Formulation and Evaluation of Novel Thiolated Intra Pocket Periodontal Composite Membrane of Doxycycline.

Localized intra-pocket, retentive, biodegradable, prolonged release thiolated membrane can provide an improved therapeutic efficacy of doxycycline at the site of action with evading off target side effects. To this end, thiolated chitosan-hyaluronic acid composite polymeric complex next-generation of the periodontal membrane was manufactured by solvent casting method. FTIR spectroscopic analysis displayed successful immobilization of thiol groups on the manufactured thiolated periodontal membrane. Moreover, XRD, DSC, AFM and TGA of the membrane confirmed the compatibility of ingredients and modifications in surface chemistry. The thiolated periodontal film was also investigated in terms of thickness, weight uniformity, water-uptake capacity, drug content, pH, entrapment efficiency, lysozymal degradation and release patterns. Also, mucoadhesion profile was explored on gingival mucosa. The immobilized thiol groups on thiolated chitosan and thiolated hyaluronate were found to be 168 ± 11 µM/g (mean ± SD, n = 3) and 189 ± 8 µM/g (mean ± SD, n = 3) respectively. Swelling capacity of the thiolated periodontal membrane was significantly ∼2-fold higher (p < 0.05) as compared to unmodified membrane. The obtained thiolated membrane depicted 3 -old higher mucoadhesive features as compared to the un-modified membrane. In vitro release kinetics indicated approximately more than 80% prolonged release within 7 days. Mechanical strength of the Thiolated bandage was also significantly ∼2-fold higher (p < 0.05) as compared to unmodified membrane. Ex-vivo retention study revealed enhanced retention of thiolated membrane as compared to unmodified membrane. In-vitro antimicrobial studies demonstrated that thiolated membrane could efficiently kill Porphyromonas gingivalis cells as compared to the native membrane. Moreover, ex-vivo biodegradation results indicated that 90% of the thiolated membrane was biodegradable in 28 days. Based on these findings, thiolated next-generation of the periodontal membrane seems to be promising for periodontitis therapy.

Development and Characterization of Novel Medicated Nanofibers Against Periodontitis.

Periodontitis is an inflammatory disease of gums involving degeneration of periodontal ligaments, creation of periodontal pocket and resorption of alveolar bone, thus disrupting the support structure of teeth causing their loosening and finally removal. Since this disease is mainly confined to the periodontal pocket, so site specific drug delivery of an antibiotic is the best suitable option. This also eradicates the demerits of oral dosing like low drug concentration reaching the target site and the various systemic side effects. In the present work, an efficient and easy technique of electrospinning has been used to develop non-woven drug loaded and biodegradable nanofiber patch with inbuilt property of high surface area to volume ratio. Hyaluronic acid (HA) has been used specifically as the polymer since it possesses remarkable properties like providing an extracellular matrix supporting tissue regeneration, anti-inflammation and mucoadhesion. A blend of this natural polymer with another polymer (Polyvinyl alcohol) has been tried since HA alone cannot be electrospun efficiently as it shows very high viscosity at very low polymer concentration. The developed formulation presented controlled release behavior with good mucoadhesive strength. The in vivo studies confirmed the maintenance of minimum inhibitory concentration (MIC) over an extended period of time in addition to a significant antiinflammatory effect. All these observations suggested that the above formulation forms a stable intra periodontal pocket drug delivery system.

[Clinical and biochemical analysis of ligature-induced periodontitis in rats].

The most common experimental model of periodontitis is a "ligature" model. However due to the complexity connected with performing on rats, modification of existing model is proposed, which differs by fixture of cotton ligature around the central incisor and not around the second molar. The purpose of research - a comparative evaluation of "peroxide" and modified by us, "ligature" models of periodontitis in rats. 2 series of experiments on 36 white Wistar rats were conducted. The animals were divided into two groups: intact rats (control) and rats with a "peroxide" model of periodontitis, which was reproduced by the addition to the diet of rats overoxidized sunflower oil (5% by weight of the feed), daily, for 45 days. "Ligature" model in rats was reproduced by applying a cotton ligature on the central incisor of the upper jaw for 14 days. Elastase activity, malondialdehyde content and catalase activity in the gums and in the blood serum was measured by biochemical methods. The degree of atrophy of the alveolar bone of the mandible was determined by morphometric method. It is found that in both models of periodontitis in rats, changes in the periodontal tissues and in the organism as a whole, is common for periodontal disease in humans. Clinically apparent inflammation of the periodontal tissues is observed, metabolic disorders in the gums, change of biochemical parameters in serum and progressive decline in the alveolar bone are determined. A comparative analysis of the two models showed that the modified "ligature" model of periodontitis in rats has several advantages over the "peroxide" model: shorter term of modeling, more pronounced clinical inflammation of periodontal tissues and faster resorption of alveolar bone.

[Impact of volatile sulphur compounds in periodontal pockets on initial periodontal therapy].

OBJECTIVE: To investigate the relationship of volatile sulphur compounds((VSC)levels in periodontal pockets with severity of periodontitis, and the impact of VSC on the result of initial periodontal therapy. METHODS: Twenty-five patients with chronic periodontitis(CP)(13 males and 12 females with average age of 35) were included in this study. Clinical periodontal parameters, plaque index, probing depth(PD), attachment loss(AL), and bleeding on probing(BOP) were recorded before and 3 months after the initial therapy. VSC levels were measured with a portable monitor in a digital score ranging from 0.0 to 5.0. All of 5 054 sites for 840 teeth were included in this study. RESULT: Before treatment the percentage of VSC-positive sites was 17.1%, 52.3% and 86.0% for shallow (PD<3 mm), moderate(PD 4-6 mm) and deep (PD>7 mm) pocket, respectively (P<0.001). In most VSC-positive sites the VSC levels were<1.0. Percentage of sites with a high VSC levels was significantly different among three groups (P<0.01). All clinical parameters in VSC-negative sites were reduced significantly following the initial therapy. The reduction of PD and AL in VSC-positive sites by treatment was less marked than that in VSC-negative sites. CONCLUSION: VSC in periodontal pockets may be a potential indicator for detecting severity of CP and a useful predictor for therapeutic success.

Development and evaluation of novel biodegradable microspheres based on poly(d,l-lactide-co-glycolide) and poly(epsilon-caprolactone) for controlled delivery of doxycycline in the treatment of human periodontal pocket: in vitro and in vivo studies.

This study reports on the development of novel biodegradable microspheres prepared by water-in-oil-water (W/O/W) double emulsion technique using the blends of poly(d,l-lactide-co-glycolide) (PLGA) and poly(epsilon-caprolactone) (PCL) in different ratios for the controlled delivery of doxycycline (DXY). Doxycycline encapsulation of up to 24% was achieved within the polymeric microspheres. Blend placebo microspheres, drug-loaded microspheres and pristine DXY were analyzed by Fourier transform infrared spectroscopy (FT-IR), which indicated no interaction between drug and polymers. Differential scanning calorimetry (DSC) on drug-loaded microspheres confirmed the polymorphism of DXY and indicated a molecular level dispersion of DXY in the microspheres. Scanning electron microscopy (SEM) confirmed the spherical nature and smooth surfaces of the microspheres produced. Mean particle size of the microspheres as measured by dynamic laser light scattering method ranged between 90 and 200 mum. In vitro release studies performed in 7.4 pH media indicated the release of DXY from 7 to 11 days, depending upon the blend ratio of the matrix. Up to 11 days, DXY concentrations in the gingival crevicular fluid were higher than the minimum inhibitory concentration of DXY against most of the periodontal pathogens. One of the developed formulations was subjected to in vivo efficacy studies in thirty sites of human periodontal pockets. Significant results were obtained with respect to both microbiological and clinical parameters up to 3 months even as compared to commercial DXY gel. Statistical analyses of the release data and in vivo results were performed using the analysis of variance (ANOVA) method.

Characterization of the antioxidant profile of human saliva in peri-implant health and disease.

OBJECTIVES: Peri-implant disease is considered to be an inflammatory disease, but many aspects of its pathogenesis remain unknown. At present, peri-implant disease is considered to be initiated and perpetuated by a small group of predominantly Gram-negative, anaerobic, or micro-aerophilic bacteria that colonize the subgingival area. Bacteria cause the observed tissue destruction directly by toxic products and indirectly by activating host defence systems, i.e. inflammation. A variety of molecular species appears in the inflamed tissues, among them are reactive species such as free radicals and reactive oxygen species (ROS). The purpose of this study was to assess levels of various antioxidants in saliva to identify differences between the saliva of patients with healthy peri-implant tissues and patients with peri-implant disease, and to examine whether the whole saliva of those with peri-implant disease conditions might have lower levels of antioxidants than that of healthy individuals. MATERIALS AND METHODS: Thirty healthy adult volunteers (14 men and 16 women) with implant-supported overdentures (Ankylos Biofunctional Implants) were selected from the group of patients from Tallinn Dental Clinic. Biochemical and clinical parameters evaluated were the following ones: the levels of urate, ascorbate, myeloperoxidase in saliva, total antioxidant status of saliva, pocket probing depth (mm), gingival index (0, 1, 2, or 3), and bleeding on probing (0 or 1). RESULTS AND CONCLUSION: Total antioxidant status (TAS) of saliva and concentration of uric acid and ascorbate, which are the main salivary antioxidants, are significantly decreased in patients with peri-implant disease. TAS in healthy subjects was 0.41+/-0.10 for resting saliva and 0.31+/-0.09 for stimulated saliva; in diseased subjects TAS was 0.19+/-0.07 and 0.12+/-0.03, respectively. In healthy subjects, the concentration of urate was 307.2+/-78.06 microM/l in resting saliva and 241.5+/-89.09 microM/l in stimulated saliva. In diseased patients, the concentration of urate was 120+/-36.13 and 91.60+/-39.35 microM/l, respectively. The concentration of ascorbate did not differ in resting and stimulated saliva. In healthy subjects, it was 2.79+/-0.81 mg/l and in diseased subjects, it was 1.54+/-0.30 mg/l. This may indicate that excessive ROS production in peri-implant disease is leading to the situation of excessive oxidative stress, which may be an important factor contributing the destruction of peri-implant tissues. The importance of these findings may be the better understanding of the processes involved in the pathogenesis of peri-implant disease and that the treatment of peri-implant disease may involve adjuvant anti-oxidants supplementation together with cumulative interceptive supportive therapy concept introduced by Mombelli & Lang.

Formulation and preliminary in vivo dog studies of a novel drug delivery system for the treatment of periodontitis.

A novel drug delivery system for the treatment of periodontitis was developed using two components. The first was tetracycline base loaded into the microtubular excipient halloysite, which was coated with chitosan to further retard drug release. Encapsulation efficiencies of 32.5% were achieved with the loading procedure, with tetracycline base showing in vitro release for up to 50 days in simulated gingival crevicular fluid. The second component developed was a vehicle for the drug loaded coated halloysite, which was primarily based on the thermoresponsive polymer, poloxamer 407. A concentration of 20% was chosen with the thermoresponsivity of the system modified using PEG 20,000 so that the mobile product at room temperature would gel by temperature rise following syringing into a periodontal pocket. Retention of the overall system in the pocket was further improved by the addition of octyl cyanoacrylate (OCA). The thermoresponsivity of the poloxamer 407 system proved to be sensitive to the presence of added excipients with the levels of PEG 20,000 and OCA requiring modification in the presence of the halloysite component. A final formulation was developed which consisted of 200 mg of halloysite double loaded with tetracycline base and coated with chitosan, suspended in 1 ml of poloxamer 407 20% (w/w), PEG 20,000 0.5% (w/w), OCA 1.0% (w/w), water to 100%, adjusted to pH 4. The syringeability of this formulation at various temperatures was evaluated to ensure ease of delivery to the periodontal pocket. A stability study was performed to examine the change in thermoresponsivity over time, with the final formulation found to be stable for at least 9 months when stored at room temperature (approximately 20 degrees C). This formulation offered ease of delivery to the periodontal pocket and sustained release of the antibiotic for up to 6 weeks. The formulation had preliminary in vivo testing performed in dogs to determine levels of drug release, antimicrobial activity and retentive ability of the product. A wound pocket creation model was developed for the purposes of the trial. The product was easy to deliver to the pockets with application times of less than 1 min. Results showed the product was retained in the pocket for up to 6 weeks with effective tetracycline levels released locally over this time period, which achieved good antibacterial activity.

Elimination of Elyzol 25% Dentalgel matrix from periodontal pockets.

Elyzo 25% Dentalgel (EDG) which is developed for use in the treatment of periodontitis is a suspension of metronidazole benzoate (40%) in a mixture of glyceryl mono-oleate (GMO) and triglyceride (sesame oil). Metronidazole can be detected in the periodontal pockets 24-36 h after application. The aim of the present study was to estimate the period of time that the gel matrix persists on periodontal pockets after 1 application of EDG. 12 patients were included in the study. From each patient, 1 sample was taken before and immediately after, and 1, 2, 3, 4, 5, 6, 8, 12 and 24 h after application. Subgingival scaling followed by absorption of gingival crevicular fluid with filter paper was used for sampling. The sampling unit was 1 tooth. Each sample was assayed for the amount of GMO and oleic acid (a degradation product of GMO) by means of high-performance liquid chromatography (HPLC) with UV detection. To allow determination of the GMO dose applied into the pockets and to estimate the recovery rate of the sampling method, 1 tooth in each patient was selected for sampling as soon as the gel had set, i.e., about 10 min after application. Only in 1 patient was a detectable amount of GMO within the pocket revealed 24 h after application. This amount was approximately 0.5% of the mean GMO dose applied around 1 tooth. GMO was found no longer than 12 h in the remaining patients.

Calcium and phosphorus content of roots exposed to the oral environment.

This study analyzed the calcium and phosphorus content of extracted tooth roots exposed to the in vivo oral environment. 20 teeth were obtained from 16 patients and divided into two groups of 10 teeth each. In group 1, the teeth had gingival probing depths of 5 mm or more, and teeth of group 2 had gingival recessions of 3 mm or more. Prior to extraction, the gingival margin location was recorded by placing a groove on the tooth surface. After extraction, the teeth were sectioned coronal-apically, air dried and coated with carbon. Energy dispersive X-ray spectra, excited in a scanning electron microscope, were analyzed to measure relative calcium and phosphorus contents and for calculation of their ratios. X-rays were collected from two positions on the sectioned root. Experimental positions were selected within the exposed portion of the roots of groups 1 and 2, and unexposed positions were selected from that portion of the same root with attached periodontal membrane. At each position, calcium and phosphorus content was measured at 4 depths into the root surface: in cementum, in dentin three-quarters of the distance to the pulp chamber, and at 2 locations in between on either side of the cemento-dentinal junction. Analysis of data demonstrated large variations in calcium and phosphorus content from surface to surface of individual teeth and from tooth to tooth in a subject. No statistically significant differences were found between experimental and unexposed locations. Calcium and phosphorus contents were greater in roots exposed to pockets when compared to roots exposed by recession at both experimental and unexposed locations.(ABSTRACT TRUNCATED AT 250 WORDS)