Potential Bone Tissue Engineering Research Articles

Improved Osteogenesis by Noggin siRNA Collagen-Bioactive Glass Composites

Collagen-bioactive glass (COL–BG) composites have significant bone regeneration ability due to the considerable osteogenic performance and desirable biological properties of collagen. In the current research, the influence of COL–BG composite scaffolds loaded with noggin siRNA on osteogenesis in MC3T3 cells have been comprehensively studied. We prepared three types of COL–BG composites scaffolds, namely, unloaded, loaded with noggin siRNA, and loaded with negative control siRNA. The effect of the aqueous extracts achieved from these scaffolds on MC3T3 cell proliferation was monitored using a CCK8 test, whilst the effect on osteogenesis was assessed with ALP (alkaline phosphatase) activity assays, quantitative real-time PCR, and Alizarin Red staining (ARS). After cells were cultured for three and five days, proliferation in the presence of the extracts of composite scaffolds was remarkably greater than in the no-scaffold blank (P < 0.05). After 14 days of cultivation, MC3T3 cells exposed to COL–BG composites scaffold loaded with noggin siRNA exhibited greater ALP performance (P < 0.05) as well as higher mRNA expression (P < 0.01) of Runx2, BSP, and ALP genes in comparison with two other composite scaffolds, whereas ARS exhibited a larger number of mineralized nodules. Overall, these outcomes illustrate that COL–BG composite scaffold loaded with noggin siRNA is effective in enhancing osteogenesis. Thus, we suggest that this material has a promising potential for bone tissue engineering and is able to locally improve bone formation in bone defects.

Zwitterionic PMCP-functionalized titanium surface resists protein adsorption, promotes cell adhesion, and enhances osteogenic activity

Titanium (Ti) has excellent biocompatibility and corrosion resistance and is widely used as a biomedical material for orthopedic implants. However, the bare Ti surface limits cell adhesion without biological activity and promotes unnecessary protein adsorption, which can activate the coagulation pathway with blood-contacting devices. To improve the antifouling and biological activity of Ti, zwitterionic poly[2-(methacryloyloxy)ethyl choline phosphate] (PMCP) was used to modify the Ti surface via surface-initiated atom transfer radical polymerization. The Ti-PMCP surface reduced bovine serum albumin and fibrinogen adsorption owing to the zwitterionic antifouling property. Ti-PMCP is involved in the unique interaction between PMCP on the Ti surface and phosphate choline on cell membranes, and therefore, the Ti-PMCP surface can promote the adhesion and proliferation of MC3T3-e1 cells and bone marrow mesenchymal cells (BMSCs). In addition, the Ti-PMCP surface was effective in promoting the osteogenic differentiation of MC3T3-e1 cells and BMSCs because the phosphate group in MCP can stimulate osteogenic signaling pathways. Therefore, the PMCP-modified Ti surface can resist protein adsorption and promote the adhesion, proliferation, and differentiation of osteoblast-related cells and has great potential in bone tissue engineering.

Features of a simvastatin-loaded multi-layered co-electrospun barrier membrane for guided bone regeneration.

A novel tri-layer membrane consisting of polycaprolactone (PCL) fibrous sheets and structured nanofibers with a gelatin (Gt) shell and a simvastatin-containing PCL core (PCL-Gt/PCL-simvastatin membrane) was prepared. The soft external layer comprised of Gt/PCL-simvastatin, the external layer of PCL and the middle layer of both microfilaments, interwoven together. The membrane was designed to promote osteoinduction and act as a barrier against cells but not against water and molecules in order to promote guided bone regeneration. The structure of the membrane was characterized by scanning electronic microscopy. The in vitro release rates of simvastatin over 32 days were determined by high-performance liquid chromatography. For in vitro biological assays, bone marrow mesenchymal stem cells and human fibroblasts were cultured on the different surfaces of the membrane. Cell adhesion, proliferation, distribution, and differentiation were examined. For in vivo testing, cranial defects were created in rabbits to assess the amount of new bone formed for each membrane. The results revealed that membranes with multi-layered structures showed good cell viability and effective osteoinductive and barrier properties. These results suggest that the novel multi-layered PCL-Gt/PCL-simvastatin membranes have great potential for bone tissue engineering.

Behavior of molybdenum lowly doped SiO2–CaO–P2O5 composite interfacing a simulated biologic medium with serum albumin

This study is focussed on the biocompatibility evaluation by probing the affinity of bovine serum albumin (BSA) towards the surface of a silica based material containing CaMoO4 and hydroxyapatite nanocrystals. The adsorption occurring upon BSA interaction with the sample surface was investigated by Scanning Electron Microscopy (SEM), Fourier Transform Infrared (FTIR) Spectroscopy and X-ray Photoelectron Spectroscopy (XPS). The elemental composition and evaluation of protein surface coverage based on information from the outermost few nanometres surface layer was achieved by XPS analysis. The ratio between carbon and nitrogen on the material surface after incubation in BSA solution was found to be reasonable close to the value obtained from the measurement of pure BSA, that proves the protein adsorption. The comparative analysis of FTIR spectra recorded from lyophilized protein and from biomaterial surface after protein adsorption was considered in order to obtain information about the secondary structure of the proteins. The protein surface coverage was also confirmed by SEM morphological characterization. The results obtained by in vitro test indicate that the nanostructured SiO2–CaO–P2O5−MoO3 system has a great potential for bone tissue engineering with respect to the ability to bind to serum protein.

Characterization and evaluation of polycaprolactone/hydroxyapatite composite scaffolds with extra surface morphology by cryogenic printing for bone tissue engineering

In this work, the polycaprolactone (PCL) scaffold with specific surface morphology was prepared by cryogenic printing technique. Biocompatible hydroxyapatite (HA) was added to the PCL matrix to enhance the surface properties and bioactivity of the scaffolds. The effects of HA on the mechanical strength and surface morphology were studied systematically. It was found that the extra surface morphology of filament enhanced the exposure of the HA particles, which induced a positive synergistic effect on surface properties. The wettability and bio-mineralization were significantly improved, and the protein adsorption capacity was 4.5 and 3.9 times than the ones of sample with 20%HA and smooth surface, respectively. Rat bone mesenchymal stem cells (rBMSCs) were used as the cell model to evaluate the cellular response to the specific surface of the scaffolds. The results showed that the scaffolds with 20–40% HA and extra surface had better proliferation ratio. The ALP assay also demonstrated that the synergistic effect was beneficial to cell osteogenic differentiation, and the ALP level was significantly higher than that of pure PCL and smooth PCL/HA composite scaffold samples at the 14th day. Therefore, the PCL/HA composite scaffold with extra surface morphology had better potential for bone tissue engineering.

A new cancellous bone material of silk fibroin/cellulose dual network composite aerogel reinforced by nano-hydroxyapatite filler

Composites materials comprised of biopolymeric aerogel matrices and inorganic nano-hydroxyapatite (n-HA) fillers have received considerable attention in bone engineering. Although with significant progress in aerogel-based biomaterials, the brittleness and low strengths limit the application. The improvements in toughness and mechanical strength of aerogel-based biomaterials are in great need. In this work, an alkali urea system was used to dissolve, regenerate and gelate cellulose and silk fibroin (SF) to prepare composite aerosol. A dual network structure was shaped in the composite aerosol materials interlaced by sheet-like SF and reticular cellulose wrapping n-HA on the surface. Through uniaxial compression, the density of the composite aerogel material was close to the one of natural bone, and mechanical strength and toughness were high. Our work indicates that the composite aerogel has the same mechanical strength range as cancellous bone when the ratio of cellulose, n-HA and SF being 8:1:1. In vitro cell culture showed HEK-293T cells cultured on composite aerogels had high ability of adhesion, proliferation and differentiation. Totally, the presented biodegradable composite aerogel has application potential in bone tissue engineering.

Osteogenesis enhancement using poly (l-lactide-co-d, l-lactide)/poly (vinyl alcohol) nanofibrous scaffolds reinforced by phospho-calcified cellulose nanowhiskers

Electrospun poly (l-lactide-co-d, l-lactide) (PLDLLA)/poly (vinyl alcohol) (PVA) nanofibers were reinforced by various contents (0–1 wt%) of phospho-calcified cellulose nanowhiskers (PCCNWs) as scaffolds in bone applications. The hydrophilicity and rate of hydrolytic degradation of PLDLLA were improved by introducing 10 wt% of PVA. PCCNWs with inherent hydrophilic properties, high aspect ratio, and large elastic modulus enhanced the hydrophilicity, accelerated the rate of degradation, and improved the mechanical properties of the nanofibrous samples. Moreover, calcium phosphate and phosphate functional groups on the surface of PCCNWs possessing act as stimulating agents for cellular activities such as proliferation and differentiation. Besides the physico-chemical properties investigation of PLDLLA/PVA-PCCNWs nanofibrous samples, their cytotoxicity was also studied and they did not show any adverse side effect. Incorporation of PCCNWs (1 wt%) into the PLDLLA/PVA nanofibrous samples showed more enzymatic activities and deposited calcium. The micrograph images of the morphology of human mesenchymal stem cells (hMSCs) cultured on the nanofibrous sample containing 1 wt% of PCCNWs after 14 days of cell differentiation revealed their high potential for bone tissue engineering.

Beneficial Influence of Water-Soluble PEG-Functionalized C60 Fullerene on Human Osteoblast Growth In Vitro.

The purpose of this study was to make an initial assessment of new PEG (polyethylene glycol)-functionalized C60 fullerene derivative for potential bone tissue engineering applications. Thus, Fourier Transform Infrared spectroscopy analysis, thermogravimetric analysis, and cyclic voltammetry measurement were performed. Moreover, cell culture experiments in vitro were carried out using normal human osteoblasts. Cell viability and proliferation were evaluated using colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test as well as by fluorescent staining. It was demonstrated that resultant derivative possessed good solubility in water, high temperature stability, and retained favorable electron accepting properties of C60 fullerene core. Most important, new fullerene derivatives at low concentrations did not exhibit cytotoxic effect and supported osteoblast proliferation compared to control. Thanks to all mentioned properties of new PEG-functionalized C60 fullerene derivative, it seems that it could be used as a component of polymer-based bone scaffolds in order to enhance their biological properties.

Porous Zirconia/Magnesia Ceramics Support Osteogenic Potential In Vitro.

Porous zirconia (ZrO2), magnesia (MgO) and zirconia/magnesia (ZrO2/MgO) ceramics were synthesised by sintering and designated as ZrO2(100), ZrO2(75)MgO(25), ZrO2(50)MgO(50), ZrO2(25)MgO(75), MgO(100) based on their composition. The ceramic samples were characterised by means of scanning electron microscopy, X-ray diffraction, energy-dispersive X-ray spectroscopy and atomic absorption spectrometry to explore the incorporation of Mg atoms into the zirconia lattice. The resulting porosity of the samples was calculated based on the composition and density. The final porosity of the cylinder-shaped ceramic samples ranged between 30 and 37%. The mechanical analysis exhibited that the Young modulus increased and the microstress decreased with increasing magnesia amount, with values ranging from 175 GPa for zirconia to 301 GPa for magnesia. The adhesion, viability, proliferation and osteogenic activity of MC3T3-E1 pre-osteoblastic cells cultured on the zirconia/magnesia ceramics was found to increase, with the magnesia-containing ceramics exhibiting higher values of calcium mineralisation. The results from the mechanical analysis, the ALP activity, the calcium and collagen production demonstrate that the zirconia/magnesia ceramics possess robust osteoinductive capacity, therefore holding great potential for bone tissue engineering.

Curdlan-Chitosan Electrospun Fibers as Potential Scaffolds for Bone Regeneration.

Polysaccharides have received a lot of attention in biomedical research for their high potential as scaffolds owing to their unique biological properties. Fibrillar scaffolds made of chitosan demonstrated high promise in tissue engineering, especially for skin. As far as bone regeneration is concerned, curdlan (1,3-β-glucan) is particularly interesting as it enhances bone growth by helping mesenchymal stem cell adhesion, by favoring their differentiation into osteoblasts and by limiting the osteoclastic activity. Therefore, we aim to combine both chitosan and curdlan polysaccharides in a new scaffold for bone regeneration. For that purpose, curdlan was electrospun as a blend with chitosan into a fibrillar scaffold. We show that this novel scaffold is biodegradable (8% at two weeks), exhibits a good swelling behavior (350%) and is non-cytotoxic in vitro. In addition, the benefit of incorporating curdlan in the scaffold was demonstrated in a scratch assay that evidences the ability of curdlan to express its immunomodulatory properties by enhancing cell migration. Thus, these innovative electrospun curdlan–chitosan scaffolds show great potential for bone tissue engineering.

Two-step strategy for constructing hierarchical pore structured chitosan–hydroxyapatite composite scaffolds for bone tissue engineering

Chitosan (CS) combined with hydroxyapatite (HA) was injected into a composite braid, and a hierarchical pore structure scaffold was obtained by freeze drying and cold atmospheric plasma (CAP) technology. The CS/HA/braid scaffold with hierarchical pore structure was analyzed and characterized by scanning electronic microscopy, Fourier transform infrared spectroscopy, true color confocal microscopy, improved liquid replacement method, and phosphate buffer solution immersion. The mechanical properties and degradation ability of the scaffold were evaluated through compression test and degradation test. Results showed that HA addition endowed the core of the scaffold with macroscopic pore sizes of 80–180 μm, and CAP treatment endowed the shell of the scaffold with microscopic pore sizes ≤10 μm. All scaffolds exhibited high porosity and swelling rates of ≥80 % and ≥300 %, respectively. The scaffold with a hierarchical pore structure had good mechanical properties and twice the degradation rate. In addition, the treated scaffold precipitated intact spherical HA crystals. Under the synergistic effect of HA and CAP treatment, scaffolds achieved 277.6 % cell viability compared with pure CS scaffold. Overall, this method was feasible for preparing bone scaffolds with hierarchical pore structure for potential bone tissue engineering.

Three-Dimensional Printing of Calcium Carbonate/Hydroxyapatite Scaffolds at Low Temperature for Bone Tissue Engineering.

Three-dimensional (3D) printing technology has been applied to fabricate bone tissue engineering scaffolds for a wide range of materials with precisely control over scaffold structures. Coral is a potential bone repair and bone replacement material. Due to the natural source limitation of coral, we developed a fabrication protocol for 3D printing of calcium carbonate (CaCO3) nanoparticles for coral replacement in the application of bone tissue engineering. Up to 80% of CaCO3 nanoparticles can be printed with high resolution using poly-l-lactide as a blender. The scaffolds were subjected to a controlled hydrothermal process for incomplete conversion of carbonate to phosphate to produce CaCO3 scaffold covered by hydroxyapatite (HA) to modify the biocompatibility and degradation of CaCO3/HA scaffolds. X-ray diffraction and Fourier transform infrared spectroscopy showed that HA was converted and attached to the surface of the scaffold, and the surface morphology and microstructure were studied using a scanning electron microscope. To confirm the bone regeneration performance of the scaffold, cell proliferation and osteogenic differentiation of MC3T3 cells on the scaffold were evaluated. In addition, in vivo experiments showed that CaCO3/HA scaffolds can promote bone growth and repairing process and has high potential in bone tissue engineering. ClinicalTrials.gov ID: SH9H-2020-A603.

Advances in Osteoporotic Bone Tissue Engineering.

The increase in osteoporotic fracture worldwide is urging bone tissue engineering research to find new, improved solutions both for the biomaterials used in designing bone scaffolds and the anti-osteoporotic agents capable of promoting bone regeneration. This review aims to report on the latest advances in biomaterials by discussing the types of biomaterials and their properties, with a special emphasis on polymer-ceramic composites. The use of hydroxyapatite in combination with natural/synthetic polymers can take advantage of each of their components properties and has a great potential in bone tissue engineering, in general. A comparison between the benefits and potential limitations of different scaffold fabrication methods lead to a raised awareness of the challenges research face in dealing with osteoporotic fracture. Advances in 3D printing techniques are providing the ways to manufacture improved, complex, and specialized 3D scaffolds, capable of delivering therapeutic factors directly at the osteoporotic skeletal defect site with predefined rate which is essential in order to optimize the osteointegration/healing rate. Among these factors, strontium has the potential to increase osseointegration, osteogenesis, and healing rate. Strontium ranelate as well as other biological active agents are known to be effective in treating osteoporosis due to both anti-resorptive and anabolic properties but has adverse effects that can be reduced/avoided by local release from biomaterials. In this manner, incorporation of these agents in polymer-ceramic composites bone scaffolds can have significant clinical applications for the recovery of fractured osteoporotic bones limiting or removing the risks associated with systemic administration.

Investigation of attributes of bourbon oil and cobalt nitrate constituted electrospun nanoscaffolds for blood compatibility and in vitro bone formation.

This work aims to fabricate scaffold using polyurethane (PU) integrated with bourbon oil (BB) and cobalt nitrate (CoNO3) using the electrospinning technique. Morphological investigation signified a fall in fibre diameter for the PU/BB and PU/BB/CoNO3 nanocomposite than the PU. Spectral analysis indicated that BB and CoNO3 were added within the PU matrix. Wettability analysis insinuated an increase in the hydrophobic nature of the PU/BB than the PU. PU/BB/CoNO3 turned to be hydrophilic due to the integration of CoNO3 in the polymer matrix. Mechanical testing of PU/BB and PU/BB/CoNO3 indicated an increase in the tensile strength of the fabricated composites. Atomic force microscopy (AFM) portrayed the reduction in the roughness of the PU/BB and PU/BB/CoNO3 compared to the PU. The coagulation studies invariably documented the improved anticoagulant behaviour and less toxic nature of the PU/BB and PU/BB/CoNO3 in comparison with the PU. Further, bone mineralization testing revealed the enhanced apatite formation of the nanocomposite. Nanocomposite scaffolds with the fore-mentioned properties hold good potential for bone tissue engineering.

Fractal Design Boosts Extrusion-Based 3D Printing of Bone-Mimicking Radial-Gradient Scaffolds.

Although extrusion-based three-dimensional (EB-3D) printing technique has been widely used in the complex fabrication of bone tissue-engineered scaffolds, a natural bone-like radial-gradient scaffold by this processing method is of huge challenge and still unmet. Inspired by a typical fractal structure of Koch snowflake, for the first time, a fractal-like porous scaffold with a controllable hierarchical gradient in the radial direction is presented via fractal design and then implemented by EB-3D printing. This radial-gradient structure successfully mimics the radially gradual decrease in porosity of natural bone from cancellous bone to cortical bone. First, we create a design-to-fabrication workflow with embedding the graded data on basis of fractal design into digital processing to instruct the extrusion process of fractal-like scaffolds. Further, by a combination of suitable extruded inks, a series of bone-mimicking scaffolds with a 3-iteration fractal-like structure are fabricated to demonstrate their superiority, including radial porosity, mechanical property, and permeability. This study showcases a robust strategy to overcome the limitations of conventional EB-3D printers for the design and fabrication of functionally graded scaffolds, showing great potential in bone tissue engineering.

A hierarchical scaffold with a highly pore-interconnective 3D printed PLGA/n-HA framework and an extracellular matrix like gelatin network filler for bone regeneration.

The ideal scaffold for bone repair should have a hierarchical pore structure and gradient degradation performance to satisfy the uniform adhesion and proliferation of cells in the scaffold at the early stage of implantation, as well as providing space for the subsequent regeneration of bone tissue. To this end, we developed a hierarchical polylactic acid glycolic acid copolymer (PLGA)/nano-hydroxyapatite (n-HA)/gelatin (Gel) (PHG) scaffold with a printed PLGA/n-HA (PH) framework and a Gel network filler for bone regeneration by the combination of 3D printing and freeze-drying technologies. The fabricated PHG scaffold features large front hole size (>1100 μm × 1100 μm) and side hole size (>500 μm) to provide sufficient open space and reliable integrated support for cell and tissue ingrowth. The gelatin network filled in the PH framework played the role of a cell holder just like an extracellular matrix (ECM) in the early stage. In vitro degradation experiments revealed that the gelatin network completely degraded within 5 weeks while the structural integrity of the framework still remained at the 32nd week. The results of cell culture confirmed that the PHG scaffold was more conducive to cell attachment. In vivo assessments in a rat femoral defect model showed that PHG scaffolds were more favored for new bone formation and achieving a tighter bond between the scaffold and the original tissues. The hierarchical PHG scaffold has great application potential in bone tissue engineering and will provide a reference for the model design of bone scaffolds.

Biomimetic polydopamine-laced hydroxyapatite collagen material orients osteoclast behavior to an anti-resorptive pattern without compromising osteoclasts' coupling to osteoblasts.

Polydopamine-assisted modification for bone substitute materials has recently shown great application potential in bone tissue engineering due to its excellent biocompatibility and adhesive properties. A scaffold material's impact on osteoclasts is equally as important as its impact on osteoblasts when considering tissue engineering for bone defect repair, as healthy bone regeneration requires an orchestrated coupling between osteoclasts and osteoblasts. How polydopamine-functionalized bone substitute materials modulate the activity of osteoblast lineage cells has been extensively investigated, but much less is known about their impact on osteoclasts. Moreover, most of the polydopamine-functionalized materials would need to additionally load a biomolecule to exert the modulation on osteoclast activity. Herein, we demonstrated that our biomimetic polydopamine-laced hydroxyapatite collagen (PDHC) scaffold material, which does not need to load additional bioactive agent, is sufficiently able to modulate osteoclast activity in vitro. First, PDHC showed an anti-resorptive potential, characterized by decreased osteoclast differentiation and resorption capacity and changes in osteoclasts' transcriptome profile. Next, cAMP response element-binding protein (CREB) activity was found to mediate PDHC's anti-osteoclastogenic effect. Finally, although PDHC altered clastokines expression pattern of osteoclasts, as revealed by transcriptomic and secretomic analysis, osteoclasts' coupling to osteoblasts was not compromised by PDHC. Collectively, this study demonstrated the PDHC material orients osteoclast behavior to an anti-resorptive pattern without compromising osteoclasts' coupling to osteoblasts. Such a feature is favorable for the net increase of bone mass, which endows the PDHC material with great application potential in preclinical/clinical bone defect repair.

The effects of alignment and diameter of electrospun fibers on the cellular behaviors and osteogenesis of BMSCs

Electrospun fiber scaffolds, due to their mimicry of bone extracellular matrix (ECM), have become an important biomaterial widely applied in bone tissue engineering in recent years. While topographic cues of electrospun membranes such as alignment and diameter played vital roles in determining cellular behaviors. Yet few researches about the effects of these two significant parameters on osteogenesis have been reported. Thus, the present work explored the influence of aligned and random poly (L-lactic acid) (PLLA) fiber matrices with diameters of nanoscale (0.6 μm) and microscale (1.2 μm), respectively, on cellular responses of bone marrow mesenchymal stem cells (BMSCs), such as cell adhesion, migration, proliferation and osteogenesis. Our results revealed that aligned nanofibers (AN) could affect cell morphology and promote the migration of BMSCs after 24 h of cell culturing. Besides, AN group was observed to possess excellent biocompatibility and have significantly improved cell growth comparing with random nanofibers. More importantly, in vitro osteogenesis researches including ALP and Alizarin Red S staining, qRT-PCR and immunofluorescence staining demonstrated that BMSCs culturing on AN group exhibited higher osteogenic induction proficiency than that on aligned microfibers (AM) and random fiber substrates (RN and RM). Accordingly, aligned nanofiber scaffolds have greater application potential in bone tissue engineering.

Human osteoblast-like SAOS-2 cells on submicron-scale fibers coated with nanocrystalline diamond films

A unique composite nanodiamond-based porous material with a hierarchically-organized submicron-nano-structure was constructed for potential bone tissue engineering. This material consisted of submicron fibers prepared by electrospinning of silicon oxide (SiOx), which were oxygen-terminated (O-SiOx) and were hermetically coated with nanocrystalline diamond (NCD) films. The NCD films were then terminated with hydrogen (H-NCD) or oxygen (O-NCD). The materials were tested as substrates for the adhesion, growth and osteogenic differentiation of human osteoblast-like Saos-2 cells. The number and the spreading area of the initially adhered cells, their growth rate during 7 days after seeding and the activity of alkaline phosphatase (ALP) were significantly higher on the NCD-coated samples than on the uncoated O-SiOx samples. In addition, the concentration of type I collagen was significantly higher in the cells on the O-NCD-coated samples than on the bare O-SiOx samples. The observed differences could be attributed to the tunable wettability of NCD and to the more appropriate surface morphology of the NCD-coated samples in contrast to the less stable, rapidly eroding bare SiOx surface. The H-NCD coatings and the O-NCD coatings both promoted similar initial adhesion of Saos-2 cells, but the subsequent cell proliferation activity was higher on the O-NCD-coated samples. The concentration of beta-actin, vinculin, type I collagen and alkaline phosphatase (ALP), the ALP activity, and also the calcium deposition tended to be higher in the cells on the O-NCD-coated samples than on the H-NCD-coated samples, although these differences did not reach statistical significance. The improved cell performance on the O-NCD-coated samples could be attributed to higher wettability of these samples (water drop contact angle less than 10°), while the H-NCD-coated samples were hydrophobic (contact angle >70°). NCD-coated porous SiOx meshes can therefore be considered as appropriate scaffolds for bone tissue engineering, particularly those with an O-terminated NCD coating.

Osteogenically-induced exosomes stimulate osteogenesis of human adipose-derived stem cells

Exosomes exhibit great therapeutic potential in bone tissue engineering. The study aimed to investigate whether the exosomes derived from human adipose-derived stem cells (hADSCs-Exos) during different time-span of osteogenic differentiation could promote osteogenesis. The appropriate concentrations of hADSCs-Exos to enhance the proliferation, migration and osteogenesis of hADSCs-Exos were also examined. PKH67 labelled hADSCs-Exos was used to detect the internalization ability of hADSCs. The osteogenic differentiation abilities of hADSCs after treatment with hADSCs-Exos was evaluated by Alizarin red staining (ARS). The proliferation and migration of hADSCs was examined by cell counting kit-8 and wound healing assay, respectively. The expression of exosomal surface markers and osteoblast-related protein of hADSCs was assessed by Western blot. PKH67-labelled exosomes were internalized by hADSCs after 4 h incubation. ARS showed that the amount of mineralized nodules in Exo1−14d group was significantly higher than that in Exo15−28d group. hADSCs-Exos could promote the proliferation and migration capacity of hADSCs. Western blot analysis showed that after hADSCs-Exos treatment, ALP and RUNX2 were significantly enhanced. Specially, the Exo1−14d group of 15 μg/mL significantly upregulated the expression of RUNX2 than the other exosomes treated groups. Our findings suggest that exosomes secreted by hADSCs during osteogenic induction for 1–14 days could be efficiently internalized by hADSCs and could induce osteogenic differentiation of hADSCs. Moreover, administration of Exo1−14d at 15 μg/mL promoted the proliferation and migration of hADSCs. In conclusion, our research confirmed that comprised of hADSCs-Exos and hADSCs may provide a new therapeutic paradigm for bone tissue engineering.