A multi-material coating containing chemically-modified apatites for combined enhanced bioactivity and reduced infection via a drop-on-demand micro-dispensing technique.

Prevention of infection and enhanced osseointegration are closely related, and required for a successful orthopaedic implant, which necessitate implant designs to consider both criteria in tandem. A multi-material coating containing 1:1 ratio of silicon-substituted hydroxyapatite and silver-substituted hydroxyapatite as the top functional layer, and hydroxyapatite as the base layer, was produced via the drop-on-demand micro-dispensing technique, as a strategic approach in the fight against infection along with the promotion of bone tissue regeneration. The homogeneous distribution of silicon-substituted hydroxyapatite and silver-substituted hydroxyapatite micro-droplets at alternate position in silicon-substituted hydroxyapatite-silver-substituted hydroxyapatite/hydroxyapatite coating delayed the exponential growth of Staphylococcus aureus for up to 24 h, and gave rise to up-regulated expression of alkaline phosphatase activity, type I collagen and osteocalcin as compared to hydroxyapatite and silver-substituted hydroxyapatite coatings. Despite containing reduced amounts of silicon-substituted hydroxyapatite and silver-substituted hydroxyapatite micro-droplets over the coated area than silicon-substituted hydroxyapatite and silver-substituted hydroxyapatite coatings, silicon-substituted hydroxyapatite-silver-substituted hydroxyapatite/hydroxyapatite coating exhibited effective antibacterial property with enhanced bioactivity. By exhibiting good controllability of distributing silicon-substituted hydroxyapatite, silver-substituted hydroxyapatite and hydroxyapatite micro-droplets, it was demonstrated that drop-on-demand micro-dispensing technique was capable in harnessing the advantages of silver-substituted hydroxyapatite, silicon-substituted hydroxyapatite and hydroxyapatite to produce a multi-material coating along with enhanced bioactivity and reduced infection.

Mechanically-enhanced three-dimensional scaffold with anisotropic morphology for tendon regeneration.

Tissue engineering has showed promising results in restoring diseased tendon tissue functions. Herein, a hybrid three-dimensional (3D) porous scaffold comprising an outer portion rolled from an electrohydrodynamic jet printed poly(ɛ-caprolactone) (PCL) fiber mesh, and an inner portion fabricated from uniaxial stretching of a heat-sealed PCL tube, was developed for tendon tissue engineering (TE) application. The outer portion included three layers of micrometer-scale fibrous bundles (fiber diameter: ~25 µm), with an interconnected spacing and geometric anisotropy along the scaffold length. The inner portion showed orientated micro-ridges/grooves in a parallel direction to that of the outer portion. Owning to the addition of the inner portion, the as-fabricated scaffold exhibited comparable mechanical properties to those of the human patellar tendon in terms of Young's modulus (~227 MPa) and ultimate tensile stress (~50 MPa). Compared to the rolled electrospun fibers, human tenocytes cultured in the tendon scaffolds showed increased cellular metabolism. Furthermore, the 3D tendon scaffold resulted in up-regulated cell alignment, cell elongation and formation of collagen type I. These results demonstrated the potential of mechanically-enhanced 3D fibrous scaffold for applications in tendon TE, with desired cell alignment and functional differentiation.

Optimizing fabrication parameters via Taguchi method for production of high yield hydroxyapatite microsphere scaffolds using Drop-on-Demand inkjet method.

Drop on demand (DOD) inkjet method is a cost-efficient way of producing hydroxyapatite (HAp) microsphere scaffolds with narrow size distribution. However, DOD fabrication parameters may influence the yield and characteristics of the microsphere scaffolds. Testing different permutations and combinations of fabrication parameters is costly and time consuming. Taguchi method could be used as a predictive tool for optimizing the key fabrication parameters to produce HAp microspheres with desired yield and properties, minimizing the number of experimental combinations to be tested. The aim of this study is to investigate the influence of the fabrication parameters on the characteristics of the microspheres formed and determine optimum parameter conditions for producing high yield HAp microsphere scaffolds with the desired properties intended to serve as potential bone substitutes. We aimed to achieve microspheres with high production yield, microsphere size of <230 µm, micropore sizes <1 µm, rough surface morphology and high sphericity. Experiments were conducted using Taguchi method with a L9 orthogonal array at three levels per parameter to determine optimum parameter values for (1) operating pressure, (2) shutter speed duration, (3) nozzle height and (4) CaCl2 concentration. Based on signal-to-noise (S/N) ratio analysis, the identified optimum parameter conditions for operating pressure, shutter speed duration, nozzle height and CaCl2 concentration to be 0.9-1.3 bar, 100 ms, 8 cm and 0.4 M respectively. The microspheres obtained had an average size of 213 µm, 0.45 µm micropore size, high sphericity index of 0.95 and high production yield of 98%. Confirmation tests and ANOVA results affirms the validity of Taguchi method in optimizing HAp microspheres with high yield, desired size, micropore size and shape. HAp microsphere scaffolds produced by optimum conditions were subjected to a 7-day in-vitro study. Cells remained viable and continued to proliferate (increased 1.2-fold) over 7 days with microspheres maintaining high cell density with cells bridging between microspheres. Alkaline phosphatase (ALP) assay increased 1.5-fold from day 1, suggesting good osteogenic potency of HAp microspheres as potential bone substitutes.

Biodegradation of polymers in managing plastic waste - A review.

The modern economy that is fast-moving and convenience centric has led to excessive consumption of plastic. This has unwittingly led to egregious accumulation of plastic waste polluting the environment. Unfortunately, present means of plastic waste management have all been proven as less than adequate; namely recycling, landfill and incineration. Recent focus on plastic waste management has seen the confluence of the developments in biodegradable polymers and microbial engineering strategy for more expedient decomposition of plastic waste at composting facilities. This review paper is an assimilation of current developments in the areas of biodegradable polymer as well as microbial strategy towards management of polymer waste. Advents in biodegradable polymers have been promising, especially with aliphatic polyesters and starch in blends or co-polymers of these. Microbial strategies have been pursued for the identification of microbial strains and understanding of their enzymatic degradation process on polymers. New insights in these two areas have been focused in improving the rate of degradation of plastic waste at composting facilities. Recent alignment of testing and certification standards is outlined to give intimate insights into the mechanisms and factors influencing biodegradation. Despite recent milestones, economic viability of composting plastic waste in mainstream waste facilities is still a distance away. As it remains that a polymer that is biodegradable is functionally inferior to conventional polymers. Rather, it requires a shift in consumer behaviour to accept less durable biodegradable plastic products, this will then lower the threshold for biodegradable polymers to become a commercial reality.

Silver-substituted hydroxyapatite inhibits Pseudomonas aeruginosa outer membrane protein F: A potential antibacterial mechanism.

Bone fractures are one of the most common injuries, and they have a big effect on population health worldwide. Traumatic bone injuries can be partially treated with implanting bone-graft substitutes, for example, hydroxyapatite (HA), a bioceramic that is similar materially to natural bones with good bioactivity and osteoconductivity. It could, however, be vulnerable to infections because of the way an HA-based bone graft is put in, which could be a weakness in the host's defense. This study incorporated silver (Ag) into hydroxyapatite (Ag-HA) and silicon-containing hydroxyapatite (AgSi-HA) discs to combat this implant-triggered infection. Further, we investigated the antibacterial activities and potential underlying mechanism against a gram-negative bacterium, Pseudomonas aeruginosa. We noticed that the rich calcium (Ca2+) content in HA discs could trigger the change in P. aeruginosa physiology that leads to the enhanced bacterial growth on non­silver incorporated HA discs. But the released Ag+ from Ag-HA and AgSi-HA discs caused significant damage to bacterial cells at a low concentration of 0.3 ppm. We also observed dramatic morphological changes of Ag-HA and AgSi-HA surface-attached bacteria cells. Finally, we identified a potential action mechanism - the surface-bound Ag+ from Ag-HA and AgSi-HA potently inhibited the outer membrane protein F (OprF) expression of P. aeruginosa. Collectively, our results indicate that incorporating silver ions into HA could contribute viably to excellent antibacterial activities against P. aeruginosa to prevent HA-based bone graft infection.

Surface Design for Antibacterial Materials: From Fundamentals to Advanced Strategies.

Healthcare-acquired infections as well as increasing antimicrobial resistance have become an urgent global challenge, thus smart alternative solutions are needed to tackle bacterial infections. Antibacterial materials in biomedical applications and hospital hygiene have attracted great interest, in particular, the emergence of surface design strategies offer an effective alternative to antibiotics, thereby preventing the possible development of bacterial resistance. In this review, recent progress on advanced surface modifications to prevent bacterial infections are addressed comprehensively, starting with the key factors against bacterial adhesion, followed by varying strategies that can inhibit biofilm formation effectively. Furthermore, "super antibacterial systems" through pre-treatment defense and targeted bactericidal system, are proposed with increasing evidence of clinical potential. Finally, the advantages and future challenges of surface strategies to resist healthcare-associated infections are discussed, with promising prospects of developing novel antimicrobial materials.

Silicon-incorporated nanohydroxyapatite-reinforced poly(ε-caprolactone) film to enhance osteogenesis for bone tissue engineering applications.

Biomaterials composed of polymers and bioceramics have great prospects to repair large and complicated bone defects. Here, we developed a composite film consisting of poly(ε-caprolactone) (PCL) and silicon-substituted hydroxyapatite (Si-HA) nanoparticles to enhance the osteogenic effects of the scaffold for bone tissue engineering applications. The results showed that the Si-HA nanoparticles obtained an even distribution in the PCL matrix, resulting in a homogeneous composite film. Compared to HA-incorporated PCL film, the addition of silicon did not cause hydrophilic alterations to the film surface. With the seeding of mouse calvarial preosteoblasts (MC3T3-E1), the cells exhibited the good behaviors of adhesion and growth on the PCL/Si-HA film. Compared to the PCL/HA films, incorporation of Si-HA nanoparticles in PCL/Si-HA films showed the increased production of alkaline phosphatase (ALP) and calcium content by MC3T3-E1 cells. These results suggested the suitability of the PCL/Si-HA composite film to elicit cellular growth and functional differentiation with the potential for bone tissue engineering applications.

Fabrication of chelate-setting α-tricalcium phosphate cement using sodium citrate and sodium alginate as mixing solution and its in vivo osteoconductivity.

Moldable and injectable calcium-phosphate cements (CPCs) are material candidates for bone replacement applications. In the present study, we examined the effectiveness of sodium alginate and sodium citrate additives to the liquid phase of CPC, in improving its handling property as well as mechanical strength. The use of these additives enhanced the handling property significantly, in terms of consistency as compared to CPC without additives due to the liquefying effect caused by the adsorption of citrate ions on the cement particles. Sodium alginate and sodium citrate were added to CPC, which was set by the chelate-bonding capability of inositol phosphate, and was composed of mainly α-tricalcium phosphate (α-TCP) phase (>90%). The compressive strength of the CPC containing sodium alginate and sodium citrate was 3.4 ± 0.3 MPa, which was significantly higher than cement without additives. Furthermore, this cement exhibited favorable osteoconductivity and bioresorbability, and remained the α-TCP phase after 4-week implantation in a pig tibiae model. These results suggested that the cement is a potential candidate as a bioresorbable paste-like artificial bone. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2361-2370, 2018.

Injectable chelate-setting hydroxyapatite cement prepared by using chitosan solution: Fabrication, material properties, biocompatibility, and osteoconductivity.

An injectable chelate-setting hydroxyapatite cement (IP6-HAp), formed by chelate-bonding capability of inositol phosphate (IP6), was developed. The effects of ball-milling duration of starting HAp powder and IP6 concentration on the material properties such as injectability and mechanical strength of the cement were examined. The cement powder was prepared by ball-milling the as-synthesized HAp powder for 5 min using ZrO2 beads with a diameter of 10 mm, followed by another 60 min with ZrO2 beads with a diameter of 2 mm, and thereafter surface-modified with 5000 ppm of IP6 solution. Injectable cement was then fabricated with this HAp powder and 2.5 mass% chitosan as a mixing solution, with a setting time of 36.3 ± 4.7 min and a compressive strength of 19.0 ± 2.1 MPa. The IP6-HAp cements prepared with chitosan showed favorable biocompatibility in vitro using an osteoblast cell model, and osteoconductivity in vivo using a pig tibia model.

Uniformly-dispersed nanohydroxapatite-reinforced poly(ε-caprolactone) composite films for tendon tissue engineering application.

Regeneration of injuries at tendon-to-bone interface (TBI) remains a challenging issue due to the complex tissue composition involving both soft tendon tissues and relatively hard bone tissues. Tissue engineering using polymeric/ceramic composites has been of great interest to generate scaffolds for tissue's healing at TBI. Herein, we presented a novel method to blend polymers and bioceramics for tendon tissue engineering application. A homogeneous composite comprising of nanohydroxyapatite (nHA) particles in poly(ε-caprolactone) (PCL) matrix was obtained using a combination of solvent and mechanical blending process. X-ray diffraction analysis showed that the as-fabricated PCL/nHA composite film retained phase-pure apatite and semi-crystalline properties of PCL. Infrared spectroscopy spectra confirmed that the PCL/nHA composite film exhibited the characteristics functional groups of PCL and nHA, without alteration to the chemical properties of the composite. The incorporation of nHA resulted in PCL/nHA composite film with improved mechanical properties such as Young's Modulus and ultimate tensile stress, which were comparable to that of the native human rotator tendon. Seeding with human tenocytes, cells attached on the PCL/nHA composite film, and after 14days of culturing, these cells could acquire elongated morphology without induced cytotoxicity. PCL/nHA composite film could also result in increased cell metabolism with prolonged culturing, which was comparable to that of the PCL group and higher than that of the nHA group. All these results demonstrated that the developed technique of combining solvent and mechanical blending could be applied to fabricate composite films with potential for tendon tissue engineering applications.

The response of osteoblasts to nanocrystalline silicon-substituted hydroxyapatite thin films.

Magnetron co-sputtering has been employed to fabricate thin nanocrystalline coatings of silicon-substituted hydroxyapatite (SiHA) of different Si compositions: 0.8 wt%, 2.2 wt%, and 4.9 wt%. A human osteoblast-like (HOB) cell model was used to study the long-term interaction between the HOB cells and coatings in vitro. Results showed that the number of cells growing on all coated titanium (Ti) samples were statistically significantly higher than on uncoated Ti. In addition, HOB cells growing on all SiHA surfaces displayed enhanced cell spreading, with extensive extracellular matrix synthesis. DNA staining revealed normal phenotype nuclear morphology for HOB cells, with several dense chromosomes surrounded by a periphery of intact nuclear membrane. Furthermore, immunofluorescent staining indicated that cells showed improved adhesion on the coated surfaces with increasing Si content, developing mature cytoskeletons with numerous distinct and well-defined actin stress fibres in the cell membranes. Results also demonstrated that the bone mineralisation process was greatest in the presence of the highest Si level (4.9 wt%). However, at very early culturing time point, cells did not attach so readily on the surface of this coating due to rapid dissolution. Thus, this work suggests that a Si content of 2.2 wt% may be the optimum loading to improve the bioactive property of HA thin films.

Controlled delivery of stromal derived factor-1α from poly lactic-co-glycolic acid core-shell particles to recruit mesenchymal stem cells for cardiac regeneration.

Stromal derived factor-1α (SDF-1α) has shown promising results in treatment of myocardial infarction (MI), via recruitment of endogenous stem cells into the injured myocardium. However, the bioactivity of this susceptible signalling chemokine is reduced significantly during the common fabrication processes of drug delivery systems, due to the exposure to organic-aqueous interfaces or elevated temperature. In this study, we developed a novel SDF-1α delivery system using coaxial electrospraying, the technique which enables fabrication of core-shell particles with minimized contact of organic-aqueous phases. The SDF-1α incorporated PLGA particles exhibited distinct core-shell structure, confirmed by transmission electron microscopy (TEM). Controlled release of SDF-1α was obtained for at least 40days, and the release rate was tailored by co-encapsulation of bovine serum albumin (BSA) into the core of the particles. The SDF-1α released from PLGA/SDF-1α and PLGA/BSA-SDF-1α particles retained its chemotactic activity, and enhanced the number of migrated mesenchymal stem cells (MSCs) by 38% and 54%, respectively, compared to basal medium used as the control. Moreover, both SDF-1α and BSA supported the proliferation of MSCs within 3days of cell culture. The SDF-1α incorporated core-shell particles developed by electrospraying technique, can be effectively employed as injectable drug delivery system for in situ cardiac regeneration.

Dual-Microstructured Porous, Anisotropic Film for Biomimicking of Endothelial Basement Membrane.

Human endothelial basement membrane (BM) plays a pivotal role in vascular development and homeostasis. Here, a bioresponsive film with dual-microstructured geometries was engineered to mimic the structural roles of the endothelial BM in developing vessels, for vascular tissue engineering (TE) application. Flexible poly(ε-caprolactone) (PCL) thin film was fabricated with microscale anisotropic ridges/grooves and through-holes using a combination of uniaxial thermal stretching and direct laser perforation, respectively. Through optimizing the interhole distance, human mesenchymal stem cells (MSCs) cultured on the PCL film's ridges/grooves obtained an intact cell alignment efficiency. With prolonged culturing for 8 days, these cells formed aligned cell multilayers as found in native tunica media. By coculturing human umbilical vein endothelial cells (HUVECs) on the opposite side of the film, HUVECs were observed to build up transmural interdigitation cell-cell contact with MSCs via the through-holes, leading to a rapid endothelialization on the PCL film surface. Furthermore, vascular tissue construction based on the PCL film showed enhanced bioactivity with an elevated total nitric oxide level as compared to single MSCs or HUVECs culturing and indirect MSCs/HUVECs coculturing systems. These results suggested that the dual-microstructured porous and anisotropic film could simulate the structural roles of endothelial BM for vascular reconstruction, with aligned stromal cell multilayers, rapid endothelialization, and direct cell-cell interaction between the engineered stromal and endothelial components. This study has implications of recapitulating endothelial BM architecture for the de novo design of vascular TE scaffolds.

Gelatin-siloxane nanoparticles to deliver nitric oxide for vascular cell regulation: synthesis, cytocompatibility, and cellular responses.

Nitric oxide (NO) is an important mediator in cardiovascular system to regulate vascular tone and maintain tissue homeostasis. Its role in vascular cell regulation makes it promising to address the post-surgery restenosis problem. However, the application of NO is constrained by its high reactivity. Here, we developed a novel NO-releasing gelatin-siloxane nanoparticle (GS-NO NP) to deliver NO effectively for vascular cell regulation. Results showed that gelatin-siloxane nanoparticles (GS NPs) could be synthesized via sol-gel chemistry with a diameter of ∼200 nm. It could be modified into GS-NO NPs via S-nitrosothiol (RSNO) modification. The synthesized GS-NO NPs could release a total of ∼0.12 µmol/mg NO sustainably for 7 days following a first-order exponential profile. They showed not only excellent cytocompatibility, but also rapid intracellularization within 2 h. GS-NO NPs showed inhibition of human aortic smooth muscle cell (AoSMC) proliferation and promotion of human umbilical vein endothelial cell (HUVEC) proliferation in a dose-dependent manner, which is an important approach to prevent restenosis. With GS-NO NP dose at 100 µg/mL, the proliferation of AoSMCs could be slowed down whereas the growth of HUVECs was significantly promoted. We concluded that GS-NO NPs could have potential to be used as a promising nano-system to deliver NO for vascular cell regulation.

Protein encapsulated core-shell structured particles prepared by coaxial electrospraying: investigation on material and processing variables.

Biodegradable polymeric particles have been extensively investigated for controlled drug delivery of various therapeutic agents. 'Coaxial' electrospraying was successfully employed in this study, to fabricate core-shell PLGA particles containing bovine serum albumin (BSA) as the model protein, and the results were also compared to particles prepared by 'emulsion' electrospraying. Two different molecular weights of PLGA were employed to encapsulate the protein. Solution properties and processing parameters were found to influence the morphology of the core-shell particles. Depending on the type of solvent used to dissolve the polymer as well as the polymer concentration and molecular weight, the mean diameter of the particles varied between 3.0 to 5.5 µm. Fluorescence microscopic analysis of the electrosprayed particles using FITC-conjugated BSA demonstrated the core-shell structure of the developed particles. The encapsulation efficiency and release behavior of BSA was influenced by shell:core feeding ratio, protein concentration, and the electrospraying method. The encapsulation efficiency of BSA within the core-shell particles of high and low molecular weight PLGA was found 15.7% and 25.1% higher than the emulsion electrosprayed particles, respectively. Moreover, the total amount of BSA released from low molecular weight PLGA particles was significantly higher than high molecular weight PLGA particles within 43 days of release studies, with negligible effect on encapsulation efficiency. The technique of coaxial electrospraying has high potential for encapsulation of susceptible protein-based therapeutic agents such as growth factors for multiple drug delivery applications.

A scalable approach to obtain mesenchymal stem cells with osteogenic potency on apatite microcarriers.

Bone tissue engineering, which relies on the interactions between stem cells and suitable scaffold materials, represents a highly desirable alternative to currently used allograft or autograft strategies for the treatment of bone defects caused by injury or disease, with one of the major challenges being to generate sufficient quantities of stem cells to bring about the intended therapeutic effect. However, conventional cell culture to achieve sufficient cell numbers faces limitations of low efficiency and diminished efficacy of stem cells due to repeated passaging. Furthermore, current microcarriers available may not be suitable for therapeutic implantation. Here, the authors featured an apatite-based microcarrier intended for bone tissue engineering applications. These apatite microcarriers have a diameter of ∼230 µm, and exhibited porous and rough surface morphology. Peaks obtained from X-ray diffractometry (XRD) corresponded to hydroxyapatite (HA) with high crystallinity. Fourier transform infrared spectrophotometry (FTIR) showed that no residues of alginate remained, and all bands observed belong to phosphate and hydroxyl groups of HA. To evaluate the cytocompatibility of these microcarriers, in vitro proliferation studies were conducted and compared with conventional monolayer as well as Cytodex 3. The authors found that human foetal mesenchymal stem cells (hfMSCs) cultured on apatite microcarriers exhibited comparable growth characteristics, achieving 1.4-fold higher live cells than Cytodex 3 over a 9-day culture period. As these microcarriers were hypothesised to offer enhanced osteogenic potency over conventional monolayer culture, alkaline phosphatase (ALP), type I collagen and osteocalcin expression of hfMSCs cultured on the apatite microcarriers were evaluated over a 12-day period. ALP expression for hfMSCs seeded on apatite microcarriers was 2.7-fold higher than that of adherent monolayer culture (p < 0.001). Additionally, type I collagen and osteocalcin expression were 1.8- and 1.5-fold higher than that of adherent monolayer culture on day 12, respectively (p < 0.001).

Biomimetic three-dimensional anisotropic geometries by uniaxial stretching of poly(ε-caprolactone) films: degradation and mesenchymal stem cell responses.

Geometric cues have been used for a variety of cell regulation and tissue regenerative applications. While the function of geometric cues is being recognized, their stability and degradation behaviors are not well known. Here, we studied the influence of degradation on uniaxial-stretch-induced poly(ε-caprolactone) (UX-PCL) ridge/groove arrays and further cellular responses. Results from accelerated hydrolysis in vitro showed that UX-PCL ridge/groove arrays followed a surface-controlled erosion, with an overall geometry remained even at ∼45% film weight loss. Compared to unstretched PCL flat surfaces and/or ridge/groove arrays, UX-PCL ridge/groove arrays achieved an enhanced morphological stability against degradation. Over the degradation period, UX-PCL ridge/groove arrays exhibited an "S-shape" behavior of film weight loss, and retained more stable surface hydrophilicity and higher film mechanical properties than those of unstretched PCL surfaces. Human mesenchymal stem cells (MSCs) aligned better toward UX-PCL ridge/groove arrays when the geometries were remained intact, and became sensitive with gradually declined nucleus alignment and elongation to the geometric degradation of ridges. We speculate that uniaxial stretching confers UX-PCL ridge/groove arrays with enhanced stability against degradation in erosive environment. This study provides insights of how degradation influences geometric cues and further cell responses, and has implications for the design of biomaterials with stability-enhanced geometric cues for long-term tissue regeneration.

Fabrication of three-dimensional porous scaffolds with controlled filament orientation and large pore size via an improved E-jetting technique.

Biodegradable polymeric scaffolds have been widely used in tissue engineering as a platform for cell proliferation and subsequent tissue regeneration. Conventional microextrusion methods for three-dimensional (3D) scaffold fabrication were limited by their low resolution. Electrospinning, a form of electrohydrodynamic (EHD) printing, is an attractive method due to its capability of fabricating high-resolution scaffolds at the nanometer/micrometer scale level. However, the scaffold was composed of randomly orientated filaments which could not guide the cells in a specific direction. Furthermore, the pores of the electrospun scaffold were small, thus preventing cell infiltration. In this study, an alternative EHD jet printing (E-jetting) technique has been developed and employed to fabricate 3D polycaprolactone (PCL) scaffolds with desired filament orientation and pore size. The effect of PCL solution concentration was evaluated. Results showed that solidified filaments were achieved at concentration >70% (w/v). Uniform filaments of diameter 20 µm were produced via the E-jetting technique, and X-ray diffraction and attenuated total reflectance Fourier transform infrared spectroscopic analyses revealed that there was no physicochemical changes toward PCL. Scaffold with a pore size of 450 µm and porosity level of 92%, was achieved. A preliminary in vitro study illustrated that live chondrocytes were attaching on the outer and inner surfaces of collagen-coated E-jetted PCL scaffolds. E-jetted scaffolds increased chondrocytes extracellular matrix secretion, and newly formed matrices from chondrocytes contributed significantly to the mechanical strength of the scaffolds. All these results suggested that E-jetting is an alternative scaffold fabrication technique, which has the capability to construct 3D scaffolds with aligned filaments and large pore sizes for tissue engineering applications.

Effect of silver content on the antibacterial and bioactive properties of silver-substituted hydroxyapatite.

The long-term success of a biomaterial used during surgery may be compromised by infection. A possible effective solution is to make the biomaterial osteoconductive and antibacterial. A range of silver-substituted hydroxyapatite (AgHA) of up to 1.1 wt. % of Ag was synthesized. AgHA displayed a rod-like morphology of dimensions ~50 nm in length and ~15 nm in width. Phase-pure AgHA was demonstrated in the X-ray diffraction patterns and Fourier transform infrared spectroscopy spectra. Comparing with hydroxyaptite (HA), 0.5AgHA exhibited a 3-log reduction in the number of bacteria. Diffusion of the entrapped Ag(+) ions towards the crystal structure surface was revealed by an increase of 6 at. % Ag in the X-ray photoelectron spectroscopy results. Furthermore, less than 0.5 ppm of Ag(+) ions being released from 0.5AgHA into the deionized water medium was evidenced from the inductively coupled plasma mass spectrometry results. AgHA produced by co-precipitation gave rise to minimal release of Ag(+) ions. It was hypothesized that the diffused surface Ag(+) ions damaged the bacteria cell membrane and impede its replication. With the culturing time, significant increase in the number of human mesenchymal stem cells (p < 0.05) was demonstrated on 0.5AgHA.

Biomimetic three-dimensional anisotropic geometries by uniaxial stretch of poly(ε-caprolactone) films for mesenchymal stem cell proliferation, alignment, and myogenic differentiation.

Anisotropic geometries are critical for eliciting cell alignment to dictate tissue microarchitectures and biological functions. Current fabrication techniques are complex and utilize toxic solvents, hampering their applications for translational research. Here, we present a novel simple, solvent-free, and reproducible method via uniaxial stretching for incorporating anisotropic topographies on bioresorbable films with ambitions to realize stem cell alignment control. Uniaxial stretching of poly(ε-caprolactone) (PCL) films resulted in a three-dimensional micro-ridge/groove topography (inter-ridge-distance: ~6 µm; ridge-length: ~90 µm; ridge-depth: 200-900 nm) with uniform distribution and controllable orientation by the direction of stretch on the whole film surface. When stretch temperature (Ts) and draw ratio (DR) were increased, the inter-ridge-distance was reduced and ridge-length increased. Through modification of hydrolysis, increased surface hydrophilicity was achieved, while maintaining the morphology of PCL ridge/grooves. Upon seeding human mesenchymal stem cells (hMSCs) on uniaxial-stretched PCL (UX-PCL) films, aligned hMSC organization was obtained. Compared to unstretched films, hMSCs on UX-PCL had larger increase in cellular alignment (>85%) and elongation, without indication of cytotoxicity or reduction in cellular proliferation. This aligned hMSC organization was homogenous and stably maintained with controlled orientation along the ridges on the whole UX-PCL surface for over 2 weeks. Moreover, the hMSCs on UX-PCL had a higher level of myogenic genes' expression than that on the unstretched films. We conclude that uniaxial stretching has potential in patterning film topography with anisotropic structures. The UX-PCL in conjunction with hMSCs could be used as "basic units" to create tissue constructs with microscale control of cellular alignment and elongation for tissue engineering applications.