ABSTRACT
The reparative and regenerative capabilities of dental pulp stem cells (DPSCs) are crucial for responding to pulp injuries, with protein phosphatase 1 (PP1) playing a significant role in regulating cellular functions pertinent to tissue healing. Accordingly, this study aimed to explore the effects of a novel cell-penetrating peptide Modified Sperm Stop 1-MSS1, that disrupts PP1, on the proliferation and odontogenic differentiation of DPSCs. Employing MSS1 as a bioportide, DPSCs were cultured and characterized for metabolic activity, cell proliferation, and cell morphology alongside the odontogenic differentiation through gene expression and alkaline phosphatase (ALP) activity analysis. MSS1 exposure induced early DPSC proliferation, upregulated genes related to odontogenic differentiation, and increased ALP activity. Markers associated with early differentiation events were induced at early culture time points and those associated with matrix mineralization were upregulated at mid-culture stages. This investigation is the first to document the potential of a PP1-disrupting bioportide in modulating DPSC functionality, suggesting a promising avenue for enhancing dental tissue regeneration and repair.
Subject(s)
Cell Differentiation , Cell Proliferation , Dental Pulp , Odontogenesis , Protein Phosphatase 1 , Stem Cells , Dental Pulp/cytology , Dental Pulp/drug effects , Stem Cells/drug effects , Stem Cells/cytology , Stem Cells/metabolism , Humans , Protein Phosphatase 1/metabolism , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Odontogenesis/drug effects , Peptides/pharmacology , Peptides/metabolism , Cells, Cultured , Alkaline Phosphatase/metabolismABSTRACT
INTRODUCTION: Tetracyclines (TCs) embrace a class of broad-spectrum antibiotics with unrelated effects at sub-antimicrobial levels, including an effective anti-inflammatory activity and stimulation of osteogenesis, allowing their repurposing for different clinical applications. Recently, sarecycline (SA)-a new-generation molecule with a narrower antimicrobial spectrum-was clinically approved due to its anti-inflammatory profile and reduced adverse effects verified with prolonged use. Notwithstanding, little is known about its osteogenic potential, previously verified for early generation TCs. MATERIALS AND METHODS: Accordingly, the present study is focused on the assessment of the response of human bone marrow-derived mesenchymal stromal cells (hBMSCs) to a concentration range of SA, addressing the metabolic activity, morphology and osteoblastic differentiation capability, further detailing the modulation of Wnt, Hedgehog, and Notch signaling pathways. In addition, an ex vivo organotypic bone development system was established in the presence of SA and characterized by microtomographic and histochemical analysis. RESULTS: hBMSCs cultured with SA presented a significantly increased metabolic activity compared to control, with an indistinguishable cell morphology. Moreover, RUNX2 expression was upregulated 2.5-fold, and ALP expression was increased around sevenfold in the presence of SA. Further, GLI2 expression was significantly upregulated, while HEY1 and HNF1A were downregulated, substantiating Hedgehog and Notch signaling pathways' modulation. The ex vivo model developed in the presence of SA presented a significantly enhanced collagen deposition, extended migration areas of osteogenesis, and an increased bone mineral content, substantiating an increased osteogenic development. CONCLUSION: Summarizing, SA is a promising candidate for drug repurposing within therapies envisaging the enhancement of bone healing/regeneration.
Subject(s)
Drug Repositioning , Hedgehogs , Humans , Animals , Osteogenesis , Cell Differentiation , Tetracyclines/pharmacology , Cells, Cultured , Bone Marrow CellsABSTRACT
Development of multifunctional 3D patches with appropriate antibacterial and biocompatible properties is needed to deal with wound care regeneration. Combining gelatin-based hydrogel with a well-known natural antibacterial honey (Manuka honey, MH) in a 3D patch can provide improved printability and at the same time provide favourable biological effects that may be useful in regenerative wound treatment. In this study, an antibacterial Manuka-Gelatin 3D patches was developed by an extrusion-based printing process, with controlled porosity, high shape fidelity, and structural stability. It was demonstrated the antibacterial activity of Manuka-Gelatin 3D patches against both gram-positive bacteria (S. epidermidis and S. aureus) and gram-negative (E. coli), common in wound infection. The 3D Manuka-Gelatin base patches demonstrated antibacterial activity, and moreover enhanced the proliferation of human dermal fibroblasts and human epidermal keratinocytes, and promotion of angiogenesis. Moreover, the ease of printing achieved by the addition of honey, coupled with the interesting biological response obtained, makes this 3D patch a good candidate for wound healing applications.
Subject(s)
Honey , Staphylococcus aureus , Humans , Gelatin , Microbial Sensitivity Tests , Escherichia coli , Wound Healing , Honey/analysis , Anti-Bacterial Agents/chemistry , Printing, Three-Dimensional , HydrogelsABSTRACT
STATEMENT OF PROBLEM: Microgap and bacterial microleakage at the implant-prosthetic abutment interface are recognized concerns for implant-supported restorations, leading to inflammation of the peri-implant tissues, with deleterious consequences for crestal bone levels. However, little is known regarding the interface established between the implant and the healing abutment or cover screw placed for the osseointegration phase. PURPOSE: The purpose of this in vitro study was to characterize the implant-cover screw and implant-healing abutment interfaces of a platform-switched implant system to determine the microgap and bacterial microleakage of the system and evaluate the biological response and functionality of an interface sealing agent. MATERIAL AND METHODS: The interfacial microgaps of the implant-healing abutment and implant-cover screw interfaces were characterized by scanning electron microscopy (n=10), and bacterial microleakage was evaluated after colonization with Enterococcus faecalis in a 30-day follow-up (n=10). The sealing efficacy and irritation potential of a silicone-based sealer were determined by using the hen's egg test on chorioallantoic membrane assay. The 2-sample t test was performed to compare means between groups, and data presented with the Kaplan-Meier method were compared statistically by using the log-rank test (α=.05). RESULTS: The interfacial microgap was less than 2.5 µm for both systems. Bacterial microleakage was noted in approximately 50% of the specimens, particularly at early time points, at both the healing abutment and cover screw interfaces. The silicone-based sealer prevented bacterial leakage in the experimental setting. CONCLUSIONS: The implant-healing abutment and implant-cover screw interfaces of the tested system, despite the low microgap, allowed for bacterial microleakage after internal colonization. The use of a nonirritating silicone-based sealing agent effectively sealed the system.
Subject(s)
Dental Implants , Osseointegration , Animals , Female , Chickens , Dental Abutments , Dental Implants/microbiology , Bacteria , Dental Implant-Abutment DesignABSTRACT
This work aims at the development and characterization of fluorapatite coatings, innovatively prepared by the hydrothermal method, aiming for enhanced osseointegration of titanium implants. Fluoride-containing coatings were prepared and characterized by scanning and transmission electron microscopy, Fourier-transform infrared spectroscopy-attenuated total reflectance, and X-ray photoelectron spectroscopy. The biological response was characterized by microtomographic evaluation and histomorphometric analysis upon orthotopic implantation in a translational rabbit experimental model. Physic-chemical analysis revealed the inclusion of fluoride in the apatite lattice with fluorapatite formation, associated with the presence of citrate species. The in vivo biological assessment of coated implants revealed an enhanced bone formation process-with increased bone-to-implant contact and bone volume. The attained enhancement of the osteogenic process may be attributable to the conjoined modulatory activity of selected fluoride and citrate levels within the produced coatings. In this regard, the production of fluorapatite coatings with citrate, through the hydrothermal method, entails a promising approach for enhanced osseointegration in implant dentistry and orthopedic applications.
ABSTRACT
Silicone-based medical devices composed of polydimethylsiloxane (PDMS) are widely used all over the human body (e.g., urinary stents and catheters, central venous catheters stents) with extreme clinical success. Nevertheless, their abiotic surfaces, being prone to microorganism colonization, are often involved in infection occurrence. Improving PDMS antimicrobial properties by surface functionalization with biosurfactants to prevent related infections has been the goal of different works, but studies that mimic the clinical use of these novel surfaces are missing. This work aims at the biofunctional assessment of PDMS functionalized with rhamnolipids (RLs), using translational tests that more closely mimic the clinical microenvironment. Rhamnolipids were covalently bonded to PDMS, and the obtained surfaces were characterized by contact angle modification assessment, ATR-FTIR analysis and atomic force microscopy imaging. Moreover, a parallel flow chamber was used to assess the Staphylococcus aureus antibiofilm activity of the obtained surfaces under dynamic conditions, and an in vitro characterization with human dermal fibroblast cells in both direct and indirect characterization assays, along with an in vivo subcutaneous implantation assay in the translational rabbit model, was performed. A 1.2 log reduction in S. aureus biofilm was observed after 24 h under flow dynamic conditions. Additionally, functionalized PDMS lessened cell adhesion upon direct contact, while supporting a cytocompatible profile, within an indirect assay. The adequacy of the biological response was further validated upon in vivo subcutaneous tissue implantation. An important step was taken towards biofunctional assessment of RLs-functionalized PDMS, reinforcing their suitability for medical device usage and infection prevention.
ABSTRACT
Controlling bacterial biofilm formation on silicone-based bloodstream catheters is of great concern to prevent related-infections. In this study, rhamnolipids (RLs), glycolipid biosurfactants, specifically a RLs mixture and the purified di-RL (RhaRhaC10:0C10:0) were covalently bonded to silicone with the intention of reaching long-lasting antibiofilm surfaces. RLs mixture and di-RL were identified by an UHPLC-MS method that also allowed the confirmation of compound isolation by automated flash chromatography. Silicone surfaces underwent air-plasma treatment, inducing reactive oxygen radicals able to promote the RLs grafting that was confirmed by contact angle, FTIR-ATR and AFM measurements. The antibiofilm activity towards different Gram positive strains was evaluated by colony forming units (CFU) count and confocal laser microscopy. In addition, protein adsorption and biocompatibility were also investigated. RLs were successfully grafted onto silicone and RLs mixture and RhaRhaC10C10:0 functionalized specimens reduced the biofilm formation over 2.3 log units against methicillin sensitive Staphylococcus aureus. Additionally, a decrease of 1 log unit was observed against methicillin resistant S. aureus and S. epidermidis. Functionalized samples showed cytocompatibility towards human dermal fibroblasts, hemocompatibility and no vascular irritation potential. The results mentioned above revealed a synergy between the antimicrobial and the anti-adhesive properties of RLs, making these compounds good candidates for the improvement of the medical devices antibiofilm properties.
Subject(s)
Anti-Infective Agents , Methicillin-Resistant Staphylococcus aureus , Anti-Bacterial Agents/pharmacology , Biofilms , Catheters/microbiology , Dimethylpolysiloxanes , Glycolipids/pharmacology , Humans , Staphylococcus epidermidisABSTRACT
BACKGROUND AND AIMS: This study aims to access the effectiveness of mid-infrared (MIR) spectroscopy on the identification of the reticular form of OLP, following the assessment of gingival crevicular fluid (GCF) and oral mucosa transudate (OMT). MATERIAL AND METHODS: The trial follows a case-control design. Samples were characterized through MIR spectroscopy and chemometric tools were applied to distinguish between case and control participants, further identifying the spectral regions with the highest contribution to the developed models. RESULTS: MIR spectroscopy was capable to discriminate between OLP patients and controls with 95.1% and 85.4% of correct predictions, regarding GCF and OMT samples, respectively. Additionally, the spectral regions mostly contributing to the successful prediction were identified, and possibly related with the distinctive presence of amino acids/proteins and oxidative stress mediators in oral biofluids, supporting the role of the immune-inflammatory activation on OLP etiology and disease course. CONCLUSION: MIR spectroscopy analysis of GCF and OMT may be regarded as an innovative, non-invasive, low cost and sensitive technique, contributing to the identification of the reticular from of OLP.
Subject(s)
Lichen Planus, Oral , Case-Control Studies , Gingival Crevicular Fluid , Humans , Mouth Mucosa , Spectrophotometry, InfraredABSTRACT
INTRODUCTION: Mineral trioxide aggregate (MTA)-based sealers are endodontic materials with widespread success in distinct clinical applications, potentially embracing direct contact with the bone tissue. Bone response to these materials has been traditionally addressed in vitro. Nonetheless, translational data are limited by the absence of native cell-to-cell and cell-to-matrix interactions that hinder the representativeness of the analysis. Ex vivo organotypic systems, relying on the culture of explanted biological tissues, preserve the cell/tissue composition, reproducing the spatial and organizational in situ complexity. This study was grounded on an innovative research approach, relying on the assessment of an ex vivo organotypic bone tissue culture system to address the osteogenic response to 3 distinct MTA-based sealers. METHODS: Embryonic chick femurs were isolated and grown ex vivo for 11 days in the presence of MTA Plus (Avalon Biomed Inc, Bradenton, FL), ProRoot MTA (Dentsply Tulsa Dental, Hohnson City, Germany), Biodentine (Septodont, Saint Maurdes Fosses, France), or AH Plus (Dentsply Sirona, Konstanz, Germany); the latter was used as a control material. Femurs were characterized by histologic, histochemical, and histomorphometric analysis. Gene expression assessment of relevant osteogenic markers was conducted by quantitative polymerase chain reaction. RESULTS: All MTA-based sealers presented an enhanced osteogenic performance compared with AH Plus. Histochemical and histomorphometric analyses support the increased activation of the osteogenic program by MTA-based sealers, with enhanced collagenous matrix deposition and tissue mineralization. Gene expression analysis supported the enhanced activation of the osteogenic program. Comparatively, ProRoot MTA induced the highest osteogenic functionality on the characterized femurs. CONCLUSIONS: MTA-based sealers enhanced the osteogenic activity within the assayed organotypic bone model, which was found to be a sensitive system for the assessment of osteogenic modulation mediated by endodontic sealers.
Subject(s)
Root Canal Filling Materials , Aluminum Compounds/pharmacology , Bone and Bones , Calcium Compounds/pharmacology , Drug Combinations , Materials Testing , Osteogenesis , Oxides/pharmacology , Root Canal Filling Materials/pharmacology , Silicates/pharmacologyABSTRACT
The development of biomaterials that mimicking the hydroxyapatite nanoparticles existent in the immature bone tissue is crucial, especially to accelerate the bone remodeling and regeneration. In this work, it was developed for the first time, hydroxyapatite nanoparticles (NPs) incorporating citrate and zinc (cit-Zn-Hap) in their composition towards a one-step hydrothermal procedure. For comparison purposes, hydroxyapatite NPs incorporating only zinc (Zn-Hap) or citrate (cit-Hap), as well as hydroxyapatite without any of these elements (Hap) were synthesised. The physicochemical characterization was carried out reveling that, the presence of zinc on hydroxyapatite (cit-Zn-Hap), reduced the size of nanoparticles, changed the phosphate environment and decreased the surface charge when compared with cit-Hap nanoparticles. The osteogenic potential of cit-Zn-Hap NPs was analysed in human bone marrow-derived stromal cells (BMSCs), in the absence of osteoinductive factors. NPs were internalized by endocytosis appearing trapped in endosomes and lysosomes scattered through the cytoplasm. Exposure to these NPs resulted in a significant induction of ALP activity, extracellular matrix mineralization, and gene expression of early and later osteogenic transcription factors, as well as of osteoblastic markers. The osteoinductive effect might be regulated, at least in part, by the increased signalling through the canonical WNT pathway. Evaluation of the cell behaviour following exposure to Zn-Hap and cit-Hap strongly suggested a synergistic effect of citrate and Zn in cit-Zn-Hap NPs towards the induction of the osteogenic commitment and functionality of BMSCs. These findings will allow the design of new biomimetic hydroxyapatite nanoparticles with great potential for bone regeneration.
Subject(s)
Bone Regeneration/drug effects , Durapatite/pharmacology , Mesenchymal Stem Cells/drug effects , Osteogenesis/drug effects , Cells, Cultured , Citrates/chemistry , Durapatite/chemistry , Gene Expression Regulation/drug effects , Humans , Mesenchymal Stem Cells/cytology , Nanotubes , Particle Size , Zinc/chemistryABSTRACT
The clinical demand for bone scaffolds as an alternative strategy for bone grafting has increased exponentially and, up to date, numerous formulations have been proposed to regenerate the bone tissue. However, most of these structures lack at least one of the fundamental/ideal properties of these materials (e.g., mechanical resistance, interconnected porosity, bioactivity, biodegradability, etc.). In this work, we developed innovative composite scaffolds, based on crosslinked chitosan with glutaraldehyde (GA), combined with different atomized calcium phosphates (CaP) granules - hydroxyapatite (HA) or biphasic mixtures of HA and ß - tricalcium phosphate (ß-TCP), with improved biomechanical behavior and enhanced biological response. This innovative combination was designed to improve the scaffolds' functionality, in which GA improved chitosan mechanical strength and stability, whereas CaP granules enhanced the scaffolds' bioactivity and osteoblastic response, further reinforcing the scaffolds' structure. The biological assessment of the composite scaffolds showed that the specimens with 0.2% crosslinking were the ones with the best biological performance. In addition, the inclusion of biphasic granules induced a trend for increase osteogenic activation, as compared to the addition of HA granules. In conclusion, scaffolds produced in the present work, both with HA granules or the biphasic ones, and with low concentrations of GA, have shown adequate properties and enhanced biological performance, being potential candidates for application in bone tissue engineering.
Subject(s)
Bone and Bones/metabolism , Chitosan/chemistry , Durapatite/chemistry , Osteoblasts/metabolism , Osteogenesis , Tissue Scaffolds/chemistry , Bone and Bones/cytology , Cell Line, Tumor , Cross-Linking Reagents/chemistry , Glutaral/chemistry , Humans , Osteoblasts/cytologyABSTRACT
PURPOSE: Eating disorders (EDs) are characterized by a persistent disturbance of eating patterns, leading to poor psychological and physical health. EDs' symptoms are diverse, but their biochemical manifestations can be identified in biofluids, as the gingival crevicular fluid (GCF). The development of a rapid and accurate analytical diagnostic technique, able to provide a wider comprehension of established biochemical abnormalities, would greatly assist EDs' management. Mid-infrared (MIR) spectroscopy gathers all the referred features, and is considered a fingerprint technique. In this pilot trial, the GCF discrimination of patients with EDs and controls was accessed through MIR spectroscopy, further elucidating the relevant spectral differences between both groups. METHODS: GCF was collected from 20 women with ED diagnosis and from age-matched controls. Principal component analysis and partial least squares discriminant analysis (PLSDA) were conducted on GCF MIR spectra. Different PLSDA models were considered to address the predictive capability regarding patient identification, sampling site, and presence of EDs. RESULTS: MIR spectroscopy was capable to discriminate GCF samples, between EDs and controls, with 84.1% of correct predictions. Regression coefficient vectors' analyses revealed that major differences were related to higher protein content in EDs. CONCLUSIONS: Whether further studies are needed to validate the attained data, GCF MIR analysis may be regarded as an innovative, fast, and low-cost technique to assist on early diagnosis and clinical follow-up of EDs' patients. LEVEL OF EVIDENCE: Level IV, case-control trial.
Subject(s)
Feeding and Eating Disorders , Gingival Crevicular Fluid , Case-Control Studies , Female , Humans , Pilot Projects , Spectrum AnalysisABSTRACT
Periodontal diseases remain a challenge due to a complex interplay of factors involving a chronic inflammatory activation and bacteria internalization in periodontal cells. In this work, chitosan-nanoparticles loaded with minocycline (MH-NPs), a tetracycline with antimicrobial and anti-inflammatory effects, were developed for in situ delivery in the periodontal milieu aiming to improve drug effectiveness. A general cytocompatibility evaluation and a detailed approach to address the cellular uptake process, trafficking pathways and the modulation of relevant inflammatory gene expression was conducted using human gingival fibroblasts. Results show that MH-NPs with an adequate cytocompatible profile can be internalized by distinct endocytic processes (macropinocytosis and clathrin-mediated endocytosis). The ability to modulate autophagy with the delivery within the same endosomal/lysosomal pathway as periodontal pathogens was observed, which increases the intracellular drug effectiveness. Porphyromonas gingivalis LPS-stimulated cultures, grown in the presence of MH-NPs, were found to express significantly reduced levels of inflammation-related markers (IL-1b, TNFα, CXCL-8, NFKB1). These nanoparticles can be potentially used in periodontal disease treatment conjoining the ability of intracellular drug targeting with significant anti-inflammatory effects.
Subject(s)
Anti-Bacterial Agents/administration & dosage , Anti-Inflammatory Agents/administration & dosage , Chitosan/chemistry , Minocycline/administration & dosage , Anti-Bacterial Agents/pharmacology , Anti-Inflammatory Agents/pharmacology , Cells, Cultured , Drug Delivery Systems , Fibroblasts/drug effects , Fibroblasts/pathology , Gingiva/cytology , Gingiva/drug effects , Gingiva/microbiology , Humans , Inflammation/drug therapy , Inflammation/pathology , Minocycline/pharmacology , Nanoparticles , Periodontal Diseases/drug therapy , Porphyromonas gingivalis/drug effectsABSTRACT
Nano-hydroxyapatite has been used as an oral care ingredient, being incorporated in several products for the treatment of dental hypersensitivity and enamel remineralisation. Despite its promising results, regulatory and safety concerns have been discussed and questioned by the European Scientific Committee on Consumer Safety (SCCS) regarding the usage of hydroxyapatite nanoparticles in oral care products. In this work, a commercially available nano-hydroxyapatite was characterized and its cytocompatibility towards human gingival fibroblasts was evaluated, as well as its irritation potential using the in vitro HET-CAM assay. All the conditions chosen in this study tried to simulate the tooth brushing procedure and the hydroxyapatite nanoparticles levels normally incorporated in oral care products. The commercial hydroxyapatite nanoparticles used in this study exhibited a rod-like morphology and the expected chemical and phase composition. The set of in vitro cytotoxicity parameters accessed showed that these nanoparticles are highly cytocompatible towards human gingival fibroblasts. Additionally, these nanoparticles did not possess any irritation potential on HET-CAM assay. This study clarifies the issues raised by SCCS and it concludes that this specific nano-hydroxyapatite is cytocompatible, as these nanoparticles did not alter the normal behaviour of the cells. Therefore, they are safe to be used in oral care products.
Subject(s)
Biocompatible Materials/toxicity , Durapatite/toxicity , Fibroblasts/drug effects , Nanoparticles/toxicity , Consumer Product Safety , Cosmetics/toxicity , Humans , Oral HealthABSTRACT
In the quest to fully comprehend the proteolytic events leading to the generation of the salivary peptidome, we have developed a method for the sequential elution of salivary peptides throughout progressive endogenous proteolysis. By screening the time-dependent changes in the salivary peptidome we can predict the activity pattern of salivary proteases responsible for such peptide fingerprint and identify susceptible protein targets. Herein, we describe a step-by-step tutorial based on a filter-aided sample preparation (FASP) method, taking advantage of the endogenous salivary proteases armamentarium (endoProteoFASP), to produce new peptides from the salivary proteins, adding to those present in the sample at the time of collection. In this protocol, the different sets of peptides retrieved after sample elution are identified following a liquid chromatography-tandem mass spectrometry approach. The likelihood of a large set of endogenous proteases (collected from several public sources) to be responsible for the generation of such peptides can be predicted by the analysis of the cleavage site specificity by Proteasix ( http://proteasix.cs.man.ac.uk /) algorithm. The attained peptidome-protease profile can be useful to elucidate the peptidome dynamics and the proteolytic events underpinning pathophysiological phenomena taking place locally within the oral cavity. This may help clinicians to diagnose oral pathologies and develop preventive therapeutic plans.
Subject(s)
Endopeptidases/analysis , Peptide Hydrolases/analysis , Proteomics/methods , Saliva/chemistry , Saliva/metabolism , Salivary Proteins and Peptides/analysis , Tandem Mass Spectrometry/methods , Chromatography, Liquid , Endopeptidases/metabolism , Humans , Peptide Hydrolases/metabolism , Salivary Proteins and Peptides/metabolism , Specimen HandlingABSTRACT
The development of biomaterials for application in advanced therapies requires thorough characterization of its biological behavior, which ultimately entails in vivo compatibility and performance assays. Electrospun fiber membranes of poly(l-lactic acid) (PLLA) and fish gelatin blends were produced and characterized, coupling the biomechanical features of PLLA with gelatin (GEL) biocompatibility. Fiber diameter was not affected by polymer blending, whereas the swelling degree increased with increasing GEL contents for values up to 566 ± 13%, behaving as a superhydrophilic material. Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) adhesion was favored in the PLLA-GEL membranes, and cell viability was not affected after 7 days in culture. Membranes were then evaluated for in vivo biocompatibility through subcutaneous implantation in a rat model, for up to 15 days. No significant differences between the biological behavior of PLLA, PLLA-GEL, and GEL electrospun membranes at 15 days postimplantation were verified, with attained inflammation scores supporting an acceptable tissue response, deeming them fit for further biological assays. This work demonstrates that fiber blends of PLLA and GEL present promising in vitro and in vivo characteristics to be explored for tissue engineering.
ABSTRACT
OBJECTIVE: This case-control study aims to evaluate the oral health status and orofacial problems in a group of outpatients with eating disorders (ED)-either anorexia nervosa (AN) or bulimia nervosa (BN)-further focusing on the influence of vomit. MATERIALS AND METHODS: Fifty-five women outpatients with AN or BN diagnosis were invited to participate, of which 33 agreed. ED outpatients and matched controls were submitted to a questionnaire and clinical oral examination. RESULTS: Multivariate analysis identified a significantly higher incidence of teeth-related complications (i.e., tooth decay, dental erosion, and self-reported dentin hypersensitivity), periodontal disease, salivary alterations (i.e., hyposalivation and xerostomia), and oral mucosa-related complications in ED outpatients. Dental erosion, self-reported dentin hypersensitivity, hyposalivation, xerostomia, and angular cheilitis were found to be highly correlated with the vomiting behavior. CONCLUSIONS: ED outpatients were found to present a higher incidence of oral-related complications and an inferior oral health status, compared to gender- and age-matched controls. Alterations verified within outpatients were acknowledged to be quite similar to those previously reported within inpatients, in both of nature and severity, thus sustaining that the cranio-maxillofacial region is significantly affected by ED, even in the early/milder forms of the condition, as expectedly verified within outpatients.
Subject(s)
Anorexia Nervosa/complications , Bulimia Nervosa/complications , Mouth Diseases/diagnosis , Mouth Diseases/etiology , Oral Health , Vomiting/complications , Adolescent , Adult , Case-Control Studies , Diagnosis, Oral , Female , Humans , Surveys and QuestionnairesABSTRACT
A novel method to process electrospun poly(l-lactic acid) (PLLA) membranes incorporating glass reinforced hydroxyapatite granules (gHA) interspacially between the polymeric fibers is reported, thus increasing the surface area for cellular interactions. gHA granules (≤150µm) electrospun together with the polymer solution, lead to an average fiber diameter of 550±150nm for pristine PLLA and 440±170nm for the composite samples. An increase of the overall porosity was observed, from 79±3% for the PLLA up to 88±5% for the hybrid samples, keeping material's wettability and mechanical properties. Bone-bonding ability showed that both samples induced HA crystal nucleation, but with a distinct pattern of mineral deposition. gHA microcomposite allows a better F-actin cytoskeleton organization during the initial adhesion and spreading, favoring cell-fibers and cell-to-cell interactions and enhanced alkaline phosphatase activity, making them potential candidates for bone healing strategies.
Subject(s)
Bone Regeneration , Bone Substitutes/chemistry , Ceramics/chemistry , Durapatite/chemistry , Osteoclasts/cytology , Polyesters/chemistry , Tissue Scaffolds/chemistry , Cell Line , Humans , Materials Testing , Tissue Engineering , WettabilityABSTRACT
Tissue engineering is constantly evolving towards novel materials that mimic the properties of the replaced injured tissue or organ. A hybrid electrospun membrane of electroactive poly(l-acid lactic) (PLLA) polymer with glass reinforced hydroxyapatite (Bonelike®) microparticles placed among the polymer fibres in a morphology like "islands in the sea" was processed. The incorporation of 60 to 80wt% Bonelike® bone grafts granules with ≤150µm into the polymer solution lead to an amorphous polymeric fibre membranes, and a decrease of the average polymer fibre diameter from 550±150nm for neat PLA down to 440±170nm for the hybrid composite. The presence of Bonelike® in the polymer mats reduced the activation energy for thermal degradation from 134kJ·mol-1, obtained for the neat PLLA membranes down to 71kJ·mol-1, calculated for the hybrid composite membranes. In vitro cell culture results suggest that the developed processing method does not induce cytotoxic effects in MG 63 osteoblastic cells, and creates an environment that enhances cell proliferation, when compared to the neat PLLA membrane. The simplicity and scalability of the processing method suggests a large application potential of this novel hybrid polymer-microparticles fibre membranes for bone regenerative medicine.
Subject(s)
Bone Regeneration/drug effects , Bone Substitutes , Durapatite , Glass/chemistry , Membranes, Artificial , Osteoblasts/metabolism , Polyesters , Animals , Bone Substitutes/chemistry , Bone Substitutes/pharmacology , Cell Line , Cell Proliferation/drug effects , Durapatite/chemistry , Durapatite/pharmacology , Mice , Polyesters/chemistry , Polyesters/pharmacology , Tissue Engineering/methodsABSTRACT
In the present work, we study the development and biological characterization of a polymethyl methacrylate (PMMA)-based minocycline delivery system, to be used as a space maintainer within craniofacial staged regenerative interventions. The developed delivery systems were characterized regarding solid state characteristics and assayed in vitro for antibacterial and anti-inflammatory activity, and cytocompatibility with human bone cells. A drug release profile allowed for an initial burst release and a more sustained and controlled release over time, with minimum inhibitory concentrations for the assayed and relevant pathogenic bacteria (i.e., Staphylococcus aureus, slime-producer Staphylococcus epidermidis and Escherichia coli) being easily attained in the early time points, and sustained up to 72 h. Furthermore, an improved osteoblastic cell response-with enhancement of cell adhesion and cell proliferation-and increased anti-inflammatory activity were verified in developed systems, compared to a control (non minocycline-loaded PMMA cement). The obtained results converge to support the possible efficacy of the developed PMMA-based minocycline delivery systems for the clinical management of complex craniofacial trauma. Here, biomaterials with space maintenance properties are necessary for the management of staged reconstructive approaches, thus minimizing the risk of peri-operative infections and enhancing the local tissue healing and early stages of regeneration.