Evaluation of a ß-calcium metaphosphate bone Graft containing bone morphogenetic protein-7 in rabbit maxillary defects.

BACKGROUND: Calcium phosphate-based materials have been widely used as bone substitutes and more recently are being exploited together with growth factors as bone tissue engineering scaffolds regulating cell behavior. The aim of this study is to evaluate the in vitro and in vivo response to a newly developed calcium metaphosphate (CMP) bone graft, with and without bone-stimulating growth factor. METHODS: Porous scaffolds of CMP were developed and extensively tested in vitro. Subsequently, CMP grafts with osteogenic protein-1 (OP-1) (test) and without OP-1 (control) were implanted into experimental rabbit maxillary bone defects. Animals were sacrificed at 2, 4, and 8 weeks, and samples were examined with microcomputed tomography (micro-CT) and processed for histomorphometric analysis. RESULTS: At 8 weeks, the scaffolds containing OP-1 induced greater bone formation (P = 0.018) than CMP alone, based on histomorphometric evaluation (percentage bone area: test: 57.1 ± 5.6; control: 49.4 ± 7.7) and micro-CT analysis (percentage bone volume density: test: 63.46 ± 5.61; control: 51.20 ± 6.71). Thus, these data indicated that both test and control CMP grafts showed a good degree of bone formation. Furthermore, the CMP materials showed signs of resorption from 4 weeks, and no graft materials were observed at 8 weeks. CONCLUSIONS: In vitro, the OP-1 loaded graft demonstrated a release profile and bioactivity over a 28-day period. In vivo testing confirmed enhanced bone formation of the OP-1 loaded graft after 8 weeks of healing.

Factors affecting survival and usefulness of implants placed in vascularized free composite grafts used in post-head and neck cancer reconstruction.

BACKGROUND: Bone-containing vascularized grafts have been used successfully to reconstruct post-cancer surgical defects. Dental implants can be placed in these bone-containing grafts to allow implant-supported prosthodontic reconstruction of these patients. PURPOSE: The aim of this study was to evaluate the survival of dental implants used in the rehabilitation of subjects treated with bone-containing vascularized grafts to compare usability of implants placed at the time of reconstruction and after healing. MATERIALS AND METHODS: A cross-sectional study was undertaken to examine survival rates of implants placed in vascularized bone-containing grafts either immediately at the time of surgical reconstruction or after 3 months healing. Other factors such as graft type, whether radiation therapy was given, and implant type were recorded. RESULTS: A total of 41 patients had 145 implants placed in 47 vascularized bone-containing flaps. Increased failure rate of implants was seen in immediately placed implants. There was also a significant increase in the number of osseointegrated implants that were prosthodontically unusable or sub-optimally placed in the immediate placement group. Radiation therapy was associated with a significant increase in failure rate. Modern implant surfaces appeared to perform better than machined/turned surfaces. Graft donor site did not influence implant survival. CONCLUSION: This study demonstrated the difficulties encountered with immediate placement of dental implants at the time of post-cancer reconstructive surgery.

The effect of smoking on periodontal bone regeneration: a systematic review and meta-analysis.

BACKGROUND: The effect of smoking on soft tissues after periodontal treatment has been extensively studied. However, little focus has been placed on the impact of smoking on bone regeneration after treatment. The aim of this review is to systematically assess the effect of smoking on bone regeneration after periodontal treatment. METHODS: A protocol was established and studies were sourced from five electronic databases. Screening, data abstraction, and quality assessment was conducted by two review authors. Prospective and retrospective clinical studies assessing bone regeneration in smokers and non-smokers after periodontal therapy were selected. In addition, arms of clinical trials comparing different interventions that reported results separately for smokers and non-smokers were also included. Primary outcome measures were based on clinical and/or radiographic indicators of bone regeneration after periodontal therapy. The review and meta-analysis followed many of the recommendations outlined in the preferred reporting items for systematic reviews and meta-analyses statement. RESULTS: Six of 10 studies included in this review concluded that smoking negatively influenced bone regeneration. A meta-analysis of a subgroup of three studies demonstrated that smoking resulted in significantly less bone gain (P = 0.03) as measured by a change in the probing bone level after the treatment of intrabony defects with guided tissue regeneration. The meta-analysis showed a standardized mean difference of -2.05 (95% confidence interval: -2.64 to -1.47) using the random-effects model. CONCLUSIONS: Smoking has a negative effect on bone regeneration after periodontal treatment. Patients should be advised that their smoking habit may result in poorer bone regeneration after periodontal treatment.

Effect of maxillary sinus floor augmentation on sinus membrane thickness in computed tomography.

BACKGROUND: Little is known about maxillary sinus compliance, i.e., the intrinsic potential of the sinus membrane to resume its homeostatic status after the surgical trauma caused by sinus floor elevation. The aim of the present study is to investigate the effect of maxillary sinus floor augmentation on sinus membrane thickness. METHODS: Within-patient comparison of computed tomographic scans before bone grafting versus 4 to 6 months after bone grafting was performed. Changes in membrane thickness were evaluated in 65 maxillary sinus floor augmentation procedures via a lateral approach in 35 patients without clinical signs of sinus pathology at any time. RESULTS: Sinus membrane thickness differed significantly before (0.8 ± 1.2 mm) versus after (1.5 ± 1.3 mm) augmentation surgery (P <0.001), with a mean increase of 0.8 ± 1.6 mm (maximum: 4.4 mm). Only 28% of augmented sinuses did not show membrane thickening. In non-augmented control sinuses, there was no evidence of membrane thickness increase. CONCLUSIONS: The results indicate that the maxillary sinus membrane, even in healthy clinical conditions, undergoes morphologic modifications after sinus floor elevation, yet membrane reactions demonstrate significant variability. Future research on the effect of augmentation surgery on maxillary sinus physiology is recommended.

Modification of thiol functionalized aptamers by conjugation of synthetic polymers.

Aptamers are known for their short in vivo circulating half-life and rapid renal clearance. Their conjugation to poly(ethylene glycol) (PEG) is a way to improve their residence in the body. Two aptamers (AptD and AptF), having a disulfide protected thiol modification on the 3' end, have been conjugated to maleimide activated PEGs of various molecular weights and structures (linear PEG20; branched PEG20 and 40; PolyPEG17, 40, and 60 kDa). The high yield coupling (70-80% in most of the cases) could be achieved using immobilized tris[2-carboxyethyl]phosphine hydrochloride (TCEP) as reducing agent at pH 4. The affinity of PEGylated AptD for its target was reduced by conjugation to linear PEG20 and branched PEG40, but not to branched PEG20 and PolyPEGs. This work demonstrates an alternative approach to PEGylation of aptamers, and that the effect of PEG on the affinity for the target varies according to the structure and conformation of the synthetic polymer.

Smoking behaviour and attitudes to periodontal health and quit smoking in patients with periodontal disease.

OBJECTIVE: The aim of this study was to assess oral health-related beliefs and attitudes, health behaviour of smokers in relation to the Transtheoretical Model (TTM) of behaviour change, willingness to have smoking cessation provided together with periodontal treatment. MATERIAL AND METHODS: Postal questionnaire was sent to 500 referred patients. Part 1 looked at attitudes and beliefs about periodontal disease, Part 2 aimed at current smokers focused on the TTM and smoking cessation. RESULTS: Response rate was 56% (n=277); 67% females, 33% males. Mean age was 44.9 years (SD 12.45); 24.5% current smokers, 30.3% past smokers, 45.5% never smokers. Fewer smokers reported "bleeding gums" (p=0.027), but more smokers reported "having loose teeth" (p=0.016). The TTM stages of change indicated that 31% of current smokers were in pre-contemplation of quitting smoking, 46% were in contemplation and 23% were in preparation. Twenty-three percent of the past smokers were in action and 77% in maintenance. Smokers showed differences in the "self-re-evaluation" (p=0.001) and "self-liberation" (p=0.015) processes of change depending on their stage of change (pre-contemplation or preparation). Nearly half (49%) of the current smokers who wanted to quit requested smoking cessation to be provided alongside their periodontal treatment. CONCLUSION: A large proportion of periodontal patient smokers may be considering quitting, and nearly half requested provision of smoking cessation intervention in conjunction with the periodontal treatment.

The influence of nicotine on granulocytic differentiation - inhibition of the oxidative burst and bacterial killing and increased matrix metalloproteinase-9 release.

BACKGROUND: Neutrophils leave the bone marrow as terminally differentiated cells, yet little is known of the influence of nicotine or other tobacco smoke components on neutrophil differentiation. Therefore, promyelocytic HL-60 cells were differentiated into neutrophils using dimethylsulfoxide in the presence and absence of nicotine (3-(1-methyl-2-pyrrolidinyl) pyridine). Differentiation was evaluated over 5 days by monitoring terminal differentiation markers (CD11b expression and formazan deposition); cell viability, growth phase, kinetics, and apoptosis; assessing cellular morphology and ultrastructure; and conformational changes to major cellular components. Key neutrophil effector functions (oxidative burst, bacterial killing, matrix metalloproteinase release) were also examined. RESULTS: Nicotine increased the percentage of cells in late differentiation phases (metamyelocytes, banded neutrophils and segmented neutrophils) compared to DMSO alone (p < 0.05), but did not affect any other marker of neutrophil differentiation examined. However, nicotine exposure during differentiation suppressed the oxidative burst in HL-60 cells (p < 0.001); inhibited bacterial killing (p < 0.01); and increased the LPS-induced release of MMP-9, but not MMP-2 (p < 0.05). These phenomena may be alpha-7-acetylcholine nicotinic receptor-dependent. Furthermore, smokers exhibited an increased MMP-9 burden compared to non-smokers in vivo (p < 0.05). CONCLUSION: These findings may partially explain the known increase in susceptibility to bacterial infection and neutrophil-associated destructive inflammatory diseases in individuals chronically exposed to nicotine.

A comparison of bone mineral density in the spine, hip and jaws of edentulous subjects.

OBJECTIVES: The aim was to investigate the relationship between bone mineral density (BMD) of the jaws (mandible and maxilla) and other skeletal sites. In addition, the influence of gender, smoking and the number of years without natural teeth were examined. MATERIALS AND METHODS: 18 edentulous patients (9 females, 9 males) with a mean age of 67.1 (sd 12.6) years had DXA scans to assess the BMD of the lumbar spine and hip, together with the ramus, body and symphysis of the mandible and the anterior of the maxilla. RESULTS: BMD values for the ramus were similar to those for the femur but significantly lower than the lumbar spine. The body and anterior mandible had higher values and the anterior maxilla lower values than both the femur and ramus. The ramus BMD showed moderately strong relationships with the standard measures of BMD in the spine and hip, but the BMD of other areas of the jaws showed no relationship with skeletal sites. The BMD for both the hip and the ramus showed an inverse relationship with increasing age. There was no statistically significant relationship between BMD of hip, spine and jaw and either years edentulous or cigarette years. (207) CONCLUSIONS: Although the ramus of the mandible may show correlation of BMD with skeletal sites, the areas of the jaws where implants may be placed do not. Therefore BMD of the skeletal sites could not be used to predict BMD of the jaws. The BMD of the jaws as measured by DXA showed no relationship with either years edentulous or cigarette smoking.

Investigation of periodontal destruction patterns in smokers and non-smokers.

BACKGROUND: Previous work has suggested that tobacco smoking has a local as well as a systemic effect on the severity of periodontal disease. OBJECTIVE: To test the hypothesis that smokers have more disease in the upper anterior region. METHODS: A retrospective stratified random sample of 49 non-smokers and 39 heavy smokers (>or=20 cigarettes/day) was obtained from a total of 3678 referred patients with adult periodontitis. Probing depth data were collected from clinical records and radiographic measurements were carried out on existing dental panoramic tomographs to assess the inter-proximal bone levels. RESULTS: The proportion of sites with "bone loss" 4.5 mm or greater was higher in smokers, the greatest difference being observed in upper anterior sites (smokers: 73.3+/-25.5%, non-smokers: 48.3+/-31.2%, p<0.001). A difference was also observed when the number of palatal sites probing 4 mm or greater in the upper anterior region was expressed as a proportion of all such sites in the mouth (smokers: 12.3+/-6.8%, non-smokers: 9.8+/-8.8%; p=0.050). CONCLUSION: The overall pattern of tissue destruction was consistent with a systemic effect of smoking. The suggestion of a marginal local effect of the smoking habit in maxillary anterior palatal sites requires further investigation.

Mechanisms of action of environmental factors--tobacco smoking.

AIM: To review the potential biological mechanisms underlying the effects of tobacco smoking on periodontitis. MAIN FINDINGS: Smoking has major effects on the host response, but there are also a number of studies that show some microbiological differences between smokers and non-smokers. Smoking has a long-term chronic effect on many important aspects of the inflammatory and immune responses. Histological studies have shown alterations in the vasculature of the periodontal tissues in smokers. Smoking induces a significant systemic neutrophilia, but neutrophil transmigration across the periodontal microvasculature is impeded. The suppression of neutrophil cell spreading, chemokinesis, chemotaxis and phagocytosis have been described. Protease release from neutrophils may be an important mechanism in tissue destruction. Tobacco smoke has been found to affect both cell-mediated immunity and humoral immunity. Research on gingival crevicular fluid has demonstrated that there are lower levels of cytokines, enzymes and possibly polymorphonuclear cells in smokers. In vitro studies have shown detrimental effects of nicotine and some other tobacco compounds on fibroblast function, including fibroblast proliferation, adhesion to root surfaces and cytotoxicity. CONCLUSION: Tobacco smoking has widespread systemic effects, many of which may provide mechanisms for the increased susceptibility to periodontitis and the poorer response to treatment.

A prospective study of single stage surgery for implant supported overdentures.

OBJECTIVES: To investigate the clinical and psychological outcomes with ball attachment supported mandibular overdentures on Branemark implants placed in a single stage surgical approach. MATERIAL AND METHODS Sixteen edentulous patients aged 32-74, with atrophic (flat) mandibular residual alveolar ridges, and a history of having difficulty coping with technically adequate mandibular dentures made within the last 2 years were recruited. Mark II Branemark implants were placed using a single stage surgical technique with healing abutments placed at time of surgery. Mandibular complete dentures were modified by relieving the areas over the abutments and a resilient liner was placed. After a period of 3 months, healing abutments were replaced with ball abutments and new complete dentures were made and the patients followed for a period of 2 years. Patients completed the General Health Questionnaire (GHQ) and answered further questions relating to the functional and psychological impact of complete denture wearing before treatment and 3 months after the dentures were provided. Patients additionally completed the GHQ 2 years after overdenture insertion. RESULTS: Six patients suffered from loss of one of their two implants in the first 2 years after placement. Three of the implants had failed to osseointegrate, which was detected at initial loading 3 months after placement. General Health Questionnaire scores were statistically significantly reduced after treatment and the improvement was maintained over 2 years. Responses to the functional and psychological questions were similarly improved. CONCLUSION: Single stage surgery led to a higher rate of implant failure than expected. Patients showed significant psychological health benefits from the treatment provided.

Cytokine release by osteoblast-like cells cultured on implant discs of varying alloy compositions.

OBJECTIVE: The aims of this study were (i). to assess the morphological features of osteo-blast-like, osteosarcoma cells (cell line SaOS-2) cultured on implant surfaces of varying alloys and (ii). to evaluate the biological activity of these cells, following their adhesion onto these surfaces. MATERIALS AND METHODS: SaOS-2 cells (6 x 104) were grown on titanium discs (diameter 30 mm), each with a surface of differing composition and roughness (commercially pure titanium, titanium-aluminium-vanadium alloy, oxide-blasted titanium and Astra-Tech special treatment titanium; the alloys are directly comparable with those used to construct implants). The cells were grown for time periods of 1, 3, 5 and 7 days, the media were collected and the cells were fixed with 2.5% glutaraldehyde. The media were then assayed (using enzyme-linked immunosorbant assay) for the levels of interleukin (IL)-1, interleukin-6, interleukin-18 and osteoprotegerin (OPG) produced by the cells. The discs, with the cells fixed on them, were viewed under scanning electron microscopy (SEM, x 2.0 k) to evaluate cell morphology. RESULTS: Following attachment, the cells changed their morphology and released local factors known to activate osteoclasts. Commercially pure titanium stimulated the cells the most and titanium-aluminium-vanadium alloy the least. All implant materials stimulated production of IL-1, IL-6, IL-18 above that produced by cells grown on Petri dishes (polystyrene). The titanium-aluminium-vanadium alloy allowed cell attachment but levels of IL-1 in this medium were significantly lower (31.5 +/- 5.2 pg/ml on same day) than cultures with pure titanium (201.8 +/- 11.5 pg/ml at day 5). The same pattern was observed with the IL-6, IL-18, and OPG with polystyrene appearing to stimulate most production of OPG. Titanium-aluminium-vanadium produced the least biological response.