The effect of a bionic bone ionic environment on osteogenesis, osteoimmunology, and in situ bone tissue engineering.

Bone, a mineralized tissue, continuously undergoes remodeling. It is a process that engages the mineralization and demineralization of the bone matrix, orchestrated by the interactions among cells and cell-secreted biomolecules under the bone ionic microenvironment (BIE). The osteoinductive properties of the demineralized organic bone matrix and many biological factors have been well-investigated. However, the impact of the bone ionic environment on cell differentiation and osteogenesis remains largely unknown. In this study, we extracted and isolated inorganic bone components (bone-derived monetite, BM) using a low-temperature method and, for the first time, investigated whether the BIE could actively affect cell differentiation and regulate osteoimmune reactions. It was evidenced that the BIE could foster the osteogenesis of human bone marrow stromal cells (hBMSCs) and promote hBMSCs mineralization without using osteogenic inductive agents. Interestingly, it was noted that BIE resulted in intracellular mineralization, evidenced by intracellular accumulation of carbonate hydroxyapatite similar to that oberved in osteoblasts cultured in osteoinductive media. Additionally, BIE was found to enhance osteogenesis by generating a favorable osteoimmune environment. In a rat calvarial bone defect model, the osteogenic capacity of BIE was evaluated using a collagen type I-impregnated BM (Col-BM) composite. It showed that Col-BM significantly promoted new bone formation in the critical-size bone defect areas. Taken together, this is the first study that investigated the influence of the BIE on osteogenesis, osteoimmunology, and in situ bone tissue engineering. The innate osteoinductive potential of inorganic bone components, both in vitro and in vivo, not only expands the understanding of the BIE on osteogenesis but also benefits future biomaterials engineering for bone tissue regeneration.

Fluoroamide-Directed Regiodivergent C-Alkylation of Nitroalkanes.

Herein, by exploiting different activation modes of fluoroamides, we achieved α- and δ-C(sp3)-H alkylation of nitroalkanes with switchable regioselectivity. Cu catalysis enabled the interception of a distal C-centered radical by a N-centered radical to couple nitroalkanes and unactivated δ-C-H bonds. In addition, imines generated in situ by fluoroamides were trapped by nitroalkanes to realize the α-C-H alkylation of amides. Both of those scalable protocols have broad substrate scopes and good functional group tolerance.

The Localized Ionic Microenvironment in Bone Modelling/Remodelling: A Potential Guide for the Design of Biomaterials for Bone Tissue Engineering.

Bone is capable of adjusting size, shape, and quality to maintain its strength, toughness, and stiffness and to meet different needs of the body through continuous remodeling. The balance of bone homeostasis is orchestrated by interactions among different types of cells (mainly osteoblasts and osteoclasts), extracellular matrix, the surrounding biological milieus, and waste products from cell metabolisms. Inorganic ions liberated into the localized microenvironment during bone matrix degradation not only form apatite crystals as components or enter blood circulation to meet other bodily needs but also alter cellular activities as molecular modulators. The osteoinductive potential of inorganic motifs of bone has been gradually understood since the last century. Still, few have considered the naturally generated ionic microenvironment's biological roles in bone remodeling. It is believed that a better understanding of the naturally balanced ionic microenvironment during bone remodeling can facilitate future biomaterial design for bone tissue engineering in terms of the modulatory roles of the ionic environment in the regenerative process.

Nanosilicate-Functionalized Polycaprolactone Orchestrates Osteogenesis and Osteoblast-Induced Multicellular Interactions for Potential Endogenous Vascularized Bone Regeneration.

Massive oral and maxillofacial bone defect regeneration remains a major clinical challenge due to the absence of functionalized bone grafts with ideal mechanical and proregeneration properties. In the present study, Laponite (LAP), a synthetic nanosilicate, is incorporated into polycaprolactone (PCL) to develop a biomaterial for bone regeneration. It is explored whether LAP-embedded PCL would accelerate bone regeneration by orchestrating osteoblasts to directly and indirectly induce bone regeneration processes. The results confirmed the presence of LAP in PCL, and LAP is distributed in the exfoliated structure without aggregates. Incorporation of LAP in PCL slightly improved the compressive properties. LAP-embedded PCL is biocompatible and exerts pronounced enhancements in cell viability, osteogenic differentiation, and extracellular matrix formation of osteoblasts. Furthermore, osteoblasts cultured on LAP-embedded PCL facilitate angiogenesis of vessel endothelial cells and alleviate osteoclastogenesis of osteoclasts in a paracrine manner. The addition of LAP to the PCL endows favorable bone formation in vivo. Based upon these results, LAP-embedded PCL shows great potential as an ideal bone graft that exerts both space-maintaining and vascularized bone regeneration synergistic effects and can be envisioned for oral and maxillofacial bone defect regeneration.

Morphometric Changes of Osteocyte Lacunar in Diabetic Pig Mandibular Cancellous Bone.

Osteocytes play an important role in bone metabolism. The interactions of osteocytes with the surrounding microenvironment can alter cellular and lacunar morphological changes. However, objective quantification of osteocyte lacunae is challenging due to their deep location in the bone matrix. This project established a novel method for the analytical study of osteocytes/lacunae, which was then used to evaluate the osteocyte morphological changes in diabetic pig mandibular bone. Eight miniature pigs were sourced, and diabetes was randomly induced in four animals using streptozotocin (STZ) administration. The mandibular tissues were collected and processed. The jawbone density was evaluated with micro-CT. Osteocyte lacunae were effectively acquired and identified using backscattered electron scanning microscopy (BSE). A significantly decreased osteocyte lacunae size was found in the diabetic group. Using the acid etching method, it was demonstrated that the area of osteocyte and lacunae, and the pericellular areas were both significantly reduced in the diabetes group. In conclusion, a standard and relatively reliable method for analyzing osteocyte/lacunae morphological changes under compromised conditions has been successfully established. This method demonstrates that diabetes can significantly decrease osteocyte/lacunae size in a pig's mandibular cancellous bone.

Vascularized bone regeneration accelerated by 3D-printed nanosilicate-functionalized polycaprolactone scaffold.

Critical oral-maxillofacial bone defects, damaged by trauma and tumors, not only affect the physiological functions and mental health of patients but are also highly challenging to reconstruct. Personalized biomaterials customized by 3D printing technology have the potential to match oral-maxillofacial bone repair and regeneration requirements. Laponite (LAP) nanosilicates have been added to biomaterials to achieve biofunctional modification owing to their excellent biocompatibility and bioactivity. Herein, porous nanosilicate-functionalized polycaprolactone (PCL/LAP) was fabricated by 3D printing technology, and its bioactivities in bone regeneration were investigated in vitro and in vivo. In vitro experiments demonstrated that PCL/LAP exhibited good cytocompatibility and enhanced the viability of bone marrow mesenchymal stem cells (BMSCs). PCL/LAP functioned to stimulate osteogenic differentiation of BMSCs at the mRNA and protein levels and elevated angiogenic gene expression and cytokine secretion. Moreover, BMSCs cultured on PCL/LAP promoted the angiogenesis potential of endothelial cells by angiogenic cytokine secretion. Then, PCL/LAP scaffolds were implanted into the calvarial defect model. Toxicological safety of PCL/LAP was confirmed, and significant enhancement of vascularized bone formation was observed. Taken together, 3D-printed PCL/LAP scaffolds with brilliant osteogenesis to enhance bone regeneration could be envisaged as an outstanding bone substitute for a promising change in oral-maxillofacial bone defect reconstruction.

Targeting Early Healing Phase with Titania Nanotube Arrays on Tunable Diameters to Accelerate Bone Regeneration and Osseointegration.

Blood coagulation and inflammation are the earliest biological responses to implant surfaces. Implant nano-surfaces can significantly impact the osseointegration through the influence on the early phase of bone regeneration. However, the interplay between blood clot property and inflammatory reaction on nanosurfaces is rarely understood. Herein, titania nanotube arrays (TNAs) with different diameters are fabricated on titanium. In vitro evaluation with the whole blood indicates that TNA with a diameter of 15 nm (TNA 15) enables noteworthy platelet activation resulting in distinct clot features compared with that of pure Ti and TNA with a diameter of 120 nm (TNA 120). Further co-culture with macrophages on the clot or in the clot-conditioned medium shows that the clot on TNA 15 downregulates the inflammation and manipulates a favorable osteoimmunomodulatory environment for osteogenesis. In vivo studies further demonstrate that TNA 15 could downregulate the inflammation-related genes while upregulating growth metabolism-related genes in an early healing hematoma. Additionally, TNA 15 promotes de novo bone formation with improved extending of osteocyte dendrites, demonstrating the desired osseointegration. These findings indicate that surface nano-dimensions can significantly influence clot formation and appropriate clot features can manipulate a favorable osteoimmunomodulatory environment for bone regeneration and osseointegration.

Epigenetic changes caused by diabetes and their potential role in the development of periodontitis.

AIMS/INTRODUCTION: Periodontal disease, a chronic inflammation induced by bacteria, is closely linked with diabetes mellitus. Many complications associated with diabetes are related to epigenetic changes. However, the exact epigenetic changes whereby diabetes affects periodontal disease remain largely unknown. Thus, we sought to investigate the role of diabetes-dependent epigenetic changes of gingival tissue in the susceptibility to periodontal disease. MATERIALS AND METHODS: We studied the effect of streptozotocin-induced diabetes in minipigs on gingival morphological and epigenetic tissue changes. Accordingly, we randomly divided six minipigs into two groups: streptozotocin-induced diabetes group, n = 3; and non-diabetes healthy control group, n = 3. After 85 days, all animals were killed, and gingival tissue was collected for histology, deoxyribonucleic acid methylation analysis and immunohistochemistry. RESULTS: A diabetes mellitus model was successfully created, as evidenced by significantly increased blood glucose levels, reduction of pancreatic insulin-producing ß-cells and histopathological changes in the kidneys. The gingival tissues in the diabetes group presented acanthosis of both gingival squamous epithelium and sulcular/junctional epithelium, and a significant reduction in the number and length of rete pegs. Deoxyribonucleic acid methylation analysis showed a total of 1,163 affected genes, of which 599 and 564 were significantly hypermethylated and hypomethylated, respectively. Immunohistochemistry staining showed that the hypomethylated genes - tumor necrosis factor-α and interleukin-6 - were positively expressed under the junctional epithelium area in the diabetes group. CONCLUSIONS: Diabetes mellitus induces morphological and epigenetic changes in periodontal tissue, which might contribute to the increased susceptibility of periodontal diseases in patients with diabetes.

Graphene oxide coated Titanium Surfaces with Osteoimmunomodulatory Role to Enhance Osteogenesis.

Graphene oxide (GO) and its derivatives are currently being explored for the modification of bone biomaterials. However, the effect of GO coatings on immunoregulation and subsequent impacts on osteogenesis are not known. In this study, GO was coated on pure titanium using dopamine. GO-coated titanium (Ti-GO) surfaces exhibited good biocompatibility, with the ability to stimulate the expression of osteogenic genes, and extracellular matrix mineralization in human mesenchymal stromal cells (hMSCs). Interestingly, it was found that GO-coated surfaces could manipulate the polarization of macrophages and expression of inflammatory cytokines via the Toll-like receptor pathway. Under physiological conditions, Ti-GO activated macrophages and induced mild inflammation and a pro-osteogenic environment, characterized by a slight increase in the levels of proinflammatory cytokines, as well as increased expression of the TGF-ß1 and oncostatin M genes. In an environment mimicking acute inflammatory conditions, Ti-GO attenuated inflammatory responses, as shown by the downregulation of proinflammatory cytokines. Conditioned medium collected from macrophages stimulated by Ti-GO played a significant stimulatory role in the osteogenic differentiation of hMSCs. In summary, GO-coated surfaces displayed beneficial immunomodulatory effects in osteogenesis, indicating that GO could be a potential substance for the modification of bone scaffolds and implants.

Estimating Some General Molecular Descriptors of Saturated Hydrocarbons.

Three general molecular descriptors, namely the general sum-connectivity index, general Platt index and ordinary generalized geometric-arithmetic index, are studied here. Best possible bounds for the aforementioned descriptors of arbitrary saturated hydrocarbons are derived under certain constraints. These bounds are expressed in terms of number of carbon atoms and number of carbon-carbon bonds of the considered hydrocarbons.

The role of organic phosphate in the spatial control of periodontium complex bio-mineralization: an in vitro study.

The periodontal structure is a particularly exquisite model of hierarchical spatial control of mineralization. Extracellular matrix control in the selective mineralization of the periodontium complex remains elusive since the extracellular matrix is a set of mineralization promoters and inhibitors. The phosphorylated proteins, which are ubiquitous in the extracellular matrix of the periodontium complex, are well-documented as primary factors in the regulation of tissue mineralization. Whether organic phosphates are key regulators in defining the interfaces between dentin, cementum, periodontal ligament and alveolar bone is an issue worthy of research. Here, we investigated the in vitro remineralization process of demineralized and dephosphorylated periodontal tissue sections. When exposed to a metastable mineralization solution, a large number of calcospherulites deposited on the surface of the dephosphorylated sections and the tissue selective mineralization were disrupted. Interestingly, on adding a dentin matrix protein-1 analogue named polyacrylic acid, the surface mineralization rate in the dephosphorylated periodontal complex reduced dramatically. In contrast, hierarchical mineralization was displayed by the demineralized section at the tissue collagen fibrillar levels in both alveolar bone and dentin regions. These results demonstrated that the organic phosphate could prevent surface mineral deposition, and the minerals could penetrate the collagen fibrils to initiate a selective and hierarchal tissue mineralization with the assistance of the dentin matrix protein-1 analogue in the periodontal complex. This study enhances our understanding of the mineralization discrepancy in the periodontal tissues, which will provide some insight into the development of biomaterials for the regeneration of soft-hard tissue interfaces.

Aberrant activation of Wnt signaling pathway altered osteocyte mineralization.

Mineralization of bone is a dynamic process, involving a complex interplay between cells, secreted macromolecules, signaling pathways, and enzymatic reactions; the dysregulation of bone mineralization may lead to serious skeletal disorders, including hypophosphatemic rickets, osteoporosis, and rheumatoid arthritis. Very few studies have reported the role of osteocytes - the most abundant bone cells in the skeletal system and the major orchestrators of bone remodeling in bone mineralization, which is owed to their nature of being deeply embedded in the mineralized bone matrix. The Wnt/ß-catenin signaling pathway is actively involved in various life processes including osteogenesis; however, the role of Wnt/ß-catenin signaling in the terminal mineralization of bone, especially in the regulation of osteocytes, is largely unknown. This research demonstrates that during the terminal mineralization process, the Wnt/ß-catenin pathway is downregulated, and when Wnt/ß-catenin signaling is activated in osteocytes, dendrite development is suppressed and the expression of dentin matrix protein 1 (DMP1) is inhibited. Aberrant activation of Wnt/ß-catenin signaling in osteocytes leads to the spontaneous deposition of extra-large mineralized nodules on the surface of collagen fibrils. The altered mineral crystal structure and decreased bonding force between minerals and the organic matrix indicate the inferior integration of minerals and collagen. In conclusion, Wnt/ß-catenin signaling plays a critical role in the terminal differentiation of osteocytes and as such, targeting Wnt/ß-catenin signaling in osteocytes may serve as a potential therapeutic approach for the management of bone-related diseases.

The influence of high-dose systemic zoledronate administration on osseointegration of implants with different surface topography.

AIM: To evaluate the influence of systemic zoledronate administration on the osseointegration of titanium implants with different surface topography in rat maxillae. METHODS: Twenty Sprague-Dawley rats were divided into two groups-test (bisphosphonate) and control (healthy). Bisphosphonate administration began three weeks prior to implant placement, and the animals received zoledronate (66 µg/kg) three times per week. Forty endosseous implants with a moderately rough (20 implants) or a turned surface (20 implants) were immediately placed bilaterally into extraction sockets of maxillary first molars. Animals were sacrificed after 14 and 28 days of healing, and en bloc specimens were harvested for histological and histomorphometric analysis. Osseointegration was quantified by measuring the percentage of bone-to-implant contact. RESULTS: Bone-to-implant contact (BIC) (mean ± SD) values of moderately rough and turned implants at day 14 in test group were 17.62 ± 6.68 and 10.69 ± 1.48, respectively, while in the control group, they were 46.36 ± 5.08 and 33.29 ± 8.89, respectively. At day 28, BIC values of moderately rough and turned implants in the test group were 25.94 ± 7.87 and 7.83 ± 4.30, respectively, while in the control group, they were 72.99 ± 6.60 and 47.62 ± 18.19, respectively. Statistically significant higher BIC values were measured on moderately rough implants compared to turned implants at 28 days, and the control group compared to the test group for both implant surfaces. Histological observations for the control and the test groups demonstrated initial bone formation around moderately rough implants not only on the surface of the parent bone, as was the case with the turned surfaced implants, but also along the implant surface itself. CONCLUSIONS: Systemic zoledronate administration negatively influences osseointegration. Osseointegration was enhanced adjacent to moderately rough compared to turned implants in both the presence and absence of systemic zoledronate administration. Therefore, topographical surface modification may partially offset the negative impact of zoledronate administration.

Effect of ovariectomy on tissue-level changes in rat maxilla.

PURPOSE: It remains unclear whether estrogen deficiency affects the ultrastructure and tissue-level mechanical properties of the maxilla. The hypothesis of this study was that the ovariectomized rat could induce tissue-level changes of the maxilla. MATERIALS AND METHODS: Twelve 3-month-old female Sprague Dawley rats were acquired and randomly divided into two groups: ovariectomized and SHAM (control) (n = 6 for each group). Three months after the ovariectomy, implants were placed; the animals were sacrificed at day 28, and then samples were collected and prepared according to the previously established protocols. Advanced analytical equipment including scanning electron microscope with energy-dispersive spectrometry, transmission electron microscope, and nanoindentation were used to analyze bone quality. RESULTS: The results showed that the mature bone areas in the ovariectomized group were significantly affected in the mineral crystal and the microstructure. The micro-mechanical properties of the mature bone were also affected, showing significantly increased hardness (H) and reduced modulus (Er) in ovariectomized rats compared with the normal rats (P < .05). Differences in H and Er in new bone areas between the normal and ovariectomized rats were less significant. CONCLUSION: Ovariectomy affected maxilla bone tissue-level quality; however, the effects mainly existed in the mature bone areas, which were characterized by higher crystalline mineralization, hardness, and modulus.

Alkanes with the First Three Maximal/Minimal Modified First Zagreb Connection Indices.

The modified first Zagreb connection index ( Z C 1 * ) is a molecular descriptor, which was initially appeared within a formula of the total electron energy of alternant hydrocarbons in 1972. In a recent paper [A. Ali, N. Trinajstic, A novel/old modification of the first Zagreb index, Mol. Inform. 37 (2018) 1800008], it was observed that the molecular descriptor Z C 1 * correlates well with the entropy and acentric factor of octane isomers. In this article, the molecules with the first three maximal Z C 1 * values as well as the first three minimal Z C 1 * values are determined from the family of all alkanes with n carbon atoms, for n ≥ 6 . This extends the main results of the aforementioned paper.

The Alkanes with Maximum Wiener Polarity Index.

The Wiener polarity index (usually denoted by W p ) of an alkane is the number of unordered pairs of carbon atoms which are separated by three carbon-carbon bonds. This topological index W p is useful for predicting the boiling points of alkanes. Deng [MATCH Commun. Math. Comput. Chem. 66 (2011) 305] proved that the maximum W p value among all alkanes, with n carbon atoms, is 3 n - 15 . The main purpose of present paper is to find all those alkanes with n carbon atoms, which attain the maximum value of W p .

Differential effect of hydroxyapatite nano-particle versus nano-rod decorated titanium micro-surface on osseointegration.

Coating materials applied for intraosseous implants must be optimized to stimulate osseointegration. Osseointegration is a temporal and spatial physiological process that not only requires interactions between osteogenesis and angiogenesis but also necessitates a favorable immune microenvironment. It is now well-documented that hierarchical nano-micro surface structures promote the long-term stability of implants, the interactions between nano-micro structure and the immune response are largely unknown. Here, we report the effects of microporous titanium (Ti) surfaces coated with nano-hydroxyapatite (HA) produced by micro-arc oxidation and steam-hydrothermal treatment (SHT) on multiple cell behavior and osseointegration. By altering the processing time of SHT it was possible to shift HA structures from nano-particles to nano-rods on the microporous Ti surfaces. Ti surfaces coated with HA nano-particles were found to modulate the inflammatory response resulting in an osteoimmune microenvironment more favorable for osteo-/angio-genesis, most likely via the activation of certain key signaling pathways (TGF-ß, OPG/RANKL, and VEGF). By contrast, Ti surfaces coated with nano-rod shaped HA particles had a negative impact on osteo-/angio-genesis and osteoimmunomodulation. In vivo results further demonstrated that Ti implant surfaces decorated with HA nano-particles can stimulate new bone formation and osseointegration with enhanced interaction between osteocytes and implant surfaces. This study demonstrated that Ti implants with micro-surfaces coated with nano-particle shaped HA have a positive impact on osseointegration. STATEMENT OF SIGNIFICANCE: Osteo-/angio-genesis are of importance during osteointegration of the implants. Recent advances unravel that immune response of macrophages and its manipulated osteoimmunomodulation also exerts a pivotal role to determine the fate of the implant. Surface nano-micro modification has evidenced to be efficient to influence osteogenesis, however, little is known links nano-microstructured surface to immune response, as well the osteoimmunomodulation. This study demonstrates that the nano-particles decorated micro-surface, compared with the nano-rods decorated micro-surface enables osteogenesis and angiogenesis concurrently that has not been investigated previously. This study also unravels that the immune response of macrophages can be manipulated by the nano-micro surface, especially the nano-dimension matters, leading to a differential effect on osteointegration. The additional knowledge obtained from this study may provide foundation and reference for future design of the coating materials for implantable materials.

The inverse Wiener polarity index problem for chemical trees.

The Wiener polarity number (which, nowadays, known as the Wiener polarity index and usually denoted by Wp) was devised by the chemist Harold Wiener, for predicting the boiling points of alkanes. The index Wp of chemical trees (chemical graphs representing alkanes) is defined as the number of unordered pairs of vertices (carbon atoms) at distance 3. The inverse problems based on some well-known topological indices have already been addressed in the literature. The solution of such inverse problems may be helpful in speeding up the discovery of lead compounds having the desired properties. This paper is devoted to solving a stronger version of the inverse problem based on Wiener polarity index for chemical trees. More precisely, it is proved that for every integer t ∈ {n - 3, n - 2,…,3n - 16, 3n - 15}, n ≥ 6, there exists an n-vertex chemical tree T such that Wp(T) = t.

Human oral microbiota and its modulation for oral health.

The oral microbiome is an important part of the human microbiome. The oral cavity contains several significantly different niches with distinct microbial communities. A wide range of microorganisms inhabit the human oral cavity, including bacteria, fungi, viruses, archaea and protozoa. These microorganisms form a complex ecological community that influences oral and systemic health. The most prevalent oral diseases, dental caries and periodontal diseases, are microbiota-associated diseases. Moreover, increasing evidences have supported that many systemic diseases are associated with disturbances in the oral ecosystem, such as diabetes, cardiovascular diseases and tumors. The current control of dental plaque-related diseases is nonspecific and is centered on the removal of plaque by mechanical means. Due to this realization about the oral microbiome, several new methods based on the modulation of the microbiome that aim at maintaining and reestablishing a healthy oral ecosystem have been developed.

The minimal-ABC trees with B1-branches.

The atom-bond connectivity index (or, for short, ABC index) is a molecular structure descriptor bridging chemistry to graph theory. It is probably the most studied topological index among all numerical parameters of a graph that characterize its topology. For a given graph G = (V, E), the ABC index of G is defined as [Formula: see text], where di denotes the degree of the vertex i, and ij is the edge incident to the vertices i and j. A combination of physicochemical and the ABC index properties are commonly used to foresee the bioactivity of different chemical composites. Additionally, the applicability of the ABC index in chemical thermodynamics and other areas of chemistry, such as in dendrimer nanostars, benzenoid systems, fluoranthene congeners, and phenylenes is well studied in the literature. While finding of the graphs with the greatest ABC-value is a straightforward assignment, the characterization of the tree(s) with minimal ABC index is a problem largely open and has recently given rise to numerous studies and conjectures. A B1-branch of a graph is a pendent path of order 2. In this paper, we provide an important step forward to the full characterization of these minimal trees. Namely, we show that a minimal-ABC tree contains neither 4 nor 3 B1-branches. The case when the number of B1-branches is 2 is also considered.