Expression of inflammatory cytokines and MMPs on replanted teeth at different extra-alveolar time: an ex vivo and in vivo study.

BACKGROUND: Immediately after the avulsed tooth is replanted, a complex inflammatory response ensues. As part of the periodontium healing process, the extracellular matrix macromolecules are essential to create the cellular environment required during healing and morphogenesis. AIM: This study was designed to evaluate the correlation between different extra-alveolar dry times and inflammatory cytokines and matrix metalloproteinases (MMPs) as part of the periodontal ligament (PDL) gene expression. DESIGN: The first phase of the study aimed testing human PDL cells ex vivo. Extracted teeth were dried for 15 and 30 min. The PDL cells were extracted and analyzed by qRT-PCR. The second phase was performed in vivo, and 36 Sprague Dawley rat first maxillary molars were extracted and replanted after 15, 30, and 60 min extra-alveolar time. We tested the levels of inflammatory cytokines and MMPS in periodontal tissue at 3, 7, and 28 days after tooth replantation. The replanted area was dissected, grounded, and analyzed by RT-PCR. RESULTS: Expressions of IL-1ß, IL-6, TNF-α, and MMP-3 and MMP-9 were significantly higher in the replanted teeth. Extended dry time had a direct correlation with induction of pro-inflammatory cytokine and MMPs in PDL cells. CONCLUSION: Our study showed that pro-inflammatory cytokines were more significantly expressed in the tissues surrounding the replanted teeth. Future research must be undertaken to additionally confirm the release of these cytokines and be focused on the inhibition of these cytokines to reduce inflammation of replanted teeth.

Expression of pituitary adenylate cyclase-activating peptide (PACAP) and PAC1 in the periodontal ligament after tooth luxation.

Pituitary adenylate cyclase-activating peptide (PACAP) is widely distributed throughout the nervous system. PACAP not only acts as a neurotransmitter but also elicits a broad spectrum of biological action via the PACAP-specific receptor, PAC1. However, no studies have investigated PACAP and PAC1 in the periodontal ligament (PDL), so we aimed to perform this investigation in rats after tooth luxation. In the PDL of an intact first molar, there are few osteoclasts and osteoblasts. However, at days 3 and 5 after luxation, large PAC1-positive cells, thought to be osteoclasts because of their expression of the osteoclast marker, tartrate-resistant acid phosphatase, were detected in appreciable numbers. Osteoblast numbers increased dramatically on day 7 after luxation, and PAC1-positive mononuclear small cells were increased at day 14, many of which expressed the osteoblast marker, alkaline phosphatase. PACAP-positive nerve fibers were rarely detected in the PDL of intact first molars, but were increasingly evident at this site on days 5 and 7 after luxation. Double-immunofluorescence analysis demonstrated the relationship between PACAP-positive nerve fibers and PAC1-positive osteoclasts/-blasts in the PDL. At 5 days after luxation, PACAP-positive nerve fibers appeared in close proximity to PAC1-positive osteoclasts. At 7 days after luxation, PACAP-positive nerve fibers appeared in close proximity to PAC1-positive osteoblasts. These results suggest that PACAP may have effects on osteoclasts and osteoblasts in the PDL after tooth luxation and thus regulate bone remodeling after these types of injury.

Increase of CGRP-containing nerve fibers in the rat periodontal ligament after luxation.

The distribution of calcitonin gene-related peptide (CGRP) was examined in the periodontal ligament (PDL) after experimental luxation injury of the rat first molar tooth. The luxational injury increased the number of CGRP-immunoreactive (IR) nerve fibers. At 3-7 days, numerous CGRP-IR nerve fibers appeared throughout the injured PDL. These nerve fibers terminated as free nerve endings within resorption cavities. Immunohistochemistry for receptor activity modifying protein 1 (RAMP1) also demonstrated that the subunit of CGRP receptor was expressed by periodontal cells adjacent to the alveolar bone in the intact and injured PDL. RAMP1-IR cells were divided into two types; small cells with single nucleus and large cells with 2-6 nuclei. After the luxational injury, both types of RAMP1-IR cells abundantly appeared within resorption cavities. As a result, the treatment increased the number of large RAMP1-IR cells at 3-7 days and small RAMP1-IR cells at 7 days. In addition, a double immunofluorescence analysis demonstrated that CGRP-IR nerve fibers were seen away from RAMP1-IR cells in the intact PDL. After the traumatic injury, however, CGRP-IR nerve fibers appeared in the close vicinity of small and large RAMP1-IR cells at 5-7 days. The morphology and distribution of RAMP1-IR cells suggest that they contain osteoblasts and osteoclasts. By affecting osteoclasts and osteoblasts, CGRP may have effects on bone remodeling in the luxated PDL.

Changes in bone mineralization pattern: a response to local stimulus in maxilla and mandible of dogs.

A pilot study was carried out in order to verify the pattern of changes in mineralization of bone in the maxillas and mandibles of dogs which had a tooth extraction or luxation. Bone mineral content was determined using computerized microdensitometry. Significant changes in patterns of mineralization were found for alveolar bone, cortical bone and trabecular bone at the sites adjacent to the area of operation. These findings suggest that the three envelopes of jaw bones of the dogs are influenced by Regional activation phenomenon (RAP). These results have important implications for the design of clinical studies of periodontium. A more detailed study should elucidate the cellular mechanisms by which these changes occur.