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Scaffold-free spheroids offer great potential as a direct supply of cells for bottom-up bone tissue engineering. However, the building of functional spheroids with both cells and bioactive signals remains challenging. Here, we engineered functional spheroids with mesenchymal stem cells (MSCs) and two-dimensional hetero-nano-layers (2DHNL) that consisted of black phosphorus (BP) and graphene oxide (GO) to create a 3D cell-instructive microenvironment for large defect bone repair. The effects of the engineered 2D materials on the proliferation, osteogenic differentiation of stem cells was evaluated in an in vitro 3D spheroidal microenvironment. Excellent in vivo support of osteogenesis of MSCs, neovascularization, and bone regeneration was achieved after transplanting these engineered spheroids into critical-sized rat calvarial defects. Further loading of osteogenic factor dexamethasone (DEX) on the 2DHNL showed outstanding in vivo osteogenic induction and bone regrowth without prior in vitro culture in osteogenic medium. The shortened overall culture time would be advantageous for clinical translation. These functional spheroids impregnated with engineered 2DHNL enabling stem cell and osteogenic factor co-delivery could be promising functional building blocks to provide cells and differential clues in an all-in-one system to create large tissues for time-effective in vivo bone repair.

Stem cell paracrine activity is implicated in cardiac repair. Linkage between secretome functionality and therapeutic outcome was here interrogated by systems analytics of biobanked human cardiopoietic cells, a regenerative biologic in advanced clinical trials. Protein chip array identified 155 proteins differentially secreted by cardiopoietic cells with clinical benefit, expanded into a 520 node network, collectively revealing inherent vasculogenic properties along with cardiac and smooth muscle differentiation and development. Next generation RNA sequencing, refined by pathway analysis, pinpointed miR‐146 dependent regulation upstream of the decoded secretome. Intracellular and extracellular integration unmasked commonality across cardio‐vasculogenic processes. Mirroring the secretome pattern, infarcted hearts benefiting from cardiopoietic cell therapy restored the disease proteome engaging cardiovascular system functions. The cardiopoietic cell secretome thus confers a therapeutic molecular imprint on recipient hearts, with response informed by predictive systems profiling., Reverse translational decoding characterized the secretome of cardiopoietic cells, a regenerative therapeutic in clinical testing. A cardiovasculogenic systems signature distinguished high from low response profiles, an imprint echoed in the restored diseased proteome. Linkage of realized outcome with secretome identity suggests paracrine centrality in regenerative fitness. Pre‐intervention secretome profiling would thus inform optimized selection of regenerative biologic candidates.

A promising strategy that emerged in tissue engineering is to incorporate two-dimensional (2D) materials into polymer scaffolds, producing materials with desirable mechanical properties and surface chemistries, which also display broad biocompatibility. Black phosphorus (BP) is a 2D material that has sparked recent scientific interest due to its unique structure and electrochemical characteristics. In this study, BP nanosheets (BPNSs) were incorporated into a cross-linkable oligo[poly(ethylene glycol) fumarate] (OPF) hydrogel to produce a new nanocomposite for bone regeneration. BPNSs exhibited a controllable degradation rate coupled with the release of phosphate in vitro. MTS assay results together with live/dead images confirmed that the introduction of BPNSs into OPF hydrogels enhanced MC3T3-E1 cell proliferation. Moreover, the morphology parameters indicated better attachments of cells in the BPNSs containing group. Immunofluorescence images as well as intercellular ALP and OCN activities showed that adding a certain amount of BPNSs to OPF hydrogel could greatly improve differentiation of pre-osteoblasts on the hydrogel. Additionally, embedding black phosphorous into a neutral polymer network helped to control its cytotoxicity, with optimal cell growth observed at BP concentrations as high as 500 ppm. These results reinforced that the supplementation of OPF with BPNSs can increase the osteogenic capacity of polymer scaffolds for use in bone tissue engineering.

Objectives: Peripheral arterial disease is a significant cause of morbidity and mortality. The goal of this study was to develop a novel treatment to promote angiogenesis. Platelets have been previously implicated in angiogenic events; however, platelet activation rather than presence of growth factors has been associated with new blood vessel formation. This study evaluated exosomes purified from activated platelets to determine their potential in promoting angiogenesis both in vitro and in vivo. Methods & Results: Under Current Good Manufacturing Practices, conditioned medium processed from apheresis platelet cultures yielded a purified exosome product (PEP) enriched for CD63, CD9 and Alix. In vitro, PEP drove HUVEC proliferation, migration, and vascular tube formation. Proteomic evaluation showed Flk1 (VEGFR2) was one of the key angiogenic proteins in PEP. HUVECs treated with Flk1 antibody neutralized PEP (anti-Flk1-PEP) showed reduced vascular tube formation compared to PEP. To evaluate in vivo angiogenesis, a rodent model of hindlimb ischemia was treated with sustained release PEP embedded in fibrin glue (PEP/TISSEEL) with blood flow evaluated by SPY angiography at 28 days post-treatment. Animals treated with PEP/TISSEEL showed improved blood flow with significant difference (p Conclusions: Exosome driven donation of Flk1 here promoted angiogenic events and was successfully translated into healing outcomes in the setting of two animal models of peripheral ischemia. This novel insight into biotherapy achievable pro-angiogenesis establishes a clinical-grade off-the-shelf exosome technology for the treatment of peripheral vascular disease.

Conduits that promote nerve regeneration are currently of great medical concern, particularly when gaps exist between nerve endings. To address this issue, our laboratory previously developed a nerve conduit from biodegradable poly(caprolactone fumarate) (PCLF) that supports peripheral nerve regeneration. The present study improves upon this work by further developing an electrically conductive, positively charged PCLF scaffold through the incorporation of graphene, carbon nanotubes (CNTs), and [2-(methacryloyloxy)ethyl]trimethylammonium chloride (MTAC) (PCLF-Graphene-CNT-MTAC) using ultraviolet (UV) induced photocrosslinking. Scanning electron microscopy, transmission electron microscopy, and atomic force microscopy were used to assess the incorporation of CNTs and graphene into PCLF-Graphene-CNT-MTAC scaffolds, which displayed enhanced surface roughness and reduced electrochemical impedance when compared to neat PCLF. Scaffolds with these surface modifications also showed improved growth and differentiation of rat pheochromocytoma 12 cells in vitro, with enhanced cell growth, neurite extension, and cellular migration. Furthermore, an increased number of neurite protrusions were observed when the conduit was electrically stimulated. These results show that the electrically conductive PCLF-Graphene-CNT-MTAC nerve scaffolds presented here support the cellular behaviors that are critical for nerve regeneration, ultimately making this material an attractive candidate for regenerative medicine applications.

Three-dimensional (3D) printing is a promising technology for tissue engineering. However, 3D-printing methods are limited in their ability to produce desired microscale features or electrochemical properties in support of robust cell adhesion, proliferation, and differentiation. This study addresses this deficiency by proposing an integrated, one-step, method to increase the cytocompatibility of 3D-printed scaffolds through functionalization leveraging conductive carbon nanotubes (CNTs). To this end, CNTs were first sonicated with water-soluble single-stranded deoxyribonucleic acid (ssDNA) to generate a negatively charged ssDNA@CNT nano-complex. Concomitantly, 3D-printed poly(propylene fumarate) (PPF) scaffolds were ammonolyzed to introduce free amine groups, which can take on a positive surface charge in water. The ssDNA@CNT nano-complex was then applied to 3D-printed scaffolds through a simple one-step coating utilizing electric-static force. This fast and facile functionalization step resulted in a homogenous and non-toxic coating of CNTs to the surface, which significantly improved the adhesion, proliferation, and differentiation of pre-osteoblast cells. In addition, the CNT based conductive coating layer enabled modulation of cell behavior through electrical stimuli (ES) leading to cellular proliferation and osteogenic gene marker expression, including alkaline phosphatase (ALP), osteocalcin (OCN), and osteopontin (OPN). Collectively, these data provide the foundation for a one-step functionalization method for simple, fast, and effective functionalization of 3D printed scaffolds that support enhanced cell adhesion, proliferation, and differentiation, especially when employed in conjunction with ES. STATEMENT OF SIGNIFICANCE: Three-dimensional (3D) printing is a promising technology for tissue engineering. However, 3D-printing methods have limited ability to produce desired features or electrochemical properties in support of robust cell behavior. To address this deficiency, the current study proposed an integrated, one-step method to increase the cytocompatibility of 3D-printed scaffolds through functionalization leveraging conductive carbon nanotubes (CNTs). This fast and facile functionalization resulted in a homogenous and non-toxic coating of CNTs to the surface, which significantly improved the adhesion, proliferation, and differentiation of cells on the 3D-printed scaffolds.

Plasmalemmal ATP sensitive potassium (K

In situ cell recruitment is a promising regenerative medicine strategy with the purpose of tissue regeneration without stem cell transplantation. This chemotaxis-based strategy is aimed at ensuring a restorative environment through the release of chemokines that promote site-specific migration of healing cell populations. Stromal cell-derived factor-1α (SDF-1α) is a critical chemokine that can regulate the migration of mesenchymal stem cells (MSCs). Accordingly, here, SDF-1α-loaded microporous oligo[poly(ethylene glycol) fumarate]/bis[2-(methacryloyloxy)ethyl] phosphate composites (SDF-1α/OPF/BP) were engineered and probed. SDF-1α/OPF/BP composites were loaded with escalating SDF-1α concentrations, namely, 0 ng/ml, 50 ng/ml, 100 ng/ml, and 200 ng/ml, and were cocultured with MSC. Scratching assay, Transwell assay, and three-dimensional migration model were utilized to assess the migration response of MSCs. Immunofluorescence staining of Runx2 and osteopontin (OPN), ELISA assay of osteocalcin (OCN) and alkaline phosphatase (ALP), and Alizarin Red S staining were conducted to assess the osteogenesis of MSCs. All SDF-1α/OPF/BP composites engendered a release of SDF-1α (>80%) during the first four days. SDF-1α released from the composites significantly promoted migration and osteogenic differentiation of MSCs documented by upregulated expression of osteogenic-related proteins, ALP, Runx2, OCN, and OPN. SDF-1α at 100 ng/ml was optimal for enhanced migration and osteogenic proficiency. Thus, designed SDF-1α/OPF/BP composites were competent in promoting the homing and osteogenesis of MSCs and thus offer a promising bioactive scaffold candidate for on-demand bone tissue regeneration.

The lack of molecular targets and targeting probes remains a major drawback for targeted imaging and drug delivery in lung cancer. In this study, we exploited in vivo phage display to identify a novel targeting probe that homes to the tumor in a K-rasLA2 mutant mouse lung cancer model. Compared with other candidate peptides selected from 5 rounds of phage display, the CRQTKN peptide homed to tumor nodules in the lung of mutant mice at higher levels. Photoacoustic tomography of mutant mice detected lung tumors via tumor homing of the near-infrared fluorescence dye-labeled CRQTKN peptide. Ex vivo photoacoustic images of isolated organs further demonstrated tumor homing of the CRQTKN peptide, whereas minimal accumulation was observed in control organs, such as the liver. Compared with untargeted liposomes and doxorubicin, doxorubicin-loaded liposomes whose surface was modified with the CRQTKN peptide more efficiently delivered doxorubicin and reduced the number or size of tumor lesions in K-rasLA2 mutant mice. Analysis of hematologic parameters and liver and kidney function showed no significant systemic side effects by the treatments. Affinity-based identification was used to detect TNF receptor superfamily member 19L (TNFRSF19L), which was upregulated in lung tumors of mutant mice, as the receptor for the CRQTKN peptide. In conclusion, these results suggest that the CRQTKN peptide is a promising targeting probe for photoacoustic-guided detection and drug delivery to lung cancer, and acts by binding to TNFRSF19L. Significance: These findings present a new tumor-targeting probe for photoacoustic-guided detection and drug delivery.

Two-dimensional (2D) materials have emerged as a new promising research topic for tissue engineering because of their ability to alter the surface properties of tissue scaffolds and thus improve their biocompatibility and cell affinity. Multiple 2D materials, such as graphene and graphene oxide (GO), have been widely reported to enhance cell adhesion and proliferation. Recently, a newly emerged black phosphorus (BP) 2D material has attracted attention in biomedical applications because of its unique mechanical and electrochemical characteristics. In this study, we investigated the synergistic effect of these two types of 2D materials on cell osteogenesis for bone tissue engineering. BP was first wrapped in negatively charged GO nanosheets, which were then adsorbed together onto positively charged poly(propylene fumarate) three-dimensional (3D) scaffolds. The increased surface area provided by GO nanosheets would enhance cell attachment at the initial stage. In addition, slow oxidation of BP nanosheets wrapped within GO layers would generate a continuous release of phosphate, an important osteoblast differentiation facilitator designed to stimulate cell osteogenesis toward the new bone formation. Through the use of 3D confocal imaging, unique interactions between cells and BP nanosheets were observed, including a stretched cell shape and the development of filaments around the BP nanosheets, along with increased cell proliferation when compared with scaffolds incorporating only one of the 2D materials. Furthermore, the biomineralization of 3D scaffolds, as well as cellular osteogenic markers, was all measured and improved on scaffolds with both BP and GO nanosheets. All these results indicate that the incorporation of 2D BP and GO materials could effectively and synergistically stimulate cell proliferation and osteogenesis on 3D tissue scaffolds.

α2-adrenoceptor (α2-AR) isoforms, abundant in sympathetic synapses and noradrenergic neurons of the central nervous system, are integral in the presynaptic feed-back loop mechanism that moderates norepinephrine surges. We recently identified that postsynaptic α2-ARs, found in the myocellular sarcolemma, also contribute to a muscle-delimited feedback control capable of attenuating mobilization of intracellular Ca2+ and myocardial contractility. This previously unrecognized α2-AR-dependent rheostat is able to counteract competing adrenergic receptor actions in cardiac muscle. Specifically, in ventricular myocytes, nitric oxide (NO) and cGMP are the intracellular messengers of α2-AR signal transduction pathways that gauge the kinase-phosphatase balance and manage cellular Ca2+ handling preventing catecholamine-induced Ca2+ overload. Moreover, α2-AR signaling counterbalances phospholipase C - PKC-dependent mechanisms underscoring a broader cardioprotective potential under sympathoadrenergic and angiotensinergic challenge. Recruitment of such tissue-specific features of α2-AR under sustained sympathoadrenergic drive may, in principle, be harnessed to mitigate or prevent cardiac malfunction. However, cardiovascular disease may compromise peripheral α2-AR signaling limiting pharmacological targeting of these receptors. Prospective cardiac-specific gene or cell-based therapeutic approaches aimed at repairing or improving stress-protective α2-AR signaling may offer an alternative towards enhanced preservation of cardiac muscle structure and function.

An internal and marginal fit between prosthesis and abutment is an important factor of the durability of dental prosthesis. In this paper, we have proposed the use of swept-source optical coherence tomography (OCT) as a novel application for non-ionized and high-resolution measurements of internal and marginal discrepancies at anatomically critical four points, such as occlusal, angle, axial, and margin during prosthesis attachment. A tooth model was fabricated by a 3D printing technique, and the dental prosthesis was designed using dental CAD software. The cross-sectional images along with intensity peak profile analysis of the sample were acquired using the OCT system for measurements of fit. The demonstrated qualitative and quantitative evaluations can be well-utilized for assessment of the internal and marginal fit of dental prosthesis.

Urinary incontinence afflicts up to 40% of adult women in the United States. Stress urinary incontinence (SUI) accounts for approximately one-third of these cases, precipitating ~200,000 surgical procedures annually. Continence is maintained through the interplay of sub-urethral support and urethral sphincter coaptation, particularly during activities that increase intra-abdominal pressure. Currently, surgical correction of SUI focuses on the re-establishment of sub-urethral support. However, mesh-based repairs are associated with foreign body reactions and poor localized tissue healing, which leads to mesh exposure, prompting the pursuit of technologies that restore external urethral sphincter function and limit surgical risk. The present work utilizes a human platelet-derived CD41a and CD9 expressing extracellular vesicle product (PEP) enriched for NF-κB and PD-L1 and derived to ensure the preservation of lipid bilayer for enhanced stability and compatibility with hydrogel-based sustained delivery approaches. In vitro, the application of PEP to skeletal muscle satellite cells in vitro drove proliferation and differentiation in an NF-κB-dependent fashion, with full inhibition of impact on exposure to resveratrol. PEP biopotentiation of collagen-1 and fibrin glue hydrogel achieved sustained exosome release at 37 °C, creating an ultrastructural "bead on a string" pattern on scanning electron microscopy. Initial testing in a rodent model of latissimus dorsi injury documented activation of skeletal muscle proliferation of healing. In a porcine model of stress urinary incontinence, delivery of PEP-biopotentiated collagen-1 induced functional restoration of the external urethral sphincter. The histological evaluation found that sustained PEP release was associated with new skeletal muscle formation and polarization of local macrophages towards the regenerative M2 phenotype. The results provided herein serve as the first description of PEP-based biopotentiation of hydrogels implemented to restore skeletal muscle function and may serve as a promising approach for the nonsurgical management of SUI.

Sarcolemmal α2 adrenoceptors (α2-AR), represented by α2A, α2B and α2C isoforms, can safeguard cardiac muscle under sympathoadrenergic surge by governing Ca2+ handling and contractility of cardiomyocytes. Cardiomyocyte-specific targeting of α2-AR would provide cardiac muscle-delimited stress control and enhance the efficacy of cardiac malfunction treatments. However, little is known about the specific contribution of the α2-AR subtypes in modulating cardiomyocyte functions. Herein, we analyzed the expression profile of α2A, α2B and α2C subtypes in mouse ventricle and conducted electrophysiological antagonist assay evaluating the contribution of these isoforms to the suppression of L-type Ca2+ current (ICaL). Patch-clamp electro-pharmacological studies revealed that the α2-agonist-induced suppression of ICaL involves mainly the α2C, to a lesser extent the α2B, and not the α2A isoforms. RT-qPCR evaluation revealed the presence of adra2b and adra2c (α2B and α2C isoform genes, respectively), but was unable to identify the expression of adra2a (α2A isoform gene) in the mouse left ventricle. Immunoblotting confirmed the presence only of the α2B and the α2C proteins in this tissue. The identified α2-AR isoform-linked regulation of ICaL in the mouse ventricle provides an important molecular substrate for the cardioprotective targeting.

Injectable polymers have attracted intensive attention in tissue engineering and drug delivery applications. Current injectable polymer systems often require free-radical or heavy-metal initiators and catalysts for the crosslinking process, which may be extremely toxic to the human body. Here, we report a novel polyhedral oligomeric silsesquioxane (POSS) based strain-promoted alkyne-azide cycloaddition (SPAAC) "click" organic-inorganic nanohybrids (click-ON) system that can be click-crosslinked without any toxic initiators or catalysts. The click-ON scaffolds supported excellent adhesion, proliferation, and osteogenesis of stem cells. In vivo evaluation using a rat cranial defect model showed outstanding bone formation with minimum cytotoxicity. Essential osteogenic alkaline phosphatase (ALP) and vascular CD31 marker expression were detected on the defect site, indicating excellent support of in vivo osteogenesis and vascularization. Using salt leaching techniques, an injectable porous click-ON cement was developed to create porous structures and support better in vivo bone regeneration. Beyond defect filling, the click-ON cement also showed promising application for spinal fusion using rabbits as a model. Compared to the current clinically used poly (methyl methacrylate) (PMMA) cement, this click-ON cement showed great advantages of low heat generation, better biocompatibility and biodegradability, and thus has great potential for bone and related tissue engineering applications.

Adenylate kinase 2 (AK2) catalyzes trans-compartmental nucleotide exchange, but the functional implications of this mitochondrial intermembrane isoform is only partially understood. Here, transgenic AK2−/− null homozygosity was lethal early in embryo, indicating a mandatory role for intact AK2 in utero development. In the adult, conditional organ-specific ablation of AK2 precipitated abrupt heart failure with Krebs cycle and glycolytic metabolite buildup, suggesting a vital contribution to energy demanding cardiac performance. Depressed pump function recovered to pre-deletion levels overtime, suggestive of an adaptive response. Compensatory upregulation of phosphotransferase AK1, AK3, AK4 isozymes, creatine kinase isoforms, and hexokinase, along with remodeling of cell cycle/growth genes and mitochondrial ultrastructure supported organ rescue. Taken together, the requirement of AK2 in early embryonic stages, and the immediate collapse of heart performance in the AK2-deficient postnatal state underscore a primordial function of the AK2 isoform. Unsalvageable in embryo, loss of AK2 in the adult heart was recoverable, underscoring an AK2-integrated bioenergetics system with innate plasticity to maintain homeostasis on demand.

Background With the enormous increment of globalization and global warming, it is expected that the number of newly evolved infectious diseases will continue to increase. To prevent damage due to these infections, the development of a diagnostic method for detecting a virus with high sensitivity in a short time is highly desired. In this study, we have developed a disposable electrode with high-sensitivity and accuracy to evaluate its performances for several target viruses. Results Conductive silicon rubber (CSR) was used to fabricate a disposable sensing matrix composed of nitrogen and sulfur-co-doped graphene quantum dots (N,S-GQDs) and a gold-polyaniline nanocomposite (AuNP-PAni). A specific anti-white spot syndrome virus (WSSV) antibody was conjugated to the surface of this nanocomposite, which was successfully applied for the detection of WSSV over a wide linear range of concentration from 1.45 × 102 to 1.45 × 105 DNA copies/ml, with a detection limit as low as 48.4 DNA copies/ml. Conclusion The engineered sensor electrode can retain the detection activity up to 5 weeks, to confirm its long-term stability, required for disposable sensing applications. This is the first demonstration of the detection of WSSV by a nanofabricated sensing electrode with high sensitivity, selectivity, and stability, providing as a potential diagnostic tool to monitor WSSV in the aquaculture industry.

Additional file 1: Figure S1 Analysis of functional groups on the surface of named materials by FT-IR infrared absorption spectra. Figure S2. Impedance analysis of the PAni-PAni/CSR sensor electrode after 5 to 20 cycles of electrodeposition. Figure S3 (A–C) The difference in thickness of the polyaniline layer deposited on CSR was observed by SEM. Figure S4. Cyclic voltammetry analysis of the Ab-N,S-GQD@AuNP-PAni-PAni/CSR sensor electrode after 2 to 50 cycles. Figure S5. Cyclic voltammetry of CSR day1 (black line) and day2 (red line). Figure S6. Comparison of the Rct values for AuNP-coated CSR (blue line) and AuNP/PAni-coated CSR (orange line). Figure S7. Detection result of WSSV using different surface areas of Ab-N,S-GQD@AuNP-PAni-PAni/CSR. Figure S8. AFM images of the bare disposable sensor electrode (A) and WSSV-bound electrode (B). Figure S9. Nyquist impedance plots of the Ab-N,S-GQD@AuNP-PAni-PAni/CSR sensor electrode before and after loading A) HEV and B) influenza virus. Figure S10. Detection results of genotype 3 HEV (A) and influenza virus A (H1N1) (B) using the Ab-N,S-GQD@AuNP-PAni/CSR sensor electrode with their corresponding antibodies attached.

Incorporation of hydroxyapatite (HA) into polymer networks is a promising strategy to enhance the mechanical properties and osteoinductivity of the composite scaffolds for bone tissue engineering. In this study, we designed a group of nanocomposite scaffolds based on cross-linkable poly(propylene fumarate) (PPF) and 30 wt % strontium-hydroxyapatite (Sr-HA) nanoparticles. Four different Sr contents [Sr:(Sr + Ca), molar ratio] in the Sr-HA particles were studied: 0% (HA), 5% (Sr5-HA), 10% (Sr10-HA), and 20% (Sr20-HA). Two-dimensional (2D) disks were prepared using a thermal crosslinking method. The structure and surface morphology of different Sr-HA and PPF/Sr-HA composites were characterized using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), and atomic force microscopy (AFM). To detect cellular responses in vitro, MC3T3-E1 cells were seeded and cultured on the different PPF/Sr-HA composite disks. Cell morphology after 24 h and 5 days were imaged using Live/Dead live cell staining and SEM, respectively. Cell proliferation was quantified using an MTS assay at 1, 4, and 7 days. Osteogenic differentiation of the cells was examined by alkaline phosphatase (ALP) staining at 10 days and quantified using ALP activity and osteocalcin assays at 7, 14, and 21 days. The sizes of the HA, Sr5-HA, Sr10-HA, and Sr20-HA particles were mainly between 10 × 20 nm and 10 × 250 nm, and these nanoparticles were dispersed or clustered in the composite scaffolds. in vitro cell studies showed that the PPF/Sr10-HA scaffold was significantly better than the other three groups (PPF/HA, PPF/Sr5-HA, and PPF/Sr20-HA) in supporting MC3T3-E1 cell adhesion, proliferation, and differentiation. PPF/Sr10-HA may, therefore, serve as a promising scaffold material for bone tissue engineering. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 631-642, 2019.

Laser transmission welding offers significant benefits over conventional welding techniques enabling single-stage rapid plastic joining. The quality of laser transmission welded products is commonly assessed by measuring the weld penetration depth, hardened weld boundary, and inspecting the formation of porosity. However, the existing methods of verification are inevitably accompanied by destruction of the specimen. Thus, non-destructive quality assessment methods for laser transmission welding have gained attention recently. Here, we demonstrated an extended industrial application of 860 nm wavelength-based spectral domain optical coherence tomography for the non-destructive inspection of the aforementioned quality parameters of laser transmission welded industrial plastic materials, i.e., polycarbonate and acrylonitrile butadiene styrene copolymer. The acquired cross-sectional resolution and volumetric and intensity profiles sufficiently contributed to the quality assessment procedure, revealing the weld boundary and porosity formation, and thus confirming the potential applicability of optical coherence tomography for the quality inspection of laser transmission welded products.

Biomimetic biomaterials require good biocompatibility and bioactivity to serve as appropriate scaffolds for tissue engineering applications. Recent developments demonstrated that the unique properties of carbon nanotubes (CNTs) can enhance neural cell growth and axon organization. We previously developed a promising nerve conduit manufactured from biodegradable polycaprolactone fumarate (PCLF) for use in peripheral nerve regeneration applications. In the present study, we fabricated conductive PCLF-CNT scaffolds using ultraviolet (UV) induced photocrosslinking. We confirmed the successful incorporation of CNTs into the PCLF-CNT scaffolds which exhibited improved surface roughness compared with plain PCLF by scanning electronic microscopy, transmission electronic microscopy, and atomic force microscopy examinations. The PCLF-CNT substrates also had reduced impedance by electrochemical measurements. Enhanced PC-12 cell growth and differentiation were observed on PCLF-CNT sheets compared with PCLF sheets, indicating the beneficial effects of embedding CNTs into PCLF. Electrical stimulation not only enhanced PC-12 cell proliferation and neurite extension, but also promoted cellular migration and intracellular connections, which are all critical cellular behaviours for nerve regeneration. Overall, this study provides a new promising strategy for using electrically conductive PCLF-CNT nerve scaffolds in regenerative medicine.

Insulin-like peptide 3 (INSL3) is a member of the relaxin/insulin superfamily and is expressed in testicular Leydig cells. Essential for fetal testis descent, INSL3 has been implicated in testicular and sperm function in adult males via interaction with relaxin/insulin-like family peptide receptor 2 (RXFP2). The INSL3 is typically prepared using chemical synthesis or overexpression in Escherichia coli followed by oxidative refolding and proteolysis. Here, we expressed and purified full-length porcine INSL3 (pINSL3) using a silkworm-based Bombyx mori nucleopolyhedrovirus bacmid expression system. Biophysical measurements and proteomic analysis revealed that this recombinant pINSL3 exhibited the correct conformation, with the three critical disulfide bonds observed in native pINSL3, although partial cleavage occurred. In cAMP stimulation assays using RXFP2-expressing HEK293 cells, the recombinant pINSL3 possessed full biological activity. This is the first report concerning the production of fully active pINSL3 without post-expression treatments and provides an efficient production platform for expressing relaxin/insulin superfamily peptides.

Mice and rats are rodent specimens commonly used in multidisciplinary research. Specifically, vasculature imaging of rodents has been widely performed in preclinical studies using various techniques, such as computed tomography, magnetic resonance imaging, and ultrasound imaging. Photoacoustic CT (PACT) is a noninvasive, nonionizing optical imaging technique derived from photoacoustic tomography and benefits from using intrinsic endogenous contrast agents to produce three-dimensional volumetric data from images. In this study, a commercial PACT device was employed to assess the cervicothoracic vasculature of mouse and rat specimens, which has rarely been examined using PACT, under two conditions with depilation and skin incision. Various blood vessels, including the common carotid artery, internal/external jugular veins, cranial vena cava, internal thoracic vein, and mammary, were identified in the acquired PACT images. The difference between the depilated and skin-incised specimens also revealed the presence of branches from certain blood vessels and specific anatomical features such as the manubrium of the sternum. This study presents detailed PACT images observing the cervicothoracic vasculature of rodent specimens and is expected to be used as a reference for various preclinical experiments on mice and rats.

Nerve regeneration after injury is a critical medical issue. In previous work, we have developed an oligo(poly(ethylene glycol) fumarate) (OPF) hydrogel incorporated with positive charges as a promising nerve conduit. In this study, we introduced cross-linkable bonds to graphene oxide and carbon nanotube to obtain the functionalized graphene oxide acrylate (GOa) and carbon nanotube poly(ethylene glycol) acrylate (CNTpega). An electrically conductive hydrogel was then fabricated by covalently embedding GOa and CNTpega within OPF hydrogel through chemical cross-linking followed by in situ reduction of GOa in l-ascorbic acid solution. Positive charges were incorporated by 2-(methacryloyloxy)ethyltrimethylammonium chloride (MTAC) to obtain rGOaCNTpega-OPF-MTAC composite hydrogel with both surface charge and electrical conductivity. The distribution of CNTpega and GOa in the hydrogels was substantiated by transmission electron microscopy (TEM), and strengthened electrical conductivities were determined. Excellent biocompatibility was demonstrated for the carbon embedded composite hydrogels. Biological evaluation showed enhanced proliferation and spreading of PC12 cells on the conductive hydrogels. After induced differentiation using nerve growth factor (NGF), cells on the conductive hydrogels were effectively stimulated to have robust neurite development as observed by confocal microscope. A synergistic effect of electrical conductivity and positive charges on nerve cells was also observed in this study. Using a glass mold method, the composite hydrogel was successfully fabricated into conductive nerve conduits with surficial positive charges. These results suggest that rGOa-CNTpega-OPF-MTAC composite hydrogel holds great potential as conduits for neural tissue engineering.

Corneal transplantation by full‐thickness penetrating keratoplasty with human donor tissue is a widely accepted treatment for damaged or diseased corneas. Although corneal transplantation has a high success rate, a shortage of high‐quality donor tissue is a considerable limitation. Therefore, bioengineered corneas could be an effective solution for this limitation, and a decellularized extracellular matrix comprises a promising scaffold for their fabrication. In this study, three‐dimensional bioprinted decellularized collagen sheets were implanted into the stromal layer of the cornea of five rabbits. We performed in vivo noninvasive monitoring of the rabbit corneas using swept‐source optical coherence tomography (OCT) after implanting the collagen sheets. Anterior segment OCT images and averaged amplitude‐scans were acquired biweekly to monitor corneal thickness after implantation for 1 month. The averaged cornea thickness in the control images was 430.3 ± 5.9 μm, while the averaged thickness after corneal implantation was 598.5 ± 11.8 μm and 564.5 ± 12.5 μm at 2 and 4 weeks, respectively. The corneal thickness reduction of 34 μm confirmed the biocompatibility through the image analysis of the depth‐intensity profile base. Moreover, hematoxylin and eosin staining supported the biocompatibility evaluation of the bioprinted decellularized collagen sheet implantation. Hence, the developed bioprinted decellularized collagen sheets could become an alternative solution to human corneal donor tissue, and the proposed image analysis procedure could be beneficial to confirm the success of the surgery., Three‐dimensional bioprinted decellularized collagen sheets were implanted into the stromal layer of the cornea of five rabbits. After implanting the collagen sheets, the rabbit corneas were monitored biweekly in vivo with a swept‐source optical coherence tomography system to assess the biocompatibility of the bioprinted implantation for 1 month through corneal thickness measurements. The small variation in the averaged corneal thickness along with evidence from hematoxylin and eosin‐stained histology images shows the potential use of the biocompatible collagen sheet.

This study was designed to find an effective way of ceramic brackets debonding through the laser irradiation on the interface between the teeth and the bracket. Various laser parameters were applied to remove orthodontic brackets. After debonding, the adhesive remnant index (ARI) and the degree of enamel fractures were examined with microscope. The laser-assisted debonding be capable of lowering the risk of enamel damage and the less adhesive residue. The results indicated that the line laser system can be successfully applied to ceramic bracket remove tool. This method is expected to be useful in the field of orthodontics.

Objective: Human mesenchymal stromal cells (MSCs) exhibit variable differentiation potential and can be divided accordingly into distinct subpopulations whose ratios vary with donor age. However, it is unknown whether the same is true in pigs. This study investigated MSC subpopulations in miniature pig and compared their characteristics in young (2 to 3 months) and adult (27 to 35 months) pigs.Methods: Osteogenic, chondrogenic, and adipogenic capacity of isolated MSCs was evaluated by von Kossa, Alcian blue, and oil red O staining, respectively. Cell surface antigen expression was determined by flow cytometry. Proliferative capacity was assessed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Expression of marker genes was detected by quantitative real-time polymerase chain reaction.Results: Porcine MSCs comprised cells with trilineage and bilineage differentiation potential (tMSCs and bMSCs, respectively) and non-differentiating stromal cells (NDSCs). The tMSC and bMSC fractions were smaller in adult than in young pigs (63.0% vs 71.2% and 11.6% vs 24.0%, respectively, p

In this report, we have examined the distance- and size-dependent localized surface plasmon resonance (LSPR) between fluorescent quantum dots (QDs) and adjacent gold nanoparticles (AuNPs) to provide a comprehensive evaluation, aiming for practical application in biosensing platform. A series of peptides with different chain lengths, connected between QDs and AuNPs is initially applied to prepare various CdSe QDs-peptide-AuNP systems to optimize LSPR signal. Separation distance between two nanoparticles of these systems before and after conjugation is also confirmed by quantum mechanical modeling and corroborated with their LSPR influenced fluorescence variations. After detailed optimizations, it can be noted that larger sized AuNPs make strong quenching of QDs, which gradually shows enhancement of fluorescence with the increment of distance and the smaller sized AuNPs. Depending on the requirement, it is possible to tune the optimized structure of the CdSe QD-peptide-AuNP nanostructures for the application. In this work, two different structural designs with different peptide chain length are chosen to construct two biosensor systems, observing their fluorescence enhancement and quenching effects, respectively. Using different structural orientation of these biosensors, two nanoconjugates has applied for detection of norovirus and influenza virus, respectively to confirm their application in sensing.

Healing of nerve injuries is a critical medical issue. Biodegradable polymeric conduits are a promising therapeutic solution to provide guidance for axon growth in a given space, thus helping nerve heal. Extensive studies in the past decade reported that conductive materials could effectively increase neurite and axon extension in vitro and nerve regeneration in vivo. In this study, graphene oxide and carbon nanotubes were covalently functionalized with double bonds to obtain crosslinkable graphene oxide acrylate (GOa) sheets and carbon nanotube poly(ethylene glycol) acrylate (CNTpega). An electrically conductive reduced GOa-CNTpega-oligo(polyethylene glycol fumarate) (OPF) hydrogel (rGOa-CNTpega-OPF) was successfully fabricated by chemically crosslinking GOa sheets and CNTpega with OPF chains followed by in situ chemical reduction in L-ascorbic acid solution. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) imaging showed homogenous distribution of GOa/CNTpega carbon content in the rGOa-CNTpega-OPF composite hydrogel, resulting in a significant increase of electrical conductivity compared with neutral OPF without carbon content. Cell studies showed excellent biocompatibility and distinguished PC12 cell proliferation and spreading on the rGOa-CNTpega-OPF composite hydrogel. Fluorescent microscopy imaging demonstrated robustly stimulated neurite development in these cells on a conductive rGOa-CNTpega-OPF composite hydrogel compared with that on neutral OPF hydrogels. These results illustrated a promising potential for the rGOa-CNTpega-OPF composite hydrogel to serve as conduits for neural tissue engineering.

Photoacoustic imaging (PAI) has gained significant attention over the last two decades as a noninvasive, nonionizing, real-time, and inexpensive medical imaging modality. By exploiting the low acoustic scattering compared to optical scattering in biological tissues, PAI can achieve high resolution while maintaining superior optical contrast in deep tissues. PAI can probe many endogenous chromophores based on their strong optical absorption contrast, including oxygenated hemoglobin, deoxygenated hemoglobin, melanin, water, and lipid. However, these intrinsic absorbers are typically activated by visible (e.g., wavelength 400≤λ≤700 nm) or infrared (λ>1000 nm) light, at which wavelengths light penetration is severely limited. Because the optimal spectral region is the near-infrared (NIR, 700≤λ≤1000 nm) window for deep-tissue imaging, additional contrast mechanisms for PAI should be found. So far, various exogenous contrast agents have been developed for this purpose including small monomers, inorganic nanoparticles (NPs), organic NPs, etc. For successful and fast clinical translation, the proposed contrast agents must be safe and degradable in vivo, and the organic nanostructures may be good candidates for this purpose. In this article, we review recent progress in developing organic nanoformulations as PAI contrast agents.

The geometrical characteristics of dental implants, such as thread depth and width, facing angle, pitch, and surface roughness, are important to assess their stability and osseointegration after implant surgery. Herein, we demonstrate the potential use of depth-resolved swept-source optical coherence tomography (SS-OCT) to assess the structural quality of widely used dental implants. The implemented SS-OCT system was centered at a wavelength of 1300 nm with a 100 nm full-width at half-maximum. Four dental implants with different structural formations fabricated using either titanium or ceramic were visualized. Qualitative assessments were performed using boundary flattening with an amplitude-profiling algorithm to emphasize and compare the thread depths, surface roughness, and inner structures of the experimental samples. Cross-sectional and volumetric OCT data clearly revealed the depth, width, and pitch of the dental implants, and especially, the quantitative assessment of axial and lateral thread depth unobtainable using conventional inspection methods was successfully conducted. The depth of thread was measured using a depth-directional intensity profile. In conclusion, the high-resolution SS-OCT system could be utilized to improve the quality assurance of dental implant products through multi-plane assessment.

A large field-of-view and fast scanning of photoacoustic microscopy (PAM) relatively have been difficult to obtain due to the water-drowned structure of the system for the transmission of ultrasonic signals. Researchers have widely studied the achievement of a waterproof scanner for dynamic biological applications with a high-resolution and high signal-to-noise ratio. This Letter reports a novel, to the best of our knowledge, waterproof galvanometer scanner-based PAM system with a successfully attainable 9.0 m m × 14.5 m m scan region, amplitude scan rate of 40 kHz, and spatial resolution of 4.9 µm. The in vivo characterization of a mouse brain in intact-skull microvascular visualization demonstrated its capability in biomedical imaging and is anticipated to be an effective technique for various preclinical and clinical studies.

The lack of molecular targets and targeting probes remains a major drawback for targeted imaging and drug delivery in lung cancer. In this study, we exploited

Dental implants are common method to replace decayed or broken tooth. As the implant treatment procedures varies according to the patients’ jawbone, bone ridge, and sinus structure, appropriate examinations are necessary for successful treatment. Currently, radiographic examinations including periapical radiology, panoramic X-ray, and computed tomography are commonly used for diagnosing and monitoring. However, these radiographic examinations have limitations in that patients and operators are exposed to radioactivity and multiple examinations are performed during the treatment. In this study, we demonstrated photoacoustic (PA) and ultrasound (US) combined imaging of dental implant that can lower the total amount of absorbed radiation dose in dental implant treatment. An acoustic resolution PA macroscopy and a clinical PA/US system was used for dental implant imaging. The acquired dual modal PA/US imaging results support that the proposed photoacoustic imaging strategy can reduce the radiation dose rate during dental implant treatment.

In this study, we developed an imaging device for preclinical and clinical application using photoacoustic effect. The photoacoustic effect is to make ultrasound images using light as its name illustrates. Light irradiated in a short time (nano seconds) is absorbed by the tissue, and the local pressure is generated by the thermo-elastic expansion. The photoacoustic effect is a safe method of imaging because it uses an endogenous chromophore in the human body without the use of an external contrast agent. In this study, the image uses an optical-resolution photoacoustic microscope (OR-PAM) which has a resolution of 12 ¥im. The conventional or-pam has the disadvantage of being slow and bulky using mechanical motors. To overcome this problem, our research team developed a photoacoustic imaging system that uses a MEMS scanner to equip a fast image capability into a compact handheld probe.