Bacteria-responsive programmed self-activating antibacterial hydrogel to remodel regeneration microenvironment for infected wound healing.

There is still an urgent need to develop hydrogels with intelligent antibacterial ability to achieve on-demand treatment of infected wounds and accelerate wound healing by improving the regeneration microenvironment. We proposed a strategy of hydrogel wound dressing with bacteria-responsive self-activating antibacterial property and multiple nanozyme activities to remodel the regeneration microenvironment in order to significantly promote infected wound healing. Specifically, pH-responsive H2O2 self-supplying composite nanozyme (MSCO) and pH/enzyme-sensitive bacteria-responsive triblock micelles encapsulated with lactate oxidase (PPEL) were prepared and encapsulated in hydrogels composed of L-arginine-modified chitosan (CA) and phenylboronic acid-modified oxidized dextran (ODP) to form a cascade bacteria-responsive self-activating antibacterial composite hydrogel platform. The hydrogels respond to multifactorial changes of the bacterial metabolic microenvironment to achieve on-demand antibacterial and biofilm eradication through transformation of bacterial metabolites, and chemodynamic therapy enhanced by nanozyme activity in conjunction with self-driven nitric oxide (NO) release. The composite hydrogel showed 'self-diagnostic' treatment for changes in the wound microenvironment. Through self-activating antibacterial therapy in the infection stage to self-adaptive oxidative stress relief and angiogenesis in the post-infection stage, it promotes wound closure, accelerates wound collagen deposition and angiogenesis, and completely improves the microenvironment of infected wound regeneration, which provides a new method for the design of intelligent wound dressings.

Membrane-anchoring selenophene viologens for antibacterial photodynamic therapy against periodontitis via restoring subgingival flora and alleviating inflammation.

Antibacterial photodynamic therapy (aPDT) has emerged as a promising strategy for treating periodontitis. However, the weak binding of most photosensitizers to bacteria and the hypoxic environment of periodontal pockets severely hamper the therapeutic efficacy. Herein, two novel oxygen-independent photosensitizers are developed by introducing selenophene into viologens and modifying with hexane chains (HASeV) or quaternary ammonium chains (QASeV), which improve the adsorption to bacteria through anchoring to the negatively charged cell membrane. Notably, QASeV binds only to the bacterial surface of Porphyromonas gingivalis and Fusobacterium nucleatum due to electrostatic binding, but HASeV can insert into their membrane by strong hydrophobic interactions. Therefore, HASeV exhibits superior antimicrobial activity and more pronounced plaque biofilm disruption than QASeV when combined with light irradiation (MVL-210 photoreactor, 350-600 nm, 50 mW/cm2), and a better effect on reducing the diversity and restoring the structure of subgingival flora in periodontitis rat model was found through 16S rRNA gene sequencing analysis. The histological and Micro-CT analyses reveal that HASeV-based aPDT has a better therapeutic effect in reducing periodontal tissue inflammation and alveolar bone resorption. This work provides a new strategy for the development of viologen-based photosensitizers, which may be a favorable candidate for the aPDT against periodontitis.

Journey of Mineral Precursors in Bone Mineralization: Evolution and Inspiration for Biomimetic Design.

Bone mineralization is a ubiquitous process among vertebrates that involves a dynamic physical/chemical interplay between the organic and inorganic components of bone tissues. It is now well documented that carbonated apatite, an inorganic component of bone, is proceeded through transient amorphous mineral precursors that transforms into the crystalline mineral phase. Here, the evolution on mineral precursors from their sources to the terminus in the bone mineralization process is reviewed. How organisms tightly control each step of mineralization to drive the formation, stabilization, and phase transformation of amorphous mineral precursors in the right place, at the right time, and rate are highlighted. The paradigm shifts in biomineralization and biomaterial design strategies are intertwined, which promotes breakthroughs in biomineralization-inspired material. The design principles and implementation methods of mineral precursor-based biomaterials in bone graft materials such as implant coatings, bone cements, hydrogels, and nanoparticles are detailed in the present manuscript. The biologically controlled mineralization mechanisms will hold promise for overcoming the barriers to the application of biomineralization-inspired biomaterials.

A telluroviologen-anchored tetraphenylporphyrin as sonosensitizer for periodontitis sonodynamic therapy.

Periodontitis is a chronic disease caused by bacteria (e.g. Porphyromonas gingivalis, P.gingivalis) that currently lacks effective non-invasive treatment options. Sonodynamic therapy (SDT) is an emerging non-invasive antimicrobial therapeutic strategy. Since ultrasonic tooth cleaning is widely used in dental treatments, SDT has significant potential for the facile implementation of treat periodontitis. However, hypoxia in periodontitis severely limits the effectiveness of traditional sonosensitizers. To address this issue, we have developed a new sonosensitizer termed as TPP-TeV, which combines the traditional sonosensitizer tetraphenylporphyrin (TPP) with a new photosensitizer telluroviologen (TeV). Under ultrasound radiation, TPP-TeV can produce numerous cationic free radicals (TPP-TeV•), which subsequently generate ROS free radicals (O2•-, •OH) efficiently via electron transfer mechanism, resulting in the effective killing of anaerobic P.gingivalis both in vivo and in vitro. As a result, the dental environment is improved, and the inhibition rate of alveolar bone loss reaches 80 %. The introduction of tellurium into the viologen molecule induces changes in its reduction potential, resulting in increased rigidity of the molecule. This modification systematically reduces the biotoxicity of our novel sonosensitizer by 75 % at 50 µM based on bacterial experiments. These promising findings could potentially establish new options for sonodynamic therapy (SDT) in periodontitis clinical treatments.

Nanoscaled Titanium Oxide Layer Provokes Quick Osseointegration on 3D-Printed Dental Implants: A Domino Effect Induced by Hydrophilic Surface.

Three-dimensional printing is a revolutionary strategy to fabricate dental implants. Especially, 3D-printed dental implants modified with nanoscaled titanium oxide layer (H-SLM) have impressively shown quick osseointegration, but the accurate mechanism remains elusive. Herein, we unmask a domino effect that the hydrophilic surface of the H-SLM facilitates blood wetting, enhances the blood shear rate, promotes blood clotting, and changes clot features for quick osseointegration. Combining computational fluid dynamic simulation and biological verification, we find a blood shear rate during blood wetting of the hydrophilic H-SLM 1.2-fold higher than that of the raw 3D-printed implant, which activates blood clot formation. Blood clots formed on the hydrophilic H-SLM demonstrate anti-inflammatory and pro-osteogenesis effects, leading to a 1.5-fold higher bone-to-implant contact and a 1.8-fold higher mechanical anchorage at the early stage of osseointegration. This mechanism deepens current knowledge between osseointegration speed and implant surface characteristics, which is instructive in surface nanoscaled modification of multiple 3D-printed intrabony implants.

Mesenchymal stem cell-derived extracellular vesicles for treatment of bone loss within periodontitis in pre-clinical animal models: a meta-analysis.

BACKGROUND: Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) represent an effective and promising strategy for periodontitis, although studies remain pre-clinical. Herein, a meta-analysis was conducted to assess the efficacy of MSC-EVs in animal models of periodontitis. METHODS: The PubMed, Web of Science, and Embase electronic databases were searched up to Dec 2022 to retrieve preclinical studies examining the use of MSC-EVs for periodontitis treatment. Meta-analyses and sub-group analyses were performed to assess the effect of MSC-EVs on Bone Volume/Total Volume (BV/TV) or the distance between the cementoenamel junction and alveolar bone crest (CEJ-ABC) in pre-clinical animal models of periodontitis. RESULTS: 11 studies published from Mar 2019 to Oct 2022 met the inclusion criteria. Overall, MSC-EVs contributed to periodontal bone regeneration in the inflammatory bone loss area due to periodontitis, as represented by a weighted mean difference (WMD) of 14.07% (95% CI = 6.73, 21.41%, p < 0.001) for BV/TV and a WMD of -0.12 mm (95% CI= -0.14, -0.11 mm, p < 0.001) for CEJ-ABC. However, sub-analysis suggested that there was no significant difference in CEJ-ABC between studies with bioactive scaffolds and studies without bioactive scaffolds (p = 0.60). CONCLUSIONS: The present study suggests that MSC-EVs may represent an attractive therapy for the treatment of inflammatory bone loss within periodontitis.

Adhesive and biodegradable polymer mixture composed of high -biosafety pharmaceutical excipients as non-setting periodontal dressing.

Periodontal dressing is a surgical dressing applied to oral wounds after periodontal surgery. Currently, all commercially available setting periodontal dressings are stiff, uncomfortable, with poor aesthetics, and need to be removed at the patient's follow-up visit, which may cause secondary damage. A periodontal dressing with soft texture, biodegradable properties, and that could balance both comfort and aesthetics is urgently desired. Hence, non-setting and degradable dressings were developed using sodium carboxymethyl cellulose, Eudragit S 100 and povidone K30, which were compared with the commercial degradable dressing Reso-pac®. The mucosal adhesion of the dressings was evaluated by lap shear tests, which indicated adequate adhesion. The in vitro swelling rates of the dressings were approximately half that of Reso-pac®, which led to less saliva adsorption and better dimensional stability. The dressings also exhibited satisfactory biocompatibility according to the results of CCK-8, Live/Dead staining, hemolysis, and subcutaneous implantation assays. Moreover, the dressing promoted the healing of full-thickness mucosal wounds in the palatal gingivae of SD rats and contributed to better therapeutic effect than Reso-pac®. Considering the multiple advantages and the pure pharmaceutical excipient formula, we anticipate that this dressing could be a promising product and may enter clinical practice in the near future.

Evaluation of dentinal tubule occlusion and pulp tissue response after using 980-nm diode laser for dentin hypersensitivity treatment.

OBJECTIVES: To evaluate the effectiveness of the 980-nm diode laser for dentinal tubule occlusion, measure the intrapulpal temperature, and investigate the dental pulp response. MATERIALS AND METHODS: The dentinal samples were randomly divided into G1-G7 groups: control; 980-nm laser irradiation (0.5 W, 10 s; 0.5 W, 10 s × 2; 0.8 W, 10 s; 0.8 W, 10 s × 2; 1.0 W, 10 s; 1.0 W, 10 s × 2). The dentin discs were applied for laser irradiation and analyzed by scanning electron microscopy (SEM). The intrapulpal temperature was measured on the 1.0-mm and 2.0-mm thickness samples, and then divided into G2-G7 groups according to laser irradiation. Moreover, forty Sprague Dawley rats were randomly divided into the laser-irradiated group (euthanized at 1, 7, and 14 days after irradiation) and the control group (non-irradiated). qRT-PCR, histomorphology, and immunohistochemistry analysis were employed to evaluate the response of dental pulp. RESULTS: SEM indicated the occluding ratio of dentinal tubules in the G5 (0.8 W, 10 s × 2) and G7 (1.0 W, 10 s × 2) were significantly higher than the other groups (p < 0.05). The maximum intrapulpal temperature rises in the G5 were lower than the standard line (5.5 ℃). qRT-PCR showed that the mRNA expression level of TNF-α and HSP-70 upregulated significantly at 1 day (p < 0.05). Histomorphology and immunohistochemistry analysis showed that, compared with the control group, the inflammatory reaction was slightly higher at the 1 and 7 days (p < 0.05) and decreased to the normal levels at 14 days (p > 0.05). CONCLUSIONS: A 980-nm laser at a power of 0.8 W with 10 s × 2 defines the best treatment for dentin hypersensitivity in terms of compromise between the efficacy of the treatment and the safety of the pulp. CLINICAL RELEVANCE: The 980-nm laser is an effective option for treating dentin sensitivity. However, we need to ensure the safety of the pulp during laser irradiation.

Mitochondria transfer reverses the inhibitory effects of low stiffness on osteogenic differentiation of human mesenchymal stem cells.

Microenvironment biophysical factors such as matrix stiffness can noticeably affect the differentiation of mesenchymal stem cells (MSCs). In this mechanobiology transduction process, mitochondria are shown to be an active participant. The present study aims to systematically elucidate the phenotypic and functional changes of mitochondria during the stiffness-mediated osteogenic differentiation. Additionally, the effect of mitochondria transfer on the osteogenesis of impaired MSCs caused by stiffness was investigated. Human periodontal ligament stem cells (PDLSCs) were used as model cells in the current study. Low stiffness restrained the cell spreading and significantly inhibited the proliferation and osteogenic differentiation of PDLSCs. Mitochondria of PDLSCs cultured on low stiffness exhibited shorter length, rounded shape, fusion/fission imbalance, ROS and mitophagy level increase, and ATP production reduction. The inhibited mitochondria function and osteogenic differentiation capacity were recovered to near-normal levels after transferring the mitochondria of PDLSCs cultured on the high stiffness. This study indicated that low matrix stiffness altered the mitochondrial morphology and induced systematical mitochondrial dysfunction during the osteogenic differentiation of MSCs. Mitochondria transfer was proved to be a feasible technique for maintaining MSCs function in vitro by reversing the osteogenesis ability.

BMSC-Derived Exosomal CircHIPK3 Promotes Osteogenic Differentiation of MC3T3-E1 Cells via Mitophagy.

Exosome-based therapy is emerging as a promising strategy to promote bone regeneration due to exosomal bioactive cargos, among which circular RNA (circRNA) has recently been recognized as the key effector. The role of exosomal circRNA derived from bone marrow mesenchymal stem cells (BMSCs) has not been well-defined. The present study aimed to clarify the regulatory function and molecular mechanism of BMSC-derived exosomal circRNA in osteogenesis. Exosomes derived from bone marrow mesenchymal stem cells (BMSC-Exos) were isolated and identified. BMSC-Exos' pro-osteogenic effect on MC3T3-E1 cells was validated by alkaline phosphatase (ALP) activity and Alizarin Red staining. Through bioinformatic analysis and molecular experiments, circHIPK3 was selected and verified as the key circRNA of BMSC-Exos to promote osteoblast differentiation of MC3T3-E1 cells. Mechanistically, circHIPK3 acted as an miR-29a-5p sponge and functioned in mitophagy via targeting miR-29a-5p and PINK1. Additionally, we showed that the mitophagy level of MC3T3-E1 cells were mediated by BMSC-Exos, which promoted the osteogenic differentiation. Collectively, our results revealed an important role for BMSC-derived exosomal circHIPK3 in osteogenesis. These findings provide a potentially effective therapeutic strategy for bone regeneration.

Influence of calcium-based desensitizing toothpastes on the bonding performance of universal adhesive.

The study evaluated the effect of calcium-based desensitizing toothpastes on the dentinal tubule occlusion and its influence on the dentin bond strength of universal adhesive. Mid-coronal dentin samples were prepared for hypersensitivity model and treated by the following calcium-based desensitizing toothpastes: no treatment (Control), Clinpro (fTCP), Pro-Relief (Pro-Argin), and Repair & Protect (Novamin). Single Bond Universal adhesive was applied in self-etch or etch-and-rinse mode. The dentinal tubule occlusion and adhesion interface were evaluated under scanning electron microscope (SEM). A double-fluorescence technique was used to examine interfacial permeability under confocal laser scanning microscopy (CLSM). The micro-tensile bond strength (µTBS) was employed, followed by the fracture interface observation. SEM showed the toothpastes occluded dentinal tubules, and the occlusion exhibited stability against acid and abrasion. Hindered resin infiltration was observed in the adhesion interface after desensitization. CLSM showed more water permeation within or under the adhesion interface in etch-and-rinse mode than self-etch mode. Desensitization decreased the µTBS in self-etch mode. When using etch-and-rinse mode, the desensitized samples presented similar µTBS to the control group. No difference in µTBS was found between the two bonding modes, except for the control group. Calcium-based desensitizing toothpastes can effectively occlude the exposed dentinal tubules with acid-resistant and abrasion-resistant stability. The desensitization reduced the dentin bond strength of the universal adhesive system in self-etch mode but did not affect the bond strength of etch-and-rinse mode.

COPI Vesicle Disruption Inhibits Mineralization via mTORC1-Mediated Autophagy.

Bone mineralization is a sophisticated regulated process composed of crystalline calcium phosphate and collagen fibril. Autophagy, an evolutionarily conserved degradation system, whereby double-membrane vesicles deliver intracellular macromolecules and organelles to lysosomes for degradation, has recently been shown to play an essential role in mineralization. However, the formation of autophagosomes in mineralization remains to be determined. Here, we show that Coat Protein Complex I (COPI), responsible for Golgi-to-ER transport, plays a pivotal role in autophagosome formation in mineralization. COPI vesicles were increased after osteoinduction, and COPI vesicle disruption impaired osteogenesis. Mechanistically, COPI regulates autophagy activity via the mTOR complex 1 (mTORC1) pathway, a key regulator of autophagy. Inhibition of mTOR1 rescues the impaired osteogenesis by activating autophagy. Collectively, our study highlights the functional importance of COPI in mineralization and identifies COPI as a potential therapeutic target for treating bone-related diseases.

Engineering exosomes for bone defect repair.

Currently, bone defect repair is still an intractable clinical problem. Numerous treatments have been performed, but their clinical results are unsatisfactory. As a key element of cell-free therapy, exosome is becoming a promising tool of bone regeneration in recent decades, because of its promoting osteogenesis and osteogenic differentiation function in vivo and in vitro. However, low yield, weak activity, inefficient targeting ability, and unpredictable side effects of natural exosomes have limited the clinical application. To overcome the weakness, various approaches have been applied to produce engineering exosomes by regulating their production and function at present. In this review, we will focus on the engineering exosomes for bone defect repair. By summarizing the exosomal cargos affecting osteogenesis, the strategies of engineering exosomes and properties of exosome-integrated biomaterials, this work will provide novel insights into exploring advanced engineering exosome-based cell-free therapy for bone defect repair.

Bonding properties of mild universal adhesives to dentin pretreated with hydroxyapatite-based desensitizing agents.

OBJECTIVES: The purpose of the study was to evaluate the effect of hydroxyapatite (HA)-based desensiti-zing agents and determine their influence on the bonding performance of mild universal adhesives. METHODS: Mid-coronal dentin samples were sectioned from human third molars and prepared for a dentin-sensitive model. According to desensitizing applications, they were randomly divided into four groups for the following treatments: no desensitizing treatment (control), Biorepair toothpaste (HA-based desensitizing toothpaste) treatment, Dontodent toothpaste (HA-based desensitizing toothpaste) treatment, and HA paste treatment. Dentin tubular occlusion and occluded area ratios were evaluated by scanning electron microscopy (SEM). Furthermore, All-Bond Universal, Single Bond Universal, and Clearfil Universal Bond were applied to the desensitized dentin in self-etch mode. The wettability and surface free energy (SFE) of desensitized dentin were evaluated by contact angle measurements. Bonded specimens were sectioned into beams and tested for micro-tensile bond strength to analyze the effect of desensitizing treatment on the bond strength to dentin of universal adhesives. RESULTS: SEM revealed that the dentin tubule was occluded by HA-based desensitizing agents, and the area ratios for the occluded dentin tubules were in the following order: HA group>Biorepair group>Dontodent group (P<0.05). Contact angle analysis demonstrated that HA-based desensitizing agents had no statistically significant influence on the wettability of the universal adhesives (P>0.05). The SFE of dentin significantly increased after treatment by HA-based desensitizing agents (P<0.05). The micro-tensile bond strength test showed that HA-based desensitizing toothpastes always decreased the µTBS values (P<0.05), whereas the HA paste group presented similar bond strength to the control group (P>0.05), irrespective of universal adhesive types. CONCLUSIONS: HA-based desensitizing agents can occlude the exposed dentinal tubules on sensitive dentin. When mild and ultra-mild universal adhesives were used for subsequent resin restoration, the bond strength was reduced by HA-based desensitizing toothpastes, whereas the pure HA paste had no adverse effect on bond strength.

Dental stem cell-derived extracellular vesicles transfer miR-330-5p to treat traumatic brain injury by regulating microglia polarization.

Traumatic brain injury (TBI) contributes to the key causative elements of neurological deficits. However, no effective therapeutics have been developed yet. In our previous work, extracellular vesicles (EVs) secreted by stem cells from human exfoliated deciduous teeth (SHED) offered new insights as potential strategies for functional recovery of TBI. The current study aims to elucidate the mechanism of action, providing novel therapeutic targets for future clinical interventions. With the miRNA array performed and Real-time PCR validated, we revealed the crucial function of miR-330-5p transferred by SHED-derived EVs (SHED-EVs) in regulating microglia, the critical immune modulator in central nervous system. MiR-330-5p targeted Ehmt2 and mediated the transcription of CXCL14 to promote M2 microglia polarization and inhibit M1 polarization. Identified in our in vivo data, SHED-EVs and their effector miR-330-5p alleviated the secretion of inflammatory cytokines and resumed the motor functional recovery of TBI rats. In summary, by transferring miR-330-5p, SHED-EVs favored anti-inflammatory microglia polarization through Ehmt2 mediated CXCL14 transcription in treating traumatic brain injury.

Polyurethane-Based Stretchable Semiconductor Nanofilms with High Intrinsic Recovery Similar to Conventional Elastomers.

Polymer semiconductors with large elastic recovery (ER) under high strain in thin film state are highly desirable for stretchable electronics. Here we report a type of stretchable semiconductor PU(DPP)x, by copolymerization of oligodiketopyrrolopyrrole-based conjugated block and hydrogenated polybutadiene flexible block via urethane linkage for intermolecular hydrogen bonding. By regulating block ratio, PU(DPP)35 with 35 wt % conjugated block exhibits high intrinsic ER > 80% under 175% strain (ε) in pseudo free-standing thin film state, comparable with commercial elastomers, and crack onset strain (COS) > 300% along with maximum hole mobility of 0.19 cm2 V-1 s-1 in organic thin film transistors to bring it to the best performing block copolymer-type stretchable semiconductors. Enhanced mobility is achieved using PU(DPP)35 as the binder for conjugated polymer PDPPT3. The 25 wt %-PDPPT3 blend displays mobility up to 1.28 cm2 V-1 s-1 along with COS ∼120%, and 10 wt %-PDPPT3 blend exhibits ER of 78% at ε = 150%, COS of ∼230%, modulus of 36.5 MPa, maximum mobility of 0.62 cm2 V-1 s-1 and no obvious degradation of mobility at ε = 150% after 100 cycles of strain. Moreover, the structural similarity enables the blend film uniform and stable microstructure against mechanical and thermal deformation. Notably, PU(DPP)35 and the blend are characterized by high mechanical performance similar to that of commercial elastomers in thin film state, and demonstrate their potential for high performance stretchable electronics.

Novel bioactive adhesive containing dimethylaminohexadecyl methacrylate and calcium phosphate nanoparticles to inhibit metalloproteinases and nanoleakage with three months of aging in artificial saliva.

OBJECTIVES: The objectives of this study were to: (1) develop a multifunctional adhesive via dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP); and (2) investigate its ability to provide metalloproteinases (MMPs) deactivation and remineralization for long-term dentin bonding durability. METHODS: DMAHDM and NACP were incorporated into Adper™ Single Bond 2 Adhesive (SB2) at mass fractions of 5% and 20%, respectively. Degree of conversion and contact angle were measured. Endogenous MMP activity of the demineralized dentin beams, Masson's trichrome staining, nano-indentation, microtensile bond strength and interfacial nanoleakage analyses were investigated after 24 h and 3 months of storage aging in artificial saliva. RESULTS: Adding DMAHDM and NACP did not compromise the degree of conversion and contact angle of SB2 (p > 0.05). DMAHDM and NACP incorporation reduced the endogenous MMP activity by 53 %, facilitated remineralization, and increased the Young's modulus of hybrid layer by 49 % after 3 months of aging in artificial saliva, compared to control. For SB2 Control, the dentin bond strength decreased by 38 %, with greater nanoleakage expression, after 3 months of aging (p < 0.05). However, DMAHDM+NACP group showed no loss in bond strength, with much less nanoleakage, after 3 months of aging (p > 0.05). SIGNIFICANCE: DMAHDM+NACP adhesive greatly reduced MMP-degradation activity in demineralized dentin, induced remineralization at adhesive-dentin interface, and maintained the dentin bond strength after aging, without adversely affecting polymerization and dentin wettability. This new adhesive has great potential to help eliminate secondary caries, prevent hybrid layer degradation, and increase the resin-dentin bond longevity.

The Osteoinductivity of Calcium Phosphate-Based Biomaterials: A Tight Interaction With Bone Healing.

Calcium phosphate (CaP)-based bioceramics are the most widely used synthetic biomaterials for reconstructing damaged bone. Accompanied by bone healing process, implanted materials are gradually degraded while bone ultimately returns to its original geometry and function. In this progress report, we reviewed the complex and tight relationship between the bone healing response and CaP-based biomaterials, with the emphasis on the in vivo degradation mechanisms of such material and their osteoinductive properties mediated by immune responses, osteoclastogenesis and osteoblasts. A deep understanding of the interaction between biological healing process and biomaterials will optimize the design of CaP-based biomaterials, and further translate into effective strategies for biomaterials customization.

Adjunctive Diode Laser Therapy and Probiotic Lactobacillus Therapy in the Treatment of Periodontitis and Peri-Implant Disease.

Periodontal and peri-implant diseases are plaque-induced infections with a high prevalence, seriously impairing people's quality of life. The diode laser has long been recommended as adjunctive therapy in treating periodontitis. However, the optimal combination of usage mode (inside or outside periodontal pocket) and application regimen (single or multiple sessions of appointment) has not been described in detail. Meanwhile, probiotic Lactobacillus is regarded as a potential adjuvant in the management of the peri-implant disease. Nonetheless, a detailed protocol for an effective probiotic application is lacking. This article aims to summarize two clinical protocols. For periodontitis, the optimal collaboration of laser usage mode and application regimen was identified. Regarding peri-implant mucositis, a combined therapy containing professional topical use and home administration of probiotic Lactobacillus was established. This updated laser protocol clarifies the relationship between the treatment mode (inside or outside the periodontal pocket) and the number of laser appointments, further refining the existing diode laser therapy. For inside pocket irradiation, a single session of laser treatment is suggested whereas, for outside pocket irradiation, multiple sessions of laser treatment provide better effects. The improved probiotic Lactobacillus therapy resulted in the disappearance of swelling of the peri-implant mucosa, a reduced bleeding on probing (BOP), and an obvious reduction and good control of plaque and pigmentation; however, probing pocket depth (PPD) had limited improvement. The current protocol should be regarded as preliminary and could be further enhanced.

PINK1 mediated mitophagy attenuates early apoptosis of gingival epithelial cells induced by high glucose.

BACKGROUND: Oxidative stress mediated by hyperglycemia damages cell-reparative processes such as mitophagy. Down-regulation of mitophagy is considered to be a susceptible factor for diabetes mellitus (DM) and its complications. However, the role of mitophagy in DM-associated periodontitis has not been fully elucidated. Apoptosis of human gingival epithelial cells (hGECs) is one of the representative events of DM-associated periodontitis. Thus, this study aimed to investigate PTEN-induced putative kinase 1 (PINK1)-mediated mitophagy activated in the process of high glucose (HG)-induced hGECs apoptosis. METHODS: For dose-response studies, hGECs were incubated in different concentrations of glucose (5.5, 15, 25, and 50 mmol/L) for 48 h. Then, hGECs were challenged with 25 mmol/L glucose for 12 h and 48 h, respectively. Apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL), caspase 9 and mitochondrial membrane potential (MMP). Subsequently, autophagy was evaluated by estimating P62, LC3 II mRNA levels, LC3 fluorescent puncta and LC3-II/I ratio. Meanwhile, the involvement of PINK1-mediated mitophagy was assessed by qRT-PCR, western blotting and immunofluorescence. Finally, hGECs were transfected with shPINK1 and analyzed by MMP, caspase 9 and annexin V-FITC apoptosis. RESULTS: The number of TUNEL-positive cells and caspase 9 protein were significantly increased in cells challenged with HG (25 mmol/L) for 48 h (HG 48 h). MMP was impaired both at HG 12 h and HG 48 h, but the degree of depolarization was more serious at HG 48 h. The autophagy improved as the amount of LC3 II increased and p62 decreased in HG 12 h. During this process, HG 12 h treatment induced PINK1-mediated mitophagy. PINK1 silencing with HG 12 h resulted in MMP depolarization and cell apoptosis. CONCLUSIONS: These results suggested that loss of the PINK1 gene may cause mitochondrial dysfunction and increase sensitivity to HG-induced apoptosis of hGECs at the early stage. PINK1 mediated mitophagy attenuates early apoptosis of gingival epithelial cells induced by high glucose.