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ABSTRACT: BACKGROUND: Composites of biodegradable polymers and bioactive ceramics are candidates for tissue-engineered scaffolds that closely match the properties of bone. We previously developed a porous, three-dimensional poly (D,L-lactide-co-glycolide) (PLAGA)/nanohydroxyapatite (n-HA) scaffold as a potential bone tissue engineering matrix suitable for high-aspect ratio vessel (HARV) bioreactor applications. However, the physical and cellular properties of this scaffold are unknown. The present study aims to evaluate the effect of n-HA in modulating PLAGA scaffold properties and human mesenchymal stem cell (HMSC) responses in a HARV bioreactor. QUESTIONS/PURPOSES: By comparing PLAGA/n-HA and PLAGA scaffolds, we asked whether incorporation of n-HA (1) accelerates scaffold degradation and compromises mechanical integrity; (2) promotes HMSC proliferation and differentiation; and (3) enhances HMSC mineralization when cultured in HARV bioreactors. METHODS: PLAGA/n-HA scaffolds (total number = 48) were loaded into HARV bioreactors for 6 weeks and monitored for mass, molecular weight, mechanical, and morphological changes. HMSCs were seeded on PLAGA/n-HA scaffolds (total number = 38) and cultured in HARV bioreactors for 28 days. Cell migration, proliferation, osteogenic differentiation, and mineralization were characterized at four selected time points. The same amount of PLAGA scaffolds were used as controls. RESULTS: The incorporation of n-HA did not alter the scaffold degradation pattern. PLAGA/n-HA scaffolds maintained their mechanical integrity throughout the 6 weeks in the dynamic culture environment. HMSCs seeded on PLAGA/n-HA scaffolds showed elevated proliferation, expression of osteogenic phenotypic markers, and mineral deposition as compared with cells seeded on PLAGA scaffolds. HMSCs migrated into the scaffold center with nearly uniform cell and extracellular matrix distribution in the scaffold interior. CONCLUSIONS: The combination of PLAGA/n-HA scaffolds with HMSCs in HARV bioreactors may allow for the generation of engineered bone tissue. CLINICAL RELEVANCE: In cases of large bone voids (such as bone cancer), tissue-engineered constructs may provide alternatives to traditional bone grafts by culturing patients' own MSCs with PLAGA/n-HA scaffolds in a HARV culture system.
Clinical Orthopaedics and Related Research 02/2013; · 2.53 Impact Factor
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ABSTRACT: Postoperative pain within the first days following musculoskeletal surgeries is a significant problem for which appropriate management correlates to positive clinical outcomes. While a variety of pain management modalities are currently used for postoperative pain, an optimal strategy has yet to be identified. Utilizing local anesthetics to convey analgesia through neural blockade represents a promising approach to alleviate postoperative pain. Unfortunately, local anesthetics are often associated with short half-lives, local tissue site reactions, and systemic toxicity. Drug delivery systems such as liposomes, microparticles, and nanoparticles have been previously utilized to extend analgesia, but these systems can easily diffuse from the injection site. In order to overcome this limitation a combination of drug delivery technologies were utilized. Ropivacaine base nanoparticles were fabricated and entrapped with dexamethasone using a chitosan thermogel delivery system in order to enhance neural blockade. Using a rat sciatic neural blockade model, this system was able to limit sensory function and motor function for up to 48 h. This approach utilized a low solubility drug, a drug action enhancer, nanoparticles, and a thermogel matrix together to yield a multi-faceted delivery system capable of providing moderate-term pain management.
Biomaterials 01/2013; · 7.40 Impact Factor
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ABSTRACT: Regenerative engineering approaches utilizing biomimetic synthetic scaffolds provide alternative strategies to repair and restore damaged bone. The efficacy of the scaffolds for functional bone regeneration critically depends on their ability to induce and support vascular infiltration. In the present study, three-dimensional (3D) biomimetic poly(lactide-co-glycolide) (PLAGA) sintered microsphere scaffolds were developed by sintering together PLAGA microspheres followed by nucleation of minerals in a simulated body fluid. Further, the angiogenic potential of vascular endothelial growth factor (VEGF)-incorporated mineralized PLAGA scaffolds were examined by monitoring the growth and phenotypic expression of endothelial cells on scaffolds. Scanning electron microscopy micrographs confirmed the growth of bone-like mineral layers on the surface of microspheres. The mineralized PLAGA scaffolds possessed interconnectivity and a compressive modulus of 402 ± 61 MPa and compressive strength of 14.6 ± 2.9 MPa. Mineralized scaffolds supported the attachment and growth and normal phenotypic expression of endothelial cells. Further, precipitation of apatite layer on PLAGA scaffolds resulted in an enhanced VEGF adsorption and prolonged release compared to nonmineralized PLAGA and, thus, a significant increase in endothelial cell proliferation. Together, these results demonstrated the potential of VEGF-incorporated biomimetic PLAGA sintered microsphere scaffolds for bone tissue engineering as they possess the combined effects of osteointegrativity and angiogenesis. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2012.
Journal of Biomedical Materials Research Part B Applied Biomaterials 08/2012; 100(8):2187-96. · 2.15 Impact Factor
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ABSTRACT: The aim of the present study was to evaluate the in vivo biocompatibility of injectable thermo gelling chitosan-ammonium hydrogen phosphate solution (chitosan-AHP) and its efficacy to deliver recombinant human bone morphogenetic protein-2 (rhBMP-2) in a bioactive form. The thermogel showed a typical foreign body response upon subcutaneous implantation surrounded by a fibrous capsule. Even at 4 and 8 weeks post implantation, significant neutrophil infiltration was observed within the gel. Chitosan-AHP gel retained most of the loaded rhBMP-2 after a small initial release. The bioactivity of the released protein was demonstrated in vitro by the increase in alkaline phosphatase activity of mouse pre osteoblast cells (MC3T3-E1). Histological and micro-computed tomography (μCT) evaluation showed evidence of ectopic bone formation upon 4 μg/mL rhBMP-2 loaded chitosan-AHP injection. The study demonstrated a neutrophil mediated local tissue response to chitosan-AHP gel and its ability to encapsulate and maintain the bioactivity of rhBMP-2.
Journal of Materials Science Materials in Medicine 06/2012; 23(9):2141-9. · 2.32 Impact Factor
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ABSTRACT: The need and the growing interest in polymers as biomaterials have led to the synthesis of new polymers with a variety of
physico-chemical properties. Biomedical application of such materials not only depends on their physical properties but also
on biocompatibility and biodegradability. Polyphosphazenes are a family of ‘hybrid inorganic–organic polymers’ with inorganic
elements in the backbone and organic side-groups. The polyphosphazenes constitute a family of greatly diverse performance
materials with a broad spectrum of properties. The present review focuses on the biodegradable polyphosphazenes, their biocompatibility,
and degradation behavior both in vitro and in vivo. This review also covers the use of biodegradable polyphosphazenes as controlled release devices.
Journal of Inorganic and Organometallic Polymers and Materials 04/2012; 16(4):365-385. · 1.45 Impact Factor
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Lakshmi S Nair
Translational research : the journal of laboratory and clinical medicine. 09/2011; 158(3):129-31.
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ABSTRACT: Utilization of polymers as biomaterials has greatly impacted the advancement of modern medicine. Specifically, polymeric biomaterials that are biodegradable provide the significant advantage of being able to be broken down and removed after they have served their function. Applications are wide ranging with degradable polymers being used clinically as surgical sutures and implants. In order to fit functional demand, materials with desired physical, chemical, biological, biomechanical and degradation properties must be selected. Fortunately, a wide range of natural and synthetic degradable polymers has been investigated for biomedical applications with novel materials constantly being developed to meet new challenges. This review summarizes the most recent advances in the field over the past 4 years, specifically highlighting new and interesting discoveries in tissue engineering and drug delivery applications.
Journal of Polymer Science Part B Polymer Physics 06/2011; 49(12):832-864. · 1.53 Impact Factor
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Advanced Functional Materials 05/2011; 21(14):2641 - 2651. · 10.18 Impact Factor
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ABSTRACT: The most successful metal implant materials currently have relatively smooth surfaces on the micron size scale, with most failures occurring after only 10 years. To move beyond this limiting time scale, texturing methods have been developed to modify the metal surface to enhance integration of the implant directly with surrounding bone. A flexible single-step ultrafast-laser texturing process has been developed that results in a surface texture that exhibits micron scale peaks and troughs with superimposed submicron and nano-scale features. The textured titanium samples remain completely hydrophilic with no measurable contact angle even after several weeks in normal atmosphere. An increase in mesenchymal stem cell number is observed over that on an untreated control titanium surface. Extensive formation of cellular bridges by stromal cells between pillars shows the favorable response of differentiated cells to the surface and the promotion of their attachment. Expression of the alkaline phosphatase and osteocalcin genes in human bone marrow cells were seen to increase on the textured surface. The development of this single-step method for creating micron, submicron, and nano-scale surface texture directly on metals makes a significant contribution to the goal of improving the integration and life span of joint replacement implants.
Journal of Biomedical Materials Research Part B Applied Biomaterials 03/2011; 97(2):299-305. · 2.15 Impact Factor
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ABSTRACT: Novel polyphosphazenes containing various vitamin substituents were synthesized and characterized, and their sensitivity to hydrolysis and pH behavior was investigated. Vitamins L1, E, and B6 were used because of their biocompatibility, their importance in a variety of biological functions, and their potential to increase the mechanical properties of the resulting polymers, thus making these materials promising candidates for hard tissue engineering scaffolds. Chlorine replacement reactions were carried out initially with the small molecule, hexachlorocyclotriphosphazene, as a model for high polymeric poly(dichlorophosphazene). Because of the steric hindrance generated by vitamin E as a substituent, co-substituted polymers were synthesized with either glycine ethyl ester or sodium ethoxide as the second substituent. Similarly, vitamin B6 was co-substituted with glycine ethyl ester or phenylalanine ethyl ester to favor biodegradability. To prevent cross-linking via multifunctional reagents, the hydroxyl groups in vitamin B6 were protected and subsequently deprotected under acidic conditions after side group linkage to the polymer backbone. The glass transition temperatures of the polymers ranged from −24.0 to 44.0 °C. Hydrolysis of the polymers in deionized water at 37 °C was used as an initial estimate of their hydrolytic sensitivity. Different solid polymers underwent 10−100% weight loss in 6 weeks with the generation of a broad pH range of 2.5−9. The weight loss during preliminary hydrolysis experiments was attributed to cleavage of the polymer backbone and/or the polymers becoming soluble in the aqueous media during hydrolytic reactions.
02/2011;
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ABSTRACT: Self-setting hydroxyapatite-biodegradable injectable composites are excellent candidates for applications in orthopaedics. We have previously demonstrated the feasibility of development of self-setting calcium-deficient nanocrystalline hydroxyapatite-polymer composites using different calcium phosphate precursors and biodegradable polyphosphazenes. This study aimed to evaluate these novel injectable composites as suitable materials for orthopaedic applications through evaluating their biomechanical properties, osteoblast cellular attachment and gene expression over time. Our studies demonstrated that the morphology of the composite groups (PNEA-CDHA, PNEA-CDSHA, PNEA(50)mPh(50)-CDHA, PNEA(50)mPh(50)-CDSHA, PNEA(50)PhPh(50)-CDHA, and PNEA(50)PhPh(50)-CDSHA) formed was similar and found to have micro- and nanoporous structures resembling trabecular bone. The osteoblast phenotypic marker of bone, alkaline phosphatase, was expressed by the cells on the surface of the composites throughout the study and was comparable to tissue-culture polystyrene (control). Furthermore, the cells seeded on the composites expressed the characteristic osteoblastic genes, such as type-I collagen, alkaline phosphatase, osteocalcin, osteopontin and bone sialoprotein, indicating osteoblast differentiation, maturation and mineralization. Within our injectable composite groups, significant gene expression levels were displayed (P < 0.05). These novel injectable biodegradable polyphosphazenes-calcium-deficient hydroxyapatites materials are promising candidates for orthopaedic applications.
Journal of Biomaterials Science Polymer Edition 01/2011; 22(4-6):733-52. · 1.69 Impact Factor
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The Journal of Bone and Joint Surgery 12/2010; 92 Suppl 2:170-9. · 3.27 Impact Factor
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ABSTRACT: The preparation of phosphazene tissue engineering scaffolds with bioactive side groups has been accomplished using the biological buffer, choline chloride. Mixed-substituent phosphazene cyclic trimers (as model systems) and polymers with choline chloride and glycine ethyl ester, alanine ethyl ester, valine ethyl ester, or phenylalanine ethyl ester were synthesized. Two different synthetic protocols were examined. A sodium hydride mediated route resulted in polyphosphazenes with a low choline content, while a cesium carbonate mediated process produced polyphosphazenes with higher choline content. The phosphazene structures and physical properties were studied using multinuclear NMR, differential scanning calorimetry (DSC), and gel permeation chromatography (GPC) techniques. The resultant polymers were then blended with PLGA (50:50) or PLGA (85:15) and characterized by DSC analysis and scanning electron microscopy (SEM). Polymer products obtained via the sodium hydride route produced miscible blends with both ratios of PLGA, while the cesium carbonate route yielded products with reduced blend miscibility. Heterophase hydrolysis experiments in aqueous media revealed that the polymer blends hydrolyzed to near-neutral pH media (∼5.8 to 6.8). The effect of different molecular structures on cellular adhesion showed osteoblast proliferation with an elevated osteoblast phenotype expression compared to PLGA over a 21-day culture period.
Biomaterials 11/2010; 31(33):8507-15. · 7.40 Impact Factor
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ABSTRACT: Synthetic biodegradable polymers serve as temporary substrates that accommodate cell infiltration and tissue in-growth in regenerative medicine. To allow tissue in-growth and nutrient transport, traditional three-dimensional (3D) scaffolds must be prefabricated with an interconnected porous structure. Here we demonstrated for the first time a unique polymer erosion process through which polymer matrices evolve from a solid coherent film to an assemblage of microspheres with an interconnected 3D porous structure. This polymer system was developed on the highly versatile platform of polyphosphazene-polyester blends. Co-substituting a polyphosphazene backbone with both hydrophilic glycylglycine dipeptide and hydrophobic 4-phenylphenoxy group generated a polymer with strong hydrogen bonding capacity. Rapid hydrolysis of the polyester component permitted the formation of 3D void space filled with self-assembled polyphosphazene spheres. Characterization of such self-assembled porous structures revealed macropores (10-100 μm) between spheres as well as micro- and nanopores on the sphere surface. A similar degradation pattern was confirmed in vivo using a rat subcutaneous implantation model. 12 weeks of implantation resulted in an interconnected porous structure with 82-87% porosity. Cell infiltration and collagen tissue in-growth between microspheres observed by histology confirmed the formation of an in situ 3D interconnected porous structure. It was determined that the in situ porous structure resulted from unique hydrogen bonding in the blend promoting a three-stage degradation mechanism. The robust tissue in-growth of this dynamic pore forming scaffold attests to the utility of this system as a new strategy in regenerative medicine for developing solid matrices that balance degradation with tissue formation.
Advanced Functional Materials 09/2010; 20(17):2743-2957. · 10.18 Impact Factor
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ABSTRACT: Tissue engineering often benefits from the use of composites to produce an ideal scaffold. We present the focused development of a novel structure that combines the biomimetic properties of nanofibers with the robust mechanical aspects of the sintered microsphere scaffold to produce a composite scaffold that demonstrates an ability to mimic the mechanical environment of trabecular bone while also promoting the phenotype progression of osteoblast progenitor cells. These composite nanofiber/microsphere scaffolds exhibited a mechanical modulus and compressive strength similar to trabecular bone and exhibited degradation resulting in a mass loss of 30% after 24 weeks. The nanofiber portion of these scaffolds was sufficiently porous to allow cell migration throughout the fibrous portion of the scaffold and promoted phenotype progression through focal adhesion kinase-mediated activation of the transcription factor Runx2, control scaffolds not containing nanofibers did not demonstrate extensive cell migration or phenotype progression. Ultimately, the focal adhesion kinase activity on the composite nanofiber/microsphere scaffolds demonstrated causality over the production of the mature osteoblast marker, osteocalcin, and the development of a calcified matrix.
Journal of Biomedical Materials Research Part A 09/2010; 95(4):1150-8. · 2.63 Impact Factor
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07/2010: pages 179 - 203; , ISBN: 9780470891315
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ABSTRACT: Advances in bone repair have focused on the minimally-invasive delivery of tissue-engineered bone (TEB). A promising injectable biopolymer of chitosan and inorganic phosphates was seeded with mesenchymal stem cells (MSCs) and a bone growth factor (BMP-2), and evaluated in a rat calvarial critical size defect (CSD). Green fluorescent protein (GFP)-labeled MSCs are used to evaluate patterns of cell viability and proliferation.
Prospective, controlled trial in an animal model.
In 30 male rats, 8-mm calvarial CSDs were created, and divided into five groups of six animals each. In the experimental groups, the defects were injected with either chitosan gel, gel loaded with MSCs (0.3 x 10(6) cells/defect), gel loaded with BMP-2 (2 microg/defect), or gel loaded with both MSC and BMP-2. In the control group, the defect was left untreated. At 4 weeks, in vivo microcomputed tomography (micro-CT) analysis was performed. At 8 weeks, calvarial specimens were examined by micro-CT, histology, and immunohistochemistry.
New areas of bone growth were seen in the defects of all treated animals. Micro-CT analysis revealed a significant (P < .001) time-dependent increase in the regeneration of bone volume and bone area in defects treated with gel/MSC/BMP-2 as compared to all other groups. Histological analysis confirmed this difference. GFP-labeled TEB was detected within the areas of new bone, indicating cell viability and contribution to new bone growth by the injected MSC.
This study demonstrates that an injectable form of TEB using a chitosan gel, MSC, and BMP-2 can enhance bone formation in a rat calvarial CSD.
The Laryngoscope 05/2010; 120(5):895-901. · 1.75 Impact Factor
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Meng Deng, Lakshmi S Nair,
Syam P Nukavarapu,
Tao Jiang,
William A Kanner,
Xudong Li,
Sangamesh G Kumbar,
Arlin L Weikel,
Nicholas R Krogman,
Harry R Allcock,
Cato T Laurencin
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ABSTRACT: Polyphosphazene-polyester blends are attractive materials for bone tissue engineering applications due to their controllable degradation pattern with non-toxic and neutral pH degradation products. In our ongoing quest for an ideal completely miscible polyphosphazene-polyester blend system, we report synthesis and characterization of a mixed-substituent biodegradable polyphosphazene poly[(glycine ethyl glycinato)(1)(phenyl phenoxy)(1)phosphazene] (PNGEG/PhPh) and its blends with a polyester. Two dipeptide-based blends namely 25:75 (Matrix1) and 50:50 (Matrix2) were produced at two different weight ratios of PNGEG/PhPh to poly(lactic acid-glycolic acid) (PLAGA). Blend miscibility was confirmed by differential scanning calorimetry, Fourier transform infrared spectroscopy, and scanning electron microscopy. Both blends resulted in higher tensile modulus and strength than the polyester. The blends showed a degradation rate in the order of Matrix2<Matrix1<PLAGA in phosphate buffered saline at 37 degrees C over 12 weeks. Significantly higher pH values of degradation media were observed for blends compared to PLAGA confirming the neutralization of PLAGA acidic degradation by polyphosphazene hydrolysis products. The blend components PLAGA and polyphosphazene exhibited a similar degradation pattern as characterized by the molecular weight loss. Furthermore, blends demonstrated significantly higher osteoblast growth rates compared to PLAGA while maintaining osteoblast phenotype over a 21-day culture. Both blends demonstrated improved biocompatibility in a rat subcutaneous implantation model compared to PLAGA over 12 weeks.
Biomaterials 03/2010; 31(18):4898-908. · 7.40 Impact Factor
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ABSTRACT: The versatility of polymers for tissue regeneration lies in the feasibility to modulate the physical and biological properties by varying the side groups grafted to the polymers. Biodegradable polyphosphazenes are high-molecular-weight polymers with alternating nitrogen and phosphorus atoms in the backbone. This study is the first of its kind to systematically investigate the effect of side group structure on the compressive strength of novel biodegradable polyphosphazene based polymers as potential materials for tissue regeneration. The alanine polyphosphazene based polymers, poly(bis(ethyl alanato) phosphazene) (PNEA), poly((50% ethyl alanato) (50% methyl phenoxy) phosphazene) (PNEA(50)mPh(50)), poly((50% ethyl alanato) (50% phenyl phenoxy) phosphazene) (PNEA(50)PhPh(50)) were investigated to demonstrate their mechanical properties and osteocompatibility. Results of mechanical testing studies demonstrated that the nature and the ratio of the pendent groups attached to the polymer backbone play a significant role in determining the mechanical properties of the resulting polymer. The compressive strength of PNEA(50)PhPh(50) was significantly higher than poly(lactide-co-glycolide) (85:15 PLAGA) (p<0.05). Additional studies evaluated the cellular response and gene expression of primary rat osteoblast cells on PNEA, PNEA(50)mPh(50) and PNEA(50)PhPh(50) films as candidates for bone tissue engineering applications. Results of the in vitro osteocompatibility evaluation demonstrated that cells adhere, proliferate, and maintain their phenotype when seeded directly on the surface of PNEA, PNEA(50)mPh(50), and PNEA(50)PhPh(50). Moreover, cells on the surface of the polymers expressed type I collagen, alkaline phosphatase, osteocalcin, osteopontin, and bone sialoprotein, which are characteristic genes for osteoblast maturation, differentiation, and mineralization.
Acta biomaterialia 12/2009; 6(6):1931-7. · 3.98 Impact Factor
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ABSTRACT: Scaffolds exhibiting biological recognition and specificity play an important role in tissue engineering and regenerative medicine. The bioactivity of scaffolds in turn influences, directs, or manipulates cellular responses. In this study, chitosan/poly(lactic acid-co-glycolic acid) (chitosan/PLAGA) sintered microsphere scaffolds were functionalized via heparin immobilization. Heparin was successfully immobilized on chitosan/PLAGA scaffolds with controllable loading efficiency. Mechanical testing showed that heparinization of chitosan/PLAGA scaffolds did not significantly alter the mechanical properties and porous structures. In addition, the heparinized chitosan/PLAGA scaffolds possessed a compressive modulus of 403.98 +/- 19.53 MPa and a compressive strength of 9.83 +/- 0.94 MPa, which are in the range of human trabecular bone. Furthermore, the heparinized chitosan/PLAGA scaffolds had an interconnected porous structure with a total pore volume of 30.93 +/- 0.90% and a median pore size of 172.33 +/- 5.89 mum. The effect of immobilized heparin on osteoblast-like MC3T3-E1 cell growth was investigated. MC3T3-E1 cells proliferated three dimensionally throughout the porous structure of the scaffolds. Heparinized chitosan/PLAGA scaffolds with low heparin loading (1.7 microg/scaffold) were shown to be capable of stimulating MC3T3-E1 cell proliferation by MTS assay and cell differentiation as evidenced by elevated osteocalcin expression when compared with nonheparinized chitosan/PLAGA scaffold and chitosan/PLAGA scaffold with high heparin loading (14.1 microg/scaffold). This study demonstrated the potential of functionalizing chitosan/PLAGA scaffolds via heparinization with improved cell functions for bone tissue engineering applications.
Journal of Biomedical Materials Research Part A 09/2009; 93(3):1193-208. · 2.63 Impact Factor
