周炎症和牙槽骨丢失的调节作用。具体目标3。确定姜黄素这种有效的抗炎/抗氧化天然膳食物质对大鼠牙周炎症和牙槽骨丢失的调节作用，这些牙周炎症和牙槽骨丢失是由致病性牙龈卟啉单胞菌、连翘单胞菌和牙齿单胞菌引起的。具体目标4。确定膳食抗炎/抗氧化剂组合作为共同补充对调节或减少大鼠牙周炎症和牙槽骨丢失的影响，牙周炎症和牙槽骨丢失是由致病性牙龈卟啉单胞菌、连翘单胞菌和牙齿单胞菌引起的。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The original Project 3 focused on polymicrobial infections inducing inflammation and bone loss using a murine calvarial model. It was designed to examine synergistic virulence effects of P. gingivalis, T. forsythensis, and T. denticola. Additionally, the aims were developed to determine the impact of acquired immune responses to infection and active immunity following immunization of the inflammation and tissue destruction. Following submission of the COBRE and prior to its funding in Sept. 2004, this research was funded as an R01 from the NIDCR. Thus, the COBRE grant provided the PI, Dr. Kesavalu, a ?Transition Award? to extend his research and contribute to submission of a 2nd R01 application. The funding provided support for a pilot study, initiated to document the ability to infect the oral cavity of rats with the polymicrobial consortium of P. gingivalis (Pg), T. denticola (Td), and T. forsythia (Tf) (Pg/Td/Tf). This was compared to the potential that incorporation of F. nucleatum (bridging and coaggregating pathogen) into this mixture (Pg/Td/Tf/Fn) would enhance the effectiveness of the infection regimen. Using an modified infection procedure, rats were challenged orally on 5 consecutive days with a 2% CMC solution containing a mixture of the microorganisms. The molar teeth were swabbed, DNA isolated from oral samples, and assessed using PCR for the presence of the bacteria. The results demonstrated seminal data that the rats were infected with the polymicrobial pathogenic consortia. Rats were euthanized, the gingival tissues collected and snap frozen in liquid nitrogen. Blood and rat jaws also were collected for serum IgG antibody and assessment of alveolar bone resorption. A significant serum antibody response to members of the polymicrobial infection was noted, supporting oral tissue infection and systemic challenge by the oral infection. Interestingly, the polymicrobial challenge with the Pg/Td/Tf/Fn consortium (5 days, 1 time during the 12 weeks), appeared to elicited substantially greater antibody. Alveolar bone loss in rats infected with these consortia demonstrated 2 primary findings. The single infection regimen with the Pg/Td/Tf challenged elicited minimal bone loss, supporting that a single challenge regimen is not sufficient to establish and maintain a disease process. However, the Pg/Td/Tf/Fn consortium, with a minimal infection challenge, induced significant bone loss, that was significantly greater than noted with any of the monoinfections, which had received the challenge multiple times during the 12 weeks. These findings provided additional preliminary data in support of an R01 application that is currently under review at the NIH. The aims of this proposal include: SPECIFIC AIM 1: To determine the effect of n-3 PUFA to modulate periodontal inflammation and alveolar bone loss induced by the pathogenic consortium P. gingivalis-T. forsythia-T. denticola in rats. SPECIFIC AIM 2. To determine the effect of a dietary antioxidant supplement ?-lipoic acid and vitamin E (?-LA/VE) to modulate periodontal inflammation and alveolar bone loss induced by the pathogenic consortium P. gingivalis, T. forsythia, and T. denticola in rats. SPECIFIC AIM 3. To determine the effect of a potent anti-inflammatory/antioxidant natural dietary substance, curcumin, to modulate periodontal inflammation and alveolar bone loss induced by the pathogenic consortium P. gingivalis, T. forsythia, and T. denticola in rats. SPECIFIC AIM 4. To determine the effect of combinations of dietary anti-inflammatories/antioxidants as co-supplements to modulate or decrease periodontal inflammation and alveolar bone loss induced by the pathogenic consortium P. gingivalis, T. forsythia, and T. denticola in rats.