Nano eggshell-based slurry as a direct pulp-capping material: In vitro characterization and histopathological assessment in an experimental animal model.

AIM: Pulp vitality is essential for tooth integrity. Following pulp exposure, choosing a suitable pulp-capping material is crucial to maintain pulp vitality. However, the reparative dentine bridge created by calcium hydroxide (Ca(OH)2 ) is generally porous and incomplete. The aim of the current study is to assess the in vitro and in vivo bioactivities of nano eggshell-based slurry (NES), using NES as a direct pulp-capping material, compared with Ca(OH)2 in rabbit animal model. METHODOLOGY: Nano eggshell powder (NE) was characterized for particle morphology, chemical composition and ion release. In vitro bioactivity was tested by immersion in simulated body fluid (SBF) for 7 days. For histopathological evaluation, 36 adult New Zealand rabbits (72 pulp exposures) were divided into nine groups (n = 8) according to the pulp-capping material (NES, Ca(OH)2 and no capping as negative control group) and the animals were sacrificed after 7, 14 or 28 days. The pulps of the two lower central incisors were exposed and then directly capped by Ca(OH)2 or NES or left untreated. The cavities were then sealed with glass ionomer cement. Teeth were collected for histopathological evaluation using an optical microscope. Pulp haemorrhage, inflammation, fibrosis and calcific bridge formation were assessed. Results were statistically analysed using anova and Tukey's tests. RESULTS: Nano eggshell particles were spherical with a 20 nm diameter and were composed mainly of calcite. Statistical analysis showed that there was a significant increase in the release of all investigated ions between days 1 and 28, except for copper. NES group showed a significantly higher release of all elements as compared to Ca(OH)2 . Environmental scanning electron microscope micrographs of NES incubated for 7 days in SBF showed the formation of HAp with a Ca/P ratio (1.686). For histopathological evaluation, the difference between groups was statistically significant. At day 28, 75% of the pulps of the Ca(OH)2 group showed mild calcific bridge in comparison with 100% moderate calcific bridge in the NES group. The NES group showed significantly less inflammation at days 7 and 28, and higher fibrosis at day 7 compared with Ca(OH)2 . CONCLUSIONS: Nano eggshell-based slurry represents a promising novel direct pulp-capping material with favourable pulp tissue response.

Effect of immersion and thermocycling in different beverages on the surface roughness of single- and multi-shade resin composites.

BACKGROUND: Resin composite restorations are highly esthetic restorations, which should have and maintain high surface polish. However, esthetic restorations are subjected to different beverages at variable temperatures, which may affect their surface roughness. This study aimed to evaluate the surface roughness of single-shade (Omnichroma) and multi-shade (Filtek Z350XT) composite materials, following aging by immersion and thermocycling in different beverages, simulating one year of clinical service. METHODS: Thirty specimens of each material were prepared and divided into 6 subgroups (n = 5). In each material, the grouping of the specimens was as follows: the first subgroup was the as-prepared specimens stored dry without immersion or thermocycling. The second, third, and fourth subgroups were immersed in saliva, tea, and red wine, respectively, for 12 days at 37 °C. The fifth and sixth subgroups were thermocycled for 10,000 cycles, in tea (the fifth between 37 and 57 °C) and in red wine (the sixth between 37 °C and12°C). The resultant surface roughness was measured by two different methods, stylus profilometer and atomic force microscopy (AFM). Intergroup comparison was performed using independent t test, while intragroup comparison was performed using one-way analysis of variance (ANOVA) followed by Tukey's post-hoc test. RESULTS: Intergroup comparison between both composites showed no statistically significant differences in all groups using the stylus profilometer roughness measurements (P>0.05), while the AFM measurements showed significant difference (P ≤ 0.05) within all storage media except the as-prepared control (P = 0.0645), where nanofilled Filtek Z350 XT showed lower nano-roughness. Intragroup comparison data were variable, depending on the material, aging conditions, and roughness assessment tool. However, the resultant average surface roughness (Ra) values in all groups did not exceed the threshold value of Ra 0.2 µm. CONCLUSIONS: Both resin composites attained and retained a clinically acceptable surface finish after immersion and thermocycling in different beverages.

Molecular Basis beyond Interrelated Bone Resorption/Regeneration in Periodontal Diseases: A Concise Review.

Periodontitis is the sixth most common chronic inflammatory disease, destroying the tissues supporting the teeth. There are three distinct stages in periodontitis: infection, inflammation, and tissue destruction, where each stage has its own characteristics and hence its line of treatment. Illuminating the underlying mechanisms of alveolar bone loss is vital in the treatment of periodontitis to allow for subsequent reconstruction of the periodontium. Bone cells, including osteoclasts, osteoblasts, and bone marrow stromal cells, classically were thought to control bone destruction in periodontitis. Lately, osteocytes were found to assist in inflammation-related bone remodeling besides being able to initiate physiological bone remodeling. Furthermore, mesenchymal stem cells (MSCs) either transplanted or homed exhibit highly immunosuppressive properties, such as preventing monocytes/hematopoietic precursor differentiation and downregulating excessive release of inflammatory cytokines. In the early stages of bone regeneration, an acute inflammatory response is critical for the recruitment of MSCs, controlling their migration, and their differentiation. Later during bone remodeling, the interaction and balance between proinflammatory and anti-inflammatory cytokines could regulate MSC properties, resulting in either bone formation or bone resorption. This narrative review elaborates on the important interactions between inflammatory stimuli during periodontal diseases, bone cells, MSCs, and subsequent bone regeneration or bone resorption. Understanding these concepts will open up new possibilities for promoting bone regeneration and hindering bone loss caused by periodontal diseases.

Effect of two artificial aging protocols on color and gloss of single-shade versus multi-shade resin composites.

The long-term color stability and gloss retention of resin composites are among the crucial factors that affect the clinical longevity of esthetic restorations, especially in anterior teeth. This study evaluated the effect of artificial aging by immersion in different storage media and thermocycling on color and gloss of dental single-shade resin composite (Omnichroma) versus multi-shade one (Filtek Z350XT). One hundred resin-composite disc-shaped specimens were used, 50 from each group, Omnichroma and Filtek Z350XT. Ten specimens from each material acted as control group (incubated in saliva). For each material, 40 specimens were divided according to the artificial-aging protocol (immersion at 37 °C for 12 days or thermocycling for 10,000 cycles) and storage media (tea, red wine). Color and gloss were measured before and after artificial aging. Color difference (∆E00) was compared with perceptibility threshold and acceptability threshold. Data were statistically analyzed; independent t test was used to compare results between two tested materials, while two-way ANOVA was used to compare results among the different immersion media within the same material. Artificial aging (immersion or thermocycling) in tea and red wine led to significant color changes and gloss reduction in both materials (P < 0.05), in contrast to control group. Red wine produced highest color differences. Both dental resin-composites; the single-shade (Omnichroma) and multi-shade (Filtek Z350XT) displayed unacceptable discoloration and gloss reduction after artificial-aging in tea and red-wine by immersion or thermocycling simulating one-year clinical-service.

Effect of Polymeric Matrix Stiffness on Osteogenic Differentiation of Mesenchymal Stem/Progenitor Cells: Concise Review.

Mesenchymal stem/progenitor cells (MSCs) have a multi-differentiation potential into specialized cell types, with remarkable regenerative and therapeutic results. Several factors could trigger the differentiation of MSCs into specific lineages, among them the biophysical and chemical characteristics of the extracellular matrix (ECM), including its stiffness, composition, topography, and mechanical properties. MSCs can sense and assess the stiffness of extracellular substrates through the process of mechanotransduction. Through this process, the extracellular matrix can govern and direct MSCs' lineage commitment through complex intracellular pathways. Hence, various biomimetic natural and synthetic polymeric matrices of tunable stiffness were developed and further investigated to mimic the MSCs' native tissues. Customizing scaffold materials to mimic cells' natural environment is of utmost importance during the process of tissue engineering. This review aims to highlight the regulatory role of matrix stiffness in directing the osteogenic differentiation of MSCs, addressing how MSCs sense and respond to their ECM, in addition to listing different polymeric biomaterials and methods used to alter their stiffness to dictate MSCs' differentiation towards the osteogenic lineage.

Hydrogels and Dentin-Pulp Complex Regeneration: From the Benchtop to Clinical Translation.

Dentin-pulp complex is a term which refers to the dental pulp (DP) surrounded by dentin along its peripheries. Dentin and dental pulp are highly specialized tissues, which can be affected by various insults, primarily by dental caries. Regeneration of the dentin-pulp complex is of paramount importance to regain tooth vitality. The regenerative endodontic procedure (REP) is a relatively current approach, which aims to regenerate the dentin-pulp complex through stimulating the differentiation of resident or transplanted stem/progenitor cells. Hydrogel-based scaffolds are a unique category of three dimensional polymeric networks with high water content. They are hydrophilic, biocompatible, with tunable degradation patterns and mechanical properties, in addition to the ability to be loaded with various bioactive molecules. Furthermore, hydrogels have a considerable degree of flexibility and elasticity, mimicking the cell extracellular matrix (ECM), particularly that of the DP. The current review presents how for dentin-pulp complex regeneration, the application of injectable hydrogels combined with stem/progenitor cells could represent a promising approach. According to the source of the polymeric chain forming the hydrogel, they can be classified into natural, synthetic or hybrid hydrogels, combining natural and synthetic ones. Natural polymers are bioactive, highly biocompatible, and biodegradable by naturally occurring enzymes or via hydrolysis. On the other hand, synthetic polymers offer tunable mechanical properties, thermostability and durability as compared to natural hydrogels. Hybrid hydrogels combine the benefits of synthetic and natural polymers. Hydrogels can be biofunctionalized with cell-binding sequences as arginine-glycine-aspartic acid (RGD), can be used for local delivery of bioactive molecules and cellularized with stem cells for dentin-pulp regeneration. Formulating a hydrogel scaffold material fulfilling the required criteria in regenerative endodontics is still an area of active research, which shows promising potential for replacing conventional endodontic treatments in the near future.

Dental Stem Cell-Derived Secretome/Conditioned Medium: The Future for Regenerative Therapeutic Applications.

Regenerative medicine literature has proposed mesenchymal stem/progenitor cell- (MSC-) mediated therapeutic approaches for their great potential in managing various diseases and tissue defects. Dental MSCs represent promising alternatives to nondental MSCs, owing to their ease of harvesting with minimally invasive procedures. Their mechanism of action has been attributed to their cell-to-cell contacts as well as to the paracrine effect of their secreted factors, namely, secretome. In this context, dental MSC-derived secretome/conditioned medium could represent a unique cell-free regenerative and therapeutic approach, with fascinating advantages over parent cells. This article reviews the application of different populations of dental MSC secretome/conditioned medium in in vitro and in vivo animal models, highlights their significant implementation in treating different tissue' diseases, and clarifies the significant bioactive molecules involved in their regenerative potential. The analysis of these recent studies clearly indicate that dental MSCs' secretome/conditioned medium could be effective in treating neural injuries, for dental tissue regeneration, in repairing bone defects, and in managing cardiovascular diseases, diabetes mellitus, hepatic regeneration, and skin injuries, through regulating anti-inflammatory, antiapoptotic, angiogenic, osteogenic, and neurogenic mediators.

Mesenchymal Stem/Progenitor Cells: The Prospect of Human Clinical Translation.

Mesenchymal stem/progenitor cells (MSCs) are key players in regenerative medicine, relying principally on their differentiation/regeneration potential, immunomodulatory properties, paracrine effects, and potent homing ability with minimal if any ethical concerns. Even though multiple preclinical and clinical studies have demonstrated remarkable properties for MSCs, the clinical applicability of MSC-based therapies is still questionable. Several challenges exist that critically hinder a successful clinical translation of MSC-based therapies, including but not limited to heterogeneity of their populations, variability in their quality and quantity, donor-related factors, discrepancies in protocols for isolation, in vitro expansion and premodification, and variability in methods of cell delivery, dosing, and cell homing. Alterations of MSC viability, proliferation, properties, and/or function are also affected by various drugs and chemicals. Moreover, significant safety concerns exist due to possible teratogenic/neoplastic potential and transmission of infectious diseases. Through the current review, we aim to highlight the major challenges facing MSCs' human clinical translation and shed light on the undergoing strategies to overcome them.

Antibacterial activity and biocompatibility of zein scaffolds containing silver-doped bioactive glass.

Composite 3D scaffolds combining natural polymers and bioceramics are promising candidates for bone tissue engineering (BTE). Zein, as a natural plant protein, offers several advantages, including biocompatibility, adequate strength properties, and low/no immunogenicity; however, it lacks bioactivity. Thus, composite zein: bioactive glass (BG) scaffolds are proposed as promising candidate for BTE applications, with silver-doping of bioactive glass providing an antibacterial effect against possible post-implantation infection. Therefore, the aim of this study was to investigate the in vitro antibacterial properties, biocompatibility, bioactivity and compressive strength of zein scaffolds containing silver-doped bioactive glass. BG nanoparticles, undoped and Ag-doped, were fabricated using the sol-gel method. 3D composite zein:BG scaffolds, containing 20 wt% BG, were prepared and their antibacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) was assessed using the disc diffusion assay. Human osteoblast-like MG-63 cells were used to evaluate the in vitro biocompatibility of the prepared scaffold groups. In addition, the compressive strength of the scaffolds was determined using uniaxial compression strength testing and the scaffold interconnected porosity was measured using helium pycnometer. Disc diffusion assay showed that only zein scaffolds containing Ag-doped sol-gel BG are antibacterially positive against E. coli and S. aureus. Pure zein scaffolds and zein scaffolds containing sol-gel-derived BG showed no negative influence on the growth of MG-63 cells, as evident by the cells' ability to survive, proliferate, and function on these scaffolds. Moreover, incorporating sol-gel-derived BG into zein scaffolds at zein:BG of 80:20 ratio showed bioactive properties with adequate porosity without affecting the scaffolds' compressive strengths, which was similar to that of trabecular bone, suggesting that the new composites have potential for BTE applications in non-loaded bearing areas.

Regenerating bone with bioactive glass scaffolds: A review of in vivo studies in bone defect models.

Large bone defects resulting from fractures and disease are a medical concern, being often unable to heal spontaneously by the body's repair mechanisms. Bone tissue engineering (BTE) is a promising approach for treating bone defects through providing a template to guide osseous regeneration. 3D scaffolds with microstructure mimicking host bone are necessary in common BTE strategies. Bioactive glasses (BGs) attract researchers' attention as BTE scaffolds as they are osteoconductive and osteoinductive in certain formulations. In vivo animal models allow understanding and evaluation of materials' performance in the complex physiological environment, being an inevitable step before clinical trials. The aim of this paper is to review for the first time published research investigating the in vivo osseous regenerative capacity of 3D BG scaffolds in bone defect animal models, to better understand and evaluate the progress and future outlook of the use of such scaffolds in BTE. The literature analysis reveals that the regenerative capacity of BG scaffolds depends on several factors; including BG composition, fabrication method, scaffold microstructure and pore characteristics, in addition to scaffold pretreatment and whether or not the scaffolds are loaded with growth factors. In addition, animal species selected, defect size and implantation time affect the scaffold in vivo behavior and outcomes. The review of the literature also makes clear the difficulty encountered to compare different types of bioactive glass scaffolds in their bone forming ability. Even considering such limitations of the current state-of-the-art, results generated from animal bone defect models provide an essential source of information to guide the design of BG scaffolds in future. STATEMENT OF SIGNIFICANCE: Bioactive glasses are at the centre of increasing research efforts in bone tissue engineering as the number of research groups around the world carrying out research on this type of biomaterials continues to increase. However, there are no previous reviews in literature which specifically cover investigations of the performance of bioactive glass scaffolds in bone defect animal models. This is the topic of the present review, in which we have analysed comprehensively all available literature in the field. The review thus fills a gap in the biomaterials literature providing a broad platform of information for researchers interested in bioactive glasses in general and specifically in the outcomes of in vivo models. Bioactive glass scaffolds of different compositions tested in relevant bone defect models are covered.

Chemical and biological evaluation of Egyptian Nile Tilapia (Oreochromis niloticas) fish scale collagen.

Collagen is considered to be one of the most useful biomaterials with different medical applications. However, collagen properties differ from one source to another. The aim of this study was to extract, purify, characterize and perform preliminary biological evaluation of type I collagen from scales of Egyptian Nile Tilapia. Pepsin-solubilized collagen (PSC) was successfully prepared from Nile Tilapia fish scale waste. Lyophilized collagen was dissolved in dilute HCl to form acidic collagen solutions (ACS) which was neutralized to form gel. To confirm the biocompatibility of the produced gel, baby hamster kidney (BHK-21) fibroblast cells were seeded onto a 3D collagen gel (0.3% and 0.5%, w/v). The results of an SDS-PAGE test showed that the extracted collagens were type I collagen, with α chain composition of (α1)2α2. Thermal analysis showed that the denaturation temperature was 32 °C. X-ray diffraction (XRD) analysis and Fourier-transform infrared spectra (FTIR) showed that the extracted collagen had a triple helix structure. Active proliferation of BHK-21 cells with no signs of toxicity was evident with both collagen gel concentrations tested. The results show that Nile Tilapia scales can be an effective source of collagen extraction that could be used as a potential biomaterial in biomedical applications.