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Abstract Asiatic acid (AA) and asiaticoside, pentacyclic triterpenoid compounds derived from Centella asiatica, are known for their biological effects in promoting type I collagen synthesis and inducing osteogenesis of stem cells. However, their applications in regenerative medicine are limited due to their low potency and poor aqueous solubility. This work aimed to evaluate the osteogenic induction activity of AA derivatives in human periodontal ligament stem cells (hPDLSCs) in vitro. Four compounds were synthesised, namely 501, 502, 503, and 506. AA was used as the control. The 502 exhibited low water solubility, while the 506 compound showed the highest. The cytotoxicity analysis demonstrated that 503 caused significant deterioration in cell viability, while other derivatives showed no harmful effect on hPDLSCs. The dimethyl aminopropyl amine derivative of AA, compound 506, demonstrated a relatively high potency in inducing osteogenic differentiation. An elevated mRNA expression of osteogenic-related genes, BMP2, WNT3A, ALP, OSX and IBSP was observed with 506. Additionally, the expression of BMP-2 protein was enhanced with increasing dose of 506, and the effect was pronounced when the Erk signalling molecule was inhibited. The 506 derivative was proposed for the promotion of osteogenic differentiation in hPDLSCs by upregulating BMP2 via the Erk signalling pathway. The 506 molecule showed promise in bone tissue regeneration.

Abstract Objectives The aim of this study was to investigate the effect of mechanical force on possible dynamic changes of the matrix proteins deposition in the PDL upon in vitro mechanical and in vivo occlusal forces in a rat model with hypofunctional conditions. Materials and methods Intermittent compressive force (ICF) and shear force (SF) were applied to human periodontal ligament stem cells (PDLSCs). Protein expression of collagen I and POSTN was analyzed by western blot technique. To establish an in vivo model, rat maxillary molars were extracted to facilitate hypofunction of the periodontal ligament (PDL) tissue of the opposing mandibular molar. The mandibles were collected after 4-, 8-, and 12-weeks post-extraction and used for micro-CT and immunohistochemical analysis. Results ICF and SF increased the synthesis of POSTN by human PDLSCs. Histological changes in the hypofunctional teeth revealed a narrowing of the PDL space, along with a decreased amount of collagen I, POSTN, and laminin in perivascular structures compared to the functional contralateral molars. Conclusion Our results revealed that loss of occlusal force disrupts deposition of some major matrix proteins in the PDL, underscoring the relevance of mechanical forces in maintaining periodontal tissue homeostasis by modulating ECM composition.

Abstract Background Light spectra have been demonstrated to result in different levels of comfort or stress, which affect plant growth and the availability of health-promoting compounds in ways that sometimes contradict one another. To determine the optimal light conditions, it is necessary to weigh the vegetable’s mass against the amount of nutrients it contains, as vegetables tend to grow poorly in environments where nutrient synthesis is optimal. This study investigates the effects of varying light conditions on the growth of red lettuce and its occurring nutrients in terms of productivities, which were determined by multiplying the total weight of the harvested vegetables by their nutrient content, particularly phenolics. Three different light-emitting diode (LED) spectral mixes, including blue, green, and red, which were all supplemented by white, denoted as BW, GW, and RW, respectively, as well as the standard white as the control, were equipped in grow tents with soilless cultivation systems for such purposes. Results Results demonstrated that the biomass and fiber content did not differ substantially across treatments. This could be due to the use of a modest amount of broad-spectrum white LEDs, which could help retain the lettuce’s core qualities. However, the concentrations of total phenolics and antioxidant capacity in lettuce grown with the BW treatment were the highest (1.3 and 1.4-fold higher than those obtained from the control, respectively), with chlorogenic acid accumulation (8.4 ± 1.5 mg g− 1 DW) being particularly notable. Meanwhile, the study observed a high glutathione reductase (GR) activity in the plant achieved from the RW treatment, which in this study was deemed the poorest treatment in terms of phenolics accumulation. Conclusion In this study, the BW treatment provided the most efficient mixed light spectrum to stimulate phenolics productivity in red lettuce without a significant detrimental effect on other key properties.

When fossil fuel substitution with biomass is viewed as a potential solution to global warming caused by greenhouse gas emissions, the demand for biomass fuel pellets has increased worldwide. Although agricultural waste is an attractive potential feedstock for fuel pellet production due to its relatively high calorific value and low cost, its excessive ash content is a major drawback. This research investigates the properties of sugarcane bagasse fuel pellets treated by dry and wet torrefaction and evaluates the economic value of selling the fuel pellets, which were priced based on their quality. It was found that the wet torrefaction could significantly reduce the ash content in the product (1% ash content at a torrefaction temperature of above 180°C), resulting in higher quality and more marketable fuel pellets. Consequently, the yield and the net present value of the production of wet torrefied fuel pellets were greater than those of dry torrefied pellets. Nevertheless, the production of fuel pellets from sugarcane bagasse treated by either process is shown to be economically viable.

Abstract The self-organizing potential of induced pluripotent stem cells (iPSCs) represents a promising tool for bone tissue engineering. Shear stress promotes the osteogenic differentiation of mesenchymal stem cells, leading us to hypothesize that specific shear stress could enhance the osteogenic differentiation of iPSCs. For osteogenesis, embryoid bodies were formed for two days and then maintained in medium supplemented with retinoic acid for three days, followed by adherent culture in osteogenic induction medium for one day. The cells were then subjected to shear loading (0.15, 0.5, or 1.5 Pa) for two days. Among different magnitudes tested, 0.5 Pa induced the highest levels of osteogenic gene expression and greatest mineral deposition, corresponding to upregulated connexin 43 (Cx43) and phosphorylated Erk1/2 expression. Erk1/2 inhibition during shear loading resulted in decreased osteogenic gene expression and the suppression of mineral deposition. These results suggest that shear stress (0.5 Pa) enhances the osteogenic differentiation of iPSCs, partly through Cx43 and Erk1/2 signaling. Our findings shed light on the application of shear-stress technology to improve iPSC-based tissue-engineered bone for regenerative bone therapy.

Objective: Basic fibroblast growth factor (bFGF) plays a pivotal role in cell proliferation, differentiation and extracellular matrix turnover in various tissues. In human dental pulp cells (HDPCs), let-7 microRNA is involved in cell proliferation and differentiation. There is little information on the effect of bFGF-induced cell proliferation on let-7 microRNA in HDPCs. This study investigated the effect of bFGF on let-7g, let-7f, and let-7i microRNAs and some of the genes involved in cell proliferation including p53 and Ki67 in HDPCs. Material and Methods: HDPCs were cultured and treated with bFGF at 0, 1, and 5 ng/mL. Cell proliferation was examined using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay at 24 and 48 hours. Additionally, gene expressions of let-7g, let-7f, let-7i microRNAs, and p53 and Ki67 were examined by quantitative real-time polymerase chain reaction at 24 hours. All experiments were performed in triplicate. Results: The results showed that let-7g, let-7f, and let-7i microRNAs were expressed in HDPCs. MTT assays showed that bFGF induced greater cell proliferation than the controls at 24 and 48 hours (p-value0.050). Conclusion: Our preliminary study showed that exogenous bFGF could decrease let-7g microRNA expression suggesting that let-7g microRNA may be involved in bFGF-induced HDPCs proliferation.

Abstract Canine mesenchymal stem cells (cMSCs) have potential applications for regenerative therapy, including the generation of insulin-producing cells (IPCs) for studying and treating diabetes. In this study, we established a useful protocol for generating IPCs from canine adipose mesenchymal stem cells (cAD-MSCs). Subsequently, in vitro preservation of pluronic F127-coated alginate (ALGPA)-encapsulated cAD-MSC-derived IPCs was performed to verify ready-to-use IPCs. IPCs were induced from cAD-MSCs with the modulated three-stepwise protocol. The first step of definitive endoderm (DE) induction showed that the cooperation of Chir99021 and Activin A created the effective production of Sox17-expressed DE cells. The second step for pancreatic endocrine (PE) progenitor induction from DE indicated that the treatment with taurine, retinoic acid, FGF2, EGF, TGFβ inhibitor, dorsomorphin, nicotinamide, and DAPT showed the significant upregulation of the pancreatic endocrine precursor markers Pdx1 and Ngn3. The last step of IPC production, the combination of taurine, nicotinamide, Glp-1, forskolin, PI3K inhibitor, and TGFβ inhibitor, yielded efficiently functional IPCs from PE precursors. Afterward, the maintenance of ALGPA-encapsulated cAD-MSC-derived IPCs with VSCBIC-1, a specialized medium, enhanced IPC properties. Conclusion, the modulated three-stepwise protocol generates the functional IPCs. Together, the encapsulation of cAD-MSC-derived IPCs and the cultivation with VSCBIC-1 enrich the maturation of generated IPCs.

Abstract In vitro manipulation of induced pluripotent stem cells (iPSCs) by environmental factors is of great interest for three-dimensional (3D) tissue/organ induction. The effects of mechanical force depend on many factors, including force and cell type. However, information on such effects in iPSCs is lacking. The aim of this study was to identify a molecular mechanism in iPSCs responding to intermittent compressive force (ICF) by analyzing the global gene expression profile. Embryoid bodies of mouse iPSCs, attached on a tissue culture plate in 3D form, were subjected to ICF in serum-free culture medium for 24 h. Gene ontology analyses for RNA sequencing data demonstrated that genes differentially regulated by ICF were mainly associated with metabolic processes, membrane and protein binding. Topology-based analysis demonstrated that ICF induced genes in cell cycle categories and downregulated genes associated with metabolic processes. The Kyoto Encyclopedia of Genes and Genomes database revealed differentially regulated genes related to the p53 signaling pathway and cell cycle. qPCR analysis demonstrated significant upregulation of Ccnd1, Cdk6 and Ccng1. Flow cytometry showed that ICF induced cell cycle and proliferation, while reducing the number of apoptotic cells. ICF also upregulated transforming growth factor β1 (Tgfb1) at both mRNA and protein levels, and pretreatment with a TGF-β inhibitor (SB431542) prior to ICF abolished ICF-induced Ccnd1 and Cdk6 expression. Taken together, these findings show that TGF-β signaling in iPSCs enhances proliferation and decreases apoptosis in response to ICF, that could give rise to an efficient protocol to manipulate iPSCs for organoid fabrication.

Intermittent compressive force influences human periodontal ligament (PDL) cell behavior that facilitates periodontal tissue regeneration. In response to mechanical stimuli, Yes-associated protein (YAP) has been recognized as a mechanosensitive transcriptional activator that regulates cell proliferation and cell fate decisions. This study aimed to investigate whether compressive forces influence cell proliferation and cell fate decisions of human PDL cells via YAP signaling. YAP expression was silenced by shRNA. The effect of YAP on cell proliferation, adipogenesis and osteogenesis of PDL cells under ICF loading were determined. Adipogenic differentiation bias upon ICF loading was confirmed by fourier-transform infrared spectroscopy (FTIR). The results revealed that ICF-induced YAP promotes osteogenesis, but it inhibits adipogenesis in PDL cells. Depletion of YAP results in PDL cells that are irresponsive to ICF and, therefore, the failure of the PDL cells to undergo osteogenic differentiation. This was shown by a significant reduction in calcium deposited in the CF-derived osteoblasts of the YAP-knockdown (YAP-KD) PDL cells. As to control treatment, reduction of YAP promoted adipogenesis, whereas ICF-induced YAP inhibited this mechanism. However, the adipocyte differentiation in YAP-KD cells was not affected upon ICF treatment as the YAP-KD cells still exhibited a better adipogenic differentiation that was unrelated to the ICF. This study demonstrated that, in response to ICF treatment, YAP could be a crucial mechanosensitive transcriptional activator for the regulation of PDL cell behavior through a mechanobiological process. Our results may provide the possibility of facilitating PDL tissue regeneration by manipulation of the Hippo-YAP signaling pathway.

Abstract Platelet-rich fibrin (PRF) promotes wound healing by providing the release of growth factors. Here, the influence of Thai and Murrah bubaline blood derived PRF on canine periodontal ligament cells (cPDLs) was investigated. PRF was prepared from Thai and Murrah buffaloes with single centrifugation. Results demonstrated that Thai bubaline blood derived PRF exhibited fiber-mesh like morphology and contained more platelet entrapment than Murrah bubaline blood derived PRF. Both bubaline PRFs were able to degrade in vitro under condition with trypsin. Thai but not Murrah bubaline blood derived PRF promoted cPDLs proliferation in serum free and 2% serum culture conditions. Correspondingly, the significant upregulation of KI67 mRNA expression was observed in those cells treated with Thai bubaline blood derived PRF. However, both Thai and Murrah bubaline blood derived PRF accelerated cell migration in an in vitro wound healing assay and facilitated cell spreading. Further, cPDLs cultured in osteogenic induction medium supplemented with Thai bubaline blood derived PRF exhibited the increased mineral deposition in vitro. Frozen Thai bubaline blood derived PRF also promoted cell proliferation, KI67 mRNA expression, cell migration, and cell spreading in cPDLs. Taken these evidence together, bubaline blood derived PRF could provide potential benefits for canine periodontal tissue healing.

Abstract The trend of regenerative therapy for diabetes in human and veterinary practices has conceptually been proven according to the Edmonton protocol and animal models. Establishing an alternative insulin-producing cell (IPC) resource for further clinical application is a challenging task. This study investigated IPC generation from two practical canine mesenchymal stem cells (cMSCs), canine bone marrow-derived MSCs (cBM-MSCs) and canine adipose-derived MSCs (cAD-MSCs). The results illustrated that cBM-MSCs and cAD-MSCs contain distinct pancreatic differentiation potential and require the tailor-made induction protocols. The effective generation of cBM-MSC-derived IPCs needs the integration of genetic and microenvironment manipulation using a hanging-drop culture of PDX1-transfected cBM-MSCs under a three-step pancreatic induction protocol. However, this protocol is resource- and time-consuming. Another study on cAD-MSC-derived IPC generation found that IPC colonies could be obtained by a low attachment culture under the three-step induction protocol. Further, Notch signaling inhibition during pancreatic endoderm/progenitor induction yielded IPC colonies through the trend of glucose-responsive C-peptide secretion. Thus, this study showed that IPCs could be obtained from cBM-MSCs and cAD-MSCs through different induction techniques. Also, further signaling manipulation studies should be conducted to maximize the protocol’s efficiency.

Periodontal ligament (PDL) cells help maintain tissue homeostasis by balancing PDL tissue inflammation and regeneration. However, the mechanisms by which interferon γ (IFNγ) modulate this process are not yet fully understood. The present study aimed to examine the effect of primed and non-primed PDL cells with IFNγ on the viability and differentiation of T lymphocytes and its functional consequences. The results showed that IFNγ-primed PDL cells possessed enhanced immunosuppression by suppressing T-lymphocyte viability and directing T-lymphocyte differentiation towards a higher T helper (Th) Th2/Th1 ratio. Suppression of T-cell viability was mainly mediated by IFNγ-inducible secreted mediators, which was prevented in the presence of direct cell contact, probably by intercellular adhesion molecule-1 (ICAM-1)-induced PI3 K-mediated transforming growth factor β1 expression in PDL cells. By contrast, ICAM-1 activation augmented IFNγ-induced IFNγ and interleukin-6 expression in PDL cells, which in turn modulated T-cell differentiation. The resulting interaction between these two cell types activated macrophage and suppressed osteoclast differentiation. In conclusion, the results have shown, for the first time to our knowledge, that primed and non-primed PDL cells with IFNγ differentially control T-cell responses via IFNγ-inducible mediators and ICAM-1-mediated direct cell contact, suggesting the role of PDL cells in shifting an inflammatory phase towards a regenerative phase.

Abstract Background Direct pulp capping is a vital pulp therapy for a pin-point dental pulp exposure. Applying a pulp capping material leads to the formation of a dentin bridge and protects pulp vitality. The aim of this study was to compare the effects of four dental materials, DyCal®, ProRoot® MTA, Biodentine™, and TheraCal™ LC in vitro. Methods Human dental pulp stem cells (hDPs) were isolated and characterized. Extraction medium was prepared from the different pulp capping materials. The hDP cytotoxicity, proliferation, and migration were examined. The odonto/osteogenic differentiation was determined by alkaline phosphatase, Von Kossa, and alizarin red s staining. Osteogenic marker gene expression was evaluated using real-time polymerase chain reaction. Results ProRoot® MTA and Biodentine™ generated less cytotoxicity than DyCal® and TheraCal™ LC, which were highly toxic. The hDPs proliferated when cultured with the ProRoot® MTA and Biodentine™ extraction media. The ProRoot® MTA and Biodentine™ extraction medium induced greater cell attachment and spreading. Moreover, the hDPs cultured in the ProRoot® MTA or Biodentine™ extraction medium migrated in a similar manner to those in serum-free medium, while a marked reduction in cell migration was observed in the cells cultured in DyCal® and TheraCal™ LC extraction media. Improved mineralization was detected in hDPs maintained in ProRoot® MTA or Biodentine™ extraction medium compared with those in serum-free medium. Conclusion This study demonstrates the favorable in vitro biocompatibility and bioactive properties of ProRoot® MTA and Biodentine™ on hDPs, suggesting their superior regenerative potential compared with DyCal® and TheraCal™.

This study aims to analyse the potential environmental impact of natural rubber (NR) condoms over its entire life cycle. The environmental performance of the production process of NR condoms is also compared to that of synthetic polyisoprene (PI) condoms. Options to reduce the environmental impact of condom production are proposed and evaluated. The potential environmental impacts are quantified by life cycle assessment (LCA). The study takes a cradle-to-grave approach, and considers six phases: 1) fresh latex production, 2) concentrated latex production, 3) condom production, 4) condom use, 5) condom transportation, and 6) condom disposal. The comparative analysis of NR and PI condom production takes a gate-to-gate scope. The functional unit is one gross condom. The results indicate that the condom production phase has the largest share (34–73%) in the environmental impact during the life cycle of NR condoms, followed by the disposal phase (20–60%). Comparing the production of NR and PI condoms reveals that the production of PI condoms results in a 1.5–2.5 times higher environmental impacts than the NR condom production, due to higher electricity consumption, especially for the compounding, dipping, and leaching processes. Reduction of electricity use is a key measure to reduce the environmental impact. Combining natural gas with electricity for the dipping process is a promising option to reduce the electricity use; it results in ~10–17% reduction in environmental impacts.

Abstract Background Current approach for diabetes treatment remained several adverse events varied from gastrointestinal to life-threatening symptoms. Regenerative therapy regarding Edmonton protocol has been facing serious limitations involving protocol efficiency and safety. This led to the study for alternative insulin-producing cell (IPC) resource and transplantation platform. In this study, evaluation of encapsulated human dental pulp-derived stem cell (hDPSC)-derived IPCs by alginate (ALG) and pluronic F127-coated alginate (ALGPA) was performed. Results The results showed that ALG and ALGPA preserved hDPSC viability and allowed glucose and insulin diffusion in and out. ALG and ALGPA-encapsulated hDPSC-derived IPCs maintained viability for at least 336 h and sustained pancreatic endoderm marker (NGN3), pancreatic islet markers (NKX6.1, MAF-A, ISL-1, GLUT-2 and INSULIN), and intracellular pro-insulin and insulin expressions for at least 14 days. Functional analysis revealed a glucose-responsive C-peptide secretion of ALG- and ALGPA-encapsulated hDPSC-derived IPCs at 14 days post-encapsulation. Conclusion ALG and ALGPA encapsulations efficiently preserved the viability and functionality of hDPSC-derived IPCs in vitro and could be the potential transplantation platform for further clinical application.

Tissues engineering has gained a lot of interest, since this approach has potential to restore lost tooth-supporting structures, which is one of the biggest challenges for periodontal treatment. In this study, we aimed to develop an in situ hydrogel that could conceivably support and promote the regeneration of lost periodontal tissues. The hydrogel was fabricated from methacrylated hyaluronic acid (MeHA). Fragment/short-chain hyaluronic acid (sHA) was incorporated in this hydrogel to encourage the bio-synergistic effects of two different molecular weights of hyaluronic acid. The physical properties of the hydrogel system, including gelation time, mechanical profile, swelling and degrading behavior, etc., were tested to assess the effect of incorporated sHA. Additionally, the biological properties of the hydrogels were performed in both in vitro and in vivo models. The results revealed that sHA slightly interfered with some behaviors of networking systems; however, the overall properties were not significantly changed compared to the base MeHA hydrogel. In addition, all hydrogel formulations were found to be compatible with oral tissues in both in vitro and in vivo models. Therefore, this HA-based hydrogel could be a promising delivery system for low molecular weight macromolecules. Further, this approach could be translated into the clinical applications for dental tissue regeneration.

Denosumab, an anti-receptor activator of nuclear factor-kappa B ligand antibody (anti-RANKL), is a fully human monoclonal antibody (mAb) available for the treatment of osteoporosis. In the present study, an anti-RANKL mAb was transiently expressed using the geminiviral expression system in Nicotiana benthamiana, and the functional activity of the plant-produced mAb was determined. The highest expression level of the plant-produced mAb was found at 8 days post-infiltration, and it was estimated to be 0.5 mg/g leaf fresh weight. The recombinant mAb from the plant crude extracts was purified by using Protein A affinity column chromatography. The plant-produced mAb demonstrated good in vitro affinity binding with human RANKL, as determined by RANKL-ELISA binding. The function of the plant-produced mAb was evaluated in vitro. CD14-positive cells isolated from human peripheral blood mononuclear cells (PBMCs) were cultured in vitro in the presence of human RANKL and macrophage-colony-stimulating factor (M-CSF) to stimulate osteoclastogenesis. The results demonstrated that plant-produced mAb could significantly decrease the number of osteoclasts compared to commercial denosumab. These results demonstrated that the plant-produced mAb has the potential to inhibit osteoclast differentiation and that it could be considered for osteoporosis treatment.

Relevant immunomodulatory effects have been proposed following allogeneic cell-based therapy with human periodontal ligament stem cells (hPDLSCs). This study aimed to examine the influence of shear stress on the immunosuppressive capacity of hPDLSCs. Cells were subjected to shear stress at different magnitudes (0.5, 5 and 10 dyn/cm2). The expression of immunosuppressive markers was evaluated in shear stress-induced hPDLSCs using qRT-PCR, western blot, enzyme activity and enzyme-linked immunosorbent assays. The effects of a shear stress-derived condition medium (SS-CM) on T cell proliferation were examined using a resazurin assay. Treg differentiation was investigated using qRT-PCR and flow cytometry analysis. Our results revealed that shear stress increased mRNA expression of IDO and COX2 but not TGF-β1 and IFN-γ. IDO activity, kynurenine and active TGF-β1 increased in SS-CM when compared to the non-shear stress-derived conditioned medium (CTL-CM). The amount of kynurenine in SS-CM was reduced in the presence of cycloheximide and ERK inhibitor. Subsequently, T cell proliferation decreased in SS-CM compared to CTL-CM. Treg differentiation was promoted in SS-CM, indicated by FOXP3, IL-10 expression and CD4+CD25hiCD127lo/− subpopulation. In conclusion, shear stress promotes kynurenine production through ERK signalling in hPDLSC, leading to the inhibition of T cell proliferation and the promotion of Treg cell differentiation.

The data contained within this article relate to a rotating rod within a stationary ring that was used to generate shear stress on cells and tissues via a medium. The geometry of the rotating rod within a stationary ring was designed to work with a 35-mm diameter culture dish. The data of the shear stress distribution are presented in terms of area-weighted average shear stress and the uniformity index, which were calculated for medium volumes of 4 and 5 ml at various rotational speeds ranging from 0 to 1000 rpm.

Abstract Background Basic fibroblast growth factor (bFGF) regulates maintenance of stemness and modulation of osteo/odontogenic differentiation and mineralization in stem cells from human exfoliated deciduous teeth (SHEDs). Mineralization in the bones and teeth is in part controlled by pericellular levels of inorganic phosphate (Pi), a component of hydroxyapatite, and inorganic pyrophosphate (PPi), an inhibitor of mineralization. The progressive ankylosis protein (gene ANKH; protein ANKH) and ectonucleotide pyrophosphatase phosphodiesterase 1 (ENPP1/ENPP1) increase PPi and inhibit mineralization, while tissue-nonspecific alkaline phosphatase (ALPL; TNAP) is a critical pro-mineralization enzyme that hydrolyzes PPi. We hypothesized that regulation by bFGF of mineralization in SHEDs occurs by modulation of Pi/PPi-associated genes. Methods Cells were isolated from human exfoliated deciduous teeth and characterized for mesenchymal stem cell characteristics. Cells were treated with bFGF, and the osteogenic differentiation ability was determined. The mRNA expression was evaluated using real-time polymerase chain reaction. The mineralization was examined using alizarin red S staining. Results Cells isolated from primary teeth expressed mesenchymal stem cell markers, CD44, CD90, and CD105, and were able to differentiate into osteo/odontogenic and adipogenic lineages. Addition of 10 ng/ml bFGF to SHEDs during in vitro osteo/odontogenic differentiation decreased ALPL mRNA expression and ALP enzyme activity, increased ANKH mRNA, and decreased both Pi/PPi ratio and mineral deposition. Effects of bFGF on ALPL and ANKH expression were detected within 24 h. Addition of 20 mM fibroblast growth factor receptor (FGFR) inhibitor SU5402 revealed the necessity of FGFR-mediated signaling, and inclusion of 1 μg/ml cyclohexamide (CHX) implicated the necessity of protein synthesis for effects on ALPL and ANKH. Addition of exogenous 10 μm PPi inhibited mineralization and increased ANKH, collagen type 1a1 (COL1A1), and osteopontin (SPP1) mRNA, while addition of exogenous Pi increased mineralization and osterix (OSX), ANKH, SPP1, and dentin matrix protein 1 (DMP1) mRNA. The effects of PPi and Pi on mineralization could be replicated by short-term 3- and 7-day treatments, suggesting signaling effects in addition to physicochemical regulation of mineral deposition. Conclusion This study reveals for the first time the effects of bFGF on Pi/PPi regulators in SHEDs and implicates these factors in how bFGF directs osteo/odontogenic differentiation and mineralization by these cells.

Torrefaction is a promising biomass thermal conversion technology to produce biochar due to its ease of operation and mild operating conditions. In this study, cassava rhizome (CR) was torrified under various sweeping gas types (nitrogen (N2), carbon dioxide (CO2), mixture gas (N2 + CO2)) and flow rates (50, 150, 250 mL/min) at 200–300 °C for 30 min. The experimental results show that fuel properties of CR were remarkably upgraded after torrefaction. Sweeping gas has less effect on fuel properties of torrified CR than torrefaction temperature. Torrefaction under CO2 atmosphere produced the biochar with minimum ash content. Torrefaction at 300 °C under 50 mL/min CO2 was recommended as the promising condition to produce biochar replacing lignite coal. Thermal properties and chemical functional groups of the derived biochar suggested that torrefaction process removed mainly oxygen and hydrogen contents which could be achieved through decarbonization (DC), dehydrogenation (DH), and deoxygenation (DO) pathways. Economic feasibility revealed that the torrefaction of CR is cost-advantage under the proposed condition.

Ethanol is a promising biofuel that can replace fossil fuel, mitigate greenhouse gas (GHG) emissions, and represent a renewable building block for biochemical production. Ethanol can be produced from various feedstocks. First-generation ethanol is mainly produced from sugar- and starch-containing feedstocks. For second-generation ethanol, lignocellulosic biomass is used as a feedstock. Typically, ethanol production contains four major steps, including the conversion of feedstock, fermentation, ethanol recovery, and ethanol storage. Each feedstock requires different procedures for its conversion to fermentable sugar. Lignocellulosic biomass requires extra pretreatment compared to sugar and starch feedstocks to disrupt the structure and improve enzymatic hydrolysis efficiency. Many pretreatment methods are available such as physical, chemical, physicochemical, and biological methods. However, the greatest concern regarding the pretreatment process is inhibitor formation, which might retard enzymatic hydrolysis and fermentation. The main inhibitors are furan derivatives, aromatic compounds, and organic acids. Actions to minimize the effects of inhibitors, detoxification, changing fermentation strategies, and metabolic engineering can subsequently be conducted. In addition to the inhibitors from pretreatment, chemicals used during the pretreatment and fermentation of byproducts may remain in the final product if they are not removed by ethanol distillation and dehydration. Maintaining the quality of ethanol during storage is another concerning issue. Initial impurities of ethanol being stored and its nature, including hygroscopic, high oxygen and carbon dioxide solubility, influence chemical reactions during the storage period and change ethanol’s characteristics (e.g., water content, ethanol content, acidity, pH, and electrical conductivity). During ethanol storage periods, nitrogen blanketing and corrosion inhibitors can be applied to reduce the quality degradation rate, the selection of which depends on several factors, such as cost and storage duration. This review article sheds light on the techniques of control used in ethanol fuel production, and also includes specific guidelines to control ethanol quality during production and the storage period in order to preserve ethanol production from first-generation to second-generation feedstock. Finally, the understanding of impurity/inhibitor formation and controlled strategies is crucial. These need to be considered when driving higher ethanol blending mandates in the short term, utilizing ethanol as a renewable building block for chemicals, or adopting ethanol as a hydrogen carrier for the long-term future, as has been recommended.

The data contained within this article relate to experimental data on mechanical behavior of an in-house cast hyperelastic Polydimethysiloxane (PDMS) based membrane (SilasticⓇ T-4, Dow Corning) for cell culture, which was used as a tool for applying tensile stress on cells and tissues. With the experimentally obtained material constants, tensile stress distribution over the membrane surface was numerically assessed using a finite element analysis (FEA). The membrane area having a uniform tensile stress distribution under different strain loading conditions of 1–20% was suggested for cell culture.

Microspace culture is promising for self-organization of induced pluripotent stem cells (iPSCs). However, the optimal size of microspaces for osteogenic differentiation is unclear. We hypothesized that a specific microspace size could facilitate self-organizing iPSC differentiation to form bone-like tissue in vitro. The objectives of this study were to investigate such effects of microspace size and to evaluate bone regeneration upon transplantation of the resulting osteogenic constructs. Dissociated mouse gingival fibroblast-derived iPSCs were plated in ultra-low-attachment microspace culture wells containing hundreds of U-bottom-shaped microwell spots per well to form cell aggregates in growth medium. The microwells had different aperture diameters/depths (400/560 μm (Elp400), 500/700 μm (Elp500), and 900/700 μm (Elp900)) (Kuraray; Elplasia). After 5 days of aggregation, cells were maintained in osteogenic induction medium for 35 days. Only cells in the Elp500 condition tightly aggregated and maintained high viability during osteogenic induction. After 10 days of induction, Elp500 cell constructs showed significantly higher gene expression of Runx2, Osterix, Collagen 1a1, Osteocalcin, Bone sialoprotein, and Osteopontin compared to constructs in Elp400 and Elp900. In methylene blue-counterstained von Kossa staining and Movat’s pentachrome staining, only Elp500 constructs showed robust osteoid formation on day 35, with high expression of type I collagen (a major osteoid component) and osteocalcin proteins. Cell constructs were transplanted into rat calvarial bone defects, and micro-CT analysis after 3 weeks showed better bone repair with significantly higher bone mineral density in the Elp500 group compared to the Elp900 group. These results suggest that microspace size affects self-organized osteogenic differentiation of iPSCs. Elp500 microspace culture specifically induces mouse iPSCs into osteoid-rich bone-like tissue possessing high bone regeneration capacity.

Indirect immobilized ligand has been shown as an effective technique to activate Notch signalling in vitro. The data presented in this article are related to the published article entitled “Indirect immobilized Jagged1 suppresses cell cycle progression and induces odonto/osteogenic differentiation in human dental pulp cells” (Manokawinchoke et al. 2017) [1]. This data article describes gene expression in indirect immobilized Jagged1 treated human dental pulp cells (hDPs) using high throughput RNA sequencing technique. These data are valuable to analyze the regulation of Notch signalling in hDPs for understanding its molecular mechanism(s). Raw RNA sequencing data were deposited in the NCBI Sequence Read Archive (SRP100068) and NCBI Gene Expression Omnibus (GSE94989).

In this work, surface disinfection and biofilm susceptibility were investigated by applying ionic silver of 0.4–1.6 µg/mL and cathodic voltage-controlled electrical treatment of 1.8 V and a current of 30 mA to Escherichia coli (E. coli) ATCC 25922 biofilm-contaminated titanium substrates. Herein, it is evident that the treatment exhibited the potential use to enhance the susceptibility of bacterial biofilms for surface disinfection. In vitro studies have demonstrated that the ionic silver treatment of 60 min significantly increased the logarithmic reduction (LR) of bacterial populations on disinfectant-treated substrates and the electrical treatment enhanced the silver susceptibility of E. coli biofilms. The LR values after the ionic silver treatments and the electric-enhanced silver treatments were in the ranges of 1.94–2.25 and 2.10–2.73, respectively. The treatment was also associated with morphological changes in silver-treated E. coli cells and biofilm-contaminated titanium surfaces. Nevertheless, the treatments showed no cytotoxic effects on the L929 mouse skin fibroblast cell line and only a slight decrease in pH was observed during the electrical polarization of titanium substrate.

Periodontal disease is the most common oral disease in dogs. Platelet-rich fibrin (PRF) is widely utilized to facilitate soft and hard tissue healing and has been proposed in periodontal healing in small animal treatment. However, the quality and amount of autologous PRF is compromised in animals with systemic diseases. The present study aimed to evaluate the efficacy of xenologous bubaline blood-derived PRF (bPRF) on periodontal tissue healing in canine periodontal defects. Split-mouth design was employed in twenty dogs diagnosed with periodontal disease. The defects were divided randomly into two groups: the open-flap debridement (OFD)-treated group and the OFD with bPRF (OFD+bPRF) application group. Results demonstrated that gingival index and periodontal probing depth decreased significantly in the OFD+bPRF group compared with those treated with OFD alone. Application of bPRF in periodontal defects also promoted fibrous tissue formation, as confirmed by the marked increase in fibrosis score. bPRF application significantly increased COL1A1 and PDGFB mRNA levels at day 14 compared with the baseline. Taking this evidence together, bPRF provided a favorable therapeutic modality in canine periodontal defects. bPRF could be an alternative biomaterial for the treatment of periodontal defects in dogs.

Nanocrystalline Co2P2O7 and carbon nanofiber (Co2P2O7/CNFs) composites with enhanced electrochemical performance were obtained by calcination after a hydrothermal process with NH4CoPO4∙H2O/bacterial cellulose precursors under an argon atmosphere. SEM images showed that the CNFs were highly dispersed on the surfaces of Co2P2O7 microplates. The diagonal size of the Co2P2O7 plates ranged from 5 to 25 µm with thicknesses on a nanometer scale. Notably, with the optimal calcining temperature, the Co2P2O7/CNFs@600 material has higher specific micropore and mesopore surface areas than other samples, and a maximal specific capacitance of 209.9 F g−1, at a current density of 0.5 A g−1. Interestingly, CNF composite electrodes can enhance electrochemical properties, and contribute to better electrical conductivity and electron transfer. EIS measurements showed that the charge–transfer resistance (Rct) of the CNF composite electrodes decreased with increasing calcination temperature. Furthermore, the Co2P2O7/CNF electrodes exhibited higher energy and power densities than Co2P2O7 electrodes.