Endogenous Matrix Metalloproteinases Research Articles

Effect of Salvadora persica on resin-dentin bond stability

BackgroundThe stability of resin–dentin interfaces is still highly questionable. The aim of this study was to evaluate the effect of Salvadora persica on resin–dentin bond durability.Materials and methodsExtracted human third molars were used to provide mid-coronal dentin, which was treated with 20% Salvadora persica extract for 1 min after acid-etching. Microtensile bond strength and interfacial nanoleakage were evaluated after 24 h and 6 months. A three-point flexure test was used to measure the stiffness of completely demineralized dentin sticks before and after treatment with Salvadora persica extract. The hydroxyproline release test was also used to measure collagen degradation by endogenous dentin proteases. Statistical analysis was performed using two-way ANOVA followed by post hoc Bonferroni test and unpaired t-test. P-values < 0.05 were considered statistically significant.ResultsThe use of Salvadora persica as an additional primer with etch-and-rinse adhesive did not affect the immediate bond strengths and nanoleakage (p > 0.05). After 6 months, the bond strength of the control group decreased (p = 0.007), and nanoleakage increased (p = 0.006), while Salvadora persica group showed no significant difference in bond strength and nanoleakage compared to their 24 h groups (p > 0.05). Salvadora persica increased dentin stiffness and decreased collagen degradation (p < 0.001) compared to their controls.ConclusionSalvadora persica extract pretreatment of acid-etched dentin preserved resin–dentin bonded interface for 6 months.Clinical significanceDurability of resin-dentin bonded interfaces is still highly questionable. Endogenous dentinal matrix metalloproteinases play an important role in degradation of dentinal collagen within such interfaces. Salvadora persica may preserve resin-dentin interfaces for longer periods of time contributing to greater clinical success and longevity of resin composite restorations.

Nitric oxide nano-reactor DNMF/PLGA enables tumor vascular microenvironment and chemo-hyperthermia synergetic therapy

BackgroundBreast cancer ranks first among malignant tumors, of which triple-negative breast cancer (TNBC) is characterized by its highly invasive behavior and the worst prognosis. Timely diagnosis and precise treatment of TNBC are substantially challenging. Abnormal tumor vessels play a crucial role in TNBC progression and treatment. Nitric oxide (NO) regulates angiogenesis and maintains vascular homeostasis, while effective NO delivery can normalize the tumor vasculature. Accordingly, we have proposed here a tumor vascular microenvironment remodeling strategy based on NO-induced vessel normalization and extracellular matrix collagen degradation with multimodality imaging-guided nanoparticles against TNBC called DNMF/PLGA.ResultsNanoparticles were synthesized using a chemotherapeutic agent doxorubicin (DOX), a NO donor L-arginine (L-Arg), ultrasmall spinel ferrites (MnFe2O4), and a poly (lactic-co-glycolic acid) (PLGA) shell. Nanoparticle distribution in the tumor was accurately monitored in real-time through highly enhanced magnetic resonance imaging and photoacoustic imaging. Near-infrared irradiation of tumor cells revealed that MnFe2O4 catalyzes the production of a large amount of reactive oxygen species (ROS) from H2O2, resulting in a cascade catalysis of L-Arg to trigger NO production in the presence of ROS. In addition, DOX activates niacinamide adenine dinucleotide phosphate oxidase to generate and supply H2O2. The generated NO improves the vascular endothelial cell integrity and pericellular contractility to promote vessel normalization and induces the activation of endogenous matrix metalloproteinases (mainly MMP-1 and MMP-2) so as to promote extravascular collagen degradation, thereby providing an auxiliary mechanism for efficient nanoparticle delivery and DOX penetration. Moreover, the chemotherapeutic effect of DOX and the photothermal effect of MnFe2O4 served as a chemo-hyperthermia synergistic therapy against TNBC.ConclusionThe two therapeutic mechanisms, along with an auxiliary mechanism, were perfectly combined to enhance the therapeutic effects. Briefly, multimodality image-guided nanoparticles provide a reliable strategy for the potential application in the fight against TNBC.Graphical

Effect of Nisin-based pretreatment solution on dentin bond strength, antibacterial property, and MMP activity of the adhesive interface

ObjectiveTo evaluate the effect of a Nisin-based dentin pretreatment solution on microtensile bond strength, antibacterial activity, and matrix metalloproteinase (MMP) activity of the adhesive interface. Materials and methods100 human molars were sectioned to expose dentin. The teeth were assigned to five groups (n = 20), according to the dentin pretreatment: 0.5%, 1.0%, or 1.5% Nisin; 0.12% chlorhexidine (positive control), and no solution (negative control), and divided into 2 subgroups: no aging, and thermomechanical aging. Specimens were etched with 37% H3PO4 for 15 s and submitted to the dentin pretreatment. Then, they were bonded with an adhesive (Adper Single Bond 2) and a resin composite for microtensile bond strength (μTBS) evaluation. Antibacterial activity against Streptococcus mutans was qualitatively examined using an agar diffusion test. Anti-MMP activity within hybrid layers was examined using in-situ zymography. Data were analyzed with two-factor ANOVA and post-hoc Tukey’s test (α = 0.050). ResultsFor μTBS, significant differences were identified for the factors “solutions” (p = 0.002), “aging” (p = 0.017), and interaction of the two factors (p = 0.002). In the absence of aging, higher μTBS was observed for the group 0.5% Nisin. In the presence of aging, all groups showed similar μTBS values. All Nisin concentrations were effective in inhibiting the growth of S. mutans. Endogenous MMP activity was more significantly inhibited using 0.5% and 1.0% Nisin (p < 0.050). Conclusion0.5% and 1.0% Nisin solutions do not adversely affect resin-dentin bond strength and exhibit a potential bactericidal effect against S. mutans. Both concentrations effectively reduce endogenous gelatinolytic activity within the hybrid layer. Clinical relevanceThe use of 0.5% and 1.0% Nisin solutions for dentin pretreatment potentially contributes to preserving the adhesive interface, increasing the longevity of composite restorations.

Photo-Triggered Cascade Therapy: A NIR-II AIE Luminogen Collaborating with Nitric Oxide Facilitates Efficient Collagen Depletion for Boosting Pancreatic Cancer Phototheranostics.

The dense extracellular matrix (ECM) in the pancreatic cancer severely hampers the penetration of nanodrugs, which causes inferior therapeutic efficacy. To address this issue, a multifunctional liposome, namely, Lip-DTI/NO, integrating a type-I photosensitizer DTITBT with glutathione (GSH) or heat-responsive nitric oxide (NO) donor S-nitroso-N-acetyl-D-penicillamine (SNAP) is constructed to deplete the tumor ECM, leading to enhanced drug delivery and consequently improved phototherapy. The loaded DTITBT possesses multiple functions including NIR-II fluorescence imaging, efficient superoxide radical (O2 •-) generation and excellent photothermal conversion efficiency, making it feasible for precisely pinpointing the tumor in the phototherapy process. Responding to the intracellular overexpressed glutathione or heat produced by photothermal effect of DTITBT, NO can be released from SNAP. Upon 808nm laser irradiation, Lip-DTI/NO could selectively induce in situ generation of peroxynitrite anion (ONOO-) in tumor after cascade processes including O2 •- production, GSH or heat-triggered NO release, and rapid reaction between O2 •- and NO. The generated ONOO- could activate the expression of endogenous matrix metalloproteinases which could efficiently digest collagen of tumor ECM, thus facilitating enhanced penetration and accumulation of Lip-DTI/NO in tumor. In vivo evaluation demonstrates the notable therapeutic efficacy via ONOO--potentiated synergistic photodynamic-photothermal therapies on both subcutaneous and orthotopic pancreatic cancer model.

Rapid Intrafibrillar Mineralization Strategy Enhances Adhesive–Dentin Interface

Biomimetic mineralization of dentin collagen appears to be a promising strategy to optimize dentin bonding durability. However, traditional postbonding mineralization strategies based on Ca/P ion release still have some drawbacks, such as being time-consuming, having a spatiotemporal mismatch, and having limited intrafibrillar minerals. To tackle these problems, a prebonding rapid intrafibrillar mineralization strategy was developed in the present study. Specifically, polyacrylic acid–stabilized amorphous calcium fluoride (PAA-ACF) was found to induce rapid intrafibrillar mineralization of the single-layer collagen model and dentin collagen at just 1 min and 10 min, as identified by transmission electron microscopy, scanning electron microscopy, and atomic force microscopy. This strategy has also been identified to strengthen the mechanical properties of demineralized dentin within a clinically acceptable timeframe. Significantly, the bonding strength of the PAA-ACF–treated groups outperformed the control group irrespective of aging modes. In addition, the endogenous matrix metalloproteinases as well as exogenous bacterial erosion were inhibited, thus reducing the degradation of dentin collagen. High-quality integration of the hybrid layer and the underlying dentin was also demonstrated. On the basis of the present results, the concept of “prebonding rapid intrafibrillar mineralization” was proposed. This user-friendly scheme introduced PAA-ACF–based intrafibrillar mineralization into dentin bonding for the first time. As multifunctional primers, PAA-ACF precursors have the potential to shed new light on prolonging the service life of adhesive restorations, with promising significance.

Novel biomimetic peptide-loaded chitosan nanoparticles improve dentin bonding via promoting dentin remineralization and inhibiting endogenous matrix metalloproteinases

ObjectiveThis study aims to synthesize novel chitosan nanoparticles loaded with an amelogenin-derived peptide QP5 (TMC-QP5/NPs), investigate their remineralization capability and inhibitory effects on endogenous matrix metalloproteinases (MMPs), and evaluate the dentin bonding properties of remineralized dentin regulated by TMC-QP5/NPs. MethodsTMC-QP5/NPs were prepared by ionic crosslinking method and characterized by dynamic light scattering method, scanning electron microscopy, transmission electron microscope, atomic force microscope, Fourier transform infrared spectroscopy, and differential scanning calorimetry. The encapsulation and loading efficiency of TMC-QP5/NPs and the release of QP5 were examined. To evaluate the remineralization capability of TMC-QP5/NPs, the mechanical properties, and the changes in structure and composition of differently conditioned dentin were characterized. The MMPs inhibitory effects of TMC-QP5/NPs were explored by MMP Activity Assay and in-situ zymography. The dentin bonding performance was detected by interfacial microleakage and microshear bond strength (μSBS). ResultsTMC-QP5/NPs were successfully synthesized, with uniform size, good stability and biosafety. The encapsulation and loading efficiency of TMC-QP5/NPs was respectively 69.63 ± 2.22% and 13.21 ± 0.73%, with a sustained release of QP5. TMC-QP5/NPs could induce mineral deposits on demineralized collagen fibers and partial occlusion of dentin tubules, and recover the surface microhardness of dentin, showing better remineralization effects than QP5. Besides, TMC-QP5/NPs significantly inhibited the endogenous MMPs activity. The remineralized dentin induced by TMC-QP5/NPs exhibited less interfacial microleakage and higher μSBS, greatly improved dentin bonding. SignificanceThis novel peptide-loaded chitosan nanoparticles improved resin-dentin bonding by promoting dentin remineralization and inactivating MMPs, suggesting a promising strategy for optimizing dentin adhesive restorations.

Effect of Different Crosslinkers on Denatured Dentin Collagen's Biostability, MMP Inhibition and Mechanical Properties.

Sound, natural dentin collagen can be stabilized against enzymatic degradation through exogenous crosslinking treatment for durable bonding; however, the effect on denatured dentin (DD) collagen is unknown. Hence, the ability of different crosslinkers to enhance/restore the properties of DD collagen was assessed. Demineralized natural and DD collagen films (7 mm × 7 mm × 7 µm) and beams (0.8 mm × 0.8 mm × 7 mm) were prepared. DD collagen was experimentally produced by heat or acid exposure, which was then assessed by various techniques. All specimens were then treated with 1 wt% of chemical crosslinker 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/n-hydroxysuccinimide (EDC/NHS) and two structurally different flavonoids-theaflavins (TF) from black tea and type-A proanthocyanidins from cranberry juice (CR) for either 30 s or 1 h. The controls were untreated. Dentin films were assessed for chemical interaction and cross-linking effect by FTIR, biostability against exogenous collagenase by weight loss (WL) and hydroxyproline release (HYP), and endogenous matrix metalloproteinases (MMPs) activity by confocal laser microscopy. Dentin beams were evaluated for tensile properties. Data were analyzed using ANOVA and Tukey's test (α = 0.05). Compared with natural collagen, DD collagen showed pronounced structural changes, altered biostability and decreased mechanical properties, which were then improved to various degrees that were dependent on the crosslinkers used, with EDC/NHS being the least effective. Surprisingly, the well-known MMP inhibitor EDC/NHS showed negligible effect on or even increased MMP activity in DD collagen. As compared with control, cross-linking induced by TF and CR significantly increased collagen biostability (reduced WL and HYP release, p < 0.05), MMP inhibition (p < 0.001) and mechanical properties (p < 0.05), regardless of denaturation. DD collagen cannot or can only minimally be stabilized via EDC/NHS crosslinking; however, the challenging substrate of DD collagen can be enhanced or restored using the promising flavonoids TF and CR.

The Venom Composition of the Snake Tribe Philodryadini: 'Omic' Techniques Reveal Intergeneric Variability among South American Racers.

Snakes of the Philodryadini tribe are included in the Dipsadidae family, which is a diverse group of rear-fanged snakes widespread in different ecological conditions, including habitats and diet. However, little is known about the composition and effects of their venoms despite their relevance for understanding the evolution of these snakes or even their impact on the occasional cases of human envenoming. In this study, we integrated venom gland transcriptomics, venom proteomics and functional assays to characterize the venoms from eight species of the Philodryadini tribe, which includes the genus Philodryas, Chlorosoma and Xenoxybelis. The most abundant components identified in the venoms were snake venom metalloproteinases (SVMPs), cysteine-rich secretory proteins (CRISPs), C-type lectins (CTLs), snake endogenous matrix metalloproteinases type 9 (seMMP-9) and snake venom serinoproteinases (SVSPs). These protein families showed a variable expression profile in each genus. SVMPs were the most abundant components in Philodryas, while seMMP-9 and CRISPs were the most expressed in Chlorosoma and Xenoxybelis, respectively. Lineage-specific differences in venom composition were also observed among Philodryas species, whereas P. olfersii presented the highest amount of SVSPs and P. agassizii was the only species to express significant amounts of 3FTx. The variability observed in venom composition was confirmed by the venom functional assays. Philodryas species presented the highest SVMP activity, whereas Chlorosoma species showed higher levels of gelatin activity, which may correlate to the seMMP-9 enzymes. The variability observed in the composition of these venoms may be related to the tribe phylogeny and influenced by their diets. In the presented study, we expanded the set of venomics studies of the Philodryadini tribe, which paves new roads for further studies on the evolution and ecology of Dipsadidae snakes.

Mineralization promotion and protection effect of carboxymethyl chitosan biomodification in biomimetic mineralization

Biomimetic mineralization emphasizes reversing the process of dental caries through bio-inspired strategies, in which mineralization promotion and collagen protection are equally important. In this study, carboxymethyl chitosan (CMC) was deemed as an analog of glycosaminoglycan for biomimetic modification of collagen, both of the mineralization facilitation and collagen protection effect were evaluated. Experiments were carried out simultaneously on two-dimensional monolayer reconstituted collagen model, three-dimensional reconstituted collagen model and demineralized dentin model. In three models, CMC was successfully cross-linked onto collagen utilizing biocompatible 1-Ethyl-3(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxy sulfosuccinimide sodium salt to achieve biomodification. Results showed that CMC biomodification increased collagen's hydrophilicity, calcium absorption capacity and thermal degradation resistance. In demineralized dentin model, the activity of endogenous matrix metalloproteinases was significantly inhibited by CMC biomodification. Furthermore, CMC biomodification significantly improved cross-linking and intrafibrillar mineralization of collagen, especially in the two-dimensional monolayer reconstituted collagen model. This study provided a biomimetic mineralization strategy with comprehensive consideration of collagen protection, and enriched the application of chitosan-based materials in dentistry.

Tension-suppressed degradation of collagen controls tissue stiffness scaling with fibrillar collagen.

Extremely soft tissues such as developing hearts or adult brain contain far less collagen than highly stiff adult tissues such as tendons, but cell and molecular mechanisms for such homeostatic differences remain unclear. We hypothesized that cell-generated or exogenous forces combine with tension-suppressed collagen degradation in order to sculpt extracellular matrix (ECM) collagen levels in tissues. For various mature mice tissues and beating embryonic chick hearts, we find collagen-sensitive second harmonic generation (SHG) image intensity scales non-linearly versus tissue stiffness, aligning well with the results from cellularized gels of collagen. Chick hearts beating at ∼5% strain maintain collagen levels until their contractile strain is suppressed by myosin-II inhibition and endogenous matrix metalloproteinases (MMPs) then degrade collagens within ∼30-60 minutes - based on SHG and mass spectrometry proteomics. Although tendons composed of oriented collagen fibrils exhibit heterogeneous strain distributions upon deformation, the addition of exogenous MMP or bacterial collagenase suppresses collagen degradation for strains within physiological limits (i.e., up to ∼5-8%). Sequestration of collagen cleavage sites by tissue strain is a likely mechanism because molecular permeation and mobility prove strain-independent whereas artificial collagen cross-links accelerate strain-dependent collagen degradation via collagen molecular unfolding. Tension-suppressed degradation of collagen thus underlies tissue stiffness scaling.

Myeloma Microenvironmental TIMP1 Induces the Invasive Phenotype in Fibroblasts to Modulate Disease Progression.

Tissue inhibitors of metalloproteinases (TIMPs) are endogenous matrix metalloproteinase inhibitors. TIMP1 is produced by cancer cells and has pleiotropic activities. However, its role and source in multiple myeloma (MM) are unclear. Here, we evaluated TIMP1 protein and mRNA levels in bone marrow (BM) plasma cells and assessed the effects of TIMP1 expression on fibroblast invasive capacity using three-dimensional spheroid cell invasion assays. TIMP1 mRNA and protein levels were elevated when patients progressed from monoclonal gammopathy of undetermined significance or smouldering myeloma to MM. Furthermore, TIMP1 levels decreased at complete response and TIMP1 protein levels increased with higher international staging. TIMP1 mRNA levels were markedly higher in extramedullary plasmacytoma and MM with t(4;14). Overall survival and post-progression survival were significantly lower in MM patients with high TIMP1 protein. Recombinant TIMP1 did not directly affect MM cells but enhanced the invasive capacity of fibroblasts; this effect was suppressed by treatment with anti-TIMP1 antibodies. Fibroblasts supported myeloma cell invasion and expansion in extracellular matrix. Overall, these results suggested that MM-derived TIMP1 induces the invasive phenotype in fibroblasts and is involved in disease progression. Further studies are required to elucidate the specific roles of TIMP1 in MM and facilitate the development of novel therapies targeting the TIMP1 pathway.

Do matrix metalloproteinase and cathepsin K inhibitors work synergistically to reduce dentin erosion?

To evaluate the effects of matrix metalloproteinase (MMP) and cathepsin K (catK) inhibitors on resistance to dentin erosion. A total of 96 dentin specimens (3×3×2 mm) were prepared and randomly assigned into four groups (n=24): deionized water (DW); 1 µM odanacatib (ODN, catK inhibitor); 1 mM 1,10-phenanthroline (PHEN, MMP inhibitor); and 1 µM odanacatib + 1 mM 1,10-phenanthroline (COM). Each group was further divided into two subgroups for the application of treatment solutions before (PRE) and after erosive challenges (POST). All specimens were subjected to four daily erosive challenges for 5 d. For each erosive challenge, the specimens in subgroup PRE were immersed in the respective solutions before cola drinks, while the specimens in subgroup POST were immersed in the respective solutions after cola drinks (the immersion duration was 5 min in both cases). All specimens were stored in artificial saliva at 37°C between erosive challenges. The erosive dentin loss (EDL) was measured by profilometry. The residual demineralized organic matrix (DOM) of specimens was removed using type VII collagenase and evaluated by profilometry. Both the EDL and thickness of the residual DOM were statistically analyzed by two-way analysis of variance (ANOVA) and Bonferroni's test (α=0.05). The surface topography and transverse sections of the specimens were observed using SEM. MMPs and catK were immunolabeled in the eroded dentin and in situ zymography was performed to evaluate the enzyme activity. Significantly lower EDL was found in the groups ODN, PHEN, and COM than in the control group (all p<0.05), while no significant difference in EDL was found among the groups ODN, PHEN, and COM (all p>0.05). The application sequence showed no significant effect on the EDL of the tested groups (p=0.310). A significantly thicker DOM was observed in the group ODN than in the control group regardless of the application sequence (both p<0.05). The treatment with ODN, PHEN, and COM inhibited the gelatinolytic activity by approximately 46.32%, 58.6%, and 74.56%, respectively. The inhibition of endogenous dentinal MMPs and catK increases the acid resistance of human dentin but without an apparent synergistic effect. The inhibition of MMPs and catK is equally effective either before or after the acid challenge.

The relationship between tissue inhibitor metalloproteinase-1 with colorectal cancer and tissue inhibitor metalloproteinase-1 as a predictor of colorectal cancer

Colorectal cancer is reported as the third most common case worldwide in men and accounts for 10% of all cancers. Matrix metalloproteinases (MMPs) comprise a family of zinc-dependent endopeptidases that affect the degradation and remodeling of extracellular components and contribute to the three Hallmarks of cancer. TIMP-1 is an endogenous MMP inhibitor with multiple functions related to the tumor microenvironment and colorectal cancer development. This study used a prospective cohort study design to prove that colorectal mucosa with high TIMP-1 expression is a prognostic factor in the incidence of colorectal cancer compared to normal/low TIMP values. This study involved 53 samples consisting of 27 polyp patients and 26 colorectal cancer patients. The bivariate analysis results showed a significant difference between TIMP-1 levels in cancer patients and colorectal polyps (p&lt;0.001; 95%CI: 33.65-67.31). The cut-off value of 66.50 obtained a sensitivity value of 80.8% and a specificity value of 77.8% (p&lt;0.001). Spearman rho correlation test found a significant positive correlation between TIMP-1 levels and cases of colorectal cancer (p&lt;0.001) of 0.680. This study shows a significant relationship between TIMP-1 levels and colorectal cancer, where TIMP-1 is a good predictor of colorectal cancer incidence.

Role of Natural Cross Linkers in Resin-Dentin Bond Durability: A Systematic Review and Meta-Analysis.

Background: The role of endogenous Matrix Metallo Proteinases in resin dentin bond deterioration over time has been well documented. The present study aimed to systematically review the literature; in vitro and ex vivo studies that assessed the outcomes of natural cross-linkers for immediate and long-term tensile bond strength were included. Methods: The manuscript search was carried out in six electronic databases—PubMed/MEDLINE, LILACS, SciELO, Cochrane, Web of Science and DOAJ, without publication year limits. Only manuscripts in English (including the translated articles) were selected, and the last search was performed in December 2020. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement was followed. Results: From the 128 potentially eligible studies, 48 full-text articles were assessed for eligibility. After eligibility assessment and exclusions, 14 studies were considered for systematic review and seven studies for meta-analysis. Amongst the selected studies for meta-analysis, three had a medium and four had a low risk of bias. Conclusions: It was evidenced by the available data that Proanthocyanidin is the most efficient natural cross-linker to date, in preserving the bond strength even after ageing.

Strategies for demineralized dentin matrix protection and its research progress in dentin-resin bonding restoration

Both pathological injuries and clinical iatrogenic operations can lead to dentin demineralization, forming demineralized dentin matrix (DDM). Dentin demineralization activates endogenous matrix metalloproteinase (MMP) and cysteine cathepsin (CC), and the mechanical properties of DDM decrease, so DDM is prone to lose its structural integrity under the action of enzymatic degradation and mechanical destruction, which in turn results in the reduction of clinical functional value of DDM in dentin-resin bonding restoration. The administrations of dentin collagen cross-linking reagents and MMP/CC inhibitors are effective strategies to protect DDM structural integrity and achieve its clinical functional value. A variety of chemically synthesized reagents and plant-derived extracts are capable of significantly improving the mechanical properties of DDM and enhancing its enzymatic tolerance. However, the cytotoxicity caused by chemically synthesized reagents and the tooth staining aroused by plant extracts have considerably affected their clinical applicability. Protecting dentin collagen while exerting antibacterial properties is a new direction for future DDM protective agent research. Accordingly, from the perspectives of cross-linking reagents, enzyme inhibitors and compounds which possess the dual proper ties, this review discusses the latest research progress in DDM protection, and looks into its application prospects in dentin-resin bonding, in an attempt to provide reference for the clinical promotion of DDM protection strategy.

Effect of Remineralisation with Nano-hydroxyapatite (nHAp) and Non-collagenous Protein Analogues (PAA and STPP) on the Micro Tensile Bond Strength of the Resin Composites to Dentin – An In vitro Study

Background: Resin-dentin bonds are less durable than enamel as they rely on organic part for mineralisation. Bond denaturation mainly takes place because of hydrolysis of polymerized resin or degradation of water-rich, resin-sparse collagen by collagenolytic endogenous matrix metalloproteinase and cysteine cathepsins. Hence, this limited stability of dentin bonding tenaciously reduces the longevity of tooth-coloured restorations. One of the strategies adopted to encounter this shortcoming is to induce biomimetic remineralisation.&#x0D; Aim: To assess the microtensile bond strength of resin composites to dentin following remineralisation using an experimental paste containing nano-hydroxyapatite (nHAp) and biomimetic analogues, Poly Acrylic Acid (PAA) and Sodium Tri Polyphosphate (STPP).&#x0D; Methodology: Nine intact freshly extracted third molar were used for this study to evaluate the microtensile bond strength after treating with three different groups; Group I with paste containing nano hydroxyapatite nHAp Group II paste containing nHAp and non-collagenous protein analogues such as (Polyacrcrylic acid and Sodium tripolyphosphate) and the control groups containing adhesive and resin composite with no additional treatment. The samples were observed over a period of 24 hours and 1 week the inter group data analysis was done using ANOVA and paired sample t test and the intra-group data analysis over period of 24 hours the post HOC Bonferroni test was used for the statistical analysis.&#x0D; Results: After 24 hours the group with nHAp group showed highest value for microtensile bond strength among other groups, however after one week the group with nHAp +NCP analogues(PAA+STPP) showed highest value for microtensile bond.&#x0D; Conclusion: The treatment with the experimental paste [nHAp+NCP] analogues (PAA+STPP)] can bring about remineralisation to improve bonding to dentin.

Cranberry Juice Extract Rapidly Protects Demineralized Dentin against Digestion and Inhibits Its Gelatinolytic Activity.

Improving the longevity of composite restorations has proven to be difficult when they are bonded to dentin. Dentin demineralization leaves collagen fibrils susceptible to enzymatic digestion, which causes breakdown of the resin–dentin interface. Therefore, measures for counteracting the enzymatic environment by enhancing dentin collagen’s resistance to degradation have the potential to improve the durability of dental composite restorations. This study aimed to evaluate the effects of polyphenol-rich extracts and a chemical cross-linker on the cross-linking interaction, resistance to digestion, and endogenous matrix metalloproteinase (MMP) activities of dentin collagen under clinically relevant conditions. Ten-µm-thick films were cut from dentin slabs of non-carious extracted human third molars. Following demineralization, polyphenol-rich extracts—including grape seed (GSE), green tea (GTE), and cranberry juice (CJE)—or chemical cross-linker carbodiimide with n-hydroxysuccinimide (EDC/NHS) were applied to the demineralized dentin surfaces for 30 s. The collagen cross-linking, bio-stabilization, and gelatinolytic activities of MMPs 2 and 9 were studied by using Fourier-transform infrared spectroscopy, weight loss, hydroxyproline release, scanning/transmission electron microscopy, and in situ zymography. All treatments significantly increased resistance to collagenase degradation and reduced the gelatinolytic MMP activity of dentin collagen compared to the untreated control. The CJE- and GSE-treated groups were more resistant to digestion than the GTE- or EDC/NHS-treated ones (p < 0.05), which was consistent with the cross-linking interaction found with FTIR and the in situ performance on the acid-etched dentin surface found with SEM/TEM. The collagen films treated with CJE showed the lowest MMP activity, followed by GSE, GTE, and, finally, EDC/NHS. The CJE-treated dentin collagen rapidly increased its resistance to digestion and MMP inhibition. An application of CJE as short as 30 s may be a clinically feasible approach to improving the longevity of dentin bonding in composite restorations.

Effect of chlorhexidine-loaded poly(amido amine) dendrimer on matrix metalloproteinase activities and remineralization in etched human dentin in vitro

To investigate the effect of chlorhexidine (CHX)-loaded carboxyl-terminated poly (amido amine) dendrimer (CHX-PAMAM-COOH) on matrix metalloproteinase (MMP) activities and remineralization in human dentin, CHX-PAMAM-COOH was prepared and characterized by Fourier-transform infrared spectroscopy. The inhibitory effects of CHX, PAMAM-COOH, and CHX-PAMAM-COOH on soluble recombinant human matrix metalloproteinase (rhMMP-2) and dentin-bound endogenous MMP activity were measured using an MMP Activity Assay Kit. In situ zymography was performed to evaluate the gelatinase activity in dentin pretreated with CHX, PAMAM-COOH, and CHX-PAMAM-COOH. The remineralization of etched dentin pretreated with CHX, PAMAM-COOH, and CHX-PAMAM-COOH was evaluated by field emission-scanning electron microscopy (SEM) and energy disperse spectroscopy (EDS) after incubation in artificial saliva for 14 days. The results of the rhMMP-2 activity assay showed that the MMP-2 activity in the CHX-PAMAM-COOH group and the CHX group decreased significantly to 5.58 ± 0.85% (P < 0.05) and 4.86 ± 1.12% (P < 0.05), respectively, but that in the PAMAM-COOH group increased significantly to 213.38 ± 0.11% (P < 0.05). The results of total MMP activity and in situ zymography showed a significant reduction in endogenous gelatinase activity in dentin in the CHX-PAMAM-COOH group and the CHX group. The SEM and EDS results showed that rod-like crystals were formed on the etched dentin surface in the PAMAM-COOH group and the CHX-PAMAM-COOH group, and their Ca/P ratios were 1.73 and 1.71, respectively. In conclusion, CHX-PAMAM-COOH can inhibit dentin-bound endogenous MMPs and induce remineralization in etched dentin simultaneously. However, it is important to note that the catalytic role of PAMAM dendrimers may have an undesired excitatory effect on MMP activity, which cannot be ignored if PAMAM dendrimers were used alone in the oral environment.

Experimental Study of Collagen Density in Reconstruction of Tracheal Defect Using Dried Amniotic Membrane

Background: Current tracheal defect management is still controversial for surgeons because there areno ideal materials to substitute trachea and many long-term complications occur in patients after trachealreconstruction. Prosthetic graft has the risks of chronic infection and post-reconstructive tracheal stenosis.Dried Amniotic Membrane (DAM) expresses several endogenous matrix metalloproteinase (MMP) genesthat can degrade extracellular matrix. Therefore, DAM is expected to be able to degrade collagen formedexcessively during the wound healing phase, hence post-reconstruction stenosis will not occur. This studyaimed to evaluate the differences of collagen density between polypropylene mesh (only) and DAM-addedpolypropylene mesh as graft in tracheal reconstruction.Methods: The animals used in this study were 36 male white rabbits, which were divided into twogroups. Both groups were wounded in the trachea area. The defects from the first group were closed usingpolypropylene mesh, while the defects from the second group were closed using DAM-added polypropylenemesh. After 30 days, the collagen density of each group was assessed.Results: In the first group there are 10 samples (55.56%) with collagen density score = 3, while in the secondgroup there are 16 samples (88.89%) with collagen density score = 2. The significance value using MannWhitney U test for differences in the collagen density score in both groups is p = 0.407.Conclusion: Tracheal reconstruction using polypropylene mesh along with DAM provides a lower collagendensity score than using polypropylene mesh alone as a graft, but it is not statistically significant.

Electrochemical immunoplatform to assist in the diagnosis and classification of breast cancer through the determination of matrix-metalloproteinase-9

Matrix metalloproteinase 9 (MMP-9) is a zinc-dependent endopeptidase that promotes angiogenesis, tumor growth, metastasis and cell invasion through the degradation of extracellular matrix. This work reports a magnetic microbeads (MBs)-based sandwich immunoassay for the amperometric determination of MMP-9 at screen-printed carbon electrodes (SPCEs). The suitable capture antibody (cAb) is immobilized onto carboxylic MBs to selectively capture the antigen which is sandwiched with a biotinylated detector antibody (biotin-dAb) further conjugated with a commercial streptavidin-horseradish peroxidase (Strep-HRP) polymer. This immunoplatform provides great analytical characteristics in terms of selectivity and sensitivity, achieving a LOD value of 2.4 pg mL−1 for standards in buffered solutions. Although this value is similar to those reported for some other approaches described so far, the method described here is simpler involving a single 30 min incubation step which makes it ideal for automation or implementation in POC devices. Moreover, the method was assayed for the accurate determination of endogenous MMP-9 in both cancer cell lysates and serum samples of patients diagnosed with different subtypes of breast cancer (BC) after a simple dilution. The results obtained show that the disposable and affordable immunoplatform developed is able not only to discriminate BC patients from healthy individuals but also to do it for the worst outcome triple negative (TNBC) subtype.