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1.
Biomacromolecules ; 22(2): 594-611, 2021 02 08.
Article in English | MEDLINE | ID: mdl-33448795

ABSTRACT

Strategies involving the inclusion of cell-instructive chemical and topographical cues to smart biomaterials in combination with a suitable physical stimulus may be beneficial to enhance nerve-regeneration rate. In this regard, we investigated the surface functionalization of poly[2-methoxy-5-(2-ethylhexyloxy)-1,4-phenylenevinylene] (MEH-PPV)-based electroconductive electrospun nanofibers coupled with externally applied electrical stimulus for accelerated neuronal growth potential. In addition, the voltage-dependent conductive mechanism of the nanofibers was studied in depth to interlink intrinsic conductive properties with electrically stimulated neuronal expressions. Surface functionalization was accomplished using 3-aminopropyltriethoxysilane (APTES) and 1,6-hexanediamine (HDA) as an alternative to costly biomolecule coating (e.g., collagen) for cell adhesion. The nanofibers were uniform, porous, electrically conductive, mechanically strong, and stable under physiological conditions. Surface amination boosted biocompatibility, 3T3 cell adhesion, and spreading, while the neuronal model rat PC12 cell line showed better differentiation on surface-functionalized mats compared to nonfunctionalized mats. When coupled with electrical stimulation (ES), these mats showed comparable or faster neurite formation and elongation than the collagen-coated mats with no-ES conditions. The findings indicate that surface amination in combination with ES may provide an improved strategy to faster nerve regeneration using MEH-PPV-based neural scaffolds.


Subject(s)
Nanofibers , Animals , Neurons , PC12 Cells , Rats , Tissue Engineering , Tissue Scaffolds
2.
Biomaterials ; 197: 119-128, 2019 03.
Article in English | MEDLINE | ID: mdl-30641263

ABSTRACT

The efficacy of cell-based therapies as an alternative to autologous bone grafts requires biomaterials to localize cells at the defect and drive osteogenic differentiation. Hydrogels are ideal cell delivery vehicles that can provide instructional cues via their composition or mechanical properties but commonly lack osteoconductive components that nucleate mineral. To address this challenge, we entrapped mesenchymal stromal cells (MSCs) in a composite hydrogel based on two naturally-derived polymers (alginate and hyaluronate) containing biomineralized polymeric microspheres. Mechanical properties of the hydrogels were dependent upon composition. The presentation of the adhesive tripeptide Arginine-Glycine-Aspartic Acid (RGD) from both polymers induced greater osteogenic differentiation of ovine MSCs in vitro compared to gels formed of RGD-alginate or RGD-alginate/hyaluronate alone. We then evaluated the capacity of this construct to stimulate bone healing when transplanting autologous, culture-expanded MSCs into a surgical induced, critical-sized ovine iliac crest bone defect. At 12 weeks post-implantation, defects treated with MSCs transplanted in composite gels exhibited significant increases in blood vessel density, osteoid formation, and bone formation compared to acellular gels or untreated defects. These findings demonstrate the capacity of osteoconductive hydrogels to promote bone formation with autologous MSCs in a large animal bone defect model and provide a promising vehicle for cell-based therapies of bone healing.


Subject(s)
Alginates/therapeutic use , Biocompatible Materials/therapeutic use , Hyaluronic Acid/therapeutic use , Hydrogels/therapeutic use , Oligopeptides/therapeutic use , Osteogenesis/drug effects , Alginates/administration & dosage , Animals , Biocompatible Materials/administration & dosage , Bone and Bones/injuries , Hyaluronic Acid/administration & dosage , Hydrogels/administration & dosage , Injections , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Oligopeptides/administration & dosage , Sheep
3.
Biomater Sci ; 6(9): 2342-2359, 2018 Aug 21.
Article in English | MEDLINE | ID: mdl-30019048

ABSTRACT

The purpose of this study was to prepare an electrically conducting poly[2-methoxy-5-(2-ethyl-hexyloxy)-1,4-phenylene vinylene] (MEH-PPV) based nanofibrous scaffold and to investigate the synergetic effect of nanofibre structure and electrical stimulation on neuronal growth for possible use in nerve repair. Nanofibres were produced by electrospinning of blended MEH-PPV with polycaprolactone (PCL) at a ratio of 20 : 80, 40 : 60, 50 : 50 and 60 : 40 (v/v). A better electrical conductivity was achieved by using core-sheath structured nanofibres of PCL (core) and MEH-PPV (sheath) produced using the coaxial electrospinning technique. The highest electrical conductivity was observed in the core-sheath nanofibres, while it increased with increasing concentration of MEH-PPV for the blended electrospun nanofibres. The biocompatibility of the electrospun nanofibres was confirmed by MTS and live-dead staining assays using 3T3 fibroblasts and a neuronal rat pheochromocytoma (PC12) cell line. Beta (III) tubulin immunochemistry showed that PC12 cells differentiated into sympathetic neurons on these porous and stiffer electrospun nanofibres coated with collagen I. Improved cell morphology and attachment on the collagen I coated electrospun meshes has been confirmed by SEM analysis. Significant enhancement in neurite formation and neurite outgrowth of PC12 cells on the conductive scaffolds under electrical potential of 500 mV cm-1 for 2 h day-1 suggests the potential use of these scaffolds for nerve repair.


Subject(s)
Nanofibers/chemistry , Polyesters/chemistry , Polymers/chemistry , Vinyl Compounds/chemistry , Adrenal Gland Neoplasms/metabolism , Animals , Cell Adhesion/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Drug Stability , Electric Conductivity , Electric Stimulation , Mice , NIH 3T3 Cells , Nanofibers/administration & dosage , PC12 Cells , Pheochromocytoma/metabolism , Polyesters/administration & dosage , Polymers/administration & dosage , Rats , Tubulin/metabolism , Vinyl Compounds/administration & dosage
4.
ACS Biomater Sci Eng ; 4(9): 3327-3346, 2018 Sep 10.
Article in English | MEDLINE | ID: mdl-33435069

ABSTRACT

In the present study, a conducting polymer, poly[2-methoxy-5-(2-ethylhexyloxy)-1,4-phenylenevinylene] (MEH-PPV) along with a biodegradable polymer poly(ε-caprolactone) (PCL) was used to prepare an electrically conductive, biocompatible, bioactive, and biodegradable nanofibrous scaffold for possible use in neural tissue engineering applications. Core-sheath electrospun nanofibers of PCL as the core and MEH-PPV as the sheath, were surface-functionalized with (3-aminopropyl) triethoxysilane (APTES) and 1,6-hexanediamine to obtain amine-functionalized surface to facilitate cell-biomaterial interactions with the aim of replacing the costly biomolecules such as collagen, fibronectin, laminin, and arginyl-glycyl-aspartic acid (RGD) peptide for surface modification. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) confirmed the formation of core-sheath morphology of the electrospun nanofibers, whereas Fourier-transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) revealed successful incorporation of amine functionality after surface functionalization. Adhesion, spreading, and proliferation of 3T3 fibroblasts were enhanced on the surface-functionalized electrospun meshes, whereas the neuronal model rat pheochromocytoma 12 (PC12) cells also adhered and differentiated into sympathetic neurons on these meshes. Under a constant electric field of 500 mV for 2 h/day for 3 consecutive days, the PC12 cells displayed remarkable improvement in the neurite formation and outgrowth on the surface-functionalized meshes that was comparable to those on the collagen-coated meshes under no electrical signal. Electrical stimulation studies further demonstrated that electrically stimulated PC12 cells cultured on collagen I coated meshes yielded more and longer neurites than those of the unstimulated cells on the same scaffolds. The enhanced neurite growth and differentiation suggest the potential use of these scaffolds for neural tissue engineering applications.

5.
Biomed Mater ; 12(3): 035001, 2017 May 10.
Article in English | MEDLINE | ID: mdl-28270638

ABSTRACT

Nanoporous adsorbents are promising materials to augment the efficacy of haemodialysis for the treatment of end stage renal disease where mortality rates remain unacceptably high despite improvements in membrane technology. Complications are linked in part to inefficient removal of protein bound and high molecular weight uraemic toxins including key marker molecules albumin bound indoxyl sulphate (IS) and p-cresyl sulphate (PCS) and large inflammatory cytokines such as IL-6. The following study describes the assessment of a nanoporous activated carbon monolith produced using a novel binder synthesis route for scale up as an in line device to augment haemodialysis through adsorption of these toxins. Small and large monoliths were synthesised using an optimised ratio of lignin binder to porous resin of 1 in 4. Small monoliths showing combined significant IS, p-CS and IL-6 adsorption were used to measure haemocompatibility in an ex vivo healthy donor blood perfusion model, assessing coagulation, platelet, granulocyte, T cells and complement activation, haemolysis, adsorption of electrolytes and plasma proteins. The small monoliths were tested in a naive rat model and showed stable blood gas values, blood pressure, blood biochemistry and the absence of coagulopathies. These monoliths were scaled up to a clinically relevant size and were able to maintain adsorption of protein bound uraemic toxins IS, PCS and high molecular weight cytokines TNF-α and IL-6 over 240 min using a flow rate of 300 ml min-1 without platelet activation. The nanoporous monoliths where haemocompatible and retained adsorptive efficacy on scale up with negligible pressure drop across the system indicating potential for use as an in-line device to improve haemodialysis efficacy by adsorption of otherwise poorly removed uraemic toxins.


Subject(s)
Acrylic Resins/chemistry , Blood Component Removal/instrumentation , Lignin/chemistry , Nanoparticles/chemistry , Renal Dialysis/instrumentation , Ultrafiltration/methods , Uremia/blood , Absorption, Physicochemical , Adsorption , Blood Component Removal/methods , Equipment Design , Equipment Failure Analysis , Humans , Materials Testing , Nanoparticles/ultrastructure , Nanopores/ultrastructure , Renal Dialysis/methods , Ultrafiltration/instrumentation , Uremia/prevention & control
6.
Sci Rep ; 6: 21154, 2016 Feb 17.
Article in English | MEDLINE | ID: mdl-26883390

ABSTRACT

The development of bulk, three-dimensional (3D), macroporous polymers with high permeability, large surface area and large volume is highly desirable for a range of applications in the biomedical, biotechnological and environmental areas. The experimental techniques currently used are limited to the production of small size and volume cryogel material. In this work we propose a novel, versatile, simple and reproducible method for the synthesis of large volume porous polymer hydrogels by cryogelation. By controlling the freezing process of the reagent/polymer solution, large-scale 3D macroporous gels with wide interconnected pores (up to 200 µm in diameter) and large accessible surface area have been synthesized. For the first time, macroporous gels (of up to 400 ml bulk volume) with controlled porous structure were manufactured, with potential for scale up to much larger gel dimensions. This method can be used for production of novel 3D multi-component macroporous composite materials with a uniform distribution of embedded particles. The proposed method provides better control of freezing conditions and thus overcomes existing drawbacks limiting production of large gel-based devices and matrices. The proposed method could serve as a new design concept for functional 3D macroporous gels and composites preparation for biomedical, biotechnological and environmental applications.

7.
Biomaterials ; 50: 140-53, 2015 May.
Article in English | MEDLINE | ID: mdl-25736504

ABSTRACT

Polymeric cryogels are efficient carriers for the immobilization of biomolecules because of their unique macroporous structure, permeability, mechanical stability and different surface chemical functionalities. The aim of the study was to demonstrate the potential use of macroporous monolithic cryogels for biotoxin removal using anthrax toxin protective antigen (PA), the central cell-binding component of the anthrax exotoxins, and covalent immobilization of monoclonal antibodies. The affinity ligand (protein A) was chemically coupled to the reactive hydroxyl and epoxy-derivatized monolithic cryogels and the binding efficiencies of protein A, monoclonal antibodies to the cryogel column were determined. Our results show differences in the binding capacity of protein A as well as monoclonal antibodies to the cryogel adsorbents caused by ligand concentrations, physical properties and morphology of surface matrices. The cytotoxicity potential of the cryogels was determined by an in vitro viability assay using V79 lung fibroblast as a model cell and the results reveal that the cryogels are non-cytotoxic. Finally, the adsorptive capacities of PA from phosphate buffered saline (PBS) were evaluated towards a non-glycosylated, plant-derived human monoclonal antibody (PANG) and a glycosylated human monoclonal antibody (Valortim(®)), both of which were covalently attached via protein A immobilization. Optimal binding capacities of 108 and 117 mg/g of antibody to the adsorbent were observed for PANG attached poly(acrylamide-allyl glycidyl ether) [poly(AAm-AGE)] and Valortim(®) attached poly(AAm-AGE) cryogels, respectively, This indicated that glycosylation status of Valortim(®) antibody could significantly increase (8%) its binding capacity relative to the PANG antibody on poly(AAm-AGE)-protien-A column (p < 0.05). The amounts of PA which remained in the solution after passing PA spiked PBS through PANG or Valortim bound poly(AAm-AGE) cryogel were significantly (p < 0.05) decreased relative to the amount of PA remained in the solution after passing through unmodified as well as protein A modified poly(AAm-AGE) cryogel columns, indicates efficient PA removal from spiked PBS over 60 min of circulation. The high adsorption capacity towards anthrax toxin PA of the cryogel adsorbents indicated potential application of these materials for treatment of Bacillus anthracis infection.


Subject(s)
Antibodies, Monoclonal/metabolism , Antigens, Bacterial/isolation & purification , Bacterial Toxins/isolation & purification , Cryogels , Immobilized Proteins/metabolism , Staphylococcal Protein A/metabolism , Acrylic Resins/chemistry , Adsorption , Animals , Buffers , Cell Death , Cell Line , Cell Survival , Cricetinae , Cryogels/metabolism , Humans , Mechanical Phenomena , Microscopy, Confocal , Porosity , Solutions , Spectroscopy, Fourier Transform Infrared , Temperature
8.
Acta Biomater ; 10(5): 1955-64, 2014 May.
Article in English | MEDLINE | ID: mdl-24468583

ABSTRACT

Recombinant proteins and cytokines are under broad preclinical and clinical investigation to promote angiogenesis, but their success is limited by ineffective delivery, lack of long-term stability and excessive cost. Mesenchymal stem/stromal cells (MSC) secrete bioactive trophic factors, and thus, may provide an effective alternative to address these challenges. Glycine-Histidine-Lysine (GHK) is a peptide fragment of osteonectin, a matricellular protein with reported proangiogenic potential. We examined the capacity of GHK to up-regulate secretion of proangiogenic factors from human MSC in culture and when covalently coupled to alginate hydrogels. GHK had no apparent cytotoxic effects on MSC in culture over a wide range of concentrations. We detected a dose-dependent increase in vascular endothelial growth factor (VEGF) concentration in media conditioned by GHK-treated MSC, which increased endothelial cell proliferation, migration and tubule formation. We covalently coupled GHK to alginate using carbodiimide chemistry, and human MSC were entrapped in alginate hydrogels to assess VEGF secretion. Similar to monolayer culture, MSC responded to GHK-modified gels by secreting increased concentrations of VEGF and basic fibroblast growth factor compared to unmodified gels. The pre-treatment of MSC with antibodies to α6 and ß1 integrins prior to entrapment in GHK-modified gels abrogated VEGF secretion, suggesting that the proangiogenic response of MSC was integrin-mediated. These data demonstrate that the proangiogenic potential of MSC can be significantly increased by the presentation of GHK with a biodegradable carrier, therefore increasing their clinical potential when used for tissue repair.


Subject(s)
Alginates/pharmacology , Hydrogels/pharmacology , Mesenchymal Stem Cells/metabolism , Oligopeptides/pharmacology , Vascular Endothelial Growth Factor A/metabolism , Cell Adhesion/drug effects , Cell Death/drug effects , Cell Movement/drug effects , Cell Survival/drug effects , Colony-Forming Units Assay , Culture Media, Conditioned/pharmacology , Endothelial Cells/cytology , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Glucuronic Acid/pharmacology , Hexuronic Acids/pharmacology , Humans , Integrins/metabolism , Mesenchymal Stem Cells/drug effects , Mitogens/pharmacology
9.
Biomaterials ; 35(11): 3558-70, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24462353

ABSTRACT

Our previous reports of interpenetrating networks (IPNs) have demonstrated drastic improvements in mechanical performance relative to individual constituent networks while maintaining viability of encapsulated cells. The current study investigated whether covalent linkage of RGD to the poly(ethylene glycol) diacrylate (PEGDA) network could improve upon cell viability and performance of agarose-PEGDA IPNs compared to unmodified IPNs (control) and to IPNs with different concentrations of physically entrapped aggrecan, providing a point of comparison to previous work. The inclusion of RGD or aggrecan generally did not adversely affect mechanical performance, and significantly improved chondrocyte viability and performance. Although both 4 and 100 µg/mL of aggrecan improved cell viability, only 100 µg/mL aggrecan was clearly beneficial to improving biosynthesis, whereas 100 µg/mL of RGD was beneficial to both chondrocyte viability and biosynthesis. Interestingly, clustering of cells within the IPNs with RGD and the higher aggrecan concentration were observed, likely indicating cell migration and/or preferred regional proliferation. This clustering resulted in a clearly visible enhancement of matrix production compared to the other IPNs. With this cell migration, we also observed significant cell proliferation and matrix synthesis beyond the periphery of the IPN, which could have important implications in facilitating integration with surrounding cartilage in vivo. With RGD and aggrecan (at its higher concentration) providing substantial and comparable improvements in cell performance, RGD would be the recommended bioactive signal for this particular IPN formulation and cell source given the significant cost savings and potentially more straightforward regulatory pathway in commercialization.


Subject(s)
Aggrecans/pharmacology , Biocompatible Materials/pharmacology , Hydrogels/pharmacology , Immobilized Proteins/pharmacology , Oligopeptides/pharmacology , Polyethylene Glycols/pharmacology , Sepharose/pharmacology , Animals , Cattle , Cell Proliferation/drug effects , Cell Shape/drug effects , Cell Survival/drug effects , Chondrocytes/cytology , Chondrocytes/drug effects , Chondrocytes/ultrastructure , DNA/metabolism , Extracellular Matrix/drug effects , Extracellular Matrix/metabolism , Glycosaminoglycans/metabolism , Hydrogels/chemical synthesis , Hydrogels/chemistry , Hydroxyproline/metabolism , Magnetic Resonance Spectroscopy , Male , Mechanical Phenomena/drug effects , Oligopeptides/chemical synthesis , Oligopeptides/chemistry , Polyethylene Glycols/chemical synthesis , Polyethylene Glycols/chemistry , Spectroscopy, Fourier Transform Infrared , Sus scrofa
10.
Tissue Eng Part B Rev ; 20(4): 277-93, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24004443

ABSTRACT

Polymeric nanofibers have potential as tissue engineering scaffolds, as they mimic the nanoscale properties and structural characteristics of native extracellular matrix (ECM). Nanofibers composed of natural and synthetic polymers, biomimetic composites, ceramics, and metals have been fabricated by electrospinning for various tissue engineering applications. The inherent advantages of electrospinning nanofibers include the generation of substrata with high surface area-to-volume ratios, the capacity to precisely control material and mechanical properties, and a tendency for cellular in-growth due to interconnectivity within the pores. Furthermore, the electrospinning process affords the opportunity to engineer scaffolds with micro- to nanoscale topography similar to the natural ECM. This review describes the fundamental aspects of the electrospinning process when applied to spinnable natural and synthetic polymers; particularly, those parameters that influence fiber geometry, morphology, mesh porosity, and scaffold mechanical properties. We describe cellular responses to fiber morphology achieved by varying processing parameters and highlight successful applications of electrospun nanofibrous scaffolds when used to tissue engineer bone, skin, and vascular grafts.


Subject(s)
Biomimetic Materials/chemistry , Nanofibers/chemistry , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Animals , Humans
11.
Tissue Eng Part A ; 19(11-12): 1349-59, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23379843

ABSTRACT

Interpenetrating network (IPN) hydrogels were recently introduced to the cartilage tissue engineering literature, with the approach of encapsulating cells in thermally gelling agarose that is then soaked in a poly(ethylene glycol) diacrylate (PEGDA) solution, which is then photopolymerized. These IPNs possess significantly enhanced mechanical performance desirable for cartilage regeneration, potentially allowing patients to return to weight-bearing activities quickly after surgical implantation. In an effort to improve cell viability and performance, inspiration was drawn from previous studies that have elicited positive chondrogenic responses to aggrecan, the proteoglycan largely responsible for the compressive stiffness of cartilage. Aggrecan was incorporated into the IPNs in conservative concentrations (40 µg/mL), and its effect was contrasted with the incorporation of chondroitin sulfate (CS), the primary glycosaminoglycan associated with aggrecan. Aggrecan was incorporated by physical entrapment within agarose and methacrylated CS was incorporated by copolymerization with PEGDA. The IPNs incorporating aggrecan or CS exhibited over 50% viability with encapsulated chondrocytes after 6 weeks. Both aggrecan and CS improved cell viability by 15.6% and 20%, respectively, relative to pure IPNs at 6 weeks culture time. In summary, we have introduced the novel approach of including a raw material from cartilage, namely aggrecan, to serve as a bioactive signal to cells encapsulated in IPN hydrogels for cartilage tissue engineering, which led to improved performance of encapsulated chondrocytes.


Subject(s)
Aggrecans/pharmacology , Cartilage/cytology , Cartilage/physiology , Hydrogels/pharmacology , Tissue Engineering/methods , Animals , Cartilage/drug effects , Cattle , Cell Survival/drug effects , Chondrocytes/cytology , Chondrocytes/drug effects , Chondrocytes/metabolism , Compressive Strength/drug effects , DNA/metabolism , Glycosaminoglycans/metabolism , Hydroxyproline/metabolism , Male , Materials Testing , Microscopy, Confocal , Sus scrofa
12.
J Mater Sci Mater Med ; 23(1): 157-70, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22116661

ABSTRACT

We recently introduced agarose-poly(ethylene glycol) diacrylate (PEGDA) interpenetrating network (IPN) hydrogels to cartilage tissue engineering that were able to encapsulate viable cells and provide a significant improvement in mechanical performance relative to its two constituent hydrogels. The goal of the current study was to develop a novel synthesis protocol to incorporate methacrylated chondroitin sulfate (MCS) into the IPN design hypothesized to improve cell viability and biosynthesis. The IPN was formed by encapsulating porcine chondrocytes in agarose, soaking the construct in a solution of 1:10 MCS:PEGDA, which was then photopolymerized to form a copolymer network as the second network. The IPN with incorporated CS (CS-IPN) (~0.5 wt%) resulted in a 4- to 5-fold increase in the compressive elastic modulus relative to either the PEGDA or agarose gels. After 6 weeks of in vitro culture, more than 50% of the encapsulated chondrocytes remained viable within the CS-modified IPN, in contrast to 35% viability observed in the unmodified. At week 6, the CS-IPN had significantly higher normalized GAG contents (347 ± 34 µg/µg) than unmodified IPNs (158 ± 27 µg/µg, P < 0.05). Overall, the approach of incorporating biopolymers such as CS from native tissue may provide favorable micro-environment and beneficial signals to cells to enhance their overall performance in IPNs.


Subject(s)
Chondrocytes/drug effects , Chondroitin Sulfates/pharmacology , Hydrogels , Polyethylene Glycols , Sepharose , Animals , Chondrocytes/cytology , Male , Swine
13.
Tissue Eng Part C Methods ; 16(6): 1533-42, 2010 Dec.
Article in English | MEDLINE | ID: mdl-20626274

ABSTRACT

A new method for encapsulating cells in interpenetrating network (IPN) hydrogels of superior mechanical integrity was developed. In this study, two biocompatible materials-agarose and poly(ethylene glycol) (PEG) diacrylate-were combined to create a new IPN hydrogel with greatly enhanced mechanical performance. Unconfined compression of hydrogel samples revealed that the IPN displayed a fourfold increase in shear modulus relative to a pure PEG-diacrylate network (39.9 vs. 9.9 kPa) and a 4.9-fold increase relative to a pure agarose network (8.2 kPa). PEG and IPN compressive failure strains were found to be 71% ± 17% and 74% ± 17%, respectively, while pure agarose gels failed around 15% strain. Similar mechanical property improvements were seen when IPNs-encapsulated chondrocytes, and LIVE/DEAD cell viability assays demonstrated that cells survived the IPN encapsulation process. The majority of IPN-encapsulated chondrocytes remained viable 1 week postencapsulation, and chondrocytes exhibited glycosaminoglycan synthesis comparable to that of agarose-encapsulated chondrocytes at 3 weeks postencapsulation. The introduction of a new method for encapsulating cells in a hydrogel with enhanced mechanical performance is a promising step toward cartilage defect repair. This method can be applied to fabricate a broad variety of cell-based IPNs by varying monomers and polymers in type and concentration and by adding functional groups such as degradable sequences or cell adhesion groups. Further, this technology may be applicable in other cell-based applications where mechanical integrity of cell-containing hydrogels is of great importance.


Subject(s)
Cartilage/growth & development , Hydrogel, Polyethylene Glycol Dimethacrylate/chemical synthesis , Polyethylene Glycols/chemistry , Polymers/chemical synthesis , Sepharose/chemistry , Tissue Engineering/methods , Animals , Biocompatible Materials/chemical synthesis , Biocompatible Materials/chemistry , Cartilage/cytology , Cartilage/physiology , Cell Survival/drug effects , Cells, Cultured , Compressive Strength , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Male , Materials Testing , Polyethylene Glycols/pharmacology , Polymers/chemistry , Polymers/pharmacology , Sepharose/pharmacology , Swine , Tissue Scaffolds/chemistry
14.
Bioresour Technol ; 99(9): 3623-9, 2008 Jun.
Article in English | MEDLINE | ID: mdl-17766105

ABSTRACT

Macroporous polymer particles containing surface epoxy groups were synthesized for immobilization of Candida rugosa lipase (CRL). The effect of incorporation of two different sets of monomers [allyl glycidyl ether (AGE) and glycidyl methacrylate (GMA)] and the effect of crosslinking density on immobilization of lipase were studied. AGE-co-EGDM polymers gave higher binding and expression of lipase than GMA-co-EGDM polymers. Optimization of immobilization parameters was done with respect to immobilization time and enzyme loading. Amongst AGE-co-EGDM polymer series, AGE-150 polymer found to give maximum lipase activity yield and therefore evaluated for temperature, pH and storage stability. Under optimum conditions, AGE-150 polymer gave 78.40% of activity yield. Immobilized lipase on AGE-150 showed a broader pH, higher temperature and excellent storage stability.


Subject(s)
Candida/enzymology , Enzymes, Immobilized/metabolism , Epoxy Compounds/metabolism , Lipase/metabolism , Methacrylates/metabolism , Polymers/metabolism , Candida/drug effects , Cross-Linking Reagents/pharmacology , Enzyme Stability/drug effects , Hydrogen-Ion Concentration , Porosity/drug effects , Spectroscopy, Fourier Transform Infrared , Temperature , Time Factors
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