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The effectiveness of using amyloid fibrillation to improve the functional qualities of soy protein had drawn growing attention. However, the relationship between protein subunits and the structural polymorphism of soy protein-derived amyloid fibrils (SAFs) was not yet completely understood. In this study, soy protein subunits were hydrolyzed to different degrees according to the different action sites of different proteases (Pepsin, Papain and Alcalase). The impact of subunits on the amyloid fibrillation of soy protein was investigated through various techniques including atomic force microscopy, thioflavin T fluorescence, 1-anililo-naphthalene-8-sulfonate, and Fourier transform infrared spectrometer. The findings showed that the α and α' subunits were associated with the formation of fibril branch chains. The degree of hydrolysis of β subunits was found to be proportional to the number of fibrils. The presence of the 11S component was identified as a necessary condition for the formation of long-rigid fibrils. Furthermore, enzymatic hydrolysis unfolded the protein structure, exposing hydrophobic groups, loosening the protein structure, and altering the proportion of parallel and antiparallel β-sheet structures. This promoted the formation of amyloid fibrils and accelerated the development of stable SAFs gel. This study advances the knowledge of the function of subunits in amyloid fibrillation., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to be declared., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Adding prefabricated seeds to the fibril formation solution can accelerate the process of fibrillation. Various variants of soy protein amyloid fibrils (SAFs) (oligomers, short fibrils, long fibrils and mature long fibrils) were employed as seeds (OL, SF, LF and MLF). These fibrils were then introduced into a solution using a homologous seeding approach. We conducted a comprehensive analysis of various aspects, including morphological characteristics, surface hydrophobicity, secondary structures, fibril conversion rate and rheological properties. The results indicated that the addition of seeds, particularly those composed of longer and more mature fibrils, played a pivotal role in accelerating the formation of rigid and extended fibrils. Mature long fibrils as seeds can shorten the formation time of long fibrils (fibril length ≥ 600 nm) from 12 to 3 h. The seeding significantly shortens the formation time of SAFs, meanwhile, this study provides insights into the intricate dynamics of amyloid fibril at homologous seeding., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In recent years, the utilization of aerogel templates in oleogels to replace animal fats has garnered considerable attention due to health concerns. This study employed a "fiber-particle core-shell nanostructure model" to combine sodium carboxymethylcellulose (CMCNa) and soy protein isolate (SPI) or SPI hydrolysate (SPIH), and freeze-dried to form aerogel template, which was then dipped into oil to induce oleogels. The results showed that adding SPIH significantly improved the physicochemical properties of oleogels. The results of ζ-potential, FTIR, and rheology demonstrated a stronger binding of SPIH to CMC-Na compared to SPI. The CMC-Na-SPIH aerogels exhibited a coarser surface and denser network structure in contrast to CMC-Na-SPI aerogels, with an oil holding capacity (OHC) of up to 84.6 % and oil absorption capacity (OAC) of 47.4 g/g. The mechanical strength of oleogels was further enhanced through chemical crosslinking. Both CMC-Na-SPI and CMC-Na-SPIH oleogels displayed excellent elasticity and reversible compressibility, with CMC-Na-SPIH oleogels demonstrating superior mechanical strength. Additionally, CMC-Na-SPIH oleogels exhibited enhanced slow release of antimicrobial substances and antioxidant properties. Increasing the content of SPI/SPIH significantly improved the mechanical strength, antioxidant capacity, and OHC of the oleogels. This research presents a straightforward and promising approach to enhance the performance of aerogel template oleogels., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to be declared., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Introduction: With amazing clinical efficacy, Yangyin Qingfei Decoction Plus (YQDP), a well-known and age-old Chinese compound made of ten Chinese botanical drugs, is utilized in clinical settings to treat a range of respiratory conditions. This study examines the impact of Yangyin Qingfei Decoction (YQDP) on lung tissue metabolic products in severe Mycoplasma pneumoniae pneumonia (SMPP) model mice and examines the mechanism of YQDP in treating MP infection using UPLC-MS/MS technology. Methods: YQDP's chemical composition was ascertained by the use of Agilent 1260 Ⅱ high-performance liquid chromatography. By using a nasal drip of 10 10 CCU/mL MP bacterial solution, an SMPP mouse model was created. The lung index, pathology and ultrastructural observation of lung tissue were utilized to assess the therapeutic effect of YQDP in SMPP mice. Lung tissue metabolites were found in the normal group, model group, and YQDP group using UPLC-MS/MS technology. Using an enzyme-linked immunosorbent test (ELISA), the amount of serum inflammatory factors, such as interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α), was found. Additionally, the protein expression of PI3K, P-PI3K, AKT, P-AKT, NF-κB, and P-NF-κB was found using Western blot. Results: The contents of chlorogenic acid, paeoniflorin, forsythrin A, forsythrin, and paeonol in YQDP were 3.480 ± 0.051, 3.255 ± 0.040, 3.612 ± 0.017, 1.757 ± 0.031, and 1.080 ± 0.007 mg/g respectively. YQDP can considerably lower the SMPP mice's lung index ( p < 0.05). In the lung tissue of YQDP groups, there has been a decrease ( p < 0.05) in the infiltration of inflammatory cells at varying concentrations in the alveoli compared with the model group. A total of 47 distinct metabolites, including choline phosphate, glutamyl lysine, L-tyrosine, 6-thioinosine, Glu Trp, 5-hydroxydecanoate, etc., were linked to the regulation of YQDP, according to metabolomics study. By controlling the metabolism of porphyrins, pyrimidines, cholines, fatty acids, sphingolipids, glycerophospholipids, ferroptosis, steroid hormone biosynthesis, and unsaturated fatty acid biosynthesis, enrichment analysis suggested that YQDP may be used to treat SMPP. YQDP can lower the amount of TNF-α and IL-6 in model group mice as well as downregulate P-PI3K, P-AKT, and P-NF-κB expression ( p < 0.05). Conclusion: A specific intervention effect of YQDP is observed in SMPP model mice. Through the PI3K/Akt/NF-κB signaling pathways, YQDP may have therapeutic benefits by regulating the body's metabolism of α-Linoleic acid, sphingolipids, glycerophospholipids, arachidonic acid, and the production of unsaturated fatty acids., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhang, Zhao, Zhang, Liang, Guan, Wang, Liu and Wu.)

Flexible membranes with ultrathin thickness and excellent mechanical properties have shown great potential for broad uses in solid polymer electrolytes (SPEs), on-skin electronics, etc. However, an ultrathin membrane (<5 μm) is rarely reported in the above applications due to the inherent trade-off between thickness and antifailure ability. We discover a protic solvent penetration strategy to prepare ultrathin, ultrastrong layered films through a continuous interweaving of aramid nanofibers (ANFs) with the assistance of simultaneous protonation and penetration of a protic solvent. The thickness of a pure ANF film can be controlled below 5 μm, with a tensile strength of 556.6 MPa, allowing us to produce the thinnest SPE (3.4 μm). The resultant SPEs enable Li-S batteries to cycle over a thousand times at a high rate of 1C due to the small ionic impedance conferred by the ultrathin characteristic and regulated ionic transportation. Besides, a high loading of the sulfur cathode (4 mg cm -2 ) with good sulfur utilization was achieved at a mild temperature (35 °C), which is difficult to realize in previously reported solid-state Li-S batteries. Through a simple laminating process at the wet state, the thicker film (tens of micrometers) obtained exhibits mechanical properties comparable to those of thin films and possesses the capability to withstand high-velocity projectile impacts, indicating that our technique features a high degree of thickness controllability. We believe that it can serve as a valuable tool to assemble nanomaterials into ultrathin, ultrastrong membranes for various applications.

Ethnopharmacological Relevance: The incidence of allergic disease is constantly increasing, but its pathogenesis is not fully understood. Saposhnikovia divaricata (SD), called 'Fangfeng' in China, not only can be used for antipyretic, analgesic and anti-inflammatory as a traditional Chinese medicine, but also as an active ingredient in about 8% prescriptions. However, its effects on type I allergy and pseudoallergy have not been clarified., Aim of the Study: To explore the treatment and potential mechanisms of SD and its major bioactive component Prim-O-glucosylcimifugin (POG) on type I allergy and pseudoallergy in vitro and in vivo., Materials and Methods: The inhibitory effect of SD decoction and POG on type I allergy and its possible mechanism were evaluated by using RBL-2H3 cells model in vitro and the passive cutaneous anaphylaxis (PCA) mouse model in vivo. The cell degranulation of RBL-2H3 cells induced by DNP-IgE/DNP-BSA and Compound 48/80 (C48/80) was investigated, and the molecules of degranulation related signaling pathway was further detected by qRT-PCR and Western Blot analysis. Meanwhile, therapeutic effect of SD Decoction and POG were evaluated using PCA models in vivo. The molecular docking technology was conducted to explore the potential mechanisms., Results: In cells model induced by DNP-IgE/DNP-BSA, the release rate of β-Hex in high dose of SD and POG groups were 43.79% and 57.01%, and the release amount of HA in high dose of SD and POG groups were 26.19 ng/mL and 24.20 ng/mL. They were significantly lower than that in the model group. Besides, SD decoction and POG could significantly inhibit intracellular Ca 2+ increasing and cell apoptosis. But there is no obvious effect on cells degranulation induced by C48/80. The molecular docking results showed that 5-O-Methylvisamioside and POG could bind with FcεRI α with stronger binding ability, but weak binding ability to Mrgprx2. Moreover, qPCR and Western blot analyses indicated that SD could down-regulate Lyn/Syk/PLCγ, MAPK and PI3K/AKT/NF-κB signal pathway to inhibit IgE-dependent cell degranulation. In mice PCA model, both SD and POG could dose-dependently attenuate the Evans Blue extravasation, paw and ear swelling induced by DNP-IgE/DNP-BSA, but no significant inhibition under the PCA models induced by C48/80., Conclusion: In conclusion, SD is effective for the therapeutic of type I allergies, suggesting that SD is a potential candidate for the treatment of type I allergy, and the underlying mechanism of these effects needs to be further studied., Competing Interests: Declaration of competing interest The authors declared that they have no conflicts of interest to this work. We declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Natural materials, such as locust mandibles and squid beaks, define significant mechanical gradients that have been attributed to the chemical gradients of their specialized structural proteins (SPs). However, the mechanism by which SPs form chemical gradients in these materials remains unknown. In this study, a highly abundant histidine-rich structural protein ( Lm MHSP) was identified in the mandible of a migratory locust ( Locusta migratoria ). Lm MHSP was proven by both in vivo and in vitro evidence to act as a core building block of the mandible with a variety of synergistic functions including chitin binding, matrix formation via liquid-liquid phase separation, chemical cross-linking, and metal coordination. Furthermore, we found that the SP gradient in the locust mandible stems from the chitin-binding activity of Lm MHSP and different microstructures of chitin scaffolds in different regions. These findings advance our understanding of the formation mechanisms of natural biomaterials and have implications for the fabrication of biomimetic materials.

Coating conventional metallic electrodes with conducting polymers has enabled the essential characteristics required for bioelectronics, such as biocompatibility, electrical conductivity, mechanical compliance, and the capacity for structural and chemical functionalization of the bioelectrodes. However, the fragile interface between the conducting polymer and the electrode in wet physiological environment greatly limits their utility and reliability. Here, a general yet reliable strategy to seamlessly interface conventional electrodes with conducting hydrogel coatings is established, featuring tissue-like modulus, highly-desirable electrochemical properties, robust interface, and long-term reliability. Numerical modeling reveals the role of toughening mechanism, synergy of covalent anchorage of long-chain polymers, and chemical cross-linking, in improving the long-term robustness of the interface. Through in vivo implantation in freely-moving mouse models, it is shown that stable electrophysiological recording can be achieved, while the conducting hydrogel-electrode interface remains robust during the long-term low-voltage electrical stimulation. This simple yet versatile design strategy addresses the long-standing technical challenges in functional bioelectrode engineering, and opens up new avenues for the next-generation diagnostic brain-machine interfaces., (© 2022 Wiley-VCH GmbH.)

With the strengthening capacity through harnessing multi-length-scale structural hierarchy, synthetic hydrogels hold tremendous promise as a low-cost and abundant material for applications demanding unprecedented mechanical robustness. However, integrating high impact resistance and high water content, yet superior softness, in a single hydrogel material still remains a grand challenge. Here, a simple, yet effective, strategy involving bidirectional freeze-casting and compression-annealing is reported, leading to a hierarchically structured hydrogel material. Rational engineering of the distinct 2D lamellar structures, well-defined nanocrystalline domains and robust interfacial interaction among the lamellae, synergistically contributes to a record-high ballistic energy absorption capability (i.e., 2.1 kJ m -1 ), without sacrificing their high water content (i.e., 85 wt%) and superior softness. Together with its low-cost and extraordinary energy dissipation capacity, the hydrogel materials present a durable alternative to conventional hydrogel materials for armor-like protection circumstances., (© 2022 Wiley-VCH GmbH.)

Flexible and stretchable light emitting devices are driving innovation in myriad applications, such as wearable and functional electronics, displays and soft robotics. However, the development of flexible electroluminescent devices via conventional techniques remains laborious and cost-prohibitive. Here, we report a facile and easily-accessible route for fabricating a class of flexible electroluminescent devices and soft robotics via direct ink writing-based 3D printing. 3D printable ion conducting, electroluminescent and insulating dielectric inks were developed, enabling facile and on-demand creation of flexible and stretchable electroluminescent devices with good fidelity. Robust interfacial adhesion with the multilayer electroluminescent devices endowed the 3D printed devices with attractive electroluminescent performance. Integrated our 3D printed electroluminescent devices with a soft quadrupedal robot and sensing units, an artificial camouflage that can instantly self-adapt to the environment by displaying matching color was fabricated, laying an efficient framework for the next generation soft camouflages., (© 2022. The Author(s).)

Hydrogels have gained intensive interest in biomedical and flexible electronics, and adhesion of hydrogels to substrates or devices is indispensable in these application scenarios. Although numerous hydrogel adhesion strategies have been developed, it is still challenging to achieve a hydrogel with robust adhesion interface through a universal yet simple method. Here, a strategy for establishing strong interfacial adhesion between various hydrogels and a wide variety of substrates (i.e., soft hydrogels and rigid solids, including glass, aluminum, PET, nylon and PDMS) even under wet conditions, is reported. This strong interfacial adhesion is realized by constructing a bioinspired mineralized transition layer through ion diffusion and subsequent mineral deposition. This strategy is not only generally applicable to a broad range of substrates and ionic pairs, but also compatible with various fabrication approaches without compromising their interfacial robustnesses. This strategy is further demonstrated in the application of single-electrode triboelectric nanogenerators (TENG), where a robust interface between the hydrogel and elastomer layers is enabled to ensure a reliable signal generation and output., (© 2022 Wiley-VCH GmbH.)

Engineering conventional hydrogels with muscle-like anisotropic structures can efficiently increase the fatigue threshold over 1000 J m -2 along the alignment direction; however, the fatigue threshold perpendicular to the alignment is still as low as ≈100-300 J m -2 , making them nonsuitable for those scenarios where isotropic properties are desired. Here, inspired by the distinct structure-properties relationship of heart valves, a simple yet general strategy to engineer conventional hydrogels with unprecedented yet isotropic fatigue resistance, with a record-high fatigue threshold over 1,500 J m -2 along two arbitrary in-plane directions is reported. The two-step process involves the formation of preferentially aligned lamellar micro/nanostructures through a bidirectional freeze-casting process, followed by compression annealing, synergistically contributing to extraordinary resistance to fatigue crack propagation. The study provides a viable means of fabricating soft materials with isotropically extreme properties, thereby unlocking paths to apply these advanced soft materials toward applications including soft robotics, flexible electronics, e-skins, and tissue patches., (© 2022 Wiley-VCH GmbH.)

Nature builds biological materials from limited ingredients, however, with unparalleled mechanical performances compared to artificial materials, by harnessing inherent structures across multi-length-scales. In contrast, synthetic material design overwhelmingly focuses on developing new compounds, and fails to reproduce the mechanical properties of natural counterparts, such as fatigue resistance. Here, a simple yet general strategy to engineer conventional hydrogels with a more than 100-fold increase in fatigue thresholds is reported. This strategy is proven to be universally applicable to various species of hydrogel materials, including polysaccharides (i.e., alginate, cellulose), proteins (i.e., gelatin), synthetic polymers (i.e., poly(vinyl alcohol)s), as well as corresponding polymer composites. These fatigue-resistant hydrogels exhibit a record-high fatigue threshold over most synthetic soft materials, making them low-cost, high-performance, and durable alternatives to soft materials used in those circumstances including robotics, artificial muscles, etc., (© 2021 Wiley-VCH GmbH.)

Dual-emission carbon dots were synthesized by one-pot hydrothermal pyrolysis of citric acid and polyethyleneimine in the presence of rhodamine B at 160 °C for 5 h. The carbon dots have an average diameter of 2.51 nm with rhodamine moiety on their surface. Two emission bands centered at 447 and 581 nm are exhibited in their fluorescence spectra excited at 360 nm, and the former is sensitive while the latter is insensitive to Hg 2+ and pH. Glutathione (GSH) can recover the fluorescence quenched by Hg 2+ . Therefore, the dual-emission carbon dots were developed as a fluorescent ratiometric probe employing the ratio of the two intensities at 447 and 581 nm (R I447/I581 ) as the signal for the determinations of pH, Hg 2+ , and GSH. In the range of 5.0-10.0, a good linear relationship between R I447/I581 and pH was built with a regression equation of R I447/I581  = 11.95-0.56 pH (R 2  = 0.998). In the range from 0.0 to 8.0 μM, an excellent linear relationship between R I447/I581 and the concentration of Hg 2+ was obtained with a calibration equation of R I447/I581  = 6.2317-0.4458c (R 2  = 0.995) and a limit of detection (LOD) of 0.24 μM. In the range from 1.0 to 10.0 μM, a linear equation, R I447/I581  = 1.9133-0.4157c (R 2  = 0.995), was calibrated between R I447/I581 and the concentration of glutathione with a LOD of 0.27 μM. The recoveries for the determinations of Hg 2+ and GSH in real samples were in the ranges of 94.6 to 103.8% and 94.3 to 104.2%, respectively. Graphical abstract Dual-emission carbon dots achieved by luminescence center modulation within one-pot synthesis for a fluorescent ratiometric probe of pH, Hg 2+ , and glutathione.

Background: With the ageing of the population and the increase of sports injuries, the number of joint injuries has increased greatly. Tissue engineering or tissue regeneration is an important method to repair articular cartilage defects. While it has recently been paid much attention to use bilayered porous scaffolds to repair both cartilage and subchondral bone, it is interesting to examine to what extent a bilayer scaffold composed of the same kind of the biodegradable polymer poly(lactide- co -glycolide) (PLGA) can restore an osteochondral defect. Herein, we fabricated bilayered PLGA scaffolds and used a rabbit model to examine the efficacy of implanting the porous scaffolds with or without bone marrow mesenchymal stem cells (BMSCs). The present manuscript reports the regenerative potential up to 24 weeks., Methods: The osteochondral defect, 4 mm in diameter and 5 mm in depth, was created in the medial condyle of each knee in 23 rabbits. The bilayered PLGA scaffolds with a pore size of 100-200 μm in the chondral layer and a pore size of 300-450 μm in the osseous layer, seeded with or without BMSCs in the chondral layer, were then transplanted into the osteochondral defect of each knee. The osteochondral defect created in the same manner was untreated to act as the control. At 12 and 24 weeks postoperatively, condyles were harvested and analyzed using histology, immunohistochemistry, real-time polymerase chain reaction, and biomechanical testing to evaluate the efficacy of osteochondral repair., Results: No joint erosion, inflammation, swelling, or deformity was observed, and all animals maintained a full range of motion. Compared with the untreated blank group, the groups implanting the bilayered scaffolds with or without cells exhibited much better resurfacing, similar to the surrounding normal tissue. The histological scores of neotissues repaired by the scaffold with cells were closer to that of normal tissue. Although the biomechanical properties of neotissues were not as good as the normal tissue, no significant difference was found between the gene levels of neotissues repaired by the scaffold with or without cells and that of the normal tissue. The repair of the osteochondral defect tends to be stable 12 weeks after implantation., Conclusions: Our bilayered PLGA porous scaffold supports long-term osteochondral repair via in vivo tissue engineering or regeneration, and its effect can be further facilitated under the scaffold seeded with allogenic BMSCs., The Translational Potential of This Article: The bilayered PLGA porous scaffold can facilitate the repair of osteochondral defects and has potential for application in osteochondral tissue engineering., (© 2019 The Authors.)

Material cues to influence cell proliferation are a fundamental issue in the fields of biomaterials, cell biology, tissue engineering, and regenerative medicine. This paper aims to investigate the proliferation of single mammal cells on micropatterned material surfaces. To this end, we prepared cell-adhesive circular microislands with 20 areas on the nonfouling background and systematically examined adhesion and proliferation behaviors of different kinds of single cells (primary stem and nonstem cells, cancer and normal cell lines) on micropatterns. On the basis of the analysis of experimental data, we found two critical areas about cell proliferation: (1) the critical spreading area of cells from almost no proliferation to confined proliferation, denoted as A P and (2) the critical spreading area of cells from confined proliferation to almost free proliferation, denoted as A FP . We further summarized the relative size relationship between these two critical areas and the characteristic areas of cell adhesion on both patterned and nonpatterned surfaces. While proliferation of single primary cells was affected by cell spreading, those cell lines, irrespective of normal and cancer cells, did not exhibit significant cell-spreading effects. As a result, this study reveals that proliferation of single cells is dependent upon spreading area, in particular for primary cells on material surfaces.

While various porous scaffolds have been developed, the focused study about which structure leads to better mechanics is rare. In this study, we designed porous scaffolds with tetragonal, hexagonal and wheel-like structures under a given porosity, and fabricated corresponding poly(lactic acid) (PLA) scaffolds with three-dimensional printing. High-resolution micro-computed tomography was carried out to calculate their experimental porosity and confirm their high interconnectivity. The theoretical and experimental compressive properties in the longitudinal direction were characterized by finite element analysis method and electromechanical universal testing system, respectively. Thereinto, the scaffold with the tetragonal structure exhibited higher mechanical strength both theoretically and experimentally. Creep and stress relaxation behaviors of the scaffolds revealed that the tetragonal scaffold had less significant viscoelasticity. Immersion dynamic mechanical analysis was performed to test their cycle-loading fatigue behaviors in the simulated body fluid at 37 °C; the tetragonal scaffold exhibited the latest fatigue beginning point at 4400 cycles, which indicated a better anti-fatigue performance; the hexagonal and wheel-like ones exhibited the middle and earliest fatigue beginning points at 3200 and 2500 cycles, respectively. What is more, cytocompatibility and histocompatibility of the scaffolds with all of the structures were confirmed by cell counting kit-8 assay in vitro and three-month subcutaneous implantation in rats in vivo. Hence, the key property difference of the three examined structures comes from their mechanics; the tetragonal structure exhibited better mechanics in the longitudinal direction examined in this study, which could be taken into consideration in design of a porous scaffold for tissue engineering and regeneration.

Congenital vertebral malformations (CVMs) are associated with human TBX6 compound inheritance that combines a rare null allele and a common hypomorphic allele at the TBX6 locus. Our previous in vitro evidence suggested that this compound inheritance resulted in a TBX6 gene dosage of less than haploinsufficiency (i.e. <50%) as a potential mechanism of TBX6-associated CVMs. To further investigate this pathogenetic model, we ascertained and collected 108 Chinese CVM cases and found that 10 (9.3%) of them carried TBX6 null mutations in combination with common hypomorphic variants at the second TBX6 allele. For in vivo functional verification and genetic analysis of TBX6 compound inheritance, we generated both null and hypomorphic mutations in mouse Tbx6 using the CRISPR-Cas9 method. These Tbx6 mutants are not identical to the patient variants at the DNA sequence level, but instead functionally mimic disease-associated TBX6 variants. Intriguingly, as anticipated by the compound inheritance model, a high penetrance of CVM phenotype was only observed in the mice with combined null and hypomorphic alleles of Tbx6. These findings are consistent with our experimental observations in humans and supported the dosage effect of TBX6 in CVM etiology. In conclusion, our findings in the newly collected human CVM subjects and Tbx6 mouse models consistently support the contention that TBX6 compound inheritance causes CVMs, potentially via a gene dosage-dependent mechanism. Furthermore, mouse Tbx6 mutants mimicking human CVM-associated variants will be useful models for further mechanistic investigations of CVM pathogenesis in the cases associated with TBX6.

This study is aimed at investigation of the osteochondral regeneration potential of bilayered PLGA/PLGA-HAp composite scaffolds with one layer made of biodegradable polymer poly(d,l-lactide- co -glycolide) (PLGA) and another layer made of PLGA polymeric matrix coated by bioactive ceramics hydroxyapatite (HAp). The composite scaffolds were fabricated by compression molding/particle leaching and plasma-treated surface deposition. The pore morphology, mechanical properties, and surface deposition of the scaffold were characterized, and the growth of bone marrow derived mesenchymal stem cells or medicinal signaling cells (MSCs) in the scaffold was verified. Thereafter, rabbit models with an artificial osteochondral defect in joint were randomized into three treatment groups: virgin bilayered scaffold, bilayered scaffold preseeded in vitro with MSCs, and untreated blank control. At 16-week postoperation, both the virgin scaffolds and cell-seeded bilayered scaffolds exhibited osteochondral repair, as verified by biomechanics analysis, histological evaluations, and Western blot. The results highlighted the potentiality of the bilayered PLGA/PLGA-HAp composite scaffold for osteochondral tissue engineering, and in particular tissue regeneration or in situ tissue induction, probably by recruiting the local cells toward chondrogenic and osteogenic differentiation in the porous biomaterials.