Green Electrospun Silk Fibroin/Galactose Chitosan Composite Nanofibrous Scaffolds for Hepatic Tissue Engineering

The electrospun nanofibrous scaffolds made of proteins and polysaccharides were thought to be able to simulate the structure of natural extracellular matrix well.Silk fibroin(SF)and chitosan(CS)are probably the most widely used natural materials in biomedical fields including liver tissue engineering for their good properties and wide variety of sources.The asialoglycoprotein receptors of hepatocyte were reported to specifically recognize and interact with galactose.In this work,a green electrospun SF/galactosylated chitosan(GC)composite nanofibrous scaffold was fabricated and characterized.The data indicated that the addition of GC greatly influenced the spinning effect of SF aqueous solution,and the average diameter of the composite nanofibers was about 520nm.Moreover,the green electrospun SF/GC nanofibrous scaffolds were demonstrated significantly enhancing the adhesion and proliferation of hepatocyte(RH35)according to our data.The present study did a useful exploration on constructing scaffolds for liver regeneration by green electrospinning,and also laid a good foundation for the further applicative research of this green electrospun scaffolds in liver tissue engineering.

Fabrication of a Bi-layer Tubular Scaffold Consisted of a Dense Nanofibrous Inner Layer and a Porous Nanoyarn Outer Layer for Vascular Tissue Engineering

Recent years, it has attracted more attentions to increase the porosity and pore size of nanofibrous scaffolds to provide the for the cells to grow into the small-diameter vascular grafts. In this study, a novel bi-layer tubular scaffold with an inner layer and an outer layer was fabricated. The inner layer was random collagen/poly （ L-lactide-co-caprolactone ） I P （ LLA- CL） ] nanofibrous mat fabricated by conventional electrospinning and the outer layer was aligned collagen/P （LLA-CL） nanoyarns prepared by a dynamic liquid dectrospinning method. Fourier transform infrared spectroscopy （FTIR） was used to characterize the chemical structure. Scanning electron microscopy （ SEM ） was employed to observe the morphology of the layers and the cross- sectioned bi-layer tubular scaffold. A liquid displacement method was employed to measure the porosities of the inner and outer layers. Stress-strain curves were obtained to evaluate the mechanical properties of the two different layers and the bi-layer membrane. The diameters of the nanofibers and the nanoyarns were （480 ± 197 ） nm and （ 19.66 ± 4.05 ） μm, respectively. The outer layer had a significantly higher porosity and a larger pore size than those of the inner layer. Furthermore, the bi-layer membrane showed a good mechanical property which was suitable as small-diameter vascular graft. The results indicated that the bi-layer tubular scaffold had a great potential application in small vascular tissue engineering.

Mineralized Composite Nanofibrous Mats for Bone Tissue Engineering

Composite nanofibrous mats consisting of poly( L-lactideco-ε-caprolactone)( PLCL) and collagen type I( COL) were fabricated by electrospinning,and ten times simulated body fluid(10SBF) were employed to mineralize nanofibrous mats. Ballshaped hydroxyapatite( HA) was deposited on the surface of nanofibrous mats in 1. 5 h at room temperature. Human fetal osteoblasts( hFob) were seeded to investigate their proliferation and differentiation on mineralized composite nanofibrous mats. The results showed that hFob grew well on mineralized composite nanofibrous mats and alkaline phosphatase( ALP) activity of hFob on mineralized composite nanofibrous mats at 14 d was much higher than that on untreated nanofibrous mats. Moreover,the expression of osteocalcin of cells on mineralized composite nanofibrous mats was also much higher than those on untreated nanofibrous mats at 7 d and 14 d. This mineralized composite nanofibrous mats may have a great potential for bone tissue engineering.

Advances in clinical applications of bioceramics in the new regenerative medicine era

In this editorial,we comment on the hard and soft tissue applications of different ceramic-based scaffolds prepared by different mechanisms such as 3D printing,sol-gel,and electrospinning.The new concept of regenerative medicine relies on biomaterials that can trigger in situ tissue regeneration and stem cell recruitment at the defect site.A large percentage of these biomaterials is ceramic-based as they provide the essential requirements of biomaterial principles such as tailored multisize porosity,antibacterial properties,and angiogenic properties.All these previously mentioned properties put bioceramics on top of the hierarchy of biomaterials utilized to stimulate tissue regeneration in soft and hard tissue wounds.Multiple clinical applications registered the use of these materials in triggering soft tissue regeneration in healthy and diabetic patients such as bioactive glass nanofibers.The results were promising and opened new frontiers for utilizing these materials on a larger scale.The same results were mentioned when using different forms and formulas of bioceramics in hard defect regeneration.Some bioceramics were used in combination with other polymers and biological scaffolds to improve their regenerative and mechanical properties.All this progress will enable a larger scale of patients to receive such services with ease and decrease the financial burden on the government.

Expert consensus on pediatric orthodontic therapies of malocclusions in children

Malocclusion,identified by the World Health Organization(WHO)as one of three major oral diseases,profoundly impacts the dental-maxillofacial functions,facial esthetics,and long-term development of~260 million children in China.Beyond its physical manifestations,malocclusion also significantly influences the psycho-social well-being of these children.Timely intervention in malocclusion can foster an environment conducive to dental-maxillofacial development and substantially decrease the incidence of malocclusion or reduce the severity and complexity of malocclusion in the permanent dentition,by mitigating the negative impact of abnormal environmental influences on the growth.Early orthodontic treatment encompasses accurate identification and treatment of dental and maxillofacial morphological and functional abnormalities during various stages of dental-maxillofacial development,ranging from fetal stages to the early permanent dentition phase.From an economic and societal standpoint,the urgency for effective early orthodontic treatments for malocclusions in childhood cannot be overstated,underlining its profound practical and social importance.This consensus paper discusses the characteristics and the detrimental effects of malocclusion in children,emphasizing critical need for early treatment.It elaborates on corresponding core principles and fundamental approaches in early orthodontics,proposing comprehensive guidance for preventive and interceptive orthodontic treatment,serving as a reference for clinicians engaged in early orthodontic treatment.

Aldehyde-Sodium Alginate and Amino-Gelatin Preparation as Soft Tissue Adhesive

Sodium alginate and gelatin are both remarkable natural biomaterials; they all have been extensively applied in tissue engineering and other relative fields,due to their low price and good biocompatibility. In this paper,we oxidized sodium alginate with sodium periodate to convert 1,2-hydroxyl groups into aldehyde groups to get aldehyde-sodium alginate( A-SA). Gelatin was modified with ethylenediamine( ED) in the presence of water-soluble1-ethyl-3( 3-dimethylaminopropyl) carbodiimide( EDC) to introduce additional amino groups to get amino-gelatin. Upon mixing the A-SA and amino-gelatin aqueous solutions together,a gel rapidly formed based on the Schiff's base reaction between the aldehyde groups in A-SA and the amino groups in amino-gelatin.Fourier transform infrared spectroscopy( FTIR) analysis confirmed the characteristic peak of Schiff's base group in the hydrogel. The gelation time measure has confirmed the gelation time is dependent on the aldehyde group content in A-SA and amino group content in amino-gelatin. The fasted hydrogel formation takes place within 30 s. The entire test suggested that this gel could be a promising candidate as soft tissue adhesive.

Study of the inflammatory activating process in the early stage of Fusobacterium nucleatum infected PDLSCs

Fusobacterium nucleatum(F.nucleatum)is an early pathogenic colonizer in periodontitis,but the host response to infection with this pathogen remains unclear.In this study,we built an F.nucleatum infectious model with human periodontal ligament stem cells(PDLSCs)and showed that F.nucleatum could inhibit proliferation,and facilitate apoptosis,ferroptosis,and inflammatory cytokine production in a dose-dependent manner.The F.nucleatum adhesin Fad A acted as a proinflammatory virulence factor and increased the expression of interleukin(IL)-1β,IL-6 and IL-8.Further study showed that Fad A could bind with PEBP1 to activate the Raf1-MAPK and IKK-NF-κB signaling pathways.Time-course RNA-sequencing analyses showed the cascade of gene activation process in PDLSCs with increasing durations of F.nucleatum infection.NFκB1 and NFκB2 upregulated after 3 h of F.nucleatum-infection,and the inflammatory-related genes in the NF-κB signaling pathway were serially elevated with time.Using computational drug repositioning analysis,we predicted and validated that two potential drugs(piperlongumine and fisetin)could attenuate the negative effects of F.nucleatum-infection.Collectively,this study unveils the potential pathogenic mechanisms of F.nucleatum and the host inflammatory response at the early stage of F.nucleatum infection.

New clinical application of digital intraoral scanning technology in occlusal reconstruction:A case report

BACKGROUND Digital intraoral scanning,although developing rapidly,is rarely used in occlusal reconstruction.To compensate for the technical drawbacks of current occlusal reconstruction techniques,such as time consumption and high technical requirements,digital intraoral scanning can be used in clinics.This report aims to provide a way of selecting the most suitable maxillo-mandibular relationship(MMR)during recovery.CASE SUMMARY A 68-year-old man with severely worn posterior teeth underwent occlusal reconstruction with fixed prosthesis using digital intraoral scanning.A series of digital models in different stages of treatment were obtained,subsequently compared,and selected using digital intraoral scanning together with traditional measurements,such as cone beam computed tomography,joint imaging,and clinical examination.Using digital intraoral scanning,the MMR in different stages of treatment was accurately recorded,which provided feasibility for deciding the best occlusal reconstruction treatment,made the treatment process easier,and improved patient satisfaction.CONCLUSION This case report highlights the clarity,recordability,repeatability,and selectivity of digital intraoral scanning to replicate and transfer the MMR during occlusal reconstruction,expanding new perspectives for its design,fabrication,and postoperative evaluation.

Osteogenic potential of human periosteum-derived progenitor cells in PLGA scaffold using allogeneic serum

The use of periosteum-derived progenitor cells （PCs） combined with bioresorbable materials is an attractive approach for tissue engineering. The aim of this study was to characterize the osteogenic differentiation of PC in 3-dimensional （3D） poly-lactic-co-glycolic acid （PLGA） fleeces cultured in medium containing allogeneic human serum. PCs were isolated and expanded in monolayer culture. Expanded cells of passage 3 were seeded into PLGA constructs and cultured in osteogenic medium for a maximum period of 28 d. Morphological, histological and cell viability analyses of three-dimensionally cultured PCs were performed to elucidate osseous synthesis and deposition of a calcified matrix. Furthermore, the mRNA expression of type Ⅰ collagen, osteocalcin and osteonectin was semi-quantitively evaluated by real-time reverse transcriptase-polymerase chain reaction （RT-PCR）. The fibrin gel immobilization technique provided homogeneous PCs distribution in 3D PLGA constructs. Live-dead staining indicated a high viability rate of PCs inside the PLGA scaffolds. Secreted nodules ofneo-bone tissue formation and the presence of matrix mineralization were confirmed by positive yon Kossa staining. The osteogenic differentiation of PCs was further demonstrated by the detection of type I collagen, osteocalcin and osteonectin gene expression. The results of this study support the concept that this tissue engineering method presents a promising method for creation of new bone in vivo.

Spatiotemporal microRNA profile in peripheral nerve regeneration:miR-138 targets vimentin and inhibits Schwann cell migration and proliferation

While the peripheral nervous system has regenerative ability,restoration of sufficient function remains a challenge.Vimentin has been shown to be localized in axonal growth fronts and associated with nerve regeneration,including myelination,neuroplasticity,kinase signaling in nerve axoplasm,and cell migration;however,the mechanisms regulating its expression within Schwann cell（SC） remain unexplored.The aim of this study was to profile the spatial and temporal expression profile of micro RNA（mi RNA） in a regenerating rat sciatic nerve after transection,and explore the potential role of mi R-138-5 p targeting vimentin in SC proliferation and migration.A rat sciatic nerve transection model,utilizing a polyethylene nerve guide,was used to investigate mi RNA expression at 7,14,30,60,and 90 days during nerve regeneration.Relative levels of mi RNA expression were determined using microarray analysis and subsequently validated with quantitative real-time polymerase chain reaction.In vitro assays were conducted with cultured Schwann cells transfected with mi RNA mimics and assessed for migratory and proliferative potential.The top seven dysregulated mi RNAs reported in this study have been implicated in cell migration elsewhere,and GO and KEGG analyses predicted activities essential to wound healing.Transfection of one of these,mi RNA-138-5 p,into SCs reduced cell migration and proliferation.mi R-138-5 p has been shown to directly target vimentin in cancer cells,and the luciferase assay performed here in rat Schwann cells confirmed it.These results detail a role of mi R-138-5 p in rat peripheral nerve regeneration and expand on reports of it as an important regulator in the peripheral nervous system.

Three-dimensional Expansion:In Suspension Culture of SD Rat's Osteoblasts in a Rotating Wall Vessel Bioreactor

Objective To study large-scale expansion of SD （Sprague-Dawley） rat＇s osteoblasts in suspension culture in a rotating wall vessel bioreactor （RWVB）. Methods The bioreactor rotation speeds were adjusted in the range of 0 to 20 rpm, which could provide low shear on the rnicrocarriers around 1 dyn/cm^2. The cells were isolated via sequential digestions of neonatal （less than 3 days old） SD rat calvaria. After the primary culture and several passages, the cells were seeded onto the microcarriers and cultivated in T-flask, spinner flask and RWVB respectively. During the culture period, the cells were counted and observed under the inverted microscope for morphology every 12 h. After 7 days, the cells were evaluated with scanning electron microscope （SEM） for histological examination of the aggregates. Also, the hematoxylin-eosin （HE） staining and alkaline phosphatase （ALP） staining were performed. Moreover, von-Kossa staining and Alizarin Red S staining were carded out for mineralized nodule formation. Results The results showed that in RWVB, the cells could be expanded by more than ten times and they presented better morphology and vitality and stronger ability to form bones. Conclusions The developed RWVB can provide the culture environment with a relatively low shear force and necessary three-dimensional （3D） interactions among cells and is suitable for osteopath expansion in vitro.

Noggin versus basic fibroblast growth factor on the differentiation of human embryonic stem cells

The difference between Noggin and basic fibroblast growth factor for the neural precursor differen- tiation from human embryonic stem cells has not been studied. In this study, 100 tJg/L Noggin or 20 IJg/L basic fibroblast growth factor in serum-free neural induction medium was used to differen- tiate human embryonic stem cells H14 into neural precursors using monolayer differentiation. Two weeks after induction, significantly higher numbers of neural rosettes formed in the Noggin-induced group than the basic fibroblast growth factor-induced group, as detected by phase contrast micro- scope. Immunofluorescence staining revealed expression levels of Nestin, [3-111 Tubulin and Sox-1 were higher in the induced cells and reverse-transcription PCR showed induced cells expressed Nestin, Sox-1 and Neurofilament mRNA. Protein and mRNA expression in the Noggin-induced group was increased compared with the basic fibroblast growth factor-induced group. Noggin has a greater effect than basic fibroblast growth factor on the induction of human embryonic stem cell differentiation into neural precursors by monolayer differentiation, as Noggin accelerates and in- creases the differentiation of neural precursors.

Applications of stem cells and bioprinting for potential treatment of diabetes

Currently, there does not exist a strategy that can reduce diabetes and scientists are working towards a cure and innovative approaches by employing stem cellbased therapies. On the other hand, bioprinting technology is a novel therapeutic approach that aims to replace the diseased or lost β-cells, insulin-secreting cells in the pancreas, which can potentially regenerate damaged organs such as the pancreas. Stem cells have the ability to differentiate into various cell lines including insulinproducing cells. However, there are still barriers that hamper the successful differentiation of stem cells into β-cells. In this review, we focus on the potential applications of stem cell research and bioprinting that may be targeted towards replacing the β-cells in the pancreas and may offer approaches towards treatment of diabetes. This review emphasizes on the applicability of employing both stem cells and other cells in 3 D bioprinting to generate substitutes for diseased β-cells and recover lost pancreatic functions. The article then proceeds to discuss the overall research done in the field of stem cell-based bioprinting and provides future directions for improving the same for potential applications in diabetic research.

CD133 and membrane microdomains:Old facets for future hypotheses

Understanding all facets of membrane microdomains in normal and cancerous cells within the digestive tract is highly important,not only from a clinical point of view,but also in terms of our basic knowledge of cellular transformation.By studying the normal and cancer stem cell-associated molecule CD133 (prominin-1),novel aspects of the organization and dynamics of polarized epithelial cells have been revealed during the last decade.Its association with particular membrane microdomains is highly relevant in these contexts and might also offer new avenues in diagnosis and/or targeting of cancer stem cells.

Numerical Simulation of Microcarrier Motion in a Rotating Wall Vessel Bioreactor

Objective To analyze the forces of rotational wall vessel （RWV） bioreactor on small tissue pieces or microcarrier particles and to determine the tracks of microcarrier particles in RWV bioreactor. Methods The motion of the microcarrier in the rotating wall vessel （RWV） bioreactor with both the inner and outer cylinders rotating was modeled by numerical simulation. Results The continuous trajectory of microcarrier particles, including the possible collision with the wall was obtained. An expression between the minimum rotational speed difference of the inner and outer cylinders and the microcarrier particle or aggregate radius could avoid collisions with either wall. The range of microcarrier radius or tissue size, which could be safely cultured in the RWV bioreactor, in terms of shear stress level, was determined. Conclusion The model works well in describing the trajectory of a heavier microcarrier particle in rotating wall vessel.

Comparison of Osteogenesis Between Two Kinds of Stem Cells from Goat Combined Calcium Phosphate Cement in Tissue Engineering

To explore the possible mechanism of osteogenesis for deciduous teeth stem cells (DTSCs) in vivo/ vitro, stem cells from goat deciduous teeth (SGDs) were firstly isolated, induced and transplanted into immunocompromised mice. The SGDs's mineralization pattern and osteogenesis were compared with bone marrow messenchymal stem cells (BMMSCs) from goats. SGDs have similar osteogenic differentiation pattern in vitro and bone-like tissue formation mechanism in vivo to BMMSCs; moreover SGDs have stronger alkaline phosphatase (ALP) gene expression and osteopontin (OPN) gene expression levels than BMMSCs; also SGDs can form more bone-like tissues than BMMSCs when cell-scaffold compounds are transplanted into immunocompromised mice. This pre-clinical study in a large-animal model confirms that DTSCs may be an appropriate source of stem cells in repairing bone defects with tissue engineering.

The role of extracellular matrix in age-related conduction disorders： a forgotten player？

Cardiovascular aging is a physiological process gradually leading to structural degeneration and functional loss of all the cardiac and vascular components. Conduction system is also deeply influenced by the aging process with relevant reflexes in the clinical side. Age-related arrhythmias carry significant morbidity and mortality and represent a clinical and economical burden. An important and unjustly unrecognized actor in the pathophysiology of aging is represented by the extracellular matrix （ECM） that not only structurally supports the heart determining its mechanical and functional properties, but also sends a biological signaling regulating cellular function and maintaining tissue homeostasis. At the biophysical level, cardiac ECM exhibits a peculiar degree of anisotropy, which is among the main determinants of the conductive properties of the specialized electrical conduction system. Age-associated alterations of cardiac ECM are therefore able to profoundly affect the function of the conduction system with striking impact on the patient clinical conditions. This review will focus on the ECM changes that occur during aging in the heart conduction system and on their translation to the clinical scenario. Potential diagnostic and therapeutical perspectives arising from the knowledge on ECM age-associated alterations are further discussed.

Periodontitis may induce gut microbiota dysbiosis via salivary microbiota

The aim of this study was to identify whether periodontitis induces gut microbiota dysbiosis via invasion by salivary microbes.First,faecal and salivary samples were collected from periodontally healthy participants(PH group,n=16)and patients with severe periodontitis(SP group,n=21)and analysed by 16S ribosomal RNA sequencing.Significant differences were observed in both the faecal and salivary microbiota between the PH and SP groups.Notably,more saliva-sourced microbes were observed in the faecal samples of the SP group.Then,the remaining salivary microbes were transplanted into C57BL6/J mice(the C-PH group and the C-SP group),and it was found that the composition of the gut microbiota of the C-SP group was significantly different from that of the C-PH group,with Porphyromonadaceae and Fusobacterium being significantly enriched in the C-SP group.In the colon,the C-SP group showed significantly reduced crypt depth and zonula occludens-1 expression.The m RNA expression levels of pro-inflammatory cytokines,chemokines and tight junction proteins were significantly higher in the C-SP group.To further investigate whether salivary bacteria could persist in the intestine,the salivary microbiota was stained with carboxyfluorescein diacetate succinimidyl ester and transplanted into mice.We found that salivary microbes from both the PH group and the SP group could persist in the gut for at least 24 h.Thus,our data demonstrate that periodontitis may induce gut microbiota dysbiosis through the influx of salivary microbes.

Stem cell microencapsulation maintains stemness in inflammatory microenvironment

Maintaining the stemness of the transplanted stem cell spheroids in an inflammatory microenvironment is challenging but important in regenerative medicine. Direct delivery of stem cells to repair periodontal defects may yield suboptimal effects due to the complexity of the periodontal inflammatory environment. Herein, stem cell spheroid is encapsulated by interfacial assembly of metal-phenolic network(MPN) nanofilm to form a stem cell microsphere capsule. Specifically, periodontal ligament stem cells(PDLSCs) spheroid was coated with FeⅢ/tannic acid coordination network to obtain spheroid@[FeⅢ-TA] microcapsules. The formed biodegradable MPN biointerface acted as a cytoprotective barrier and exhibited antioxidative, antibacterial and anti-inflammatory activities, effectively remodeling the inflammatory microenvironment and maintaining the stemness of PDLSCs. The stem cell microencapsulation proposed in this study can be applied to multiple stem cells with various functional metal ion/polyphenol coordination, providing a simple yet efficient delivery strategy for stem cell stemness maintenance in an inflammatory environment toward a better therapeutic outcome.

Patient-specific induced pluripotent stem cells as“disease-in-adish”models for inherited cardiomyopathies and channelopathies–15 years of research

Among inherited cardiac conditions,a special place is kept by cardiomyopathies(CMPs)and channelopathies(CNPs),which pose a substantial healthcare burden due to the complexity of the therapeutic management and cause early mortality.Like other inherited cardiac conditions,genetic CMPs and CNPs exhibit incomplete penetrance and variable expressivity even within carriers of the same pathogenic deoxyribonucleic acid variant,challenging our understanding of the underlying pathogenic mechanisms.Until recently,the lack of accurate physiological preclinical models hindered the investigation of fundamental cellular and molecular mechanisms.The advent of induced pluripotent stem cell(iPSC)technology,along with advances in gene editing,offered unprecedented opportunities to explore hereditary CMPs and CNPs.Hallmark features of iPSCs include the ability to differentiate into unlimited numbers of cells from any of the three germ layers,genetic identity with the subject from whom they were derived,and ease of gene editing,all of which were used to generate“disease-in-a-dish”models of monogenic cardiac conditions.Functionally,iPSC-derived cardiomyocytes that faithfully recapitulate the patient-specific phenotype,allowed the study of disease mechanisms in an individual-/allele-specific manner,as well as the customization of therapeutic regimen.This review provides a synopsis of the most important iPSC-based models of CMPs and CNPs and the potential use for modeling disease mechanisms,personalized therapy and deoxyribonucleic acid variant functional annotation.