Lithium chloride stimulates bone formation in extraction socket repair in rats.

PURPOSE: Previous evidence shows that lithium chloride (LiCl), a suppressor of glycogen synthase kinase-3ß (GSK-3ß), may enhance bone formation in several medical and dental conditions. Thus, the purpose of the current study was to assess the effects of LiCl on extraction socket repair in rats. METHODS: Thirty rats were randomly assigned into a control group (administration of water; n = 15) or a LiCl group (administration of 150 mg/kg of LiCl; n = 15). LiCl and water were given every other day, starting at 7 days before the extraction of upper first molars until the end of each experiment period. Histological sections from five rats per group were obtained at 10, 20, and 30 days post-extractions. Histometrical analysis of newly formed bone (NB) and the levels of tartrate-resistant acid phosphatase (TRAP)-stained cells were evaluated at 10, 20, and 30 days post-extractions. Immunohistochemical staining for receptor activator of nuclear factor kappa-Β ligand (RANKL), osteoprotegerin (OPG), bone sialoprotein (BSP), osteocalcin (OCN), and osteopontin (OPN) was assessed at 10 days post-extractions. RESULTS: The LiCl group had a greater proportion of NB than the control group at 20 days (P < 0.05). At 30 days, the rate of TRAP-stained cells was lower in the LiCl group than in the control group (P < 0.05). At 10 days, the LiCl group presented stronger staining for OPG, BSP, OPN, and OCN, when compared to the control group (P < 0.05). CONCLUSION: Systemic LiCl enhanced extraction socket repair, stimulated an overall increase in bone formation markers, and restricted the levels of TRAP in rats.

Strontium ranelate improves alveolar bone healing in estrogen-deficient rats.

BACKGROUND: This study evaluated the impact of strontium ranelate on tooth-extraction wound healing in estrogen-deficient and estrogen-sufficient rats. METHODS: Ninety-six Wistar rats (90 days of age) were allocated into one of the following groups: sham-surgery+water (estrogen-sufficient); ovariectomy+water (estrogen-deficient), sham-surgery+strontium ranelate (625 mg/kg/d) (strontium/estrogen-sufficient); ovariectomy+strontium ranelate (625 mg/kg/d) (strontium/estrogen-deficient). Water or strontium ranelate were administrated from the 14th day post-ovariectomy/sham surgery until euthanasia. Maxillary first molars were extracted at 21 days after sham/ovariectomy surgery. Rats were euthanized at 10, 20, and 30 days post-extractions. The following parameters were analyzed inside tooth-extraction wound: proportion of newly formed bone (bone healing/BH), number of cells stained for tartrate-resistant acid phosphatase (TRAP) and immunohistochemical staining for five bone metabolism-related markers (osteocalcin [OCN], osteopontin [OPN], bone sialoprotein [BSP], osteoprotegerin [OPG] and receptor activator of NF-КB ligand [RANKL]). RESULTS: The estrogen-deficient group presented lower BH than all other groups at 20 and 30 days post-extraction (P < 0.05). The number of TRAP-stained cells was higher in the estrogen-deficient group than in estrogen-sufficient group at 30 days post-extraction (P < 0.05). The strontium /estrogen-sufficient group exhibited stronger staining for OCN, when compared to the estrogen-sufficient and estrogen-deficient groups (P < 0.05). Both strontium ranelate-treated groups presented higher staining of OPN and BSP than both untreated groups (P < 0.05). The strontium/estrogen-sufficient group demonstrated stronger staining for OPG than the estrogen-deficient group (P < 0.05). The estrogen-sufficient group and both groups treated with strontium ranelate showed lower expression of RANKL than the estrogen-deficient group (P < 0.05). CONCLUSIONS: Strontium ranelate benefited BH and the expression of bone markers in tooth-extraction wound in estrogen-deficient rats whereas its benefits in estrogen-sufficient rats were modest.

Lithium chloride improves bone filling around implants placed in estrogen-deficient rats.

OBJECTIVE: This study evaluated the ability of lithium chloride (LiCl) to increase bone filling (BF) around threaded titanium implants inserted in estrogen-deficient rats and, thein-vitro effects of this drug on osteoblast-like cell viability, proliferation, mineralization and expression of bone-related markers. DESIGN: In vivo: Rats received sham surgery plus water (Estrogen-sufficient group), ovariectomy plus water (Estrogen-deficient group) or ovariectomy plus LiCl (150 mg/kg/every other day) (LiCl/estrogen-deficient group). On the 21st day after ovariectomy/sham surgeries, a threaded titanium implant was inserted in the rat tibia. BF and the number of TRAP + cells were assessed at 10, 20 and 30 days after implant placement. In vitro: Osteosarcoma SAOS-2 cells were exposed to 0, 0.01, 0.05, and 0.1 mM of LiCl; cell proliferation, viability, mineralization (alizarin red staining) and gene expressions of RUNX-2, OCN, OPN, BSP and ALP (Real Time PCR) were estimated in the cultures. RESULTS: In vivo: The estrogen-sufficient and LiCl/estrogen-deficient groups demonstrated higher percentages of BF, within the limits of implant threads, than the estrogen-deficient group at 20 and 30 days (p < 0.05). The number of TRAP + cells was lower in LiCl/estrogen-deficient than in the estrogen-deficient group at all experimental times (p < 0.05). In vitro: Cell cultures exposed to LiCl (0.01 or 0.05 mM) exhibited larger areas of mineralized matrix than the non-exposed cultures (p < 0.05) and demonstrated the highest expressions of the genes investigated. CONCLUSION: LiCl treatment improved BF around threaded titanium implants inserted in estrogen-deficient rats and stimulated matrix mineralization and overexpression of bone-formation markers in osteoblastic cells in culture.

Might smoking assuage the pro-inflammatory effect of diabetes in periodontal sites?

OBJECTIVES: This study evaluated the effects of type 2 diabetes mellitus (DM), smoking, and these two factors combined on gingival crevicular fluid levels and ratios of pro-/anti-inflammatory cytokines. Associations between cytokines with each other and with key periodontal pathogens in periodontal sites under the challenge of one or both of these risk factors were also assessed. METHODS: A total of 102 subjects with periodontitis were included in this cross-sectional study and assigned to one of the following groups: non-diabetic non-smokers (control group, n = 25), non-smokers with DM (DM group, n = 30), non-diabetic smokers (S group, n = 26), and smokers with DM (S + DM group, n = 21). The levels of 13 pro-inflammatory (IFN-γ, TNF-α, MIP-1α, GM-CSF, IL-1ß, IL-2, IL-6, IL-7, IL-8, IL-12, IL-17, IL-21, and IL-23) and 5 anti-inflammatory (IL-4, IL-5, IL-10, IL-13, and TGF-ß) cytokines were assessed in healthy and diseased sites, using multiplex immunoassay. Ratios of pro-/anti-inflammatory cytokines were obtained in all possible permutations. The levels of 7 key periodontal pathogens were evaluated by qPCR. RESULTS: Overall, the ratios of pro-/anti-inflammatory cytokines were higher in healthy and diseased sites of the DM group and in healthy sites of the S + DM group, and lower in diseased sites of the S group, compared with the control (p < .05). The proportion of the pro-inflammatory cytokines in relation to the 18 cytokines studied was higher in the DM group and lower in the S group, whereas the proportion of the anti-inflammatory cytokines was lower in both diabetic groups and higher in the S group, compared to the control (p < .05). A cluster of six common cytokines (IL-4, IL-5, IL-12, IL-13, IL-21, and IL-23) was observed in the diseased sites of all groups studied. Eight common cytokines (IL-4, IL-5, IL-12, IL-13, IL-17, IL-21, IL-23, and IFN-γ) grouped closely in the healthy sites of both diabetic groups. Significant associations between pathogens and cytokines occurred mainly in the diseased sites of the S + DM group (p < .05). CONCLUSION: Diabetes mellitus induced an overall pro-inflammatory state, while smoking mainly stimulated immunosuppression in periodontal sites. When the two risk factors overlapped, smoking seemed to partially assuage the hyperinflammatory effect of DM.

Lithium chloride assuages bone loss in experimental periodontitis in estrogen-deficient rats.

OBJECTIVES: Evidence shows that lithium, a medication commonly used for bipolar disorder treatment, presents bone anabolic activity. This study evaluated the effects of lithium chloride on periodontitis-induced bone loss (BL) and on intact alveolar bone during estrogen sufficiency and deficiency. MATERIALS AND METHODS: Rats (24/group) received sham surgery plus water (estrogen-sufficient group), ovariectomy plus water (estrogen-deficient group), sham surgery plus lithium chloride (150 mg/kg/every other day) (lithium/estrogen-sufficient group), or ovariectomy plus lithium chloride (lithium/estrogen-deficient group). One first mandibular molar received ligature, while the contralateral molar was left unligated. BL and trabecular bone area (TBA) were assessed in the furcation bone at 10, 20, and 30 days after ligature placement. Histochemical staining for TRAP and immunohistochemical staining for osteocalcin, osteopontin, osteoprotegerin, and RANKL were evaluated at 30 days after ligature placement. RESULTS: At 10 days, the estrogen-deficient group presented the highest BL (0.115 ± 0.026), while the lithium/estrogen-deficient group (0.048 ± 0.024) presented the lowest BL in the ligated teeth (p < 0.05). At 20 and 30 days, the estrogen-deficient group exhibited significantly higher BL than all the other groups (p < 0.05). The ligated teeth of the lithium/estrogen-sufficient group presented the highest TBA while those of the estrogen-deficient group presented the lowest TBA at 10 and 30 days (p < 0.05). Unligated teeth of lithium-treated groups had stronger staining for osteocalcin and osteopontin than the estrogen-deficient group (p < 0.05). Ligated and unligated teeth of the estrogen-deficient group exhibited lower expression of osteoprotegerin than the other groups (p < 0.05). Lithium-treated groups exhibited generally higher staining of RANKL than the untreated groups (p < 0.05). Unligated teeth in both estrogen-sufficient groups presented lower TRAP expression than both estrogen-deficient groups (p < 0.05). CONCLUSIONS: Lithium chloride reduced ligature-induced BL in estrogen-deficient rats and yielded an overall greater trabecular area and overexpression of bone markers in alveolar bone under normal and deficient estrogen states. CLINICAL RELEVANCE: Lithium chloride may be a promising agent to assuage alveolar bone loss related to periodontitis, especially in osteoporotic conditions.

Effects of strontium ranelate on ligature-induced periodontitis in estrogen-deficient and estrogen-sufficient rats.

BACKGROUND AND OBJECTIVES: Strontium ranelate is a medication indicated for the treatment of osteoporosis that presents concomitant anti-resorptive and osteoanabolic dual biological activity. However, the effects of strontium ranelate on alveolar bone have been poorly explored. Furthermore, to date, there are no data on the effects of this medication on alveolar bone loss (BL) during conditions of estrogen deficiency. Therefore, the aim of this study was to evaluate the effects of strontium ranelate on ligature-induced periodontitis in estrogen-deficient and estrogen-sufficient rats. METHODS: Ninety-six rats were assigned to one of the following groups: sham-surgery + water (estrogen-sufficient; n = 24); ovariectomy + water (estrogen-deficient; n = 24), sham-surgery + strontium ranelate (ranelate/estrogen-sufficient; n = 24) and; ovariectomy + strontium ranelate (ranelate/estrogen-deficient; n = 24). The rats received strontium ranelate or water from the 14th day after ovariectomy until the end of the experiment. On the 21st day after ovariectomy, one first mandibular molar received a ligature, while the contralateral tooth was left unligated. Eight rats per group were killed at 10, 20, and 30 days after ligature placement. Bone loss (BL) and trabecular bone area (TBA) were analyzed in the furcation area of ligated and unligated teeth at all experimental times by histometry. Tartrate-resistant acid phosphatase (TRAP) positive cells and immunohistochemical staining for osteocalcin (OCN), osteopontin (OPN), osteoprotegerin (OPG), and receptor activator of NF-КB ligand (RANKL) were assessed in the ligated teeth at 30 days after ligature placement. RESULTS: At 10 and 30 days, ligated teeth of the estrogen-deficient group exhibited higher BL, when compared to all other groups (P < .05). At 10 days, TBAs were higher in the unligated teeth of strontium ranelate-treated groups, when compared to those of untreated groups (P < .05). At 30 days, the ligated teeth of the estrogen-deficient group exhibited lower TBA than the other groups (P < .05). There were no differences among groups regarding the number of TRAP-stained cells (P < .05). The strontium ranelate-treated groups exhibited lower expressions of OCN and RANKL than the untreated groups (P < .05). The estrogen-sufficient group presented higher staining for OPG than both treated and untreated estrogen-deficient groups (P < .05). CONCLUSIONS: Strontium ranelate prevented ligature-induced BL in an estrogen-deficiency condition and, to a certain extent, increased TBA in the presence and absence of periodontal collapse in states of estrogen deficiency and estrogen sufficiency. Furthermore, strontium ranelate also affected the expression of bone markers, appearing to have acted predominantly as an anti-resorptive agent.