Injectable Tissue Engineering Scaffold Research Articles

CT-Visible Microspheres Enable Whole-Body In Vivo Tracking of Injectable Tissue Engineering Scaffolds.

Targeted delivery and retention are essential requirements for implantable tissue-engineered products. Non-invasive imaging methods that can confirm location, retention, and biodistribution of transplanted cells attached to implanted tissue engineering scaffolds will be invaluable for the optimization and enhancement of regenerative therapies. To address this need, an injectable tissue engineering scaffold consisting of highly porous microspheres compatible with transplantation of cells is modified to contain the computed tomography (CT) contrast agent barium sulphate (BaSO4). The trackable microspheres show high x-ray absorption, with contrast permitting whole-body tracking. The microspheres are cellularized with GFP+ Luciferase+ mesenchymal stem cells and show in vitro biocompatibility. In vivo, cellularized BaSO4-loaded microspheres are delivered into the hindlimb of mice where they remain viable for 14 days. Co-registration of 3D-bioluminescent imagingand µCT reconstructions enable the assessment of scaffold material and cell co-localization. The trackable microspheres are also compatible with minimally-invasive delivery by ultrasound-guided transthoracic intramyocardial injections in rats. These findings suggest that BaSO4-loaded microspheres can be used as a novel tool for optimizing delivery techniques and tracking persistence and distribution of implanted scaffold materials. Additionally, the microspheres can be cellularized and have the potential to be developed into an injectable tissue-engineered combination product for cardiac regeneration.

Alginate hydrogels containing different concentrations of magnesium-containing poly(lactic-co-glycolic acid) microspheres for bone tissue engineering

The easy loss of crosslinking ions in alginate can result in structural collapse and loss of its characteristics as a bone scaffold. A novel injectable tissue engineering scaffold containing poly(lactic-co-glycolic acid) (PLGA) microspheres and alginate was fabricated to improve alginate’s physiochemical and biological properties. MgCO3 and MgO were loaded at a 1:1 ratio into PLGA microspheres to form biodegradable PLGA microspheres containing magnesium (PMg). Subsequently, different concentrations of PMg were mixed into a Ca2+ suspension and employed as crosslinking agents for an alginate hydrogel. A pure Ca2+ suspension was used as the alginate crosslinking agent in the control group. The influence of PMg on the physiochemical properties of the injectable scaffolds, including the surface morphology, degradation rate, Mg2+ precipitation concentration, and the swelling rate, was investigated. MC3T3-E1 cells were seeded onto the hydrogels to evaluate the effect of the resultant alginate on osteoblastic attachment, proliferation, and differentiation. The physicochemical properties of the hydrogels, including morphology, degradation rate, and swelling ratio, were effectively tuned by PMg. Inductively coupled plasma-optical emission spectroscopy results showed that, in contrast to those in pure PMg, the magnesium ions (Mg2+) in alginate hydrogel containing PMg microspheres (Alg-PMg) were released in a dose-dependent and slow-releasing manner. Additionally, Alg-PMg with an appropriate concentration of PMg not only improved cell attachment and proliferation but also upregulated alkaline phosphatase activity, gene expression of osteogenic markers, and related growth factors. These findings indicate that PMg incorporation can regulate the physicochemical properties of alginate hydrogels. The resultant hydrogel promoted cell attachment, matrix mineralization, and bone regeneration. The hydrogel described in this study can be considered a promising injectable scaffold for bone tissue engineering.

Additive manufacturing of hierarchical injectable scaffolds for tissue engineering

We present a 3D-printing technology allowing free-form fabrication of centimetre-scale injectable structures for minimally invasive delivery. They result from the combination of 3D printing onto a cryogenic substrate and optimisation of carboxymethylcellulose-based cryogel inks. The resulting highly porous and elastic cryogels are biocompatible, and allow for protection of cell viability during compression for injection. Implanted into the murine subcutaneous space, they are colonized with a loose fibrovascular tissue with minimal signs of inflammation and remain encapsulation-free at three months. Finally, we vary local pore size through control of the substrate temperature during cryogenic printing. This enables control over local cell seeding density in vitro and over vascularization density in cell-free scaffolds in vivo. In sum, we address the need for 3D-bioprinting of large, yet injectable and highly biocompatible scaffolds and show modulation of the local response through control over local pore size. Statement of SignificanceThis work combines the power of 3D additive manufacturing with clinically advantageous minimally invasive delivery. We obtain porous, highly compressible and mechanically rugged structures by optimizing a cryogenic 3D printing process. Only a basic commercial 3D printer and elementary control over reaction rate and freezing are required. The porous hydrogels obtained are capable of withstanding delivery through capillaries up to 50 times smaller than their largest linear dimension, an as yet unprecedented compression ratio. Cells seeded onto the hydrogels are protected during compression. The hydrogel structures further exhibit excellent biocompatibility 3 months after subcutaneous injection into mice.We finally demonstrate that local modulation of pore size grants control over vascularization density in vivo. This provides proof-of-principle that meaningful biological information can be encoded during the 3D printing process, deploying its effect after minimally invasive implantation.

Biopolymeric hydrogels − nanostructured TiO2 hybrid materials as potential injectable scaffolds for bone regeneration

The present work aims at development of novel hybrid materials from genipin crosslinked collagen or collagen/chitosan hydrogels containing various types of TiO2 nanoparticles characterized with different anatase/rutile ratios. Collagen and chitosan were selected as hydrogel components since they are biopolymers being, like collagen, the major compound present in extracellular matrix or exhibit structural similarity to glycosaminoglycans, like chitosan. TiO2 nanoparticles were introduced to the hydrogel matrices to improve their mechanical properties as well as bioactivity. A series of twelve novel hybrid materials were prepared and their physicochemical, mechanical and biological properties were evaluated.It was found that TiO2 nanostructures introduced to the hydrogels have significant influence on the swelling properties of the synthesized hybrids and their impact is strongly dependent on the type of matrices. The surfaces of hybrid materials were found to be more hydrophilic than these of corresponding hydrogel matrix. It was also observed that, the storage modulus values of the hybrids based on collagen-chitosan hydrogel are comparable to these for plain hydrogels what indicates that the mechanical properties of the materials obtained are satisfactory for possible biomedical application. The in vitro cell culture studies have shown that prepared materials are biocompatible as they can support mitochondrial activity of MEFs as well as MG-63 cells. In vitro experiments performed under simulated body fluid (SBF) conditions have revealed that all studied TiO2 nanoparticles present in hydrogel matrices, regardless of anatase/rutile ratio, successfully induced formation of apatite-like structures. The hybrid materials developed here are promising candidates for preparation of bioactive, injectable scaffolds for tissue engineering.

Fibrin gel as an injectable biodegradable scaffold and cell carrier for tissue engineering.

Due to the increasing needs for organ transplantation and a universal shortage of donated tissues, tissue engineering emerges as a useful approach to engineer functional tissues. Although different synthetic materials have been used to fabricate tissue engineering scaffolds, they have many limitations such as the biocompatibility concerns, the inability to support cell attachment, and undesirable degradation rate. Fibrin gel, a biopolymeric material, provides numerous advantages over synthetic materials in functioning as a tissue engineering scaffold and a cell carrier. Fibrin gel exhibits excellent biocompatibility, promotes cell attachment, and can degrade in a controllable manner. Additionally, fibrin gel mimics the natural blood-clotting process and self-assembles into a polymer network. The ability for fibrin to cure in situ has been exploited to develop injectable scaffolds for the repair of damaged cardiac and cartilage tissues. Additionally, fibrin gel has been utilized as a cell carrier to protect cells from the forces during the application and cell delivery processes while enhancing the cell viability and tissue regeneration. Here, we review the recent advancement in developing fibrin-based biomaterials for the development of injectable tissue engineering scaffold and cell carriers.

A comparison of physicochemical properties of sterilized chitosan hydrogel and its applicability in a canine model of periodontal regeneration

Chitosan has previously been exploited as a scaffold in tissue engineering processes. To avoid infection, chitosan must be sterilized prior to contact with bodily fluids or blood. Previous research has shown that autoclaved chitosan solution lead to decreased molecular weight, dynamic viscosity, and rate of gelling. We prepared a thermosensitive chitosan hydrogel using autoclaved chitosan powder (121°C, 10min) and β-glycerophosphate (chitosan-PA/GP) and compared the physicochemical properties and biocompatibility in vitro with autoclaved chitosan solution/GP hydrogel. The chitosan-PA/GP hydrogel had a shortened gelation time, higher viscosity, increased water absorption, appropriate degradation time, porous structure, and no obvious cytotoxicity on human periodontal ligament cells. Scanning electron microscopy demonstrated that the cells exhibited a normal morphology. The chitosan-PA/GP hydrogel promoted periodontal tissue regeneration in dog class III furcation defects. The chitosan-PA/GP thermosensitive hydrogel displayed suitable physicochemical properties and biocompatibilities and represents a promising candidate as an injectable tissue engineering scaffold.

3D culture of tonsil-derived mesenchymal stem cells in poly(ethylene glycol)-poly(L-alanine-co-L-phenyl alanine) thermogel.

Poly(ethylene glycol)-poly(L-alanine-co-L-phenyl alanine) (PEG-PAF) aqueous solutions undergo sol-to-gel transition as the temperature increases. The transition is driven by the micelle aggregation involving the partial dehydration of the PEG block and the partial increase in β-sheet content of the PAF block. Tonsil-tissue-derived mesenchymal stem cells (TMSCs), a new stem cell resource, are encapsulated through the sol-to-gel transition of the TMSC-suspended PEG-PAF aqueous solutions. The encapsulated TMSCs are in vitro 3D cultured by using induction media supplemented with adipogenic, osteogenic, or chondrogenic factors, where the TMSCs preferentially undergo chondrogenesis with high expressions of type II collagen and sulfated glycosaminoglycan. As a feasibility study of the PEG-PAF thermogel for injectable tissue engineering, the TMSCs encapsulated in hydrogels are implanted in the subcutaneous layer of mice by injecting the TMSC suspended PEG-PAF aqueous solution. The in vivo studies also prove that TMSCs undergo chondrogenesis with high expression of the chondrogenic biomarkers. This study suggests that the TMSCs can be an excellent resource of MSCs, and the thermogelling PEG-PAF is a promising injectable tissue engineering scaffold, particularly for chondrogenic differentiation of the stem cells.

Shear Thinning Three-Dimensional Colloidal Assemblies of Chitosan and Poly(lactic acid) Nanoparticles

In this study, new materials capable of reversible self-assembly, based on concentrated negatively charged poly(lactic acid) nanoparticles and chitosan, a natural polycationic polymer, were successfully fabricated. Electrostatic interactions between oppositely charged components along with weaker interactions led to the formation of a 3D network. The resulting macroscopic assemblies were characterized by dynamic mechanical measurements, and the influences of various parameters such as chitosan/poly(lactic acid) weight ratio, duration and temperature of the mixture, and molecular weight or chitosan degree of acetylation were studied. Our results showed that the mechanical properties of assemblies were highly dependent on the nanoparticle solid content and chitosan/nanoparticle ratio. In particular, at an optimum weight ratio the colloidal assemblies exhibited remarkable high elastic moduli (about 300 kPa) for a particle solid content of 18% w/w. Thanks to the weak and reversible nature of the interactions, these materials exhibited shear thinning properties, and could instantly recover their cohesiveness at rest. The mode of interactions between PLA particles and chitosan was shown to be in part due to electrostatic interactions, but the cross-linking of chitosan-covered particles was attributed to hydrogen bonding. These materials could be envisaged as good candidates for injectable scaffolds for tissue engineering, taking advantage of the biocompatibility and bioactivity of both components. However, some issues concerning temperature stability must be resolved before applying these colloidal assemblies to cell growth in physiological conditions.

High density type I collagen gels for tissue engineering of whole menisci

This study investigates the potential of high density type I collagen gels as an injectable scaffold for tissue engineering of whole menisci, and compares these results with previous strategies using alginate as an injectable scaffold. Bovine meniscal fibrochondrocytes were mixed with collagen and injected into micro-computed tomography-based molds to create 10 and 20mgml−1 menisci that were cultured for up to 4weeks and compared with cultured alginate menisci. Contraction, histological, confocal microscopy, biochemical and mechanical analysis were performed to determine tissue development. After 4weeks culture, collagen menisci had preserved their shape and significantly improved their biochemical and mechanical properties. Both 10 and 20mgml−1 menisci maintained their DNA content while significantly improving the glycosaminoglycan and collagen content, at values significantly higher than the alginate controls. Collagen menisci matched the alginate control in terms of the equilibrium modulus, and developed a 3- to 6-fold higher tensile modulus than alginate by 4weeks. Further fibrochondrocytes were able to reorganize the collagen gels into a more fibrous appearance similar to native menisci.

Cell Therapy for Skin Wound Using Fibroblast Encapsulated Poly(ethylene glycol)-poly(l-alanine) Thermogel

As a new application of a thermogel, a poly(ethylene glycol)-b-poly(L-alanine) (PEG-L-PA) gel encapsulating fibroblasts was investigated for wound healing. The fibroblasts were encapsulated by the temperature sensitive sol-to-gel transition of the polymer aqueous solution. Under the in vitro three-dimensional (3D) cell culture condition, the PEG-L-PA thermogel was comparable with Matrigel for cell proliferation and was significantly better than Matrigel for collagen types I and III formation. After confirming the excellent 3D microenvironment of the PEG-L-PA thermogel for fibroblasts, in vivo wound healing was investigated by injecting the cell-suspended polymer aqueous solution on incisions of rat skin, where the cell-encapsulated gel was formed in situ. Compared with the phosphate buffered saline treated system and the cell-free PEG-L-PA thermogel, the cell-encapsulated PEG-L-PA thermogel not only accelerated the wound closure but also improved epithelialization and the formation of skin appendages such as keratinocyte layer (epidermis), hair follicles, and sebaceous glands. The results demonstrate the potential of thermogels for cell therapy as an injectable tissue-engineering scaffold.

Hydroxyapatite Pickering PolyHIPEs And “Thermo”HIPEs As Injectable Scaffolds For Tissue Engineering

ABSTRACTTwo types of porous materials derived from emulsion templates are described as potential scaffolds for tissue engineering. Novel oil-in-water particle stabilised, Pickering High Internal Phase Emulsions (HIPE) stabilised with hydroxyapatite (HAp) nanoparticles were prepared and polymerised to form stable polyHIPEs. By adding a water soluble glycidyl methacrylate (GMA) derivatised dextran as monomer to the continuous aqueous phase of the HIPEs, these Pickering-HIPEs stabilised by nontoxic biocompatible HAp nanoparticles, can be used as templates to manufacture interconnected high porosity macroporous hydrogels. A second type of emulsion templated “poly”HIPE was prepared without the need for covalent crosslinking chemistry which was replaced by a thermally-induced non-covalent scaffold forming process (thermoHIPE). These scaffolds form close to body temperature and potentially offer a new approach to the formation of injectable scaffolds for tissue engineering.

Modification of an Injectable Chitosan Scaffold by Blending with NaHCO3 to Improve Cytocompatibility

The chitosan/β-glycerophosphate (C/GP) system, which allows injection at room temperature and gel-formation at body temperature, can be considered as a candidate for injectable scaffolds. To improve the cytocompatiblity of the C/GPhydrogel, we used NaHCO3 as buffer to neutralize chitosan solution and lower the GP concentration from 400 to 135 mM. The chitosan/β-glycerophosphate//NaHCO3 (C/GP/NaHCO3) system was liquid before injection and became a three-dimensional porous hydrogel at body temperature, as revealed by scanning electron microscopy (SEM). The C/GP/NaHCO3 hydrogels showed higher cytocompatibility with rat bone marrow-derived mesenchymal stem cells (rBMSCs) in vitro compared to C/GP hydrogels. Our results indicated that the C/GP/NaHCO3 system was suitable as an injectable scaffold for tissue engineering.

Controlled gelation and degradation rates of injectable hyaluronic acid-based hydrogels through a double crosslinking strategy

Various biodegradable hydrogels have been employed as injectable scaffolds for tissue engineering and drug delivery. We report a double-crosslinking strategy of biocompatible and biodegradable hydrogels derived from aminated and oxidized hyaluronic acid (HA) with genipin (GP), a compound naturally derived from the gardenia fruit. Fast gelation is attributed to the Schiff-base reaction between amino and aldehyde groups of polysaccharide derivatives, and the subsequent crosslinking with GP results in ideal biodegradability and mechanical properties. The gelation time, morphology, equilibrium swelling, compressive modulus and degradation of double-crosslinked hydrogels were examined. The double crosslinked hydrogels were examined in vivo via subcutaneous injection into a mouse model. Histological results indicated favourable biocompatility, as revealed by an absence of neutrophils and macrophages. These studies demonstrate that double-crosslinked HA hydrogels are potentially useful as injectable, biodegradable hydrogels in tissue-engineering applications.

In Situ Forming Hydrogels Based on Tyramine Conjugated 4-Arm-PPO-PEO via Enzymatic Oxidative Reaction

Over the past decades, hydrogels have been widely studied as biomaterials for various biomedical applications like implants, drugs and cell delivery carriers because of their high biocompatibility, high water contents and excellent permeability for nutrients and metabolites. Especially, in situ forming hydrogel systems have received much attention because of their easy application based on minimal invasive techniques. Chemical cross-linking systems fabricated using enzymatic reactions have various advantages, such as high biocompatibility and easy control of reaction rates under mild condition. In this study, we report enzyme-triggered injectable and biodegradable hydrogels composed of Tetronic-tyramine conjugates. The Tetronic-tyramine conjugates were synthesized by first reacting Tetronic with succinic anhydride and subsequent conjugation with tyramine using DCC/NHS as coupling reagents. The chemical structure of Tetronic-succinic anhydride-tyramine (Tet-SA-TA) copolymer was characterized by (1)H NMR and FTIR. The hydrogels were prepared from a Tet-SA-TA solution above 3 wt % in the presence of horseradish peroxidase (HRP) and H(2)O(2) under physiological conditions. Their mechanical property, gelation time, swelling ratio and degradation time were evaluated at different polymer, HRP, and H(2)O(2) concentrations. In addition, a cyto-compatibility study was performed using the MC3T3-E1 cell line. In the cytotoxicity test, it was clear that the Tet-SA-TA hydrogel had no apparent cytotoxicity except for the hydrogel formed with 0.25 wt % H(2)O(2) due to the cytotoxicity of residual H(2)O(2). In conclusion, the obtained results demonstrated that the Tet-SA-TA hydrogel has great potential for use as an injectable scaffold for tissue engineering and as a drug carrier for controlled drug delivery systems.

The preparation and cytocompatibility of injectable thermosensitive chitosan/poly(vinyl alcohol) hydrogel

In order to investigate the strength, structure and cell cytocompatibility of injectable thermosensitive chitosan (CS)/poly(vinyl alcohol) (PVA) composite hydrogel, chitosan hydrochloride solution was transferred to a neutral pH and mixed with different proportions of PVA, then the gelation time and strength of these different hydrogels were tested and spatial structures were observed under a scanning electron microscopy (SEM) after freeze-drying. The cytocompatibility of the hydrogels was evaluated through cytotoxicity test and three-dimensional culture with bone marrow mesenchymal stem cells. The results showed that the CS/PVA solution kept in liquid state at low temperature (0-4 degrees C) and turned into transparent elastomer about 15-20 min at 37 degrees C. Gelation time was prolonged, the strength increased and porous structure became dense with the PVA content increased in the mixed hydrogel. The cytotoxicity grades of these gels were from 0 to 1. Rabbit bone marrow mesenchymal stem cells could survive and proliferate in the gel within 3 weeks, and the gel had good cytocompatibility. It was concluded that thermosensitive CS/PVA composite hydrogel not only has interpenetrating network structure and better mechanical strength, but also has good cytocompatibility, and may be used as an injectable scaffold for tissue engineering.

Experimental study on self-assembly of KLD-12 peptide hydrogel and 3-D culture of MSC encapsulated within hydrogel in vitro

To synthesize KLD-12 peptide with sequence of AcN-KLDLKLDLKLDL-CNH(2) and trigger its self-assembly in vitro, to encapsulate rabbit MSCs within peptide hydrogel for 3-D culture and to evaluate the feasibility of using it as injectable scaffold for tissue engineering of IVD. KLD-12 peptide was purified and tested with high performance liquid chromatography (HPLC) and mass spectroscopy (MS). KLD-12 peptide solutions with concentrations of 5 g/L, 2.5 g/L and 1 g/L were triggered to self-assembly with 1xPBS in vitro, and the self-assembled peptide hydrogel was morphologically observed. Atomic force microscope (AFM) was employed to examine the inner structure of self-assembled peptide hydrogel. Mesenchymal stem cells (MSCs) were encapsulated within peptide hydrogel for 3-D culture for 2 weeks. Calcein-AM/PI fluorescence staining was used to detect living and dead cells. Cell viability was observed to evaluate the bioactivity of MSCs in KLD-12 peptide hydrogel. The results of HPLC and MS showed that the relative molecular mass of KLD-12 peptide was 1467.83, with a purity quotient of 95.36%. KLD-12 peptide at 5 g/L could self-assemble to produce a hydrogel, which was structurally integral and homogeneous and was able to provide sufficient cohesion to retain the shape of hydrogel. AFM demonstrated that the self-assembly of KLD-12 peptide hydrogel was successful and the assembled material was composed of a kind of nano-fiber with a diameter of 30-40 nm and a length of hundreds of nm. Calcein-AM/PI fluorescence staining revealed that MSCs in KLD-12 peptide hydrogel grew well. Cell activity detection exhibited that the A value increased over the culture time. It is concluded that KLD-12 peptide was synthesized successfully and was able to self-assemble to produce nano-fiber hydrogel in vitro. MSCs in KLD-12 peptide hydrogel grew well and proliferated with the culture time. KLD-12 peptide hydrogel can serve as an excellent injectable material of biological scaffolds in tissue engineering of IVD.

Chitosan microparticles as injectable scaffolds for tissue engineering

The use of chitosan microparticles as injectable carriers for cell transplantation represents a promising alternative to avoid the drawbacks of the implantation of other forms of three-dimensional (3D) scaffolds seeded with cells. In this study, a 3D construct is obtained in vitro by combining chitosan microparticles crosslinked with genipin and goat bone marrow stromal cells (GBMCs). Cell viability and the morphology of GBMCs were evaluated after culture for 7 and 14 days. Our results show the feasibility of chitosan microparticles as potential injectable scaffolds for tissue engineering and regenerative medicine.

Temperature‐Induced Hydrogels Through Self‐Assembly of Cholesterol‐Substituted Star PEG‐b‐PLLA Copolymers: An Injectable Scaffold for Tissue Engineering

AbstractPartially cholesterol‐substituted 8‐arm poly(ethylene glycol)‐block‐poly(L‐lactide) (8‐arm PEG‐b‐PLLA‐cholesterol) has been prepared as a novel star‐shaped, biodegradable copolymer derivative. The amphiphilic 8‐arm PEG‐b‐PLLA‐cholesterol aqueous solution (polymer concentration, above 3 wt%) exhibits instantaneous temperature‐induced gelation at 34 °C, but the virgin 8‐arm PEG‐b‐PLLA does not, irrespective of concentration. Moreover, an extracellular matrix (ECM)‐like micrometer‐scale network structure has been created with favorable porosity for three‐dimensional proliferation of cells inside the hydrogel. This network structure is mainly attributed to specific self‐assembly between cholesterol groups. The 10 and 20 wt% hydrogels are eroded gradually in phosphate buffered saline at 37 °C over the course of a month, and after that the gel becomes completely dissociated. Moreover, L929 cells encapsulated into the hydrogel are viable and proliferate three‐dimensionally inside the hydrogels. Thus, in‐vitro cell culture studies demonstrate that 8‐arm PEG‐b‐PLLA‐cholesterol is a promising candidate as a novel injectable cellular scaffold.

Synthesis, characterization, and in vitro cell culture viability of degradable poly(N‐isopropylacrylamide‐co‐5,6‐benzo‐2‐methylene‐1,3‐dioxepane)‐based polymers and crosslinked gels

Poly(N-isopropylacrylamide-co-5,6-benzo-2-methylene-1,3-dioxepane) (poly(NIPAAm-co-BMDO)) was synthesized by atom transfer radical polymerization (ATRP) and reversible addition-fragmentation chain transfer (RAFT) polymerization. Using UV-vis spectroscopy, the lower critical solution temperature (LCST) of poly(NIPAAm) and poly(NIPAAm-co-BMDO) copolymers were measured, varying with respect to the amount of incorporated BMDO. This material is degradable and possesses a LCST above room temperature and below body temperature, making it a potential candidate for use as an injectable tissue engineering scaffold to enhance fracture repair. ATRP and RAFT enabled preparation of polymers with control over molecular weight up to M(n) = 50,000 g/mol and M(w)/M(n) < 1.2. Degradation studies were performed in basic solution and in complete Dulbecco's modified Eagle medium. The cytotoxicity of the material and its degradation products were analyzed by in vitro cell culture analyses, including cytotoxicity live/dead and CyQUANT cell proliferation assays. Crosslinked scaffolds with degradable units within the polymer backbone and at the crosslinking sites were prepared using an ester-containing diacrylate crosslinker. Furthermore, incorporation of a GRGDS peptide sequence improved cell attachment to the gels. Controlled/living radical polymerization techniques allow for precise control over macromolecular structure and are poised to become powerful tools for tissue engineering scaffold synthesis.

Thermosensitive and Cell-Adhesive Pluronic Hydrogels for Human Adipose-Derived Stem Cells

Pluronic F127 has received increasing attention over many years as drug delivery systems, biomaterials, and hydrogels for tissue engineering. In this study, we synthesized temperature-sensitive and cell-adhesive triblock F127 copolymers, in which Arg-Gly-Asp (RGD) peptide ligand was grafted to Pluronic F127-4-methacryloxyethyl trimellitic anhydride (4-META) to obtain F127-META-RGD. The chemical structures of the F127-META-RGD block copolymers were confirmed by FTIR, 1H and 13C NMR, and GPC. The resultant F127-META-RGD showed very similar thermosensitive behaviors to F127 and F127-META. The critical micelle temperature (CMT) of the F127 copolymers decreased in the order of F127 &lt; F127-META &lt; F127-META-RGD, whereas the particle size followed an opposite trend. Interactions between the F127 copolymers and adipose-derived stem cells (ASC) were evaluated in terms of cell adhesion and proliferation on the hydrogel. These thermosensitive RGD-grafted Pluronic hydrogels that display the enhanced cell adhesiveness, are expected to be useful as a functional injectable scaffold for tissue engineering.