Periodontal disease has been implicated in the progression of various systemic diseases, including chronic kidney disease (CKD). Recent evidence suggests that infection of Porphyromonas gingivalis , a major periodontal pathogen, may also contribute to vascular calcification in patients with CKD. In the present study, antibody array analysis of serum samples from CKD mice administered with oral P. gingivalis revealed significant alterations in protein expression profiles, with notable interleukin-7 (IL-7) upregulation. We demonstrated that P. gingivalis infection enhances the inorganic phosphate-induced calcification of vascular smooth muscle cells (VSMCs), a pathological process that is characteristically accelerated in CKD. Notably, IL-7 expression was significantly upregulated in the P. gingivalis -stimulated calcification of VSMCs. Moreover, IL-7 knockdown in VSMCs markedly attenuated the P. gingivalis -stimulated calcification of VSMCs and suppressed the expression of osteogenic markers, including alkaline phosphatase and Runt-related transcription factor 2. These findings suggest that IL-7 plays a crucial role in P. gingivalis -stimulated vascular calcification, potentially providing new therapeutic targets for preventing vascular calcification in CKD patients with periodontal infection.

Introduction
Materials and Methods
    1. Bacterial strain and growth conditions
    2. Induction of CKD in the mice model
    3. Antibody array assay
    4. Data acquisition and analysis
    5. Enzyme-linked immunosorbent assay
    6. Cell culture and P. gingivalis infection
    7. Gene knockdown by small interfering RNA
    8. Induction and quantification of calcification
    9. Alizarin red S staining
    10. Quantitative real-time reversetranscription-polymerase chain reaction
    11. Western blot analysis
    12. Statistical analysis
Results
    1. Oral administration of P. gingivalis alters serumprotein expression profiles in a CKD mouse model
    2. Expression of IL-7 in P. gingivalis -infected vascularcalcification of VSMCs
    3. Validation of IL-7 knockdown efficiency in VSMCs
    4. Effect of IL-7 silencing on P. gingivalis -stimulatedVSMC calcification and osteogenic markerexpression
Discussion
Funding
Conflicts of Interest
References

• Hyun-Joo Park(Department of Oral Physiology, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea, Periodontal Disease Signaling Network Research Center (MRC), School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea, Dental and Life Science Institute, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea)
• Mi-Kyoung Kim(Department of Oral Physiology, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea)
• Yeon Kim(Department of Oral Physiology, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea, Periodontal Disease Signaling Network Research Center (MRC), School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea, Dental and Life Science Institute, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea)
• Soo-Kyung Bae(Dental and Life Science Institute, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea, Department of Dental Pharmacology, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea)
• Hyung Joon Kim(Department of Oral Physiology, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea, Periodontal Disease Signaling Network Research Center (MRC), School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea, Dental and Life Science Institute, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea)
• Moon-Kyoung Bae(Department of Oral Physiology, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea, Periodontal Disease Signaling Network Research Center (MRC), School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea, Dental and Life Science Institute, School of Dentistry, Pusan National University, Yangsan 50612, Republic of Korea) Corresponding author