Preparation of molecule-responsive microsized hydrogels via photopolymerization for smart microchannel microvalves.

Microdevices designed for practical environmental pollution monitoring need to detect specific pollutants such as dioxins. Bisphenol A (BPA) has been widely used as a monomer for the synthesis of polycarbonate and epoxy resins. However, the recent discovery of its high potential ability to disrupt human endocrine systems has made the development of smart systems and microdevices for its detection and removal necessary. Molecule-responsive microsized hydrogels with ß-cycrodextrin (ß-CD) as ligands are prepared by photopolymerization using a fluorescence microscope. The molecule-responsive micro-hydrogels show ultra-quick shrinkage in response to target BPA. Furthermore, the flow rate of a microchannel is autonomously regulated by the molecule-responsive shrinking of their hydrogels as smart microvalves.

Responsive behavior of tumor-marker-imprinted hydrogels using macromolecular cross-linkers.

Tumor-marker-imprinted hydrogels having lectin and antibody molecules as ligands for a tumor-specific marker glycoprotein were strategically prepared by biomolecular imprinting using minute amounts of low-molecular-weight or high-molecular-weight cross-linkers. The tumor-marker-imprinted hydrogels shrank gradually in response to a target glycoprotein, because their apparent cross-linking density increased owing to simultaneous complex formation of lectin and antibody ligands with a target glycoprotein after their ligands dynamically recognized the glycoprotein. The swelling ratio of the tumor-marker-imprinted hydrogel using high-molecular-weight cross-linker with an optimal chain length decreased more drastically than that using a low-molecular-weight cross-linker, but the hydrogel using the cross-linker with a chain that was too long did not exhibit tumor-marker responsive behavior. This paper focuses on the effect of the molecular weight of cross-linkers on the responsive behavior of tumor-marker-imprinted hydrogels having lectin and antibody molecules as ligands. The cross-linker chain length was an important factor in determining the dynamic glycoprotein recognition and responsive behavior of the biomolecule-imprinted hydrogels.

Design of molecularly imprinted hydrogels with thermoresponsive drug binding sites.

Drug delivery systems (DDS) regulate the spatiotemporal distribution of drugs in vivo to maximize efficacy and minimize side effects. Stimuli-responsive hydrogels, which exhibit a drastic change in volume in response to external stimuli such as temperature and pH, have attracted considerable interest as drug reservoirs for self-regulating DDS, as stimuli-responsive changes in the network size can regulate drug diffusion. However, such hydrogels have the disadvantage of leaking drugs even in the absence of stimulation. Proteins such as hemoglobin have dynamic molecular binding sites that modify their binding capacities by their conformational changes induced when an effector molecule binds to allosteric sites. Such dynamic binding sites are useful for loading drugs into reservoirs because their conformational changes can be used to control drug loading and release. In this study, we prepared thermoresponsive hydrogels with a controlled drug binding capacity to design drug reservoirs capable of both suppressing drug leakage below the transition temperature and accelerating drug release above it. Dynamic molecular binding sites were created by molecular imprinting that used 4,4'-diaminodiphenyl sulfone (dapsone) as the model drug, ß-cyclodextrin (CD) as the ligand, and N-isopropylacrylamide as the primary monomer. The molecularly imprinted (MIP) and nonimprinted (NIP) hydrogels with CD ligands, as well as the poly(N-isopropylacrylamide) (PNIPAAm) hydrogels without CD ligands, drastically shrunk above their transition temperature because of the PNIPAAm major chains changing conformation from a hydrophilic random coil to a hydrophobic globule as temperature increased. Because the MIP hydrogel has dynamic molecular binding sites, it absorbs a larger amount of dapsone than the NIP hydrogels in an aqueous solution below the transition temperature. The amount of dapsone adsorbed into the MIP hydrogel significantly decreased with increasing temperatures above 37 °C, despite the fact that the hydrophobic interaction between the polymer chains and dapsone became strong. The decrease in dapsone adsorption capability of the MIP hydrogel is due to a conformational change from a swollen to a shrunken state as temperature increases. The MIP hydrogel suppressed drug leakage below its transition temperature due to the high binding capacity of dynamic binding sites, but accelerated the drug release above its transition temperature due to the collapse of dynamic molecular binding sites, in contrast to the drug release behavior of general PNIPAAm-based hydrogels. Thus, the thermoresponsive MIP hydrogels with dynamic molecular binding sites regulated drug release in response to a change in temperature.

Amphiphilic Liquid Crystalline Polymer Micelles That Exhibit a Phase Transition at Body Temperature.

Liquid crystalline polymers (LCPs), which exhibit unique structures and properties intermediate between those of liquids and solids, are widely utilized as functional and advanced materials for fabricating optical devices and high-performance fibers. This utility stems from their ability to abruptly change their organized structures and mobilities at their liquid crystalline-isotropic phase transition temperatures, similar to the properties of biological membranes. Despite these numerous potential applications of LCPs, no study on their use in medical applications such as drug delivery has been reported. In the present study, we synthesized amphiphilic side-chain LCPs (LCP-g-OEGs, where OEG is oligo(ethylene glycol)) for medical applications, where the LCP-g-OEGs undergo a nematic-isotropic phase transition at body temperature. The LCP-g-OEGs formed micelles with a diameter of approximately 130 nm in aqueous media. The micelles were stable and did not dissociate in aqueous media even when the temperature exceeded the nematic-isotropic phase transition temperature (TNI). Although the release of a dye as a model drug from micelles was suppressed at temperatures lower than TNI, their dye release was drastically enhanced at temperatures higher than TNI. The LCP-g-OEG micelles regulated dye release reversibly in accordance with stepwise changes in temperature, without undergoing dissociation, differing from the behavior of standard temperature-responsive micelles. The temperature-responsive dye release behavior is induced by dramatic changes in their well-organized and dynamic structures as a result of the nematic-isotropic phase transition. These results demonstrate that the LCP-g-OEG micelles have a lot of medical applications as reversibly stimuli-responsive drug carriers.

Photoresponsive behaviour of zwitterionic polymer particles with photodimerizable groups on their surfaces.

Polymer particles with precise diameters have been used as building blocks for fabricating well-defined and nanostructured materials. Polymer particles as building blocks for medical applications require both easily spatiotemporal manipulation and good biocompatibility. In this study, we designed zwitterionic polymer particles with photodimerizable groups on their surfaces and used ultraviolet (UV) light irradiation to photo-assemble them in aqueous media. After synthesizing zwitterionic polymer particles with diameters ranging from 100-200 nm via soap-free emulsion polymerization, maleimide moieties as photodimerizable groups were introduced onto the particle surfaces. UV light irradiation to an aqueous dispersion of zwitterionic polymer particles with photodimerizable groups induced their photo-assembling because interparticle bonding forms by photodimerization of the photodimerizable groups on the particle surfaces. The zwitterionic surface of their particle-assembled films effectively suppressed protein adsorption, cell adhesion, and platelet adhesion. The photoresponsive behaviour and bioinert surface of the zwitterionic polymer particles with photodimerizable groups indicate that they have several potential applications as bioinert building blocks for designing well-defined and nanostructured biomaterials used in biosensors, bioseparation and cell culture, and for modifying and repairing biomaterial surfaces in situ.

The oral health of children in a rural area of the Lao People's Democratic Republic.

The lack of information on oral health in Laos makes it difficult to estimate the need and methods for preventing oral disease. This study identified problems concerning the oral health of Lao children. The study subjects were 59 school children who lived in Pakkading District. Dental caries, gingivitis malocclusions, temporomandibular joint (TMJ) disorders, dental plaque, and calculus were examined. We observed an average of 1.6 decayed, missing, and filled teeth (DMFT) and 4.1 decayed and filled deciduous teeth (dft) per child. 25.4% had gingivitis scores from 16 to 20 on the papillary, marginal, and attached (PMA) index; 29.6% had one or more occlusal abnormality; and 0% had signs of TMJ disorders. 93.5% of the children had at least one buccal or lingual tooth surface with plaque covering more than two thirds of the surface; 32.6% had dental calculus. Oral health promotion programs for children should prioritise prevention and treatment of caries. It is likely that the high rate of gingivitis in Lao children is due mainly to unsuccessful plaque control in daily life. In addition to descriptive epidemiological studies of dental diseases in other areas, the influence of sociological and behavioural factors on oral health should be analyzed epidemiologically to promote child health.

Basal jawed vertebrate phylogeny inferred from multiple nuclear DNA-coded genes.

BACKGROUND: Phylogenetic analyses of jawed vertebrates based on mitochondrial sequences often result in confusing inferences which are obviously inconsistent with generally accepted trees. In particular, in a hypothesis by Rasmussen and Arnason based on mitochondrial trees, cartilaginous fishes have a terminal position in a paraphyletic cluster of bony fishes. No previous analysis based on nuclear DNA-coded genes could significantly reject the mitochondrial trees of jawed vertebrates. RESULTS: We have cloned and sequenced seven nuclear DNA-coded genes from 13 vertebrate species. These sequences, together with sequences available from databases including 13 jawed vertebrates from eight major groups (cartilaginous fishes, bichir, chondrosteans, gar, bowfin, teleost fishes, lungfishes and tetrapods) and an outgroup (a cyclostome and a lancelet), have been subjected to phylogenetic analyses based on the maximum likelihood method. CONCLUSION: Cartilaginous fishes have been inferred to be basal to other jawed vertebrates, which is consistent with the generally accepted view. The minimum log-likelihood difference between the maximum likelihood tree and trees not supporting the basal position of cartilaginous fishes is 18.3 +/- 13.1. The hypothesis by Rasmussen and Arnason has been significantly rejected with the minimum log-likelihood difference of 123 +/- 23.3. Our tree has also shown that living holosteans, comprising bowfin and gar, form a monophyletic group which is the sister group to teleost fishes. This is consistent with a formerly prevalent view of vertebrate classification, although inconsistent with both of the current morphology-based and mitochondrial sequence-based trees. Furthermore, the bichir has been shown to be the basal ray-finned fish. Tetrapods and lungfish have formed a monophyletic cluster in the tree inferred from the concatenated alignment, being consistent with the currently prevalent view. It also remains possible that tetrapods are more closely related to ray-finned fishes than to lungfishes.

Pervaporative dehydration characteristics of an ethanol/water azeotrope through various chitosan membranes.

The permeation and separation characteristics of an ethanol/water azeotrope through chitosan membranes of different molecular weights and degrees of deacetylation during pervaporation were investigated. The normalized permeation rate decreased with increasing molecular weight up to 90 kDa, but at over 90 kDa, the rate increased. On the other hand, the water/ethanol selectivity increased with increasing molecular weight up to 90 kDa but decreased at over 90 kDa. With increasing degree of deacetylation, the water/ethanol permselectivity increased significantly, but the normalized permeation rate decreased. The characteristics of chitosan membranes are discussed based on their chemical and physical structures such as the contact angle, density, degree of swelling, and glass transition temperature.

Dehydration of an ethanol/water azeotrope through alginate-DNA membranes cross-linked with metal ions by pervaporation.

To obtain high dehydration membranes for an ethanol/water azeotrope, dried blend membranes prepared from mixtures of sodium alginate (Alg-Na) and sodium deoxyribonucleate (DNA-Na) were cross-linked by immersing in a methanol solution of CaCl2 or MaCl2. In the dehydration of an ethanol/water azeotropic mixture by pervaporation, the effects of immersion time in methanol solution of CaCl2 or MaCl2 on the permeation rate and water/ethanol selectivity through Alg-DNA/Ca(2+) and Alg-DNA/Mg(2+) cross-linked membranes were investigated. Alg-DNA/Mg(2+) cross-linked membrane immersed for 12h in methanol solution of MaCl2 exhibited the highest water/ethanol selectivity. This results from depressed swelling of the membranes by formation of a cross-linked structure. However, excess immersion in solution containing cross-linker led to an increase in the hydrophobicity of cross-linked membrane. Therefore, the water/ethanol selectivity of Alg-DNA/Mg(2+) cross-linked membranes with an excess immersion in cross-linking solution was lowered. The relationship between the structure of Alg-DNA/Ca(2+) and Alg-DNA/Mg(2+) cross-linked membranes and their permeation and separation characteristics during pervaporation of an ethanol/water azeotropic mixture is discussed in detail.

Biomolecule-sensitive hydrogels.

Stimuli-sensitive hydrogels have attracted considerable attention as intelligent materials in the biochemical and biomedical fields, since they can sense environmental changes and induce structural changes by themselves. In particular, biomolecule-sensitive hydrogels that undergo swelling changes in response to specific biomolecules have become increasingly important because of their potential applications in the development of biomaterials and drug delivery systems. This article provides an overview of the important and historical research regarding the synthesis and applications of glucose-sensitive hydrogels which exhibit swelling changes in response to glucose concentration. Enzymatically degradable hydrogels and antigen-sensitive hydrogels are also described in detail as protein-sensitive hydrogels that can respond to larger biomolecules. The synthetic strategies of other biomolecule-sensitive hydrogels are summarized on the basis of molecular imprinting and specific interaction. The biomolecule-sensitive hydrogels reviewed in this paper are expected to contribute significantly to the exploration and development of newer generations of intelligent biomaterials and self-regulated drug delivery systems.

A pilot study on antiplaque effects of mastic chewing gum in the oral cavity.

BACKGROUND: Chemical plaque control is a useful aid in mechanical oral hygiene, and various chemical agents have been evaluated as antiplaque agents. It has been shown that mastic chewing gum has antibacterial effects on Helicobacter pylori. In this study, the antiplaque effect of mastic chewing gum was investigated. METHODS: Twenty dental students who were both systemically and periodontally healthy participated in this study. The effects of mastic gum were assessed from 2 double-blinded, randomized studies. In the first trial, after mechanical toothbrushing, the inhibitory effect of mastic gum on bacteria in saliva following its use was compared to a placebo gum. Saliva samples were collected at the end of 1, 2, 3, and 4 hours; diluted; inoculated onto 10% horse blood chocolate agar plates; and cultured anaerobically at 37 degrees C for 48 hours. The total number of bacterial colonies on each plate was calculated (n = 20). In the second trial, the effects of mastic gum on de novo plaque formation on tooth surfaces and gingival inflammation were evaluated over a 7-day period without mechanical oral hygiene following random use of either mastic or placebo chewing gum. The degree of plaque accumulation and gingival inflammation were compared between the 2 groups (n = 10). RESULTS: The total number of bacterial colonies was significantly reduced during the 4 hours of chewing mastic gum compared to the placebo gum (P < 0.05, Student t test). The mastic group showed a significantly reduced plaque index (2.69 +/- 0.29 versus 3.15 +/- 0.24; P = 0.001, Student t test) and gingival index (0.44 +/- 0.15 versus 0.66 +/- 0.23, P = 0.021, Student t test) compared to the placebo group. CONCLUSION: These results suggest that mastic chewing gum is a useful antiplaque agent in reducing the bacterial growth in saliva and plaque formation on teeth.

The influence of controlled occlusal overload on peri-implant tissue. part 4: a histologic study in monkeys.

PURPOSE: The purpose of this study was to observe, after removing occlusal trauma and conducting plaque control, possible macroscopic and histologic changes in peri-implant tissue that had deteriorated resulting from experimental peri-implantitis, and to investigate the necessity for treatment procedures for peri-implantitis. MATERIALS AND METHODS: Four monkeys (Macaca fascicularis) in good general health were used in this experiment. Three months after the second premolar and the first molar were extracted from the right mandible, 2 IMZ experimental implants were placed in each monkey. After a 3-month osseointegration period, a second surgery was conducted, followed by making an impression for fabrication of the prosthesis. Excessive occlusal height of the prosthesis was adjusted to 250 microm, and the experiment was continued for 8 weeks after placement of the prosthesis. Three models were created: (1) A superstructure with an excessive occlusal height was used for 8 weeks without any brushing (positive control, model P); (2) after the first 4 weeks with a prosthesis with excessive occlusal height and no brushing, the superstructure was removed and not used for the last 4 weeks while brushing was conducted (experimental model, model E); and (3) for 8 weeks, a prosthesis with an appropriate occlusal height was used with brushing (negative control, model N). RESULTS: When these 3 models were compared with each other, macroscopic findings indicated inflammation only in model P. Mobility of implants was not seen in any model. Histopathologic observations revealed a slight difference between model E and model P in terms of the degree of inflammatory cell infiltration in the connective tissue. DISCUSSION: No difference was found in the degree of bone resorption. Partial tearing was observed at the contact region between epithelial tissue and implant surfaces. CONCLUSIONS: (1) The contact between implants and epithelial or connective tissue is fragile; (2) inflammation and occlusion must be controlled more prudently than in the case of natural teeth; and (3) once peri-implantitis has progressed, the control of occlusion and inflammation is probably not sufficient to promote the healing mechanism.

Synthesis of glucose-responsive bioconjugated gel particles using surfactant-free emulsion polymerization.

Bioconjugated gel particles that have complexes composed of lectin concanavalin A (ConA) and 2-glucosyloxyethyl methacrylate (GEMA) were synthesized by the surfactant-free emulsion copolymerization of N,N-diethylaminoethyl methacrylate (DEAEMA), poly(ethylene glycol) dimethacrylate (PEGDMA), GEMA, and modified-ConA with polymerizable groups. The resultant gel particles having GEMA-ConA complexes (GEMA-ConA gel particles) were colloidally stable in a phosphate buffer solution and had a diameter of approximately 750nm. Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) measurements implied that GEMA-ConA gel particles have core-shell structures consisting of a hydrophobic core of DEAEMA and a hydrophilic shell of GEMA and PEGDMA containing ConA. GEMA-ConA gel particles underwent a change in size in response to glucose in a phosphate buffer solution. The swelling ratio of GEMA-ConA gel particles gradually increased with an increase in the glucose concentration. On the other hand, the swelling ratio of GEMA-ConA gel particles remained unchanged in a phosphate buffer solution containing galactose. The glucose-responsive swelling of GEMA-ConA gel particles was induced by the dissociation of GEMA-ConA complexes acting as reversible cross-links, because free glucose behaved as an inhibitor of GEMA-ConA complexes. These results indicate that GEMA-ConA gel particles can recognize glucose selectively and undergo changes in size in response to the glucose concentration. The smart functions of glucose-responsive gel particles can provide tools for constructing self-regulated drug delivery systems and sensor systems useful for treating diabetes.

Dehydration from alcohols by polyion complex cross-linked chitosan composite membranes during evapomeation.

This study describes the dehydration of an ethanol/water azeotrope during evapomeation using polyion complex cross-linked chitosan composite (q-Chito-PEO acid polyion complex/PES composite) membranes, constructed from quaternized chitosan (q-Chito) and poly(ethylene oxydiglycolic acid) (PEO acid) on a porous poly(ether sulfone) (PES) support. Both the q-Chito/PES composite and the q-Chito-PEO acid polyion complex/PES composite membranes showed high water permselectivity for an ethanol/water azeotrope. Both the permeation rate and the water permselectivity of the q-Chito/PES composite membranes were enhanced by increasing the degree of quaternization of the chitosan molecule because the affinity of the q-Chito/PES composite membranes for water was increased by introducing a quaternized ammonium group into the chitosan molecule. q-Chito-PEO acid polyion complex/PES composite membranes prepared from an equimolar ratio of carboxylate groups in the PEO acid versus quaternized ammonium groups in the q-Chito showed the maximum separation factor for water permselectivity without lowering the permeation rate. With an increasing molecular weight of PEO acid, the separation factor for water permselectivity increased, but the permeation rate almost did not change. The mechanism responsible for the separation of an ethanol/water azeotrope through the q-Chito-PEO acid polyion complex/PES composite membranes was analyzed by the solution-diffusion model. The permeation rate, separation factor for water permselectivity, and evapomeation index of q-Chito-PEO acid 400 polyion complex/PES composite membrane with an equimolar ratio of carboxylate groups in PEO acid 400 and ammonium groups in q-Chito were 3.5 x 10(-1) kg/(m(2) hr), 6300, and 2205, respectively, and very high membrane performance. The separation factor for water permselectivity for aqueous solutions of n-propyl and isopropyl alcohol was also maximized at an equimolar ratio of carboxylate groups and ammonium groups and was greater than that for an ethanol/water azeotrope. The above results were discussed from the viewpoint of the physical and chemical structure of the q-Chito-PEO acid polyion complex/PES composite membranes and the permeants.

Performance of butyrylcellulose membranes for benzene/cyclohexane mixtures containing a low benzene concentration by pervaporation.

Butyrylcellulose (BuCell) with different degrees of butyrylation was synthesized as a membrane material for the separation of benzene/cyclohexane (Bz/Chx) mixtures. A BuCell membrane with a degree of butyrylation of 2.3 showed high benzene/cyclohexane selectivity for Bz/Chx mixtures by pervaporation. Both the permeation rate and the benzene/cyclohexane selectivity of the BuCell membrane increased with increasing benzene concentration in the feed mixture. The increase in the permeation rate resulted from an increase in the swelling of the membrane, and the increase in the benzene/cyclohexane selectivity can be attributed to an increase in the diffusion selectivity. With increasing degree of butyrylation of BuCell, the permeation rate increased; on the other hand, the benzene/cyclohexane selectivity decreased slightly. This result can qualitatively be explained by the degree of swelling, the density, and the contact angle of the BuCell membranes. The permeation and separation mechanism of Bz/Chx mixtures through BuCell membranes by pervaporation is discussed on the basis of the solution-diffusion model, which is typically applied for permeation through dense, nonporous membranes.

Preparation of reversibly glucose-responsive hydrogels by covalent immobilization of lectin in polymer networks having pendant glucose.

Glucose-responsive hydrogels were prepared by copolymerization of a monomer having a pendant glucose with modified lectin (concanavalin A (ConA)) having vinyl groups. Swelling behavior of ConA-copolymerized glucosyloxyethyl methacrylate (GEMA) hydrogels was discussed from the viewpoint of their synthetic condition and structures. The swelling ratio of the ConA-copolymerized GEMA hydrogels was strongly dependent on the glucose concentration in a buffer solution. As free glucose resulted in the dissociation of the complex between ConA and pendant glucose in the networks and the cross-linking density in the hydrogel decreased, the hydrogels swelled gradually in the presence of free glucose. Leak of ConA from the ConA-entrapment hydrogel and ConA-copolymerized hydrogel was examined in order to discuss the reversible changes of their glucose-responsive behavior. During swelling in the presence of free glucose, ConA leaked out of the ConA-entrapment GEMA hydrogel but did not out of the ConA-copolymerized GEMA hydrogel. As a result, the ConA-copolymerized GEMA hydrogel showed the reversible swelling changes in response to a stepwise change in the glucose concentration. This study revealed that covalent immobilization of ConA in the GEMA networks is very important for development of reversibly glucose-responsive hydrogels.

Dehydration of an ethanol/water azeotrope by novel organic-inorganic hybrid membranes based on quaternized chitosan and tetraethoxysilane.

To control swelling of quaternized chitosan (q-Chito) membranes, mixtures of q-Chito as an organic component and tetraethoxysilane (TEOS) as an inorganic component were prepared using the sol-gel reaction, and novel q-Chito/TEOS hybrid membranes were formed. In the separation of an ethanol/water azeotrope by pervaporation, the effect of TEOS content on the water/ethanol selectivity of q-Chito/TEOS hybrid membranes was investigated. Hybrid membranes containing up to 45 mol % TEOS exhibited higher water/ethanol selectivity than the q-Chito membrane. This resulted from depressed swelling of the membranes by formation of a cross-linked structure. However, introduction of excess TEOS led to greater swelling of the hybrid membranes. Therefore, the water/ethanol selectivity of the hybrid membranes containing more than 45 mol % TEOS was lower than that of the q-Chito membrane. The relationship between the structure of q-Chito/TEOS hybrid membranes and their permeation and separation characteristics during pervaporation of an ethanol/water azeotrope is discussed in detail.

Osteocalcin, deoxypyridinoline and interleukin-1beta in peri-implant crevicular fluid of patients with peri-implantitis.

The aim of the present study was to analyze the levels of osteocalcin, deoxypyridinoline (Dpd) and interleukin-1beta as markers of bone metabolism in peri-implant crevicular fluid (PICF) from peri-implantitis patients. PICF was sampled from a total of 34 endosseous titanium implants from 16 patients; nine females (mean age 52.8, range 40-62 years) and seven males (mean age 56.0, range 36-66 years). The implants had been in place for a period of 9-112 months (mean; 35.8 months) since the loading. These sites were categorized as six peri-implantitis, eight peri-implant mucositis and 20 healthy implant. PICF volume from peri-implantitis sites was significantly higher than mucositis and healthy implant sites (P < 0.01). Osteocalcin levels in PICF from mucositis sites were significantly higher than healthy implants (P < 0.05), whereas peri-implantitis sites were not significantly different from either mucositis or healthy implant sites. Dpd could not be detected in any of the samples examined. IL-1beta levels in PICF from peri-implantitis sites were significantly higher than levels from peri-implant mucositis (P < 0.05) and healthy implant sites (P < 0.01). In conclusion, osteocalcin in PICF may reflect increased local bone turnover around implants. Further, IL-1beta should be a useful marker for peri-implant inflammation.