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21.
Immunohistochemical localization of components of the insulin-like growth factor system in human permanent teeth 总被引:1,自引:0,他引:1
There is growing evidence that the insulin-like growth factor (IGF) system plays an important role in the biology of oro-dento-facial tissues and organs, including the development, homeostasis and regeneration of the periodontium. To obtain basic data on the occurrence and distribution of IGF components in human permanent teeth we immunohistochemically investigated 25 extracted, decalcified and paraffin-embedded teeth using mono and polyclonal antibodies against the ligands IGF-I and -II, the IGF1 receptor (IGF1R) and all six IGF binding proteins (IGFBP-1 to -6). In the extracellular matrix (ECM) of the adhering periodontal ligament (PDL), immunoreactivity for IGF-I, -II and IGFBP-1 and -6 was observed. PDL fibroblasts showed immunostaining for the IGF1R. For the cementum, in the acellular cementum only IGF-II could be detected, while outer cementum layers with inserting Sharpey's fibers reacted with all antibodies applied except for IGFBP-4 and -6. In the pulp, mainly fibrotic areas and areas around denticles were immunoreactive for IGF-I, IGFBP-1, -3, -5 and -6. Predentin and odontoblastic processes were stained for IGF-I and IGFBP-3. The spatially oriented occurrence of components of the IGF system in human permanent teeth indicates that specific functions of the IGFs may be localized in particular tissue compartments. In the cementum, several IGF components were found indicating roles in tissue homeostasis or attachment. The PDL may function as a reservoir for IGFs probably bound to ECM components. PDL fibroblasts could then respond in a paracrine manner. In the pulp, the IGF system may be involved in odontoblast biology, fibrosis and denticle formation. 相似文献
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Dr. M. Wolf S. Lossdörfer R. Craveiro W. Götz A. Jäger 《Journal of orofacial orthopedics》2013,74(5):420-434
Objective
Recent studies have shown that periodontal ligament (PDL) cells interact with macrophages from the immune system during orthodontically induced repair of periodontal tissue. Hypothesizing that high-mobility group box 1 (HMGB1) protein is released by mechanically stressed PDL cells into the extracellular space and has a role in mediating the local immune response by acting as an “alarmin”, this study was performed to further elucidate these cellular interactions, with a special focus on the impact of proinflammatory mediators secreted by PDL cells on macrophage physiology.Materials and methods
The study included an in vivo part in which orthodontic stress was induced in rats and their PDL analyzed for expression of HMGB1 by immunohistochemistry after 5 days of tooth movement. In the in vitro part, human PDL cells were subjected to compressive loading, followed by stimulating human macrophages with conditioned supernatants of these stressed PDL cells and analyzing how mediators that had been released by these cells into the medium would impact macrophage physiology. Assays for macrophage migration and osteoclast differentiation were used in addition to immunohistochemistry, enzyme-linked immunosorbent assays, and western blotting.Results
Induction of mechanical stress was found to upregulate HMGB1 expression both in vivo and in vitro. At the same time, translocation HMGB1 from nuclei into cytoplasm was observed. Culturing macrophages in conditioned PDL cell medium was associated with enhanced chemotactic migration and osteoclast differentiation. Addition of anti-HMGB1 antibodies to inhibit HMGB1 in the conditioned medium was found to significantly attenuate these effects. A less marked increase of migration and osteoclast differentiation by macrophages was observed after isolated addition of HMGB1, at its observed pathological concentration, to nonconditioned medium.Conclusion
This study clearly indicates an immunomodulatory potential of human PDL cells via release of mediators, including HMGB1 protein. Our finding that these mediators modify the migration and differentiation of macrophages as a function of periodontal repair during orthodontic treatment broadens the theoretical basis toward developing interventional strategies to avoid orthodontically induced root resorption. 相似文献24.
In vivo differentiation of human periodontal ligament cells leads to formation of dental hard tissue
Dr. M. Wolf S. Lossdörfer N. Abuduwali R. Meyer S. Kebir W. Götz A. Jäger 《Journal of orofacial orthopedics》2013,74(6):494-505
Objective
Following trauma, periodontal disease, or orthodontic tooth movement, residual periodontal ligament (PDL) cells at the defect site are considered mandatory for successful regeneration of the injured structures. Recent developments in tissue engineering focus, as one pillar, on the transplantation of PDL cells to support periodontal regeneration processes. Here, we examined the ability of osteogenically predifferentiated PDL cells to undergo further osteoblastic or cementoblastic differentiation and to mineralize their extracellular matrix when transplanted in an in vivo microenvironment.Materials and methods
Using collagen sponges as carriers, osteogenically predifferentiated human PDL cells were transplanted subcutaneously into six immunocompromised CD-1® nude mice. Following explantation after 28 days, osteogenic and cementogenic marker protein expression was visualized immunohistochemically.Results
After 28 days, transplanted PDL cells revealed both cellular, cytoplasmatic and extracellular immunoreactivity for the chosen markers alkaline phosphatase, osteopontin, PTH-receptor 1, and osteocalcin. Specific osteogenic and cementoblastic differentiation was demonstrated by RUNX2 and CEMP1 immunoreactivity. Early stages of mineralization were demonstrated by calcium and phosphate staining.Conclusion
Our results reinforce the previously published reports of PDL cell mineralization in vivo and further demonstrate the successful induction of specific osteogenic and cementogenic differentiation of transplanted human PDL cells in vivo. These findings reveal promising possibilities for supporting periodontal remodeling and regeneration processes with PDL cells being potential target cells with which to influence the process of orthodontically induced root resorption. 相似文献25.
This study examined the histological changes and possible effects of intermittent parathyroid hormone (PTH) (1-34) treatment
during the early and late phase of periodontal repair in a rat model of tooth root resorption. In a total of 70 animals, which
either received intermittent PTH(1-34) systemically or sham injections for up to 70 days after discontinuation of an orthodontic
force, histological characteristics were correlated to time-dependent distinct expression patterns of osteoprotegerin and
receptor activator of nuclear factor kappaB ligand by PDL cells in the former compression and tension side of tooth movement
by means of immunohistochemistry and histomorphometrical analysis. The balance of these key regulators of bone remodeling
was demonstrated to be shifted in favor of hard tissue repair by intermittent PTH administration, which was demonstrated to
exert anabolic effects in several cell culture and animal experiments as well as in humans, in the late phase of repair. These
data indicate a role for PDL cells as potent regulators of periodontal repair by modifying the local microenvironment and
point to the anabolic potential of an intermittent PTH administration to support these reparative processes. 相似文献
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Background and Objective: Although periodontal ligament cells display several osteoblastic traits, their phenotypic expression is still not well established. It remains a matter of debate whether they resemble a terminally differentiated cell type or an intermediate maturation state that potentially can be directed towards a fibroblastic or an osteoblastic phenotype. Material and Methods: To explore the characteristics of periodontal ligament cells in greater detail, fourth‐passage, sixth‐passage and eighth‐passage human periodontal ligament cells were cultured for up to 3 wk. Ki‐67, alkaline phosphatase, osteocalcin, osteoprotegerin and receptor activator of nuclear factor‐κB ligand (RANKL) mRNA expression was quantified by real‐time polymerase chain reaction. Furthermore, the cellular response to fibroblast growth factor‐2 and bone morphogenetic protein‐7 was examined in first‐passage and fourth‐passage cells. Dermal fibroblasts (1BR.3.G) and osteoblast‐like cells (MG63) served as reference cell lines. Results: Proliferation decreased over time and was highest in fourth‐passage cells. The expression of differentiation parameters, osteoprotegerin and RANKL increased with culture time and was higher in fourth‐passage cells than in cells of later passages. The RANKL/osteoprotegerin ratio increased steadily until day 21. Administration of fibroblast growth factor‐2 enhanced cell numbers in both passages, whereas alkaline phosphatase and osteocalcin production remained unchanged. By contrast, exposure of periodontal ligament cells to bone morphogenetic protein‐7 resulted in a reduction of cell number in the first and fourth passages, whereas the production of alkaline phosphatase and osteocalcin was enhanced. In dermal fibroblasts, differentiation parameters did not respond to both stimuli. MG63 cells behaved similarly to periodontal ligament cells. Conclusion: These results indicate that subculture affects the phenotypic expression of human periodontal ligament cells with respect to the characteristics that these cells share with osteoblasts. Furthermore, the periodontal ligament cell phenotype can be altered by fibroblastic and osteoblastic growth factors. 相似文献
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Immunohistochemical localization of components of the insulin-like growth factor-system in human deciduous teeth 总被引:1,自引:0,他引:1
To investigate the occurrence of components of the insulin-like growth factor (IGF) system during the resorption process of shedding human deciduous teeth, we investigated sections of 13 decalcified and paraffin-embedded deciduous teeth immunohistochemically with antibodies against IGF-I and -II, six IGF binding proteins (IGFBPs 1-6) and the IGF receptors IGF1R and IGF2R. The teeth were in different stages of resorption and all showed reparative cementum formation. It was found that acellular extrinsic fiber cementum, reversal lines and reparative cellular intrinsic fiber cementum were immunoreactive for both IGFs and various IGFBPs. Therefore, in human deciduous teeth, all subgroups of cementum, but not dentine, may represent sources of components of the IGF system. Odontoclasts did not carry IGFs or the IGF1R, but IGFBPs and the IGF2R. Therefore, these cells, in contrast to osteoclasts, may not respond to IGFs, but may be involved in the release and sequestration of IGFs from cementum during the resorption process. In contrast to odontoclasts, cementoblasts and periodontal ligament (PDL) fibroblasts carried IGF1R. The influence of the IGF system on the function of these cells with respect to periodontal matrix turnover and cementogenesis is discussed. On the behalf of the IGFBP immunoreactivities found, the PDL extracellular matrix can be considered to be a reservoir for IGF system components, where binding proteins may regulate IGF distribution and activity. 相似文献
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Kraus D Jäger A Abuduwali N Deschner J Lossdörfer S 《Clinical oral investigations》2012,16(2):611-618
Periodontal ligament (PDL) cells have been associated with the regulation of periodontal repair processes by the differential
expression of osteoprotegerin and RANKL in response to intermittent parathyroid hormone (PTH) resulting in a modified activity
of bone-resorbing osteoclasts. Here, we examined the intracellular signaling pathways that PDL cells use to mediate the PTH(1–34)
effect on osteoprotegerin production and hypothesized that those would be dependent on the cellular maturation stage. Two
stages of confluence served as a model for cellular maturation of 5th passage human PDL cells from six donors. Intermittent
PTH(1–34) (10−12 M) and PTH(1–31), the latter lacking the protein kinase C (PKC) activating domain, induced a significant decrease of osteoprotegerin
production in confluent cultures, whereas the signal-specific fragments PTH(3–34) and PTH(7–34), which both are unable to
activate protein kinase A (PKA), had no effect. The addition of the PKA inhibitor H8 antagonized the PTH(1–34) effect, whereas
the PKC inhibitor RO-32-0432 did not. In pre-confluent, less mature cultures, intermittent PTH(1–34) resulted in a significant
increase of osteoprotegerin. Similar results were obtained when PTH(1–31) substituted for PTH(1–34) as opposed to a lack of
an effect of PTH(3–34) and PTH(7–34). Likewise, in confluent cultures, H8 inhibited the PTH(1–34) effect in pre-confluent
cultures contrasted by RO-32-0432 which had no effect. These findings indicate that PTH(1–34) signaling targeting osteoprotegerin
production in PDL cells involves a PKA-dependent pathway. The PTH(1–34) effect is dependent on cell status, whereas intracellular
signal transduction is not. Clinical trials will have to prove whether those in vitro data are of physiological relevance
for interference strategies. 相似文献
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