Hyaluronic acid enhances cell migration, viability, and mineralized tissue-specific genes in cementoblasts.

BACKGROUND/OBJECTIVES: It has been repeatedly demonstrated that cementum formation is a crucial step in periodontal regeneration. Hyaluronic acid (HA) is an important component of the extracellular matrix which regulates cells functions and cell-cell communication. Hyaluronic acid/derivatives have been used in regenerative periodontal therapy, but the cellular effects of HA are still unknown. To investigate the effects of HA on cementoblast functions, cell viability, migration, mineralization, differentiation, and mineralized tissue-associated genes and cementoblast-specific markers of the cementoblasts were tested. MATERIALS AND METHODS: Cementoblasts (OCCM-30) were treated with various dilutions (0, 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128) of HA and examined for cell viability, migration, mineralization, and gene expressions. The mRNA expressions of osteocalcin (OCN), runt-related transcription factor 2 (Runx2), bone sialoprotein (BSP), collagen type I (COL-I), alkaline phosphatase (ALP), cementum protein-1 (CEMP-1), cementum attachment protein (CAP), and small mothers against decapentaplegic (Smad) -1, 2, 3, 6, 7, ß-catenin (Ctnnb1) were performed with real-time polymerase chain reaction (RT-PCR). Total RNA was isolated on days 3 and 8, and cell viability was determined using MTT assay on days 1 and 3. The cell mineralization was evaluated by von Kossa staining on day 8. Cell migration was assessed 2, 4, 6, and 24 hours following exposure to HA dilutions using an in vitro wound healing assay (0, 1:2, 1:4, 1:8). RESULTS: At dilution of 1:2 to 1:128, HA importantly increased cell viability (p < .01). HA at a dilution of 1/2 increased wound healing rates after 4 h compared to the other dilutions and the untreated control group. Increased numbers of mineralized nodules were determined at dilutions of 1:2, 1:4, and 1:8 compared with control group. mRNA expressions of mineralized tissue marker including COL-I, BSP, RunX2, ALP, and OCN significantly improved by HA treatments compared with control group both on 3 days and on 8 days (p < .01). Smad 2, Smad 3, Smad 7, and ß-catenin (Ctnnb1) mRNAs were up-regulated, while Smad1 and Smad 6 were not affected by HA administration. Additionally, HA at dilutions of 1:2, 1:4, and 1:8 remarkably enhanced CEMP-1 and CAP expressions in a dilution- and time-dependent manner (p < .01). CONCLUSIONS: The present results have demonstrated that HA affected the expression of both mineralized tissue markers and cementoblast-specific genes. Positive effects of HA on the cementoblast functions demonstrated that HA application may play a key role in cementum regeneration.

Periodontitis severity in obstructive sleep apnea patients.

OBJECTIVES: This cross-sectional study investigated the stages of periodontitis in obstructive sleep apnea (OSA) patients and risk factors associated with periodontitis severity among them. MATERIALS AND METHODS: A total of 194 patients underwent a polysomnography/polygraphy and were referred to periodontal examination. According to apnea-hypopnea index (AHI), patients were classified as mild OSA (AHI < 15) and moderate to severe OSA (AHI ≥ 15), whereas periodontitis severity was determined by the clinical attachment level (CAL) according to the recent Classification of Periodontal Diseases and Conditions. Patients were grouped into two categories: stages 1 and 2, and stages 3 and 4. RESULTS: Higher AHI values were reported in OSA patients exhibiting periodontitis stages 3 and 4 compared to OSA patients with periodontitis stages 1 and 2 (p = 0.043) and the non-periodontitis group (p = 0.044). A positive correlation was found between AHI and mean CAL (r = 0.215; p = 0.004), and between AHI and plaque scores (r = 0.292; p < 0.001). Following a multivariable regression analysis, AHI was a significant predictor of mean CAL (ß = 0.169; p = 0.031), explaining 16.4% of variability in mean CAL (adjusted R2 = 0.164; p < 0.001). Older patients had higher odds for an increased mean CAL (ß = 0.266; p = 0.001), as well as patients smoking or formerly smoking (ß = 0.305; p < 0.001) whereas visiting a dental medicine doctor once a year or more often was associated with a decreased mean CAL (ß = - 0.182; p = 0.02). CONCLUSIONS: OSA was associated with severe stages of periodontitis along with increased age, smoking, low frequency of dental visits, and poor oral hygiene. CLINICAL RELEVANCE: Screening for periodontitis is recommended for patients with more severe forms of OSA.

ASSOCIATION BETWEEN PERIODONTITIS AND LIVER DISEASE.

Recent clinical and scientific evidence confirms the negative impact of long-term periodontitis on the clinical course and progression of various liver diseases. Periodontitis is a chronic, slow-progressing infectious disease of the tooth supporting tissues caused mainly by the gram-negative bacteria Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola and Tannerella forsythia. These specific pathogens can be easily translocated from oral cavity to the intestine. Disruption of the intestine microbiota composition by orally derived periodontal pathogenic bacteria has recently been suggested to be a causal mechanism between periodontitis and liver disease. Furthermore, both diseases have the ability to induce an inflammatory response and lead to the creation of inflammatory mediators through which they may influence each other. Recent epidemiologic studies have demonstrated that individuals with liver cirrhosis have considerably poorer periodontal clinical parameters than those without cirrhosis. Periodontal therapy in cirrhosis patients favorably modulates oral and gut microbiome, the course of systemic inflammation, cirrhosis prognostic factors, and cognitive function. Therefore, future clinical researches should be focused on detailed examination of the biological mechanisms, strength and direction of the association between advanced liver disease and periodontitis.

Cigarette smoking and its consequences on periodontal health in teenagers: a cross-sectional study.

OBJECTIVES: Smoking has been extensively investigated as a risk factor for periodontal disease and many studies have confirmed it. The aim of this study was to show data from Croatia on periodontal health among high school students, with the focus on association of tobacco use and oral hygiene habits, and the periodontal parameters. METHODS: Pocket probing depth (PPD), bleeding on probing, supragingival calculus, and oral hygiene habits were recorded for each examinee out of 517 high school students. PPD was measured at 4 interproximal sites on all first molars and central incisors (index teeth). RESULTS: There were 34.6% smokers among subjects, who started smoking on average at the age of 14. More than half of the smokers (55.3%) smoked 1-10 cigarettes per day and intensity of smoking increased with age. PPD was significantly greater (p = 0.012) in smokers (1.69 mm) than in non-smokers (1.59 mm). In non-smoking subjects, increased values of PPD were in direct proportion to the reduced frequency of brushing, while there was no difference in smokers. The number of cigarettes smoked and duration of smoking had a great influence on PPD. Smokers had on average more teeth with supragingival calculus than non-smokers, while non-smokers had more bleeding on probing, and the difference was statistically significant (p < 0.001). CONCLUSIONS: This study indicated that cigarette smoking was associated with decreased periodontal health even in this young population. Irregular oral hygiene was associated with decreased periodontal health only in non-smokers. Therefore, it should be important to inform young smokers about the negative effects of cigarette smoking on periodontal health, to provide consultations and reinforce smoking prevention measures.

Biomaterials and regenerative technologies used in bone regeneration in the craniomaxillofacial region: Consensus report of group 2 of the 15th European Workshop on Periodontology on Bone Regeneration.

BACKGROUND AND AIMS: To review the regenerative technologies used in bone regeneration: bone grafts, barrier membranes, bioactive factors and cell therapies. MATERIAL AND METHODS: Four background review publications served to elaborate this consensus report. RESULTS AND CONCLUSIONS: Biomaterials used as bone grafts must meet specific requirements: biocompatibility, porosity, osteoconductivity, osteoinductivity, surface properties, biodegradability, mechanical properties, angiogenicity, handling and manufacturing processes. Currently used biomaterials have demonstrated advantages and limitations based on the fulfilment of these requirements. Similarly, membranes for guided bone regeneration (GBR) must fulfil specific properties and potential biological mechanisms to improve their clinical applicability. Pre-clinical and clinical studies have evaluated the added effect of bone morphogenetic proteins (mainly BMP-2) and autologous platelet concentrates (APCs) when used as bioactive agents to enhance bone regeneration. Three main approaches using cell therapies to enhance bone regeneration have been evaluated: (a) "minimally manipulated" whole tissue fractions; (b) ex vivo expanded "uncommitted" stem/progenitor cells; and (c) ex vivo expanded "committed" bone-/periosteum-derived cells. Based on the evidence from clinical trials, transplantation of cells, most commonly whole bone marrow aspirates (BMA) or bone marrow aspirate concentrations (BMAC), in combination with biomaterial scaffolds has demonstrated an additional effect in sinus augmentation and horizontal ridge augmentation, and comparable bone regeneration to autogenous bone in alveolar cleft repair.

Therapeutic Manuka Honey as an Adjunct to Non-Surgical Periodontal Therapy: A 12-Month Follow-Up, Split-Mouth Pilot Study.

Periodontitis is recognized as one of the most common diseases worldwide. Non-surgical periodontal treatment (NSPT) is the initial approach in periodontal treatment. Recently, interest has shifted to various adjunctive treatments to which the bacteria cannot develop resistance, including Manuka honey. This study was designed as a split-mouth clinical trial and included 15 participants with stage III periodontitis. The participants were subjected to non-surgical full-mouth therapy, followed by applying Manuka honey to two quadrants. The benefit of adjunctive use of Manuka honey was assessed at the recall appointment after 3, 6, and 12 months, when periodontal probing depth (PPD), split-mouth plaque score (FMPS), split-mouth bleeding score (FMBS), and clinical attachment level (CAL) were reassessed. Statistically significant differences between NSPT + Manuka and NSPT alone were found in PPD improvement for all follow-up time points and CAL improvement after 3 and 6 months. These statistically significant improvements due to the adjunctive use of Manuka amounted to (mm): 0.21, 0.30, and 0.19 for delta CAL and 0.18, 0.28, and 0.16 for delta PPD values measured after 3, 6, and 12 months, respectively. No significant improvements in FMPS and FMBS were observed. This pilot study demonstrated the promising potential of Manuka honey for use as an adjunct therapy to nonsurgical treatment.

Risk factors of severe periodontitis in kidney transplant recipients: A case-control study.

BACKGROUND: Kidney transplant recipients (KTRs) represent a vulnerable group of patients who develop a number of comorbidities. Severe periodontitis (SP) is associated with the most common chronic systemic diseases including kidney diseases. The objective of this study was to explore the risk factors for SP in KTRs. METHODS: In this study, KTRs were divided into those with or without periodontitis and in relation to the severity of periodontitis. A comprehensive medical and periodontal examination was performed and evaluated. Multivariate logistic regression was performed to examine possible risk factors for SP among KTRs. RESULTS: A total of 100 KTRs were included in the analysis, of which 87% had periodontitis. Significant predictors of periodontitis were older age (OR = 1.07, 95% CI [1.01, 1.13], p = 0.016) and lower skeletal muscle mass (OR = 0.88, 95% CI [0.78, 0.99], p = 0.035). When examining periodontitis severity, predictors of SP (n = 21, 24%) were increased levels of uric acid (OR = 1.01, 95% CI [1.00, 1.02], p = 0.022) and dental plaque (OR = 1.04, 95% CI [1.01, 1.07], p = 0.013). In the subset analysis that included only KTRs with measured advanced glycation end products (AGE) (n = 47), 34% (n = 16) had SP. The predictors of SP were AGE (OR = 3.89, 95% CI [1.28, 11.82], p = 0.017) and dental plaque (OR = 1.07, 95% CI [1.01, 1.13], p = 0.028). CONCLUSIONS: KTRs with SP had significantly higher uric acid levels and AGE, which may contribute to the systemic health status of this patient population.

The proteome and gene expression profile of cementoblastic cells treated by bone morphogenetic protein-7 in vitro.

AIM: Regenerative periodontal therapy is often unpredictable and limited. Cementum regeneration is necessary for the proper repair of a periodontal ligament. The precise mechanism how bone morphogenetic protein-7 (BMP7) induces differentiation and mineralization of cementoblasts remains undetermined. The purpose of this study was to evaluate the effect of BMP7 on early proteome and gene expression profile of cementoblastic OCCM.30 cells in vitro. MATERIALS AND METHODS: Immortalized murine cementoblasts (OCCM.30) were exposed to BMP7 and evaluated for: (1) proliferation; (2) mineralization; (3) early proteome profile using liquid chromatography-mass spectrometry (LC-MS); and (4) gene expression by quantitative RT-PCR. RESULTS: Bone morphogenetic protein-7 increased the cell proliferation at 24 h and 48 h, while higher doses suppressed the cell proliferation at 48 h. BMP7 induced the mineralization of cementoblasts following 8 days of therapy. Using LC-MS we identified 1117 proteins from the cell lysate. Many belonged to extracellular matrix formation such as PCPE1, collagens, annexins and integrin receptors. RT-PCR analyses revealed a BMP7 dose-dependent upregulation of BMP1, TGFß1, osterix, osteoprotegerin, procollagen I and II, PCPE1, and noggin, while BMP6 and chordin expression were decreased. The high BMP7 dose down regulated most of the genes 24 h following therapy. CONCLUSION: Bone morphogenetic protein-7 promotes differentiation and mineralization of cementoblasts via inducing PCPE1 and BMP1 responsible for processing of type I collagen.

Effect of bone morphogenetic protein-7 on gene expression of bone morphogenetic protein-4, dentin matrix protein-1, insulin-like growth factor-I and -II in cementoblasts in vitro.

Formation of root cementum is a crucial moment in the development of the periodontium. Cells that produce the cementum are named cementoblasts and they posses some unique characteristics, which differentiates them from osteoblasts. Bone morphogenetic proteins (BMPs) are crucial regulators of both bone and tooth formation. In animal studies BMPs have shown to induce periodontal regeneration, however the molecular mechanism as how BMP-7 induces cementogenesis is largely unknown. We have investigated how BMP-7 regulates gene expression of BMP-4, Dentin matrix protein-1 (DMP-1), Insulin-like growth factor-I (IGF-I) and -II (IGF-II) in cementoblasts. BMP-7 induced proliferation, and mineralized nodule formation of cementoblasts. Our results show that gene expression was influenced by the BMP-7 concentration used, with 75 ng/mL generally down-regulating gene expression at 6 hours and then up-regulating after 24 hours. The 300 ng/mL concentration had an opposite effect while the 150 ng/mL concentration generally up-regulated gene expression after 6 hours and then after 24 hours maintained this up-regulation or had no effect compared to control, depending on the examined gene. The results show that BMP-7 down-regulated BMP-4 expression in cementoblasts but still up-regulated DMP-1 gene expression suggesting that BMP-7 can, in a paracrine manner, functionally substitute for BMP-4. Furthermore, it seems that BMP-7 exerts its effect more through the IGF-II than the IGF-I pathway as shown by an up-regulation of IGF-II and down-regulation of IGF-I. These results suggest that a combination of BMP-7/IGF-II could have a potential therapeutical significance in inducing cementogenesis and periodontal regeneration.

Heparan Sulfate Glycosaminoglycan Is Predicted to Stabilize Inflammatory Infiltrate Formation and RANKL/OPG Ratio in Severe Periodontitis in Humans.

Since chronically inflamed periodontal tissue exhibits extracellular matrix (ECM) degradation, the possible alternative to standard periodontitis treatment is to restore ECM by supplementing its components, including heparan sulfate glycosaminoglycan (HS GAG). Supplementation of the degraded ECM with synthetic derivatives of HS GAGs has been shown to be effective for periodontal tissue regeneration in experimental animal models of periodontitis. However, the potential of HS GAG supplementation for the treatment of periodontal disease in humans is still unknown. Here, we used a statistical model to investigate the role of HS GAG on inflammatory infiltrate formation and alveolar bone resorption in humans with severe periodontitis. The model was based on data from immunofluorescence staining (IF) of human gingiva samples, and reconstruction of a subset of HS GAG -related proteins from STRING reactome database. According to predictions, increased expression of native HS GAG might stabilize the accumulation of gingival inflammatory infiltrate (represented by the general inflammatory cell marker CD45) and alveolar bone resorption (represented by Receptor Activator of Nuclear ΚΒ ligand (RANKL) and osteoprotegerin (OPG) ratio) but could not restore them to healthy tissue levels. Therefore, supplementation of native HS GAG may be of limited benefits for the treatment of sever periodontitis in humans.

Medication Prescribing Practices in Croatian Dental Offices and Their Contribution to National Consumption.

OBJECTIVES: The objective of this study was to analyse the pattern of medication prescribing among dental practitioners in the Republic of Croatia and to compare it with general medical practice patterns at the national level. METHODS: Data on drug prescriptions were obtained from the Croatian Health Insurance Fund. The number of dentist prescriptions, the cost of medicine denominated in the national currency (Croatian Kuna [HRK]), and the number of packages and days prescribed have been included in the analysis. RESULTS: Results indicate that there was an increase in medication prescribing(+5.7%) by dentists, with antimicrobials comprising the majority, whereas national medical antimicrobial prescribing had slightly decreased. Antibiotics accounted for 80% of all dentists' prescriptions, with penicillins being the most commonly prescribed. In particular, amoxicillin with clavulanic acid accounted for 56.4% of all antibiotics prescribed. Such broad-spectrum antibiotics were prescribed more frequently than those of narrow-spectrum. Antibiotics were followed by nonsteroidal anti-inflammatory drugs, with prescribing frequency for ibuprofen increasing by 75%. CONCLUSION: Current trends show an increase in the overall prescription rate for all medications prescribed by dentists. The largest increase was observed for the broad-spectrum amoxicillin with clavulanic acid, and ibuprofen. As the apparent widespread use of broad-spectrum antibiotics by dentists in Croatia is in contrast to national and international recommendations for antibiotic stewardship, there is a need for further prospective investigation and possible provider education and guidelines.

Effectiveness of Autologous Tissue Grafts on Soft Tissue Ingrowth in Patients Following Partial Root Extraction with Socket Shield: A Retrospective Analysis of a Case Series.

PURPOSE: There is little knowledge about healing patterns for the socket with an intentionally retained root fragment: a socket shield. The clinical observation is soft tissue ingrowth next to the socket shield. The aim of this study was to evaluate the effectiveness of autologous grafting matrices in preventing soft tissue ingrowth. MATERIALS AND METHODS: Patient data from a private clinic were searched for sockets with a socket shield left to heal with blood clot or grafted with autologous materials: autologous platelet-rich fibrin (PRF), scraped particulate bone, cortical tuberosity bone plate, or particulate dentin and covered with PRF membranes. The included sites were exposed by the flap 4 months after the first surgery, and soft tissue ingrowth depth and width next to the root fragment were measured by a scaled probe and documented. RESULTS: Evaluation of 34 sites showed the greatest depth of soft tissue ingrowth in the nongrafted sockets (6.0 ± 0.0 mm). Grafting with PRF plugs (depth of 2.3 ± 0.2 mm) or particulate bone (depth of 2.7 ± 0.6 mm) decreased soft tissue ingrowth. Grafting with particulate dentin or cortical tuberosity bone plate resulted in a soft tissue ingrowth depth of only 1 mm, yielding the best clinical outcome. Radiography confirmed those findings. CONCLUSION: Autologous dentin particulate or tuberosity cortical bone plate is most effective for preventing soft tissue ingrowth.

Treatment of Intrabony Defects with a Combination of Hyaluronic Acid and Deproteinized Porcine Bone Mineral.

BACKGROUND: this study evaluates the clinical outcomes of a novel approach in treating deep intrabony defects utilizing papilla preservation techniques with a combination of hyaluronic acid (HA) and deproteinized porcine bone mineral. METHODS: 23 patients with 27 intrabony defects were treated with a combination of HA and deproteinized porcine bone mineral. Clinical attachment level (CAL), pocket probing depth (PPD), gingival recession (REC) were recorded at baseline and 6 months after the surgery. RESULTS: At 6 months, there was a significant CAL gain of 3.65 ± 1.67 mm (p < 0.001) with a PPD reduction of 4.54 ± 1.65 mm (p < 0.001), which was associated with an increase in gingival recession (0.89 ± 0.59 mm, p < 0.001). The percentage of pocket resolution based on a PPD ≤4 mm was 92.6% and the failure rate based on a PPD of 5 mm was 7.4%. CONCLUSIONS: the present findings indicate that applying a combined HA and xenograft approach in deep intrabony defects provides clinically relevant CAL gains and PPD reductions compared to baseline values and is a valid new approach in treating intrabony defects.

The Impact of the COVID-19 Pandemic on Dental Education: An Online Survey of Students' Perceptions and Attitudes.

PURPOSE: Dental education institutions worldwide experienced disruptive changes amid the COVID-19 pandemic, with a rapid switch to the online learning format. Thus, this study aimed to assess the impact of the COVID-19 pandemic on dental education and evaluates the perceptions and attitudes of students towards the introduction of online learning in the School of Dental Medicine in Zagreb, Croatia. METHODS: A survey was conducted on a population of undergraduate students. It was comprised of perceptions and attitudes of students on the impact of the COVID-19 pandemic on their psychoemotional status, changes introduced in the educational system, and online learning in particular. RESULTS: Of the 352 students that completed the survey, 66.2% of students reported being psychoemotionally affected by the lockdown. The most significant impact of the switch from in-person to online learning was observed in terms of missing contact with lecturers (60.3%) and peers (90.3%) and loss of practical courses, regarding which 65% of students agreed that they could not be compensated. While only 36.1% reported that online teaching fully met their expectations, the majority of the students (61.9%) agreed that online lectures were as valuable as in-person lectures and that the theoretical courses could be carried out online in the future as well (69.9%). CONCLUSIONS: Students reported relative satisfaction with changes in the learning format and a positive attitude towards online learning; however, several challenges and obstacles were identified.

Identification of bone morphogenetic protein 4 in the saliva after the placement of fixed orthodontic appliance.

BACKGROUND: This study was conducted in order to explore the effects of orthodontic tooth movement (OTM) on the changes of salivary proteome. This prospective observational pilot study recruited 12 healthy teenage boys with malocclusion treated with a fixed orthodontic appliance and 6 appropriate control participants. Saliva samples were collected a day before and at 0, 2, 7, and 30 days after initialization of treatment, corresponding to the initial, lag, and post-lag phases of OTM. Pooled samples were analyzed by liquid chromatography-mass spectrometry, ELISA, and Western blotting. To date, there is no published data on the presence of BMP molecules or their antagonists in the saliva or in the gingival cervical fluid related to orthodontic conditions. RESULTS: A total of 198 identified saliva proteins were classified based on their functional characteristics. Proteins involved in bone remodeling were observed exclusively 30 days post appliance placement, including bone morphogenetic protein 4 (BMP4), a BMP antagonist BMP-binding endothelial regulator, insulin-like growth factor-binding protein 3, cytoskeleton-associated protein 4, and fibroblast growth factor 5. Based on the analysis of protein interactions, BMP4 was found to have a central position in this OTM-related protein network. CONCLUSIONS: The placement of a fixed orthodontic appliance induced occurrence of proteins involved in bone remodeling in the saliva at a time corresponding to the post-lag period of OTM. Limitations of this study include a relatively small sample size, limited time of monitoring patients, and the lack of interindividual variability assessment.

Is periodontitis and its treatment capable of changing the quality of life of a patient?

While clinical indicators, or so-called surrogate outcomes in periodontology, allow us to assess periodontal health and evaluate outcomes of periodontal therapy from a clinician's perspective, they cannot be used to evaluate patients' subjective perceptions of their health status or satisfaction with the received treatment. These can be assessed through patient-based outcomes, such as quality of life. The aim of this paper is to give an overview of the multidimensional concept of oral health-related quality of life (OHRQoL), its measures and association with plaque-induced, inflammatory periodontal diseases. Periodontitis and its clinical consequences, such as tooth loss, have a considerable negative effect on OHRQoL, while periodontal treatment and alleviation of the symptoms can lead to improvement in OHRQoL. Implant rehabilitation of missing teeth also seems to positively influence OHRQoL.

Evaluation of Periodontal Parameters in Patients with Early Stage Chronic Lymphocytic Leukemia.

OBJECTIVE: To assess periodontal conditions in patients with early stage CLL and to compare it with the periodontal status of age matched healthy controls and to analyze the relationship between periodontal and hematological parameters in CLL patients. MATERIALS AND METHODS: 60 subjects were examined: 30 patients with CLL Rai 0 (test group) and 30 age-matching healthy individuals (control group). The exclusion criteria were: presence of other systemic disease or condition (e.g. diabetes), history of treatment for periodontitis, use of antibiotics during the last 3 months, use of medications. Socio-demographic data were obtained by means of a questionnaire. Participants with at least 8 teeth underwent a full mouth examination assessing API, PBI, PPD, REC and CAL. Medical data for CLL patients were collected from the patients' records, while hematological data were obtained from the hemogram. RESULTS: Difference between groups was statistically significant for age, number of teeth and frequency of dental checkups (p<0.05). Patients with CLL had significantly higher average values of periodontal indices (API 0.81±0.18; PBI 2.72±0.68; PPD 3.40±0.53; REC 1.95±0.87, CAL 4.37±0.80) compared to the control group (API 0.69±0.15; PBI 1.91±0.45; PPD 2.51±0.40; REC 0.99±0.54; CAL 3.00±0.58). The correlation coefficients between age and periodontal indices showed statistically significance between age and REC (r=0.357; p<0.01), and age and CAL (r=0.295; p<0.05). Age was not statistically significant covariate for CAL (F=2.205; p>0.05), only for REC (F=4.601; p<0.05). After the removal of the statistical effect of age, the difference in REC between CLL and control group remained statistically significant (F=19.732; p<0.01; eta(2)=0.287). Statistically significant association between periodontal and hematological parameters in CLL patients was not found (p>0.05). CONCLUSION: The results of this study showed that patients with CLL had worse periodontal status compared to healthy subjects. Causal relationship between periodontal and hematological parameters was not proved.

Severe Generalized Periodontitis in a Patient with an Aplastic Anemia: a 5 Year Follow-up Case Report.

Aplastic anemia is a hematological disorder characterized by pancytopenia. This case report presents a young patient with untreated periodontitis associated with hematological disorders, and cyclosporine therapy. During 2 consecutive days, periodontal therapy which consisted of nonsurgical therapy supplemented with an antibiotic treatment and antifibrinolytic therapy was performed. Commercial microbiological PCR tests and periodontitis IL-1 polymorphism risk test were performed. Following the periodontal therapy, the inflammation was resolved and the patient's occlusion was restored by means of removable partial dentures. After the 5 year follow-up, the patient still remained with shallow probing depths although there was inadequate compliance during the maintenance phase. Aplastic anemia increases the risk of onset of severe forms of periodontitis that can be additionally complicated with cyclosporine therapy. In such patients, periodontal therapy must be supplemented with antibiotics.

Periodontal Disease and its Association with Angiographically Verified Coronary Artery Disease.

PURPOSE: The aim of this research was to investigate the association of chronic and aggressive periodontitis with the severity of coronary artery disease which was angiographically verified. MATERIAL AND METHODS: Subjects were selected among the hospitalized patients at the University Hospital Centre Zagreb who had coronary angiography done because of the chest pain. Thorough clinical examination included periodontal indices and clinical and socio-demographic characteristics of participants. Subjects were divided in two test groups, acute coronary syndrome (ACS) and stable coronary artery disease (CAD), and the control group with no significant CAD. Data were analyzed using Kruskal-Wallis and Pearson's Chi-Square test. RESULTS: From 106 subjects, 66 (62.3%) were hospitalized for ACS, 22 (20.7%) had stable CAD and only 18 (17.0%) had no significant CAD. Only 26 (24.5%) out of 106 patients were never smokers (p<0.05). Chronic periodontitis was the most common finding with 68.2% in ACS group and 54.5% in stable CAD group, while healthy patients without periodontitis (72.6%) were dominant in the control group (p<0.001). Stable CAD group had the highest mean probing depth (PD) 3.92±1.16, gingival recession (GR) 1.34±0.78, clinical attachment level (CAL) 4.60±1.41 and bleeding on probing (BOP) 45.98±26.19 values, whereas ACS group had mean PD value of 3.77±0.91, GR 1.11±0.66, CAL 4.32±1.08 and BOP 41.30±22.09, and no significant CAD group had mean PD value of 3.27±0.97, GR 0.69±0.37, CAL 3.62±1.04 and BOP 26.39±13.92 (p<0.05). CONCLUSION: Periodontitis was shown to be associated with angiographically verified coronary artery disease. Physical inactivity, poor oral hygiene and periodontal inflammation were observed in patients with ACS and stable CAD.

In vitro release of metal ions from a gold-platinum alloy in saliva-simulated conditions.

The aim of this study was to evaluate the behavior of high-noble gold-platinum alloy samples (18 + 8 Rafinerija plemenitih kovina, Zagreb, Hrvatska), in a phosphate buffer pH 6.0. The aim of the study was to determine the type and amount of released metal ions from the alloy, and to evaluate how the time of exposure to the phosphate buffer pH 6.0 influences the release of metal ions from the alloy. Manufacturer samples 8.0 x 6.5 x 1.0 mm in size, with a surface area of 133 mm2 were used. The release of metal ions from the tested alloy was measured in ten time periods (after 1, 2, 3, 4, 5, 6, 7, 14, 21 and 30 days). Six samples were used (n = 6) for each time period that the alloy was in the solution, which means that a total of 60 samples was analyzed. Solutions, in which the samples were placed, were analyzed with the inductively coupled plasma atomic emission spectrometer (ICP-AES, JY 50P, Jobin-Ywon, France). It was found that four metal ions were released; zinc ions (Zn), chromium ions (Cr), copper ions (Cu) and iron ions (Fe). The most released ions from the gold-platinum alloy were those of zinc (Zn), and the least released were those of iron (Fe) (p < 0.01). The results of the study show that the time the tested alloy spent in the phosphate buffer pH 6.0 solution has an effect on the release of ions (p < 0.01).