A multiplex immunoassay demonstrates reductions in gingival crevicular fluid cytokines following initial periodontal therapy.
OBJECTIVE
Cytokines and chemokines play an important role in the pathogenesis of periodontal diseases. The objective of this study was to quantitatively assess the effect of initial periodontal therapy on gingival crevicular fluid levels of a comprehensive panel of cytokines and chemokines, including several less extensively studied mediators.


METHODS
Clinical examinations were performed and gingival crevicular fluid samples obtained from six subjects with generalized severe chronic periodontitis prior to initial periodontal therapy and at re-evaluation (6-8 weeks). Four diseased and two healthy sites were sampled in each subject. Twenty-two gingival crevicular fluid mediators were examined using a multiplex antibody capture and detection platform. Statistical analyses were performed by fitting mixed effects linear models to log-transformed gingival crevicular fluid values.


RESULTS
Gingival crevicular fluid interleukin (IL)-1alpha and IL-1beta were the only cytokines to differ in initially diseased vs. initially healthy sites. Following initial therapy, 13 of the 16 detectable cytokines and chemokines decreased significantly in diseased sites, including IL-1alpha, IL-1beta, IL-2, IL-3, IL-6, IL-7, IL-8, IL-12 (p40), CCL5/regulated on activation, normally T cell expressed and secreted (RANTES), eotaxin, macrophage chemotactic protein-1, macrophage inflammatory protein-1alpha and interferon-gamma. At healthy sites, only three of the 16 mediators were significantly altered following therapy.


CONCLUSIONS
This is the first study, to our knowledge, to evaluate such an extensive panel of gingival crevicular fluid mediators within the same sample prior to and following initial therapy. The results confirm that periodontal therapy effectively reduces pro-inflammatory cytokines and chemokines, including less well-described mediators that may be important in initiation and progression of periodontitis. The multiplex assay will prove useful for future gingival crevicular fluid studies.
Thunell
D H
DH
Department of Periodontics, College of Dentistry, The University of Iowa, Iowa City, IA, USA.
Tymkiw
K D
KD
Johnson
G K
GK
Joly
S
S
Burnell
K K
KK
Cavanaugh
J E
JE
Brogden
K A
KA
Guthmiller
J M
JM
D000328:Adult; D000368:Aged; D018932:Chemokine CCL2 / Q000032:analysis; D054405:Chemokine CCL3 / Q000032:analysis; D018946:Chemokine CCL5 / Q000032:analysis; D018925:Chemokines / Q000032:analysis; D019742:Chemokines, CC / Q000032:analysis; D055113:Chronic Periodontitis / Q000628:therapy*; D016207:Cytokines / Q000032:analysis*; D005500:Follow-Up Studies; D005883:Gingival Crevicular Fluid / Q000737:chemistry*; D005884:Gingival Hemorrhage / Q000628:therapy; D005889:Gingival Recession / Q000628:therapy; D006801:Humans; D018836:Inflammation Mediators / Q000032:analysis; D007371:Interferon-gamma / Q000032:analysis; D018664:Interleukin-12 / Q000032:analysis; D053582:Interleukin-1alpha / Q000032:analysis; D053583:Interleukin-1beta / Q000032:analysis; D007376:Interleukin-2 / Q000032:analysis; D007377:Interleukin-3 / Q000032:analysis; D015850:Interleukin-6 / Q000032:analysis; D015851:Interleukin-7 / Q000032:analysis; D016209:Interleukin-8 / Q000032:analysis; D008875:Middle Aged; D017622:Periodontal Attachment Loss / Q000628:therapy; D010514:Periodontal Pocket / Q000628:therapy; D010865:Pilot Projects
C111835:CCL2 protein, human; C483024:CCL5 protein, human; D018932:Chemokine CCL2; D054405:Chemokine CCL3; D018946:Chemokine CCL5; D018925:Chemokines; D019742:Chemokines, CC; D016207:Cytokines; D018836:Inflammation Mediators; D053582:Interleukin-1alpha; D053583:Interleukin-1beta; D007376:Interleukin-2; D007377:Interleukin-3; D015850:Interleukin-6; D015851:Interleukin-7; D016209:Interleukin-8; D018664:Interleukin-12; D007371:Interferon-gamma