Bacteroides gingivalis activates mouse spleen cells to produce a factor that stimulates resorptive activity of osteoclasts in vitro

Oral microorganisms are generally considered to be responsible for the initiation of inflammatory periodontal disease, but the mechanism by which they cause bone resorption has not yet been established. We report here that the periodontopathic bacterium B. gingivalis activates mouse spleen cells to produce a factor that strongly stimulates osteoclast-mediated mineral resorption. Resorption was measured as the release of ⁴⁵Ca from prelabeled fetal mouse long bones in vitro .     

Interleukin 1 beta and prostaglandin E are involved in the response of periodontal cells to mechanical stress in vivo and in vitro.

Cytokines are local mediators released by cells of the immune system in response to stimulation by a variety of agents. These polypeptides may interact directly or indirectly with bone cells. The objectives of this study were (1) to localize prostaglandin E (PGE) and the cytokine interleukin-1 beta (IL-1 beta) in the periodontal ligament after the application of mechanical force to teeth in vivo and (2) to determine the effects of mechanical stress or IL-1 beta (or the two in combination) on PGE synthesis and bone resorption by fibroblasts in the human periodontal ligament (PDL). In 24 female cats, one maxillary canine was tipped distally by 80 gm force for 12 hours, 24 hours, or 7 days. PGE and IL-1 beta were localized immunohistochemically in serial jaw sections, and semiquantitation of cellular-staining intensity was done by microphotometry. Unstressed periodontal ligament cells stained mildly for PGE and IL-1 beta, but the staining intensity increased significantly in sites of tension. Human periodontal ligament fibroblasts were preincubated with mechanical stress and/or IL-1 beta in the presence or absence of indomethacin for 1 hour. Then the media were replaced by BGJb (Fitton-Jackson modification) medium (GIBCO), and incubation was continued for 4, 8, or 24 hours in conditioned media. PGE concentrations in conditioned media were determined by radioimmunoassay, and bone-resorbing activity in conditioned media was assessed by 45Ca release from prelabeled neonatal mouse calvaria. The conditioned media derived from cells stimulated by mechanical stress plus IL-1 beta caused significantly more bone resorption than the conditioned media obtained from cells that had been treated by each factor alone. The addition of indomethacin did not inhibit bone resorption completely. These results demonstrate that periodontal ligament cells respond to mechanical stress by increased production of PGE, and that IL-1 beta enhances this response.     

Dental follicle cell-conditioned medium enhances the formation of osteoclast-like multinucleated cells

An influx of mononuclear cells and the subsequent increase of osteoclasts around tooth germs suggests that the dental follicle (DF) regulates or influences bone resorption required for tooth eruption. In order to study the effects of DF cell products on osteoclast formation during tooth eruption, a conditioned medium (CM) was created in which DF cells were added to mouse bone marrow cultures. Tartrate-resistant acid phosphatase (TRAP)-positive osteoclast-like multinucleated cells were formed in the presence of 10 nM 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. The CM, dose-dependently, stimulated the formation of TRAP-positive cells in the presence of 1,25(OH)2D3 for 14 days culture. The number of these cells decreased due to degradation in the control culture. A semi-solid methylcellulose assay in the presence of CM showed little expression of colony-stimulating activity. These results suggest that the DF cells of a developing tooth produce factor(s) that enhance osteoclast formation and bone resorption necessary for tooth eruption.     

Human gingival fibroblasts secrete non-dialyzable, prostanoid-independent products which stimulate bone resorption in vitro

Conditioned media from human gingival fibroblasts stimulate release of radioactive calcium, stable calcium and inorganic phosphate from mouse calvarial bones in organ culture. In addition, human gingival fibroblast-conditioned media decrease the hydroxyproline content in the bones. Stimulation of mineral mobilization could be reduced by calcitonin, bisphosphonates and glucocorticoids, but not by cyclooxygenase inhibitors. The production of stimulating factors by fibroblasts was unaffected by indomethacin, meclofenamic acid and naproxen. Dialysis of human gingival fibroblast-conditioned media showed that the product(s) had a molecular weight larger than 3000 daltons. Bone resorbing activity in human fibroblast-conditioned media was heat-labile. These data show that human gingival fibroblasts can secrete non-prostanoid products with the ability to stimulate osteoclastic bone resorption in vitro .     

Effects of parathyroid hormone and cytokines on prostaglandin E synthesis and bone resorption by human periodontal ligament fibroblasts

Cultured human periodontal ligament fibroblasts showed synergistic elevations in the synthesis of prostaglandin E and production of cAMP by the administration of parathyroid hormone and cytokines (interleukin 1 alpha, -1 beta, or tumour necrosis factor-alpha). Unstimulated conditioned media derived from these fibroblasts contained bone-resorbing activity. In addition, conditioned media generated by cytokine-or parathyroid hormone-treated fibroblasts showed further increases in bone-resorbing activity. The effects were additive when the hormone was combined with either one of the cytokines in stimulating bone resorption. These findings suggest that the effect of parathyroid hormones and cytokines together on bone resorption can be mediated in part by human periodontal ligament fibroblasts via PGE production and subsequent PGE action on the osteoclasts.     

Alveolar bone destruction in the immunosuppressed rat

The placement of a ligature around the second maxillary molar of the conventional rat caused osteoclastic bone resorption and simultaneously, alveolar bone formation. The number of peripheral mononuclear cells in the blood and lymphoblastic transformation of spleen cells in response to concanavalin A increased. Cyclophosphamide (CY), an immunosuppressive agent, given shortly after placing the ligature suppressed the lymphoid reactions, spleen size, and bone formation and enhanced bone destruction. CY given in higher doses also suppressed the number of PMN cells. Septicemia developed in several of these animals. Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli were isolated from the blood and/or ligature. Antibiotics prevented bone destruction. Without placing a ligature, the high dose of CY did not result in bone loss.
These findings suggest that 1) bone destruction of the ligature-treated rat is of bacterial origin, 2) CY suppresses proliferation of osteoblasts but does not seem to interfere with the activity of osteoclasts, and 3) suppression of the host defenses greatly facilitates bone destruction.     

牙龈卟啉菌表面相关物质的提取、纯化及骨吸收和免疫特性研究

目的 :通过对牙龈卟啉菌Pg表面相关物质 (SAM)的提取、纯化 ,研究其潜在的骨吸收活性 ,以揭示pg的SAM在牙周病过程中的致病作用。方法 :1)细菌培养。 2 )SAM粗品的制备。Sephacryl-S2 0 0凝胶过滤、冻干、纯化。3)DEAE -Sephacel阴离子交换层析。 4 )SAM介导的小鼠颅骨吸收试验。结果 :1)细菌培养得到冻干的细菌 4 .1g。2 )分离SAM得到粗品 0 .16g。Sephacryl -S2 0 0凝胶过滤纯化SAM粗品 ,冻干后得到较纯的SAM约为 0 .0 2 2 1g。DEAE -Sephacel阴离子交换层析进一步纯化SAM获得SAM纯品 3mg。 3)纯化的SAM骨吸收活性鉴定。骨吸收作用可通过体外小鼠颅骨培养基中Ca2 + 浓度的增加而间接证实。SAM的骨吸收活性随着浓度的增加而增加 ,与对照组相比 ,P <0 .0 5。SAM具有免疫原性。结论 :SAM在骨吸收方面具有极大的潜能。提示SAM在牙周病骨损害方面起到重要的作用     

The bone resorbing activity released by gingival fibroblasts isolated from patients with periodontitis is independent of interleukin-1

Supernatants from gingival fibroblast cultures obtained from 14 patients with periodontal disease contained factor(s) capable of stimulating bone resorption in vitro, as assessed by the release of 45Ca from neonatal mouse calvariae. The possibility that the factor(s) was interleukin-1 alpha (IL-1 alpha), IL-1 beta or prostaglandin E2 (PGE2) was next investigated. The human fibroblast conditioned media (HFCM) stimulated PGE2 biosynthesis in bone. The stimulatory effect by HFCM on 45Ca release, however, was not affected by blocking prostaglandin biosynthesis with indomethacin. In contrast, 45Ca release induced by IL-1 alpha, IL-1 beta, thrombin and bradykinin was significantly reduced by indomethacin, whereas the effects of PTH and PTHrP were unaffected by indomethacin. The concentration of PGE2 in HFCM was too low to be solely responsible for the 45Ca release response. In addition, the amount of bone resorbing activity produced by the gingival fibroblasts was unaffected by cyclo-oxygenase inhibitors. Similar to IL-1 alpha and IL-1 beta, the stimulatory effect of HFCM was inhibited by gamma-interferon. HFCM did not stimulate cyclic AMP formation in the mouse calvarial bones. Antisera which specifically blocked human IL-1 alpha or IL-1 beta induced 45Ca release, and the specific IL-1 receptor antagonistic protein, did not inhibit the stimulatory effect of HFCM. These data show that gingival fibroblasts secrete bone resorbing factor(s) which is not due to IL-1 and which stimulates bone resorption by a prostaglandin- and cyclic AMP-independent mechanism.     

Osteoporosis and edentulous jaws

PURPOSE: To investigate the influence of (1) osteoporosis on resorption of edentulous and (2) whether osteoporosis enhances implant loss. MATERIALS AND METHODS: MEDLINE, EMBASE, and Cochrane Central Register of Controlled Trials (CENTRAL) databases were searched for association of osteoporosis and bone resorption/loss of implants. Inclusion and exclusion criteria were defined and quality assessment of the remaining studies after exclusion was performed. The Cochrane approach for cohort studies was applied. RESULTS: Methodologic assessment of the eligible publications using the Cochrane approach resulted in the inclusion of 11 publications in the final evaluation. Most studies dealt with the association between the extent of resorption or atrophy of edentulous jaws and the systemic (femoral, lumbar spine, metacarpal) bone mineral content (BMC) or bone mineral density (BMD) status. Four studies reported a significant association between the extent of resorption and the skeletal BMC and/or BMD. Four studies evaluated the effect of endosseous implants on mandibular BMC or BMD changes and the association between systematic BMD statuses and loss of implants. These studies revealed no association between systemic BMD status, mandibular BMD status, bone quality, and implant loss. CONCLUSIONS: Although no firm conclusions could be drawn regarding the effect of osteoporosis on resorption of edentulous jaws, with or without implants, due to the different parameters applied in the various studies, the use of endosseous implants in osteoporosis patients is not contraindicated.     

Involvement of PGE synthesis in the effect of intermittent pressure and interleukin-1 beta on bone resorption

Human periodontal ligament (PDL) fibroblasts, cultured from extracted healthy premolars, and a cloned osteogenic cell line (MC3T3-E1) were used in this study to determine the effect of intermittent pressure on bone resorption. Cells (1 x 10(5] were incubated with BGJb medium in the presence or absence of the following factors: intermittent negative (-30 g/cm2) or positive (30 g/cm2) hydrostatic pressure and interleukin-1 beta (IL-1 beta, 1 ng/mL), for 24 h. Conditioned media (CM) generated from cultures of either cell types were used for prostaglandin E (PGE) assay, bone resorption assay, and assessment of osteoclast (OC)-like cell formation. Unstimulated PDL fibroblasts or MC3T3-E1 cells produced measurable amounts of PGE and bone-resorbing activity as measured by 45Ca released from mouse calvaria and OC-like cells. IL-1 beta-treated cells showed significantly elevated levels of PGE, bone resorption, and OC-like cell formation, as compared with unstimulated cells. Intermittent positive pressure (IPP) alone stimulated PGE production, but the resultant CM did not stimulate bone resorption or OC-like cell formation when IPP was applied to either cell type. The application of IPP, together with IL-1 beta in CM, caused a slight increase in the number of alpha-like cells, as compared with that of IL-1 beta-treated CM in both cell types. On the other hand, direct application of IPP on mouse bone-marrow cultures significantly increased the number of OC-like cells. This effect was additive in combination with either CM from unstimulated cells or exogenous addition of PGE2.(ABSTRACT TRUNCATED AT 250 WORDS)     

Extracellular ATP is a key modulator of alveolar bone loss in periodontitis

Periodontal diseases are initiated by pathogenic bacterial biofilm activity that induces a host inflammatory cells immune response, degradation of dento gingival fibrous tissue and its detachment from root cementum. It is well accepted, that osteoclastic alveolar bone loss is governed exclusively through secretion of proinflammatory cytokines. Nevertheless, our findings suggest that once degradation of collagen fibers by MMPs occurs, a drop of cellular strains cause immediate release of ATP from marginal gingival fibroblasts, cell deformation and influx of Ca + 2. Increased extracellular ATP (eATP) by interacting with P2 × 7 purinoreceptors, present on fibroblasts and osteoblasts, induces generation of receptor activator of nuclear factor kB ligand (RANKL) that further activates osteoclastic alveolar bone resorption and bone loss. In addition, increased eATP levels may amplify inflammation by promoting leukocyte recruitment and NALP3-inflammasome activation via P2 × 7. Then, the inflammatory cells secrete cytokines, interleukin IL-1, TNF and RANKL that further trigger alveolar bone resorption. Moreover, eATP can be secreted from periodontal bacteria that may further contribute to inflammation and bone loss in periodontitis. It seems therefore, that eATP is a key modulator that initiates the pathway of alveolar bone resorption and bone loss in patients with periodontal disease. In conclusion, we propose that strain release in gingival fibroblasts aligned on collagen fibers, due to activity of MMP, activates release of ATP that triggers the pathway of alveolar bone resorption in periodontitis. We predict that by controlling the eATP interaction with its cellular purinoreceptors will reduce significantly bone loss in periodontitis.     

Heterogeneity of bone resorbing factors produced by unstimulated murine osteoblasts in vitro and in response to stimulation by parathyroid hormone and mononuclear cell factors

The bone resorbing activity of factors released from monolayer cultures of osteoblasts (OB) was examined by measurement of calcium released by neonatal mouse calvaria in vitro. Unstimulated conditioned media (CM) were found to contain significant bone resorbing activity, which was partially inhibited by indomethacin, dexamethasone and nordihydroguaiaretic acid. Ultrafiltration of CM (molecular weight cut-off of 5000) revealed bone resorbing activity in the filtrate and retentate. Fractionation of the CM by high-performance liquid chromatography revealed four major peaks of bone resorbing activity. Stimulation of the OB by mononuclear cell factor and parathyroid hormone significantly increased the synthesis and/or release of these factors with a relatively greater increase of lipid-soluble, low molecular-weight activity. These results suggested an important role for relatively small non-popular mediators in hormonally stimulated bone resorption.     

Role of interleukin-1 and prostaglandin in in vitro bone resorption induced by Actinobacillus actinomycetemcomitans lipopolysaccharide

Lipopolysaccharide (Y4 LPS) isolated from Actinobacillus actinomycetemcomitans strain Y4 induced bone resorption in BALB/c mouse calvaria organ culture. The calcium release from LPS-low responsive C3H/HeJ mouse calvaria by Y4 LPS was very low. Indomethacin almost completely inhibited prostaglandin E₂ (PGE₂) production by Y4 LPS-stimulated BALB/c mouse calvaria, but did not suppress interleukin-1 (IL-1) release from the calvaria, and partially suppressed the bone resorption. Dexamethasone strongly inhibited the PGE₂ and IL-1 production by Y4 LPS-stimulated BALB/c mouse calvaria. as well as Y4 LPS-induced bone resorption. Dexamethasone inhibited expression of membrane IL-1 on osteoblastic cells stimulated with Y4 LPS, but indomethacin did not. Furthermore, anti-IL-1 serum partially suppressed the calcium release from Y4 LPS-stimulated BALB/c mouse calvaria. These results suggest that both PGE₂ and IL-1 participate in Y4 LPS-induced bone resorption in vitro .     

Interleukin-2 stimulates osteoclastic activity; Increased acid production and radioactive calcium release

Recombinant human interleukin-2 (IL-2) was studied to determine effects on acid production by individual osteoclasts in situ on mouse calvarial bones. This analysis was performed using a microspectrofluorimetric technique to quantify acid production in individual cells. Radioactive calcium release was determined using calvarial bones in a standard tissue culture system. This allowed us to correlate changes in acid production with a measure of bone resorption. IL-2 stimulated acid production and bone resorbing activity. Both effects were inhibited by calcitonin. No stimulation of bone resorption occurred when IL-2-containing test media was incubated with a specific anti-IL-2 antibody and ultrafiltered. Our data demonstrated a correlation between acid production and bone resorbing activity in mouse calvaria exposed to parathyroid hormone (PTH). The data obtained from cultured mouse calvaria exposed to IL-2 demonstrated similar stimulatory effects to those seen during PTH exposure. These data suggest that calvaria exposed to IL-2 in vitro have increased osteoclastic acid production corresponding with increased bone resorption.     

Role of free radicals in bone resorption

In order to examine the role of the oxygen-derived free radicals in bone metabolism, the mouse calvaria organ culture system and the mouse bone marrow culture system were used. The activity of bone resorption stimulated by 1 alpha, 25(OH)2 vitamin D3 was inhibited by the addition of catalase, an eliminating agent of H2O2. Catalase also significantly suppressed the osteoclast-like cell formation induced by 1 alpha, 25(OH)2 vitamin D3 in a dose-dependent manner, whereas the agent had no effect on the mitogenic activity of the bone marrow-derived stromal cells. This suppression was reversed by adding H2O2 to the system. The effect of catalase was limited to the early stage precursor of the osteoclast-like cells. Moreover, when superoxide dismutase (SOD), which activates the production of H2O2, was added to the bone marrow culture, the number of osteoclast-like cells increased significantly in a dose-dependent manner. Finally, the source of the oxygen-derived free radicals was examined by measuring the fluorescence of 2.7-dichlorofluorescein (DCF), the obtained result indicated that the osteoclast-like cells could produce almost the equivalent amount of H2O2 as could the macrophages. These results strongly suggest that the oxygen-derived free radicals, particularly H2O2, may regulate bone resorption, promoting the differentiation of osteoclast precursor cells to osteoclasts.     

Role of prostaglandin in the formation of osteoclasts induced by capsular-like polysaccharide antigen of Actinobacillus actinomycetemcomitans strain Y4

We found no reports that capsular-like polysaccharide antigen purified from Actinobacillus actinomycetemcomitans either induces osteoclastic bone resorp-tion in mouse organ cultures or promotes osteoclast formation in mouse marrow cultures. In contrast, capsular-like polysaccharide antigen purified from A. actinomycetemcomitans strain Y4 induced bone resorption in mouse organ culture. To examine the mechanism of bone resorption induced by A. actinomycetemcomitans , mouse bone marrow cells were cultured with A. actinomycetemcomitans strain Y4 capsular-like polysaccharide antigen. A. actinomycetemcomitans strain Y4 capsular-like polysaccharide antigen stimulated osteoclast-like cell formation in mouse bone marrow cultures. However, the polysaccharide of A. actinomycetemcomitans lipopolysaccharide did not induce the formation of osteoclast-like cells. Indomethacin inhibited osteoclast-like cell formation mediated by A. actinomycetemcomitans strain Y4 capsular-like polysaccharide antigen in a dose-dependent manner. There was a good correlation between the number of osteoclast-like cells formed in the marrow culture and the amount of prostaglandin E2 released into the culture media. When mouse bone marrow cells were cultured with prostaglandin E2 during the culture periods, many osteoclast-like cells were formed. These results indicate that prostaglandin E2 is involved in the mechanism of the formation of osteoclast-like cells mediated by A. actinomycetemcomitans strain Y4 capsular-like polysaccharide antigen. A. actinomycetemcomitans strain Y4 capsular-like polysaccharide antigen may play an important role in inflammatory bone resorption by promoting osteoclast formation in periodontal disease.     

Raloxifene reduces the risk of local alveolar bone destruction in a mouse model of periodontitis combined with systemic postmenopausal osteoporosis

OBJECTIVEPeriodontitis is characterized by local inflammation leading to tooth loss and severe destruction of alveolar bone. Raloxifene is a selective estrogen receptor modulator (SERM) that halts estrogen deficiency-induced systemic bone loss in postmenopausal osteoporosis without the side effects of cancer in breast and uterus. In this study, we examined the effects of raloxifene on alveolar bone mass in a mouse model with estrogen deficiency-induced periodontitis.METHODSPeriodontitis was induced by the injection of lipopolysaccharide (LPS) into the lower gingiva in ovariectomized (OVX) mice, and the alveolar bone and femur bone mineral density (BMD) were analyzed by dual-energy X-ray absorptiometry. To explore the direct osteoclast inhibitory effect of raloxifene, a co-culture system for osteoclast formation and organ culture of alveolar bone was established.RESULTSWhen OVX mice were treated with raloxifene, the bone loss in both alveolar bone and femur were abrogated. Interleukin 1 and/or LPS stimulated the osteoclast formation and bone-resorbing activity; however, raloxifene did not show any inhibitory effect on the osteoclast formation or function. In vivo local injection of raloxifene also did not prevent bone resorption in a mouse model of periodontitis. However, the systemic treatment of raloxifene using a mini-osmotic pump did prevent the loss of BMD of alveolar bone induced by LPS.CONCLUSIONThese results suggest that the SERM raloxifene systemically maintain alveolar bone mass in a mouse model of periodontitis with osteoporosis. Increasing the alveolar bone mass by SERMs treatment in patients with postmenopausal osteoporosis may be a useful approach to preventing the destruction of alveolar bone in late-onset periodontitis.     

Cyclical tensile force on periodontal ligament cells inhibits osteoclastogenesis through OPG induction

The periodontal ligament (PDL) maintains homeostasis of periodontal tissue under mechanical tensile-loading caused by mastication. Occlusal load inhibits atrophic alveolar bone resorption. Previously, we discovered that continuous compressive force on PDL cells induced osteoclastogenesis-supporting activity, with up-regulation of RANKL. We hypothesized that, unlike compression, cyclical tensile force up-regulates OPG expression in PDL cells via TGF-beta up-regulation, and does not induce osteoclastogenesis-supporting activity. PDL cells were mechanically stimulated by cyclical tensile force in vitro. The conditioned media of PDL cells that had been subjected to cyclical tensile force inhibited osteoclastogenesis. Cyclical tensile force up-regulated not only RANKL mRNA expression, but also OPG mRNA expression in PDL cells. Tensile force up-regulated TGF-beta expression in PDL cells as well. Administration of neutralizing antibodies to TGF-beta inhibited OPG up-regulation under cyclical tensile-force stimulation in a dose-dependent manner. Additionally, the osteoclastogenesis-inhibitory effect of the conditioned media of PDL cells under cyclical tensile force was partially rescued by the administration of TGF-beta neutralizing antibodies. In conclusion, tensile force inhibited the osteoclastogenesis-supporting activity of PDL cells by inducing the up-regulation of OPG via TGF-beta stimulation.     

Inhibition of bone DNA and collagen production by surface-associated material from bacteria implicated in the pathology of periodontal disease.

Gentle extraction of oral bacteria implicated in the pathogenesis of periodontal disease, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, or Eikenella corrodens, with saline removes the extracellular components while leaving the bacteria intact. This readily-solubilized surface-associated material (SAM) has been demonstrated to significantly inhibit DNA and collagen synthesis by murine calvaria at concentrations as low as 10 ng/ml. DNA and collagen synthesis in isolated calvarial osteoblasts were also inhibited by these SAM preparations with similar dose responses. The inhibitory effect of these bacterial expolymers was blocked by 1 microM indomethacin. The potent inhibitory actions on bone synthesis of the SAM from these bacteria may contribute to the alveolar bone loss found in patients with periodontal disease.     

Production of matrix-degrading enzymes and inhibition of osteoclast-like cell differentiation by fibroblast-like cells from the periodontal ligament of human primary teeth

Clinically, the most apparent difference between the primary and permanent dentitions is the physiologic loss of the primary tooth by root resorption. Root resorption is associated with loss of integrity of the periodontal ligament (PDL), followed by recruitment of resorptive cells that remove root structure. We therefore cultured primary dentition PDL fibroblasts (PPDL cells) to investigate in vitro their production of matrix metalloproteinases (MMPs) and tissue inhibitors of MMP (TIMPs), and the effects of soluble factors produced by these cells on osteoclast-like cell differentiation. These studies demonstrate that PPDL cells in vitro have a heterogeneous morphology, and they constitutively synthesize 92-kDa gelatinase, 72-kDa gelatinase, and 53/57-kDa procollagenase as well as TIMP-1, -2, and a third inhibitor of matrix metalloproteinase, as determined by substrate gel zymography and immunoblot analysis. Compared with PDL cells from the permanent dentition, PPDL cells generally produced a greater amount of collagenase but similar amounts of the gelatinases and inhibitors. PPDL cells were treated with pro-inflammatory cytokines to determine their effect on the expression of matrix-degrading enzymes and inhibitors. Interleukin-1alpha and tumor necrosis factor-alpha enhanced the constitutive expression of proteinases but not that of inhibitors in PPDL cells. Conditioned media from PPDL cell lines inhibited the differentiation of osteoclast-like cells in mouse bone marrow cultures. These findings indicate that PPDL cells may modulate the cascade of root resorption both by their regulated production of proteinases and inhibitors and by synthesis of unknown soluble factor(s) that may regulate osteoclast development.