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The activity of matrix metalloproteinases (MMPs) plays a crucial role in the aging of the resin-dentin interface. The in situ action of MMP-2 and MMP-9 has been confirmed in the process of dentin-collagen degradation. However, the involvement of dental pulp cells in MMP secretion as a response to oxidative stress induced by contact with resin monomers has not been fully elucidated. Myricetin (MYR), like proanthocyanidin (PAC), has antioxidant properties and may help prevent extracellular matrix degradation. The objective was to evaluate the effect of MYR on the MMP expression and activity in response to reactive oxygen species (ROS) increase induced by triethylene glycol dimethacrylate (TEGDMA) in human odontoblast-like cells (hOLCs). hOLCs differentiated from dental pulp mesenchymal stem cells were exposed to TEGDMA released from dentin blocks using a barrier model with transwell inserts for 18, 24, and 36 h. Intracellular oxidation was evaluated using the 2',7'-dichlorofluorescein probe. The effect of 600 μM MYR on the MMP-2 and MMP-9 expression was determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The extracellular MMP levels were quantified using enzyme-linked immunosorbent assay, and their activation by means of a proteolytic fluorometric assay. The results were analyzed by one-way analysis of variance and Tukey's post hoc test, p ≤ 0.05. TEGDMA exposure increased intracellular ROS and upregulated MMP-2 and MMP-9 mRNA in hOLCs (p < 0.001). The levels of MMPs increased significantly 24 h after TEGDMA exposure (p = 0.013). These secreted proteases exhibited high activation ability. MYR reduced ROS production and downregulated MMP expression and activity at both mRNA and protein levels, similar to the effect found for PAC, which was used as a control. A relationship was observed between MMP-2 and MMP-9 expression, secretion, and early activation with ROS increase due to TEGDMA exposure. MYR showed potential as a therapeutic strategy to control MMP expression and modulate redox imbalance, offering a protective effect on cellular response., (© 2025 Wiley Periodicals LLC.)

Methacrylic acid (MAA)-based biomaterials promote a vascularized host response without the addition of exogenous factors such as cells or growth factors. We presume that materials containing MAA favor an alternative foreign body response, rather than the conventional fibrotic response. Here, we characterize selected aspects of the response to two different forms of MAA-a coating, which can be used to prevascularize the subcutaneous tissue for subsequent therapeutic cell delivery or an injectable hydrogel, which can be used to vascularize and deliver cells simultaneously. We show that the MAA-coating quickly vascularized the subcutaneous space compared to an uncoated silicone tube, and after 14 days of prevascularization, the tissue surrounding the MAA-coated tube presented fewer immune cells than the uncoated control. We also compared the host response to a MAA-PEG (polyethylene glycol) hydrogel at day 1, with pancreatic islets in immune-compromised SCID/bg mice and immune-competent Balb/c mice. The Balb/c mouse presented a more inflammatory response with increased IFN-γ production as compared to the SCID/bg. Together with previously published data, this work contributes to a further understanding of tissue responses to a biomaterial in different forms as used for cell delivery., (© 2023 Wiley Periodicals LLC.)

Studies have long sought to develop engineered heart tissue for the surgical correction of structural heart defects, as well as other applications and vascularization of this tissue has presented a challenge. Recent studies suggest that vascular cells and a vascular network may have regenerative effects on implanted cardiomyocytes (CM) and nearby heart tissue separate from perfusion of oxygen and nutrients. The goal of this study was to test whether vascular cells or a formed vascular network in a fibrin-based hydrogel would alter the proliferation of human iPSC-derived CM. First, vascular network formation in a slowly degrading PEGylated fibrin hydrogel was optimized by altering the cell ratio of human umbilical vein endothelial cells to human dermal fibroblasts, the inclusion of growth factors, and the total cell concentration. An endothelial to fibroblast ratio of 5:1 and a total cell concentration of 1.1 × 10 6 cells/mL without additional growth factors generated robust vascular networks while minimizing the number of cells required. Using this optimized system, human iPSC-derived CM were cultured on hydrogels without vascular cells, hydrogels with unorganized encapsulated vascular cells, or hydrogels with encapsulated vascular cells organized into networks for 7 days. CM proliferation and gene expression were assayed following 7 days of culture on the hydrogels. The presence of vascular cells in the hydrogel, whether unorganized or in vascular networks, significantly increased CM proliferation compared to an acellular hydrogel. Hydrogels with unorganized vascular cells resulted in lower CM maturity evidenced by decreased expression of cardiac troponin t (TNNT2), myosin light chain 7, and phospholamban compared to hydrogels without vascular cells and hydrogels with vascular networks. Altogether, this study details a robust method of forming rudimentary vascular networks in a fibrin-based hydrogel and shows that a hydrogel containing endothelial cells and fibroblasts can induce proliferation in adjacent CM, and these cells do not hinder CM gene expression when organized into a vascular network., (© 2023 Wiley Periodicals LLC.)

Polyetheretherketone (PEEK) is a promising material for use in orthopedic implants, but its bio-inert character and lack of antibacterial activity limit its applications in bone repair. In the present study, considering the advantages of PEEK in self-initiated graft polymerization and of hydrogels in bone tissue engineering, we constructed a hydrogel coating (GPL) consisting of Gelatin methacryloyl (GelMA), methacrylamide-modified ε-poly-l-lysine (ε-PLMA) and Laponite on PEEK through UV-initiated crosslinking. The coating improved the hydrophilicity of PEEK, and the coating degraded slowly so that approximately 80% was retained after incubation in PBS for 8 weeks. In vitro studies revealed that as compared to culturing on PEEK, culturing on PEEK-GPL led to enhanced viability and adhesion of cultured human umbilical cord Wharton's jelly-derived mesenchymal stem cells (hWJ-MSCs). Due to the synergistic effect of the micron-scale three-dimensional surface and Laponite, PEEK-GPL exhibited a significantly improved induction of osteogenic differentiation of hWJ-MSCs compared to PEEK, as demonstrated by increased alkaline phosphatase activity, matrix mineralization, and expression of osteogenesis-related genes. Furthermore, PEEK-GPL showed antibacterial activity upon contact with Staphylococcus aureus and Escherichia coli, and this activity would be maintained before complete degradation of the hydrogel because the ε-PLMA was cross-linked covalently into the coating. Thus, PEEK-GPL achieved both osteogenesis and infection prevention in a single simple step, providing a feasible approach for the extensive use of PEEK in bone implants., (© 2023 Wiley Periodicals LLC.)

The decrease in contractility in myocardium adjacent (border zone; BZ) to a myocardial infarction (MI) is correlated with an increase in reactive oxygen species (ROS). We hypothesized that injection of a thermoresponsive hydrogel, with ROS scavenging properties, into the MI would decrease ROS and improve BZ function. Fourteen sheep underwent antero-apical MI. Seven sheep had a comb-like copolymer synthesized from N-isopropyl acrylamide (NIPAAm) and 1500 MW methoxy poly(ethylene glycol) methacrylate, (NIPAAm-PEG1500), injected (20 × 0.5 mL) into the MI zone 40 min after MI (MI + NIPAAm-PEG1500) and seven sheep were MI controls. Cardiac MRI was performed 2 weeks before and 6 weeks after MI + NIPAAm-PEG1500. BZ wall thickness at end systole was significantly higher for MI + NIPAAm-PEG1500 (12.32 ± 0.51 mm/m 2 MI + NIPAAm-PEG1500 vs. 9.88 ± 0.30 MI; p = .023). Demembranated muscle force development for BZ myocardium 6 weeks after MI was significantly higher for MI + NIPAAm-PEG1500 (67.67 ± 2.61 mN/m 2 MI + NIPAAm-PEG1500 vs. 40.53 ± 1.04 MI; p < .0001) but not significantly different from remote myocardium or BZ or non-operated controls. Levels of ROS in BZ tissue were significantly lower in the MI + NIPAAm-PEG1500 treatment group (hydroxyl p = .0031; superoxide p = .0182). We conclude that infarct injection of the NIPAAm-PEG1500 hydrogel with ROS scavenging properties decreased ROS and improved contractile protein function in the border zone 6 weeks after MI., (© 2020 Wiley Periodicals, Inc.)

A current approach in bone tissue engineering is the implantation of polymeric scaffolds that promote osteoblast attachment and growth as well as biomineralization. One promising polymer is oligo[poly(ethylene glycol) fumarate] (OPF), a polyethylene glycol-based material that is biocompatible, injectable, and biodegradable, but in its native form does not support robust bone cell attachment or growth. To address this issue, this study evaluated the osteoconductivity of bis[02-(methacryloyloxy)ethyl] phosphate (BP) functionalized OPF hydrogels (OPF-BP) using MC3T3-E1 pre-osteoblast cells, both before and after enzymatic mineralization with a calcium solution. The inclusion of negatively charged functional groups allowed for the tailored uptake and release of calcium, while also altering the mechanical properties and surface topography of the hydrogel surface. In cell culture, OPF-BP hydrogels with 20 and 30% (w/w) BP optimized osteoblast attachment, proliferation, and differentiation after a 21-day in vitro period. In addition, the OPF-BP30 treatment, when mineralized with calcium, exhibited a 128% increase in osteocalcin expression when compared with the non-mineralized treatment. These findings suggest that phosphate functionalization and enzymatic calcium mineralization can act synergistically to enhance the osteoconductivity of OPF hydrogels, making this processed material an attractive candidate for bone tissue engineering applications., (© 2019 Wiley Periodicals, Inc.)

Promising cell therapies using mesenchymal stem cells (MSCs) is proposed for stroke patients. Therefore, we aimed to efficiently accumulate human MSC (hMSC) to damaged brain area to improve the therapeutic effect using poly(ethylene glycol) (PEG)-conjugated phospholipid (PEG-lipid) carrying an oligopeptide as a ligand, specific for E-selectin which is upregulated on activated endothelial cells under hypoxia-like stroke. Here we synthesized E-selectin-binding oligopeptide (ES-bp) conjugated with PEG spacer having different molecular weights from 1 to 40 kDa. We found that ES-bp can be immobilized onto the hMSC surface through PEG-lipid without influence on cell growth and differentiation into adipocytes and osteocytes, respectively. It is also possible to control the immobilization of ES-bp on hMSC surface (<10 8 ES-bp per cell). Immobilized ES-bp can be continuously immobilized at the outside of cell membrane when PEG-lipids with PEG 5 and 40 kDa were used. In addition, the modified hMSC can specifically attach onto E-selectin-immobilized surface as a model surface of activated endothelium in human blood, indicating the sufficient number of immobilized ES-bp onto hMSC. Thus, this technique is one of the candidates for hMSC accumulation to cerebral infarction area. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1779-1792, 2019., (© 2019 Wiley Periodicals, Inc.)

In this study, a novel 3D printed porous titanium cage (3D printed cage) with interconnected pores inside was designed and manufactured. Observations by scanning electron microscopy showed that the inside of the 3D printed cage had an octahedral porous structure, with the pores uniform in size and connected to each other. The mechanical properties analysis found that the Young's modulus and compressive strength of the porous structure were close to those of the bone structure, and the overall stiffness was slightly higher than that of the polyether ether ketone (PEEK) material, but was significantly lower than that of the titanium alloy solid module. Animal experiments indicated that the new 3D printed cage was equivalent to PEEK cage in fusion time. At 3 months, the new bone grew into the cage through the pores of the new 3D printed cage surface, which had a high bone contact rate. These results demonstrate that the 3D printed porous titanium cage has good biocompatibility and osseointegration, and has a potential clinical value as bone implants. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2019., (© 2019 Wiley Periodicals, Inc.)

Compositions of resin composite exhibit cytotoxicity, especially Triethylene-glycol-dimethacrylate (TEGDMA), yet the underlying mechanisms and its relationship with filler content are poorly understood. Here, specimens of five composites (VITA LC, VITA ZETA, Z350, Filtek P60, and AP-X), containing different filler size and weight, were immersed into culture medium for 72 h. After TEGDMA quantification, the resin composite eluates were used to incubate HGFs. Cellular viability was evaluated. Total reactive oxygen species (ROS) and mitochondrial ROS were detected to assess oxidative stress. Adenosine triphosphate and cytochrome c oxidase (CcO) activity, mitochondrial membrane potential and morphology, mitochondrial biogenesis regulators were analyzed to evaluate mitochondrial functions. Results showed that TEGDMA release negatively correlated to filler size and weight of tested composites. Although cell viability reduction was not significant, total and mitochondrial ROS production showed a positive relationship with the amount of TEGDMA in composite eluates. Furthermore, the expression of mitochondrial biogenesis markers and mitochondrial fusion protein, were markedly elevated in TEGDMA rich eluates, especially in VITA-LC group, shown as elongated mitochondrial morphology and aberrant mitochondrial functions. Overall, TEGDMA could elute easier from those resin composites with less filler content and cause oxidative stress in HGFs via mitochondria dysregulation. These data can be instructive to optimize the synthesis of resin composites from the perspective of biocompatibility. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1132-1142, 2019., (© 2019 Wiley Periodicals, Inc.)

Improved thromboresistance of mechanical valves is desired to decrease the risk of thromboembolism and thrombosis and reduce the dosage of anticoagulation with a vitamin K antagonist (e.g., warfarin). For several mechanical valves, design-related features are responsible for their improved thromboresistance. However, it remains unclear whether material-related features provide a practical level of thromboresistance to mechanical valves. Here, we studied the effect of a bileaflet valve made of poly(ether ether ketone) (PEEK) with a poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC)-grafted surface (PEEK-g-PMPC). PMPC is a well-known thromboresistant polymeric material. A short-term (<26 h) porcine aortic valve replacement model using neither an anticoagulant nor an antiplatelet agent showed that the PEEK-g-PMPC valve opened and closed normally with an allowable transvalvular gradient. Unlike an untreated PEEK valve, no thrombus formed on the PEEK-g-PMPC valves on gross anatomy examination in addition to the absence of traveled thrombi in the kidney and lung tissues. Material (PEEK-g-PMPC)-related thromboresistance appeared to decrease the risk of thromboembolism and thrombosis for patients with mechanical valves. However, thromboresistance of the PEEK-g-PMPC valve requires improvement because fibrous fouling was still observed on the leaflet. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1052-1063, 2019., (© 2019 Wiley Periodicals, Inc.)

This article presents the preparation of matrices from two new families of fumaric copolymers and the effect of structural differences on their physicochemical and biological behavior. Diisopropyl fumarate (DIPF) and poly(ethylene glycol) methyl ether methacrylate (OEGMA) or N-isopropylacrylamide (NIPAM) were copolymerized by conventional radical and RAFT polymerization to obtain lineal or start architectures, respectively. These copolymers were characterized by spectroscopic (FTIR and 1 H-NMR) and chromatographic methods. The thermal stability was studied by thermal gravimetric analysis, showing some differences in relation to the architecture and chemical nature of copolymers. SEM morphological analysis demonstrated that the surface of the matrices derived from OEGMA exhibited an irregular and rough surface, while DIPF-NIPAM copolymers presented smooth surface with nearly no significant porosity. The matrix obtained of both kinds of copolymers presented no cytotoxicity as it was evaluated using a model of macrophages on culture. Moreover, cell growth was better on the surfaces of the DIPF-NIPAM matrices, suggesting a good interaction with this matrix and its potential application as matrices for tissue engineering. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 195-203, 2019., (© 2018 Wiley Periodicals, Inc.)

Collagen I interactions with integrins α 1 and α 2 are known to support human mesenchymal stem cell (hMSC) osteogenesis. Nonetheless, elucidating the relative impact of specific integrin interactions has proven challenging, in part due to the complexity of native collagen. In the present work, we employed two collagen-mimetic proteins-Scl2-2 and Scl2-3- to compare the osteogenic effects of integrin α 1 versus α 2 signaling. Scl2-2 and Scl2-3 were both derived from Scl2-1, a triple helical protein lacking known cell adhesion, cytokine binding, and matrix metalloproteinase sites. However, Scl2-2 and Scl2-3 were each engineered to display distinct collagen-based cell adhesion motifs: GFPGER (binding integrins α 1 and α 2 ) or GFPGEN (binding only integrin α 1 ), respectively. hMSCs were cultured within poly(ethylene glycol) (PEG) hydrogels containing either Scl2-2 or Scl2-3 for 2 weeks. PEG-Scl2-2 gels were associated with increased hMSC osterix expression, osteopontin production, and calcium deposition relative to PEG-Scl2-3 gels. These data indicate that integrin α 2 signaling may have an increased osteogenic effect relative to integrin α 1 . Since p38 is activated by integrin α 2 but not by integrin α 1 , hMSCs were further cultured in PEG-Scl2-2 hydrogels in the presence of a p38 inhibitor. Results suggest that p38 activity may play a key role in collagen-supported hMSC osteogenesis. This knowledge can be used toward the rational design of scaffolds which intrinsically promote hMSC osteogenesis. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2594-2604, 2018., (Copyright © 2018 Wiley Periodicals, Inc.)

Nanoparticle (NP) interactions with cellular systems are influenced by both NP physico-chemical properties and the presence of surface-bound proteins that are adsorbed in biological environments. Here, we characterize cellular responses to silver nanoparticles (AgNPs) functionalized with poly(di(ethylene glycol) methyl ether methacrylate-co-oligo(ethylene glycol) methyl methacrylate) (poly(MeO 2 MA x -co-OEGMA y )) brushes with tunable hydrophobicity and explore how these responses are modulated by the presence or absence of serum proteins. Poly(MeO 2 MA x -co-OEGMA y ) with variable composition (5-10% OEGMA) was fabricated to elicit differential hydrophobicity at 37°C for AgNPs capped with these copolymers. The increase in Ag-(MeO 2 MA x -co-OEGMA y ) surface hydrophobicity from (x:y) = 90:10 to (x:y) = 95:5 led to enhanced cytotoxicity of L-929 fibroblasts and a concomitant increase in cell uptake and reactive oxygen species generation in the presence of serum proteins. These responses were attenuated significantly in serum-free environments. Broad inhibition of PI3 kinase-mediated endocytosis reduced both cell uptake and cytotoxicity in the presence or absence of serum proteins. In contrast, selective inhibition of clathrin- and caveolae-mediated endocytosis markedly decreased cell uptake and cytotoxicity in response to Ag-(MeO 2 MA 95 -co-OEGMA 5 ) exclusively in the presence of serum proteins, whereas cell responses to the more hydrophilic Ag-(MeO 2 MA 90 -co-OEGMA 10 ) were less affected by the inhibition of these pathways with or without serum proteins. This study demonstrates an important role for both NP surface hydrophobicity and the presence of serum proteins in directing cell uptake and subsequent cellular responses, which we suggest has broad application in the design of polymer-functionalized NPs for specific biological outcomes. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1061-1071, 2018., (© 2017 Wiley Periodicals, Inc.)

In this study, a series of poly(ethylene glycol)/collagen (PEG/Col) double network (DN) hydrogel is fabricated from PEG and Col. Results of the compressive strength test indicate that the strength and toughness of these DN hydrogels are significantly enhanced. The fracture strength of PEG/Col DN hydrogels increases by 9- to 12-fold compared with that of PEG single network (SN) hydrogel, and by 36- to 48-fold compared with that of Col SN hydrogel. Taking advantage of both PEG and Col building blocks, the PEG/Col DN hydrogels possess a strengthened skeleton. Moreover, the water-storage capability and favorable biocompatibility of Col are effectively maintained. Given that the DN hydrogels can provide the appropriate environment for the adhesion, growth, and proliferation of MC3T3-E1 cells, PEG/Col DN hydrogels have potential as a load-bearing tissue repair material. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 192-200, 2018., (© 2017 Wiley Periodicals, Inc.)

A family of pH-responsive terpolymers composed of methacrylic acid (MAA), N-vinyl pyrrolidone (NVP), and poly(ethylene glycol) monomethylether monomethacrylate (PEGMMA) have been developed and evaluated for their pH-responsive swelling behavior, protein-loading capabilities, and cytocompatibility. These terpolymer hydrogels, designated as P((MAA-co-NVP)-g-EG), were synthesized with varying PEG chain lengths and monomer feed ratios. The incorporation of MAA into the terpolymer structure was quantified with potentiometric titration. Equilibrium and dynamic swelling studies confirmed the pH-responsive behavior of the hydrogel, with the system remaining collapsed/complexed in acidic pH conditions and swollen/decomplexed in neutral pH conditions. The ability of the hydrogels to partition protein into the swollen network was assessed for two model proteins of varying molecular weight: insulin and porcine growth hormone. Finally, the cytocompatibility of the hydrogels in the presence of two model intestinal cell lines was investigated and confirmed minimal cytotoxicity at and below 2.5 mg/mL. The developed P((MAA-co-NVP)-g-EG) hydrogels exhibit unique properties that could potentially be utilized for drug delivery and separation applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1243-1251, 2017., (© 2017 Wiley Periodicals, Inc.)

Inhibiting liposome uptake by macrophages using polyethylene glycol (PEG) surface modifications is a widely used approach for extending the half-life of liposomes circulating in the blood. However, the biological effects of PEGylated liposomes on macrophages have not yet been thoroughly investigated. The purpose of this study was to examine the effects of PEGylated phosphatidylserine-containing liposomes (PEG-PSLs) on the expression of two inflammation-associated cytokines, tumor necrosis factor-α (TNF-α) and transforming growth factor-β (TGF-β), in the murine macrophage-like cell line RAW 264.7. Previous studies have demonstrated that PSLs inhibit TNF-α secretion and enhance TGF-β synthesis in macrophages by mimicking apoptotic cells. We found that PEGylation differentially affected the TNF-α and TGF-β levels. The PSL-mediated inhibitory effect on TNF-α secretion was enhanced by PEGylation, and PEG-PSLs decreased TGF-β levels compared with non-PEGylated PSLs. Fluorescence-activated cell sorting analysis demonstrated that 1% PEGylation disturbed the incorporation of PSLs into macrophages. The interference of uptake is thought to extend the binding interaction between PS to PS receptors for PSL-mediated inhibition of TNF-α expression. Together, these findings indicate that PEG-PSLs can prevent TNF-α secretion without increasing TGF-β levels in macrophages, and they support the potential clinical use of PEG-PSLs as anti-inflammatory agents with a relatively low potential to induce tissue fibrosis. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1479-1486, 2017., (© 2017 Wiley Periodicals, Inc.)

Perioperative and latent infections are leading causes of revision surgery for orthopaedic devices resulting in significant increased patient care, comorbidities, and attendant costs. Identifying biomaterial surfaces that inherently resist biofilm adhesion and bacterial expression is an important emerging strategy in addressing implant-related infections. This in vitro study was designed to compare biofilm formation on three biomaterials commonly employed in spinal fusion surgery-silicon nitride (Si 3 N 4 ), polyetheretherketone (PEEK), and a titanium alloy (Ti6Al4V-ELI) -using one gram-positive and one gram-negative bacterial species. Disc samples from various surface treated Si 3 N 4 , PEEK, and Ti6Al4V were inoculated with 10 5 CFU/mm 2 Staphylococcus epidermidis (ATCC®14990™) or Escherichia coli (ATCC ® 25922™) and cultured in PBS, 7% glucose, and 10% human plasma for 24 and 48 h, followed by retrieval and rinsing. Vortexed solutions were diluted, plated, and incubated at 37 °C for 24 to 48 h. Colony forming units (CFU/mm 2 ) were determined using applicable dilution factors and surface areas. A two-tailed, heteroscedastic Student's t-test (95% confidence) was used to determine statistical significance. The various Si 3 N 4 samples showed the most favorable bacterial resistance for both bacilli tested. The mechanisms for the bacteriostatic behavior of Si 3 N 4 are likely due to multivariate surface effects including submicron-topography, negative charging, and chemical interactions which form peroxynitrite (an oxidative agent). Si 3 N 4 is a new biomaterial with the apparent potential to inhibit biofilm formation. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1521-1534, 2017., (© 2017 Wiley Periodicals, Inc.)

Poly(ethylene glycol) (PEG) hydrogels provide a versatile platform to develop cell instructive materials through incorporation of a variety of cell adhesive ligands and degradable chemistries. Synthesis of PEG gels can be accomplished via two mechanisms: chain and step growth polymerizations. The mechanism dramatically impacts hydrogel nanostructure, whereby chain polymerized hydrogels are highly heterogeneous and step growth networks exhibit more uniform structures. Underpinning these alterations in nanostructure of chain polymerized hydrogels are densely-packed hydrophobic poly(methyl methacrylate) or poly(acrylate) kinetic chains between hydrophilic PEG crosslinkers. As cell-material interactions, such as those mediated by integrins, occur at the nanoscale and affect cell behavior, it is important to understand how different modes of polymerization translate into nanoscale mechanical and hydrophobic heterogeneities of hydrogels. Therefore, chain- and step-growth polymerized PEG hydrogels with macroscopically similar macromers and compliance (for example, methacrylate-functionalized PEG (PEGDM), M W  = 10 kDa and norbornene-functionalized 4-arm PEG (PEGnorb), M W  = 10 kDa) were used to examine potential nanoscale differences in hydrogel mechanics and hydrophobicity using atomic force microscopy (AFM). It was found that chain-growth polymerized network yielded greater heterogeneities in both stiffness and hydrophobicity as compared to step-growth polymerized networks. These nanoscale heterogeneities impact cell-material interactions, particularly human mesenchymal stem cell (hMSC) adhesion and spreading, which has implications in use of these hydrogels for tissue engineering applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1112-1122, 2017., (© 2017 Wiley Periodicals, Inc.)

Tissue-engineered three-dimensional (3D) cancer models employing biomimetic hydrogels as cellular scaffolds provide contextual in vitro recapitulation of the native tumor microenvironment, thereby improving their relevance for use in cancer research. This study reports the use of poly(ethylene glycol)-fibrinogen (PF) as a suitable biosynthetic hydrogel for the 3D culture of three breast cancer cell lines: MCF7, SK-BR-3, and MDA-MB-231. Modification of the matrix characteristics of PF hydrogels was achieved by addition of excess poly(ethylene glycol) diacrylate, which resulted in differences in Young's moduli, degradation behavior, release kinetics, and ultrastructural variations in scaffold microarchitecture. Cancer cells were maintained in 3D culture with high viability within these hydrogels and resulted in cell-type dependent morphological changes over time. Cell proliferation and 3D morphology within the hydrogels were visualized through immunofluorescence staining. Finally, spatial heterogeneity of colony area within the hydrogels was quantified, with peripheral cells forming colonies of higher area compared to those in the interior regions. Overall, PF-based hydrogels facilitate 3D culture of breast cancer cells and investigation of cellular behavior in response to varying matrix characteristics. PF-based cancer models could be potentially used in future investigations of cancer biology and in anti-cancer drug-testing applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 236-252, 2017., (© 2016 Wiley Periodicals, Inc.)

Poly-ether-ether-ketone (PEEK) is currently introduced as an alternative material for orthopedic implants due to its biocompatibility and low elastic modulus compared to titanium. Also, a sulphonation treatment can functionalize PEEK to embed therapeutical substances. The objective of this work was to functionalize a PEEK film to incorporate novel lactam-based antibiofilms compounds. PEEK samples were functionalized by sulphuric acid treatment and then dissolved in dimethylsulfoxide, where lactams were added to be incorporated into the polymer. A dip-coating technique was used to synthesize a thin film on a glass-based substrate. The degree of sulfonation (DS) and the incorporation of lactams into sulphonated PEEK (sPEEK) were analyzed by Fourier transform infrared spectroscopy, nuclear magnetic resonance, thermogravimetric analysis (TGA), and scanning electron microscopy. A DS of 65% was obtained and TGA curves confirmed the presence of SO 3 H and lactams in the sPEEK structure. The growth of Streptococcus mutans biofilm decreased on sPEEK surface containing lactams when compared to sPEEK free of lactams. That indicated the antibiofilm activity of those compounds was maintained after incorporation into sPEEK. Planktonic growth analysis showed no long distant effects of sPEEK containing lactams, indicating that no systemic effects should be expected upon clinical uses of medical devices produced with lactam-treated sPEEK. Results revealed that inclusion of lactams into sPEEK represents a good alternative for the production of biomaterials resistant to bacterial accumulation. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3015-3020, 2016., (© 2016 Wiley Periodicals, Inc.)

Amphiphilic block copolymer methoxy polyethyleneglycol-polycaprolactone (mPEG-PCL) has attracted interest in the biomedical field, due to its water solubility and biodegradability. Nevertheless, the blood safety of mPEG-PCL copolymers has not been investigated in detail. Because mPEG-PCL copolymers introduced in vivo would inevitably interact with blood tissue, an investigation of possible interactions of mPEG-PCL with key blood components is crucial. We studied the effects of two mPEG-PCL copolymer solutions on blood coagulation, the morphology and lysis of human red blood cells (RBCs), the structure of plasma fibrinogen, complement activation, and platelet aggregation. We found that higher concentrations of the mPEG-PCL copolymers impaired blood clotting, and the copolymers had little impact on the morphology or lysis of RBCs. From the spectroscopy results, the copolymers affected the local microstructure of fibrinogen. The copolymers significantly activated the complement system in a concentration-dependent way. At higher concentrations, the copolymers impaired platelet aggregation, which may have been mediated by an inhibition of the arachidonic acid pathway. These findings provide important information that may be useful for the molecular design and biomedical applications of mPEG-PCL copolymers. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 802-812, 2016., (© 2016 Wiley Periodicals, Inc.)

A microporous hydrogel scaffold was developed from hyperbranched poly(glycidol) (HPG) and poly(ethylene oxide) (PEO) using electron beam (e-beam) induced cross-linking for tissue engineering applications. In this study, HPG was synthesized from glycidol using trimethylol propane as a core initiator and cross-linked hydrogels were made using 0, 10, 20, and 30% HPG with respect to PEO. The effects of %-HPG on the swelling ratio, cross-linking density, mechanical properties, morphology, degradation, and cytotoxicity of the hydrogel scaffolds were then investigated. Increasing the HPG content increased the pore size of the hydrogel scaffold, as well as the porosity, elongation at break, degree of degradation and swelling ratio. In contrast, the presence of HPG decreased the cross-linking density of the hydrogel. There was no significant difference in compressive modulus and tensile strength of all compositions. The pore size of hydrogel scaffolds could be easily tailored by controlling the content of HPG in the polymer blend. Evaluation of the cytotoxicity demonstrated that HPG/PEO hydrogel scaffold has potential for use as a matrix for cellular attachment and proliferation. These results indicate that cross-linked HPG/PEO hydrogel can function as a potential material for tissue engineering scaffolds. Moreover, a facile method to prepare hydrogel microporous scaffolds for tissue engineering by e-beam irradiation was developed., (© 2015 Wiley Periodicals, Inc.)

Expanded polytetrafluoroethylene (ePTFE) grafts were coated on the luminal surface with a cell-adhesive fluorosurfactant (FSP) polymer to promote endothelialization, followed by ethanol hydration to degas the pores and subsequent cell-adhesive, enzymatically degradable poly(ethylene glycol)-based hydrogel incorporation into the graft interstices to accommodate potential smooth muscle cell integration in the graft wall. The FSP coating on ePTFE was stable as demonstrated by a significantly reduced receding water contact angle on FSP-coated ePTFE (14.5 ± 6.4°) compared to uncoated ePTFE (105.3 ± 4.5°, P < 0.05) after ethanol exposure. X-ray photoelectron spectroscopy analysis of the same surfaces confirmed FSP presence. Localization of the FSP and hydrogel within the ePTFE graft construct was assessed using fluorescently labeled polymers, and demonstrated hydrogel infiltration throughout the thickness of the graft wall, with FSP coating limited to the lumen and adventitial surfaces. FSP at the luminal surface on dual-coated grafts was able to bind endothelial cells (EC) (98.7 ± 23.1 cells/mm(2) ) similar to fibronectin controls (129.4 ± 40.7 cells/mm(2) ), and significantly higher than uncoated ePTFE (31.6 ± 19 cells/mm(2,) P < 0.05). These results indicate that ePTFE grafts can be simultaneously modified with two different polymers that have potential to directly address both endothelialization and intimal hyperplasia. Such a construct is a promising candidate for an off-the-shelf synthetic graft for small-diameter graft applications., (© 2015 Wiley Periodicals, Inc.)

The use of polyetheretherketone (PEEK) composites in the trauma plating system, total replacement implants, and tissue scaffolds has found great interest among researchers. In recent years (2008 afterward), this type of composites has been examined for suitability as substitute material over stainless steel, titanium alloys, ultra high molecular weight polyethylene, or even biodegradable materials in orthopedic implant applications. Biomechanical and bioactivity concepts were contemplated for the development of PEEK orthopedic implants and a few primary clinical studies reported the clinical outcomes of PEEK-based orthopedic implants. This study aims to review and discuss the recent concepts and contribute further concepts in terms of biomechanical and bioactivity challenges for the development of PEEK and PEEK composites in orthopedic implants., (© 2015 Wiley Periodicals, Inc.)

Encapsulating chondrocytes in synthetic and degradable hydrogels for cartilage tissue engineering enables tuning of scaffold degradation, but provides no biological cues. Culture medium that recapitulates the physiological osmolarity of the interstitial fluid in cartilage can enhance matrix synthesis in the short term, but long-term benefits remain to be determined. This study investigates the long-term effect of culture medium osmolarity on tissue synthesis using chondrocytes isolated from three skeletally mature bovine donors encapsulated in degradable poly(ethylene glycol) hydrogels. The cell-laden hydrogels were cultured up to 4 weeks in standard chondrocyte-specific medium (330 mOsm) or medium adjusted by sucrose or salts (NaCl and KCl) to reach a physiological osmolarity (400 mOsm). Neocartilaginous matrix synthesis and matrix catabolism were evaluated by quantitative and immunofluorescence methods. Hydrogel degradation kinetics of acellular constructs were not affected by medium osmolarity or osmolyte. Matrix composition was predominantly aggrecan and collagen type II for all conditions. One day after encapsulation, total collagen accumulated in the constructs was increased by 80-90% in 400 mOsm medium, regardless of osmolyte. However, this effect did not persist, and at 4 weeks, total collagen synthesized and released to the medium was more than three times higher in 330 mOsm medium. Medium osmolarity had minimal effects on sulfated glycosaminoglycan content and did not affect catabolic activity. These findings suggest that culture medium at physiological osmolarities may not be beneficial for long-term chondrocyte culture in degradable hydrogels, but that initially culturing chondrocytes at a higher osmolarity may enhance early tissue deposition., (© 2014 Wiley Periodicals, Inc.)

The ethylene glycol-based dimethacrylates are commonly used in biomaterials and dental restorative materials as a cross-linking agent. In this study, toxic effect of triethylene glycol dimethacrylate (TEGDMA) and poly(ethylene glycol) dimethacrylates (PEG-DMAs) with various ethylene glycol repeating units was investigated in terms of cytotoxicity, oxidative stress, and the expression of co-stimulatory factors in human leukemia cell line (THP-1 cells) to verify the effect of ethylene glycol repeating units. Note that the 1-octanol/water partition coefficient of PEG-based dimethacrylates decreased with increasing the ethylene glycol repeating units, indicating that the hydrophilicity of PEG-DMAs increased with ethylene glycol repeating units. The toxic effect of PEG-DMAs such as cytotoxicity, oxidative stress, and the expression of CD86 in treated THP-1 cells are reduced with increasing the ethylene glycol repeating units in PEG-DMAs. However, the expression of CD54 in treated THP-1 cells was not influenced with the ethylene glycol repeating units and the maximal expression level of CD54 was observed at the concentration range of 2-4 mM for all samples. Accordingly, hydrophilic character of PEG-DMAs with long ethylene glycol chains definitely alleviates the some toxic aspect of PEG-based DMAs. This finding would provide important insight into the design of new biomaterials and dental materials with superior biocompatibility., (© 2014 Wiley Periodicals, Inc.)

This review focuses on the challenges associated with the design and development of injectable hydrogels of synthetic origin based on FDA approved blocks, such as polyethylene glycol (PEG) and poly(ɛ-caprolactone) (PCL). An overview of recent studies on inverse thermosensitive PEG/PCL hydrogels is provided. These systems have been proposed to overcome the limitations of previously introduced degradable thermosensitive hydrogels [e.g., PEG/poly(lactide-co-glycolic acid) hydrogels]. PEG/PCL hydrogels are advantageous due to their higher gel strength, slower degradation rate and availability in powder form. Particularly, triblock PEG/PCL copolymers have been widely investigated, with PCL-PEG-PCL (PCEC) hydrogels showing superior gel strength and slower degradation kinetics than PEG-PCL-PEG (PECE) hydrogels. Compared to triblock PEG/PCL copolymers, concentrated solutions of multiblock PEG/PCL copolymers were stable due to their slower crystallization rate. However, the resulting hydrogel gel strength was low. Inverse thermosensitive triblock PEG/PCL hydrogels have been mainly applied in tissue engineering, to decrease tissue adherence or, in combination with bioactive molecules, to promote tissue regeneration. They have also found application as in situ drug delivery carriers. On the other hand, the wide potentialities of multiblock PEG/PCL hydrogels, associated with the stability of their water-based solutions under storage, their higher degradation time compared to triblock copolymer hydrogels and the possibility to insert bioactive building blocks along the copolymer chains, have not been fully exploited yet. A critical discussion is provided to highlight advantages and limitations of currently developed themosensitive PEG/PCL hydrogels, suggesting future strategies for the realization of PEG/PCL-based copolymers with improved performance in the different application fields., (© 2014 Wiley Periodicals, Inc.)

We developed a novel antibacterial implant by forming a hydroxyapatite (HAp) film on polyetheretherketone (PEEK) substrate, and then immobilizing silver ions (Ag(+) ) on the HAp film based on the chelate-bonding ability of inositol phosphate (IP6). First, the PEEK surface was modified by immersion into concentrated sulfuric acid for 10 min. HAp film was formed on the acid-treated PEEK via the soft-solution process using simulated body fluid (SBF), urea, and urease. After HAp coating, specimens were immersed into IP6 solution, and followed by immersion into silver nitrite solution at concentrations of 0, 0.5, 1, 5 or 10 mM. Ag(+) ions were immobilized on the resulting HAp film due to the chelate-bonding ability of IP6. On cell-culture tests under indirect conditions by Transwell, MC3T3-E1 cells on the specimens derived from the 0.5 and 1 mM Ag(+) solutions showed high relative growth when compared with controls. Furthermore, on evaluation of antibacterial activity in halo test, elution of Ag(+) ions from Ag(+) -immobilized HAp film inhibited bacterial growth. Therefore, the above-mentioned results demonstrated that specimens had both biocompatibility and strong antibacterial activity. The present coating therefore provides bone bonding ability to the implant surface and prevents the formation of biofilms in the early postoperative period., (© 2014 Wiley Periodicals, Inc.)

Organic-inorganic hybrid materials based on zirconia and polyethylene glycol (PEG) have been synthesized via sol-gel method in the present study. Those materials, still in the sol phase, have been used to coat a titanium grade 4 (Ti-4) substrate to improve its biological properties. Dip-coating technique has been used to obtain thin films. PEG, a biocompatible polymer, used as the organic phase, has been incorporated with different percentages in an inorganic zirconium-based matrix. Those hybrids have been characterized by Fourier transform infrared spectroscopy (FTIR) to detect interactions between the two phases. The films have been examined using SEM to detect morphological changes with PEG percentages. The potential applications of the hybrid coatings in biomedical field have been evaluated by bioactivity and cytotoxicity tests. The coated titanium was immersed in simulated body fluid (SBF) for 21 days and the hydroxyapatite deposition on its surface was subsequently evaluated, as that feature can be used as an index of bone-bonding capability. SEM equipped with energy dispersive spectrometer (EDS) was used to examine hydroxyapatite formation. NIH 3T3 mouse embryonic fibroblast cells were seeded on specimens to evaluate cells-materials interactions and cell vitality was inspected using WST-8 Assay., (© 2014 Wiley Periodicals, Inc.)

Poly(ethylene glycol) (PEG) hydrogels are one of the most extensively utilized biomaterials systems due to their established biocompatibility and highly tunable properties. It is widely acknowledged that traditional acrylate-derivatized PEG (PEGDA) hydrogels are susceptible to slow degradation in vivo and are therefore unsuitable for long-term implantable applications. However, there is speculation whether the observed degradation is due to hydrolysis of endgroup acrylate esters or oxidation of the ether backbone, both of which are possible in the foreign body response to implanted devices. PEG diacrylamide (PEGDAA) is a polyether-based hydrogel system with similar properties to PEGDA but with amide linkages in place of the acrylate esters. This provides a hydrolytically-stable control that can be used to isolate the relative contributions of hydrolysis and oxidation to the in vivo degradation of PEGDA. Here we show that PEGDAA hydrogels remained stable over 12 weeks of subcutaneous implantation in a rat model while PEGDA hydrogels underwent significant degradation as indicated by both increased swelling ratio and decreased modulus. As PEGDA and PEGDAA have similar susceptibility to oxidation, these results demonstrate for the first time that the primary in vivo degradation mechanism of PEGDA is hydrolysis of the endgroup acrylate esters. Additionally, the maintenance of PEGDAA hydrogel properties in vivo indicates their suitability for long-term implants. These studies serve to elucidate key information about a widely used biomaterial system to allow for better implantable device design and to provide a biostable replacement option for PEGDA in applications that require long-term stability., (© 2014 Wiley Periodicals, Inc.)

Functionalization of polymer surfaces has been recognized as a valuable tool to improve their properties that significantly influence cellular behaviors, such as adhesion, proliferation, migration, and differentiation. In stem cell-mediated bone tissue engineering, surface multifunctionalization of polymeric scaffolds with cell-adhesive, osteoconductive, and osteoinductive biomolecules is a critical strategy to improve such properties. However, the traditional surface modification techniques such as physical deposition/adsorption, chemical modification, grafting, and plasma techniques have great limitations for immobilization of multiple bioactive molecules due to multistep procedures. Recently, a universal technique based on mussel-inspired self-polymerization of dopamine is developed for multifunctional coatings in a simple way. In our study, we used this newly developed technique to incorporate three biomolecules, cell adhesion-promoting (K)16 GRGDSPC peptides, osteoconductive hydroxyapatite (HAp) nanoparticles, and osteoinductive bone morphogenetic protein-2-derived P24 peptides, to functionalize poly(lactide-co-glycolide) (PLGA)-[Asp-PEG]n scaffolds, and the effects on biological behaviors of co-cultured rabbit-derived bone marrow stromal cells in vitro were investigated. The results showed (K)16 GRGDSPC, HAp, and P24 could be immobilized onto the scaffolds through predeposition of polydopamine (pDA) ad-layer, and the surface-modified scaffolds were noncytotoxic as well as the virgin scaffold. The pDA-assisted codeposition of (K)16 GRGDSPC, HAp, and P24 on the scaffold surfaces significantly promoted cell adhesion, proliferation, osteodifferentiation, and mineralization in vitro with synergistic effects. Taken together, the functionalized PLGA-[Asp-PEG]n polymeric scaffolds achieved significantly elevated affinity, osteoconductive and osteoinductive ability, and may be a potentially promising bone graft substitute for bone repair., (© 2014 Wiley Periodicals, Inc.)

Polyetheretherketone (PEEK) films were oxygen plasma treated to increase surface free energy and characterized by X-ray photoelectron microscopy, atomic force microscopy, and water contact angles. A parallel plate flow chamber was used to measure Staphylococcus epidermidis, Staphylococcus aureus, and U-2 OS osteosarcomal cell-line adhesion to the PEEK films in separate monocultures. In addition, bacteria and U-2 OS cells were cocultured to model competition between osteoblasts and contaminating bacteria for the test surfaces. Plasma treatment of the surfaces increased surface oxygen content and decreased the hydrophobicity of the materials, but did not lead to a significant difference in bacterial or U-2 OS cell adhesion in the monocultures. In the S. epidermidis coculture experiments, the U-2 OS cells adhered in greater numbers on the treated surfaces compared to the untreated PEEK and spread to a similar extent. However, in the presence of S. aureus, cell death of the U-2 OS occurred within 10 h on all surfaces. The results of this study suggest that oxygen plasma treatment of PEEK may maintain the ability of osteoblast-like cells to adhere and spread, even in the presence of S. epidermidis contamination, without increasing the risk of preoperative bacterial adhesion. Therefore, oxygen plasma-treated PEEK remains a promising method to improve implant surface free energy for osseointegration., (© 2014 Wiley Periodicals, Inc.)

Healing articular cartilage defects remains a significant clinical challenge because of its limited capacity for self-repair. While delivery of autologous chondrocytes to cartilage defects has received growing interest, combining cell-based therapies with growth factor delivery that can locally signal cells and promote their function is often advantageous. We have previously shown that PEG thiol-ene hydrogels permit covalent attachment of growth factors. However, it is not well known if embedded chondrocytes respond to tethered signals over a long period. Here, chondrocytes were encapsulated in PEG hydrogels functionalized with transforming growth factor-beta 1 (TGF-β1) with the goal of increasing proliferation and matrix production. Tethered TGF-β1 was found to be distributed homogenously throughout the gel, and its bioactivity was confirmed with a TGF-β1 responsive reporter cell line. Relative to solubly delivered TGF-β1, chondrocytes presented with immobilized TGF-β1 showed significantly increased DNA content, and GAG and collagen production over 28 days, while maintaining markers of articular cartilage. These results indicate the potential of thiol-ene chemistry to covalently conjugate TGF-β1 to PEG to locally influence chondrocyte function over 4 weeks. Scaffolds with other or multiple tethered growth factors may prove broadly useful in the design of chondrocyte delivery vehicles for cartilage tissue engineering applications., (© 2014 The Authors. Journal of Biomedical Materials Research Part A Published by Wiley Periodicals, Inc.)

Although triethylene glycol dimethacrylate (TEGDMA), a resin monomer widely used in dental practice, has been shown to have cytotoxic effects on eukaryotic cells, little is known about how the oral environment influences the cytotoxicity of this biomaterial. The aim of this study was to evaluate eukaryotic cell reaction to TEGDMA in terms of the production of reactive oxygen species (ROS), the expression of Bax, the disturbance of mitochondrial membrane potential (MMP), and the occurrence of apoptosis in an in vitro coculture model of human gingival fibroblasts (HGFs) and Streptococcus mitis strain in presence of saliva. We found that S. mitis and saliva reduced the production of ROS (from 2.2 to 1.8 fold), the occurrence of apoptosis (from 11.3 to 4.7%), and the decrease of MMP (from 0.75 to 0.9 fold) induced by TEGDMA treatment. Addition of N-acetylcysteine, a well known antioxidant, improved cell viability in all experimental conditions. The results obtained in this study suggest that the presence of S. mitis and saliva in the periodontal environment could protect cells against TEGDMA toxicity. These results, shedding more light on the biological and molecular events that occur in conjuction with TEGDMA treatment in vitro in a coculture model that mimics the environment of the oral cavity, confirm the key role played by oral bacteria and saliva in preventing toxic events that can occur in vivo in HGFs., (© 2013 Wiley Periodicals, Inc.)

Sodium chloride and polyethylene glycol (PEG) were used as water-soluble porogens for the formation of porous polycaprolactone (PCL) scaffolds. The main purpose was to prepare and evaluate in vitro efficacy of highly interconnected, three-dimensional, porous polymeric scaffolds, as obtained from the combined particulate and polymer leaching techniques. Microscopic analysis confirmed the high interconnectivity of the pores and relatively uniform pore size of 378-435 μm. The PCL scaffolds were further characterized for their density and pore characteristics, water absorption and flow behaviors, and mechanical properties and the potential for their use as bone scaffolding materials was evaluated in vitro using mouse calvaria-derived preosteoblastic cells (MC3T3-E1). Evidently, the use of PEG as the secondary porogen not only improved the interconnectivity of the pore structures but also resulted in the PCL scaffolds that exhibited much better support for the proliferation and differentiation of the cultured bone cells.

Aromatic poly(ether ether ketone) (PEEK) is a super engineering plastic, which has good mechanical properties and is resistant to physical and chemical stimuli. We have, therefore, attempted to use PEEK in cardiovascular devices. Synthetic cardiovascular devices require both high hemocompatibility and anti-inflammatory activity in addition to the mechanical properties. We modified the PEEK surface by photoinduced and self-initiated graft polymerization with 2-methacryloyloxyethyl phosphorylcholine (MPC; PMPC-grafted PEEK) for obtaining good antithrombogenicity. Polymerization was carried out on the surface of PEEK under radiation of ultraviolet (UV) light during which we controlled monomer concentrations, temperatures, and UV intensities. The biological performance of the PMPC-grafted PEEK was examined and compared with that of unmodified PEEK. With increase in the thickness of the PMPC layer, the amount of fibrinogen adsorption decreased significantly in comparison to that in the case of unmodified PEEK. When placed in contact with human platelet-rich plasma, surface of the PMPC-grafted PEEK clearly showed inhibition of platelet adhesion and activation. Also, bacterial adhesion was reduced dramatically on the PMPC-grafted PEEK. Thus, the PMPC grafting on PEEK improved the antithrombogenicity., (Copyright © 2013 Wiley Periodicals, Inc.)

Bone regenerative medicine, based on the combined use of cells and scaffolds, represents a promising strategy in bone regeneration. Hydrogels have attracted huge interests for application as a scaffold for minimally invasive surgery. Collagen and oligo(poly(ethylene glycol)fumarate) (OPF) hydrogels are the representatives of two main categories of hydrogels, that is, natural- and synthetic-based hydrogels. With these the optimal cell-loading (i.e., cell distribution inside the hydrogels) method was assessed. The cell behavior of both bone marrow- and adipose tissue-derived mesenchymal stem cells (BM- and AT-MSCs) in three loading methods, which are dispersed (i.e., homogeneous cell encapsulation, D), sandwich (i.e., cells located in between two hydrogel layers, S), and spheroid (i.e., cell pellets encapsulation, Sp) loading in two hydrogel systems (i.e., collagen and OPF), was compared. The results suggested that the cell behavior was influenced by the hydrogel type, meaning cells cultured in collagen hydrogels had higher proliferation and osteogenic differentiation capacity than in OPF hydrogels. In addition, AT-MSCs exhibited higher proliferation and osteogenic properties compared to BM-MSCs. However, no difference was observed for mineralization among the three loading methods, which did not approve the hypothesis that S and Sp loading would increase osteogenic capacity compared to D loading. In conclusion, D and Sp loading represents two promising cell loading methods for injectable bone substitute materials that allow application of minimally invasive surgery for cell-based regenerative treatment., (Copyright © 2013 Wiley Periodicals, Inc.)

Electrospun scaffolds are widely used in tissue engineering; however, a common problem is the poor cell infiltration because of the small pore size and tightly packed structure of these fibrous scaffolds. To address this issue, a novel technique was developed to fabricate electrospun silk fibroin (SF) scaffolds with rather macropores and high porosity using electrospraying-generated PEO microparticles as porogen. The morphology and pore size of MPES scaffolds were evaluated by scanning electron microscopy. It was revealed that MPES scaffold had a relatively loose structure with an increase of mean pore size (i.e., approx. 30 μm of MPES vs. approx. 5 μm of traditional electrospun scaffolds (TES) and porosity (i.e., 95% vs. 84% of TES). Culture of mouse 3T3 fibroblast in TES and MPES scaffold revealed that both scaffolds could support cell attachment, spread and proliferation. Yet, cell inflitration in vitro under the static culture condition only occurred in the MPES scaffold. Subcutaneous implantation of scaffolds in rats further confirmed that the tissue ingrowth was more efficient in the MPES scaffold compared to TES scaffold. Thus, the use of PEO microparticles as porogen was a feasible and effective method for creating macroporous electrospun SF scaffold, which provided an alternative to address the limitation of cell infiltration associated with electrospun fibrous scaffold., (Copyright © 2013 Wiley Periodicals, Inc., a Wiley Company.)

Three-dimensional tissue engineered constructs provide a platform to examine how the local extracellular matrix (ECM) contributes to the malignancy of cancers such as human glioblastoma multiforme. Improved resolution of how local matrix biophysical features impact glioma proliferation, genomic and signal transduction paths, as well as phenotypic malignancy markers would complement recent improvements in our understanding of molecular mechanisms associated with enhanced malignancy. Here, we report the use of a gelatin methacrylate (GelMA) platform to create libraries of three-dimensional biomaterials to identify combinations of biophysical features that promote malignant phenotypes of human U87MG glioma cells. We noted key biophysical properties, namely matrix density, crosslinking density, and biodegradability, that significantly impact glioma cell morphology, proliferation, and motility. Gene expression profiles and secreted markers of increased malignancy, notably VEGF, MMP-2, MMP-9, HIF-1, and the ECM protein fibronectin, were also significantly impacted by the local biophysical environment as well as matrix-induced deficits in diffusion-mediated oxygen and nutrient biotransport. Overall, this biomaterial system provides a flexible platform to explore the role biophysical factors play in the etiology, growth, and subsequent invasive spreading of gliomas., (Copyright © 2013 Wiley Periodicals, Inc., a Wiley Company.)

In this study, PEGylated nanoparticles quercetin drug delivery vehicles were investigated as carriers for anticancer drugs induced programed cell death (PCD). PEG2000-DPSE-coated quercetin nanoparticles were prepared and tumor cell killing efficacy was studied on glioma C6 cells and assayed for cell survival, apoptosis, or necrosis. The levels of ROS production and mitochondrial membrane potential (ΔΨm) were determined. Western blot assayed p53, p-p53, cytochrome C, and caspase proteins expression were also studied. Results indicate that PEG2000-DPSE-QUE-NPS showed dose-dependent cytotoxicity to C6 glioma cells and enhanced ROS accumulation induced upregulation of p53 protein, which was accompanied with an increase in cytochrome c and caspase-3 protein levels. These results support the hypothesis that quercetin nanoparticles-coated PEG2000-DPSE remarkably enhanced anticancer effect of induced programed cell death on C6 glioma cells. Overall, PEG2000-DPSE-coated quercetin nanoparticles showed promising potential as a drug carrier for cancer therapy., (Copyright © 2013 Wiley Periodicals, Inc.)

A poly (lactide-co-glycolide) (PLGA) scaffold filled with fibrin gel, mesenchymal stem cells (MSCs) and poly(ethylene oxide)-b-poly (L-lysine) (PEO-b-PLL)/pDNA-TGF-β1 complexes was fabricated and applied in vivo for synchronized regeneration of cartilage and subchondral bone. The PEO-b-PLL/pDNA-TGF-β1 complexes could transfect MSCs in vitro to produce TGF-β1 in situ and up regulate the expression of chondrogenesis-related genes in the construct. The expression of heterogeneous TGF-β1 in vivo declined along with the prolongation of implantation time, and lasted for 3 and 6 weeks in the mRNA and protein levels, respectively. The constructs (Experimental group) of PLGA/fibrin gel/MSCs/(PEO-b-PLL/pDNA-TGF-β1 complexes) were implanted into the osteochondral defects of rabbits to restore the functional cartilages, with gene-absent constructs as the Control. After 12 weeks, the Experimental group regenerated the neo-cartilage and subchondral bone with abundant deposition of glycosaminoglycans (GAGs) and type II collagen. The regenerated tissues had good integration with the host tissues too. By contrast, the defects were only partially repaired by the Control constructs. qRT-PCR results demonstrated that expression of the chondrogenesis-marker genes in the Experimental group was significantly higher than that of the Control group, and was very close to that of the normal cartilage tissue., (Copyright © 2013 Wiley Periodicals, Inc.)

Temperature-sensitive hydrogels are attractive alternatives to porous cell-seeded scaffolds and is minimally invasive through simple injection and in situ gelling. In this study, we compared the performance of two types of temperature-sensitive hydrogels on chondrocytes encapsulation for the use of tissue engineering of cartilage. The two hydrogels are composed of methoxy poly(ethylene glycol)- poly(lactic-co-valerolactone) (mPEG-PVLA), and methoxy poly(ethylene glycol)-poly(lactic- co-glycolide) (mPEG-PLGA). Osmolarity and pH were optimized through the manipulation of polymer concentration and dispersion medium. Chondrocytes proliferation in mPEG-PVLA hydrogels was observed as well as accumulation of GAGs and collagen. On the other hand, chondrocytes encapsulated in mPEG-PLGA hydrogels showed low viability and chondrogenesis. Also, mPEG-PVLA hydrogel, which is more hydrophobic, retained physical integrity after 14 days while mPEG-PLGA hydrogel underwent full degradation due to faster hydrolysis rate and more pronounced acidic self-catalyzed degradation. The mPEG-PVLA hydrogel can be furthered tuned by manipulation of molecular weights to obtain hydrogels with different swelling and degradation characteristics, which may be useful as producing a selection of hydrogels compatible with different cell types. Taken together, these results demonstrate that mPEG-PVLA hydrogels are promising to serve as three-dimensional cell carriers for chondrocytes and potentially applicable in cartilage tissue engineering., (Copyright © 2013 Wiley Periodicals, Inc.)

In vivo bone regeneration of chitosan-poly(ethylene oxide) (PEO) hydrogel in rat carlvarial defects was evaluated by using both human bone marrow-derived stromal cells (hMSCs) and recombinant human bone marrow protein-2 (rhBMP-2) for 4 and 8 weeks. In situ chitosan-PEO hydrogel was fabricated by mixing the precursor solutions of both chitosan-acrylate and PEO-thiol. Fabrication of the injectable hydrogels was modulated from within a minute to hours by controlling the temperature and pHs of the precursor solution. Gel swellings were dependent on the conditions of pHs and temperatures of the precursor solutions, showing higher gel swelling in basic water than in either acidic or neutral water. The compression strengths and in vitro degradation of hydrogels were also evaluated by controlling the concentrations of both precursor solutions and lysozyme, respectively, by referencing to the morphology of the control hydrogel with no enzyme added. Hydrogels showed sustained release of rhodamine-B over time. After implantation of the injectable hydrogels in rat calvarial defects for 4 and 8 weeks, in vivo bone regenerations were compared with by evaluating the degrees of new bone formations with Soft X-ray, microcomputed tomography, and histological stainings of hematoxylin and eosine Y and Masson's trichrome. Degrees of in vivo bone regeneration were controlled by encapsulating in advance either hMSCs, rhBMP-2, or both in the precursor solutions of the hydrogel. The defect implanted with hydrogel only showed higher amount of bone tissue regeneration than that of the control defect site. The defect sites with hydrogel containing both hMSCs and rhBMP-2 demonstrated highest amount of bone tissue regeneration among the samples., (Copyright © 2012 Wiley Periodicals, Inc.)

Osseointegration of surface-modified polyetheretherketone (PEEK) implants was studied in vivo. A total of 18 cylinder-shaped PEEK implants were inserted in the femurs of nine New Zealand rabbits; half were coated with nanocrystalline hydroxyapatite (nanoHA) and half were uncoated controls. Healing time was 6 weeks. Samples were retrieved with the implant and surrounding tissue, processed to cut and ground sections, and analyzed histomorphometrically. The implant surfaces were analyzed with optical interferometry, scanning electron microscopy (SEM), atomic force microscopy, and X-ray photoelectron spectroscopy (XPS). NanoHA-coated PEEK surfaces had lower height deviation (Sa) than controls [mean ± SD: 0.41 μm (± 0.14) vs. 0.96 μm (± 0.28)]. SEM images showed the nanoHA crystals as a thin layer on the polymer surface. XPS analysis of the coated implants showed a Ca/P ratio of 1.67. Histomorphometry indicated that the nanoHA-coated implants had more bone-to-implant contact [16% (± 4.7) vs. 13% (± 9.3)] and more bone area [52% (± 9.5) vs. 45% (± 11.9)]. We found no difference between smooth nanoHA-coated cylinder-shaped PEEK implants and uncoated controls. However, higher mean bone-to-implant contact indicated better osseointegration in the coated implants than in the uncoated controls. The large number of lost implants was interpreted as a lack of primary stability due to implant design., (Copyright © 2012 Wiley Periodicals, Inc.)

With the help of a silane coupling agent, poly(ethylene glycol) (PEG), a well-biocompatable agent, was grafted onto the surface of ultrahigh molecular weight polyethylene (UHMWPE) by ultraviolet initiation. Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy analysis proved the success of PEG grafting. Water contact angle measurement showed that the modified UHMWPE was obviously improved in surface hydrophilicity and thermogravimetric analysis result showed that its thermostability did not decline even it was pretreated by strong acids. Then, the protein adsorption of the modified UHMWPE was investigated using three model proteins including bovine serum albumin, lysozyme, and fibrinogen. Rabbit blood was used to study the platelet adhesion on the surface of modified UHMWPE. The results indicated that the quantity of protein adsorption on the modified UHMWPE grafted PEG reduced apparently for all the model proteins while there was some specific differences or exceptions among them. It was ascribed to the changed surface chemical composition, surface hydrophilicity and surface topography after modification. The adhesive ability of blood platelets on the modified surface of UHMWPE decreased after PEG grafting. Owing to the improved resistance to fibrinogen adsorption and platelet adhesion, the surface modification might endow the UHWMPE surface better anticoagulation ability according to clotting mechanism., (Copyright © 2012 Wiley Periodicals, Inc.)

Polyetheretherketone (PEEK) is a high performance polymer, with high melting temperature and high resistance to wear. PEEK biomedical devices are typically manufactured to produce nonflexible structures. In this study, we fabricated flexible PEEK scaffolds from multifilament and monofilament yarns, using weaving technologies. Scaffolds were compared for structural and mechanical properties, and assessed for in vitro biological response to L929 mouse fibroblast cells. PEEK scaffolds were found to support fibroblast cell attachment and proliferation, with similar cell numbers to a polyethylene terephthalate scaffold. The large pores (261-280 μm) of the monofilament scaffold prevented pore coverage by cells, confining cells to filaments, whereas the smaller pores (81-100 μm) of the multifilament scaffold permitted partial pore coverage. Poor cell adhesion, due to large filament curvature angles, created a checkered pattern on the woven surface, a previously undocumented phenomenon. The multifilament scaffold was found to be lighter, thinner, and less porous, with better mechanical properties (load at break: 657 N, elastic recovery: 66%, burst strength: 492 N) than the monofilament scaffold (load at break: 534 N, elastic recovery: 30%, burst strength: 401 N). Results indicate that flexible PEEK woven structures may find application as tissue engineering scaffolds, particularly for engineering soft tissues., (Copyright © 2012 Wiley Periodicals, Inc.)

Pluronic-immobilized nanofibrous meshes were tailored for thermally induced incorporation of dexamethasone. A diblock copolymer composed of poly(e-caprolactone)-poly (ethyleneglycol) (NH(2)) (PCL-PEG (NH(2))) was electrospun to a nanofibrous mesh, and Pluronic was subsequently surface-immobilized on the mesh in aqueous phase. Surface-wettability analysis and (1)H NMR spectroscopy confirmed surface-decoration of nanofibrous meshes with Pluronic moieties depending on the blend ratios of PCL-PEG(NH(2)). Fluorescently-labeled micelles were incorporated in the nanofibrous meshes by temperature modulation and showed attenuated release profiles of the micelles were for 1 month. The suppression degree of drug-loaded micelle releases was proportional to the blend ratio of PCL-PEG(NH(2)). Dexamethasone was formulated into micellar nanoaggregates, and the dexamethasone micelles-loaded nanofibrous meshes were used for antiproliferation studies of smooth muscle cells. Flow cytometric analysis of the arrested cells at a G(0)-G(1) phase revealed that the dexamethasone micelles-loaded nanofibrous meshes effectively controlled proliferation of the smooth muscle cells when cells were cultivated with the nanofibrous meshes. The antiproliferation effects of the nanofibrous meshes were closely correlated to the release profiles of the micelles from the nanofibrous meshes with different blend ratios. Thus, dexamethasone-incorporated nanofibrous meshes can be potentially used for treatments of restenosis after percutaneous transluminal coronary angioplasty., (Copyright © 2012 Wiley Periodicals, Inc.)

Titanium dioxide (TiO(2)), known to exhibit good biocompatibility, is applied in this study as a thin film formed onto polyetheretherketone (PEEK) substrate, which has been widely used in spinal interbody fusion cages. For successful deposition, an arc ionplating (AIP) technique was applied to deposit TiO(2) at low deposition temperature without damaging PEEK substrate, while providing satisfactory film adhesion. This study systematically investigates the effects of TiO(2) thin film phase composition and surface characteristics, controlled by using different target current and substrate bias, on osteoblast compatibility. Experimental results showed that anatase phase (A-TiO(2)) and/or rutile phase (R-TiO(2) ) TiO(2) coatings, respectively, can be prepared in appropriate deposition conditions. Overall, the TiO(2)-coated PEEK presented better osteoblast compatibility than the bare PEEK material in terms of cell adhesion, cell proliferation, and cell differentiation abilities, as well as osteogenesis performance (as determined by levels of osteopontin, osteocalcin, and calcium content). Surface roughness and hydrophilicity of the AIP-TiO(2) films were found to be responsible for significant osteoblast cell growth. It is also noticeable that the R-TiO(2) exhibited better osteoblast compatibility than the A-TiO(2) due to the presence of negatively charged hydroxyl groups on R-TiO(2) (110) surface in nature., (Copyright © 2012 Wiley Periodicals, Inc.)

A model in vitro system was developed for eliciting classical (M1) activation of surface-adherent murine macrophages, which was then used to study the interaction of the M1 macrophages with Staphylococcus epidermidis. Glass substrata were first covalently grafted with a mixture of methoxy- and biotin-terminated silanated polyethylene glycol. Interferon (IFN)-γ and/or lipopolysaccharide (LPS), ligands known to induce the highly microbicidal M1 activation state in macrophages, were biotinylated and immobilized by way of a streptavidin intermediate to the biotin-PEG base substratum. Assessment of mouse bone marrow-derived macrophage (BMDM) interleukin (IL)-12(p40) and nitric oxide response to the fabricated surfaces confirmed that the model system achieved activation of adherent macrophage: IFN-γ-presenting surfaces primed cells for M1 activation, LPS-presenting surfaces elicited innate activation, and surface presenting a combination of IFN-γ and LPS induced M1 activation. The phagocytic and microbicidal capacity of activated, surface-adherent BMDM was evaluated using S. epidermidis, a bacterial species prevalent in implant-associated infections. Results indicate that M1 activation of implant-adherent macrophages trends towards diminishing their phagocytic capacity, but enhances their microbicidal capacity for S. epidermidis., (Copyright © 2012 Wiley Periodicals, Inc.)

The biodegradable polylactide/poly(ethylene glycol) (PLA/PEG) hybrid membranes were fabricated via electrospinning of PLA/PEG solution. Their structures and properties were investigated by scanning electron microscopy, differential scanning calorimetry, and water contact angle. In vitro hydrolytic degradation showed that PEG content influenced the degradation rate of the PLA/PEG hybrid mats. The mechanical property was measured by tensile test and the result revealed that the addition of PEG had an obvious plasticization on PLA matrix. In-vitro biocompatibility was investigated by culturing cell on the scaffolds and MTT assay. The results indicated that the cell could attach and proliferate on the membranes, so confirmed that the PLA/PEG hybrid membrane had good biocompatibility, and it could be a promising biomaterial for tissue engineering applications., (Copyright © 2011 Wiley Periodicals, Inc.)