Biomedical subjects

Kristi S Anseth

Publications and source records attributed to Kristi S Anseth.

At least 55 records · Page 3Linked to original sources

PubMed2004

Crosslinking density influences chondrocyte metabolism in dynamically loaded photocrosslinked poly(ethylene glycol) hydrogels.

In approaches to tissue engineer articular cartilage, an important consideration for in situ forming cell carriers is the impact of mechanical loading on the cell composite structure and function. Photopolymerized hydrogel scaffolds based on poly(ethylene glycol) (PEG) may be synthesized with a range of crosslinking densities and corresponding macroscopic properties. This study tests the hypothesis that changes in the hydrogel crosslinking density influences the metabolic response of encapsulated chondrocytes to an applied load. PEG hydrogels were formulated with two crosslinking densities that resulted in gel compressive moduli ranging from 60 to 670 kPa. When chondrocytes were encapsulated in these PEG gels, an increase in crosslinking density resulted in an inhibition in cell proliferation and proteoglycan synthesis. Moreover, when the gels were dynamically loaded for 48 h in unconfined compression with compressive strains oscillating from 0 to 15% at a frequency of 1 Hz, cell proliferation and proteoglycan synthesis were affected in a crosslinking-density-dependent manner. Cell proliferation was inhibited in both crosslinked gels, but was greater in the highly crosslinked gel. In contrast, dynamic loading did not influence proteoglycan synthesis in the loosely crosslinked gel, but a marked decrease in proteoglycan production was observed in the highly crosslinked gel. In summary, changes in PEG hydrogel properties greatly affect how chondrocytes respond to an applied dynamic load.

Animals↗

PubMed2003

Manipulations in hydrogel chemistry control photoencapsulated chondrocyte behavior and their extracellular matrix production.

In engineering a cell-carrier to support cartilage growth, hydrogels provide a unique, largely aqueous environment for 3-dimensional chondrocyte culture that facilitates nutrient transport yet provides an elastic framework dictating tissue shape and supporting external loads. Although the gel environment is often >90% water, we demonstrate that slight variations in hydrogel chemistry control gel degradation, evolving macroscopic properties, and ultimately the secretion and distribution of extracellular matrix molecules. Specifically, biodegradable poly(ethylene glycol)-co-poly(lactic acid) hydrogels were fabricated via photopolymerization. When chondrocytes were photoencapsulated in these gels, changes in the poly(ethylene glycol)-co-poly(lactic acid) repeat unit ratio from 19 to 7 increased total collagen synthesis 2.5-fold after 6 weeks in vitro. Furthermore, the ratio of collagen to glycosaminoglycans varied from glycosaminoglycan-rich, 0.33 +/- 0.13, to collagen-rich, 4.58 +/- 1.21, depending on gel chemistry and in vitro versus in vivo culture environment. By tuning scaffold chemistry, and subsequently, gel structure and degradation behavior, we can better guide tissue evolution and development.

Animals↗

PubMed2003

Histocompatibility of photocrosslinked polyanhydrides: a novel in situ forming orthopaedic biomaterial.

Cell-polymer interactions in subcutaneous and bony tissue were examined for a novel class of in situ forming and surface eroding polyanhydride networks. Specifically, photopolymerized disks of several polyanhydride compositions were implanted subcutaneously in rats, and the tissue was analyzed for an inflammatory response. The compositions elicited varied histological responses, ranging from highly active cell layers to moderate fibrous capsules, depending on the degrading polymer composition. Furthermore, one composition was photopolymerized in a model orthopaedic defect in the proximal tibia. The feasibility of photopolymerizing the methacrylated monomers in situ and the adherence of the photocrosslinked polyanhydride to the medullary canal were examined.

Anhydrides↗

PubMed2003

Controlling the spatial distribution of ECM components in degradable PEG hydrogels for tissue engineering cartilage.

In developing a scaffold to support new tissue growth, the degradation rate and mass loss profiles of the scaffold are important design parameters. In this study, hydrogels were prepared by copolymerizing a degradable macromer, poly(lactic acid)-b-poly(ethylene glycol)-b-poly(lactic acid) endcapped with acrylate groups (PEG-LA-DA) with a nondegradable macromer, poly(ethylene glycol) dimethacrylate (PEGDM). The resulting hydrogels exhibited a range of degradation behavior and mass loss profiles. Chondrocytes were photoencapsulated in gels formulated with 50:50, 25:75, and 15:85 (mol % PEGDM: mol % PEG-LA-DA) and cultured for 6 weeks in vitro. The neocartilaginous tissue formed was examined biochemically and histologically. After 6 weeks, the DNA content in gels with 75 and 85% degradable crosslinks was nearly twice that of the DNA content in the 50% gels. The total collagen content was significantly higher in the 85% gel [2.4 +/- 0.8% wet weight (ww)] compared to the 50% gel (0.22 +/- 0.29% ww). In examining the neocartilaginous tissue with immunohistochemistry, type II collagen was localized in the pericellular region in the 50% gel; however, when increased degradation was incorporated into the gel, type II collagen was found throughout the neotissue. In summary, the important role of hydrogel degradation in controlling and influencing the deposition and distribution of extracellular matrix molecules was demonstrated and quantified.

Animals↗

PubMed2003

An initial investigation of photocurable three-dimensional lactic acid based scaffolds in a critical-sized cranial defect.

Degradable polymer networks formed by the photoinitiated polymerization of multifunctional monomers have great potential as in situ forming materials, especially for bone tissue engineering. In this study, one specific chemistry was analyzed with respect to bone formation in a critical-sized defect model with and without adsorbed osteoinductive growth factors present. The scaffolds degraded in approximately 8 months and possessed an elastic modulus similar to that of trabecular bone. A porous scaffold fabricated with approximately 80% porosity and pore diameters ranging from 45 to 150 mm was implanted in a critical-sized cranial defect in rats. When implanted alone, the scaffolds were filled primarily with fibrous tissue after 9 weeks with only mild inflammation at the defect site. When the scaffolds released osteoinductive growth factors, statistically more bone filled the scaffold. For instance, 65.8+/-9.4% (n=5) of the defects were filled with radiopaque tissue in the osteoinductive releasing scaffolds, whereas only 24.2+/-7.4% (n=5) of the defects were filled in the untreated defects 9 weeks after implantation. These results illustrate not only the benefits of delivering osteoinductive factors when developing synthetic bone grafts, but the potential of these materials for supporting the infiltration and development of bone in large defects.

Animals↗

PubMed2003

Photoinitiated crosslinked degradable copolymer networks for tissue engineering applications.

Diethylene glycol was used to initiate the ring opening polymerization of D,L-lactide and epsilon -caprolactone, as well as combinations of the two monomers. Esterification of the oligomer end groups with methacryloyl chloride led to divinyl terminated macromers that were reacted via photoinitiated polymerizations to produce crosslinked networks. The lactic and/or caproic acid repeat units can be hydrolyzed under physiological conditions, leading to degradable networks that may be useful for tissue engineering applications. Specifically, methacryloyl terminated poly(lactic acid-co-caproic acid) diethylene glycol based oligomers were prepared and characterized by 1H NMR. The number of ester linkages was kept constant while the ratio of lactic:caproic acid segments was varied. These macromers were then photopolymerized at 450 nm using a visible light initiating system to produce crosslinked degradable networks. The kinetics of the polymerizations were followed by DSC, and the dynamic mechanical behavior was monitored as a function of temperature to obtain the T(g) for each network composition. 1mm thick disks were subjected to hydrolytic degradation in an aqueous phosphate buffer solution at a pH=7.4 and 37 degrees C. The changes in the compressive modulus, as well as the % mass loss as a function of time, were recorded. Cellular compatibility was determined by seeding primary rat calverial osteoblast cells onto the disks and characterizing the cell morphology using scanning electron microscopy.

Animals↗

PubMed2003

Gene delivery in tissue engineering: a photopolymer platform to coencapsulate cells and plasmid DNA.

PURPOSE: Toward the ultimate goal of developing an engineered tissue capable of mimicking complex natural healing processes, we have designed a photopolymer platform that enables simultaneous encapsulation of cells and plasmid DNA in degradable hydrogels. Photopolymerization enables spatial and temporal control of gel formation under physiological conditions, but the presence of photoinitiator radicals poses challenges for DNA photoencapsulation. METHODS: The effects of photoinitiating conditions (ultraviolet light and photoinitiator radicals) on plasmid DNA were studied. Protection methods were identified. Plasmid DNA was photoencapsulated in photocrosslinked hydrogels, and the quantity and quality of the released DNA were assessed. Plasmid DNA was simultaneously entrapped (coencapsulated) with cells in hydrogels to assess in situ transfection. RESULTS: Experiments showed that in the absence of other species, plasmid DNA was sensitive to photoinitiator radicals, but the addition of transfection agents and/or antioxidants greatly reduced DNA damage by radicals. Encapsulated plasmid DNA was released from degradable, photocrosslinked hydrogels in active forms (supercoiled and relaxed plasmids) with an overall -60% recovery. Released DNA was capable of transfecting both plated and encapsulated cells. Encapsulated cells expressed the encoded gene of the coencapsulated plasmid as the polymer degraded. CONCLUSIONS: This photopolymerization platform allows for the creation of engineered tissues with enhanced control of cell behavior through the spatially and temporally controlled release of plasmid DNA.

DNA↗

PubMed2003

Copolymerization of photocrosslinkable anhydride monomers for use as a biodegradable bone cement.

A multifunctional anhydride monomer, methacrylated sebacic anhydride (MSA), was synthesized and copolymerized with methacrylic anhydride via photoinitiated polymerization to form highly crosslinked, degradable networks. This material system was investigated as a potential degradable bone cement. Several aspects were examined, including curing characteristics, degradation rates and mechanical properties. These copolymer networks reached high double-bond conversions on clinically acceptable time scales (<5 min). Furthermore, these divinyl monomer copolymerizations exhibit features of classical crosslinking polymerizations, including autoacceleration, autodeceleration and limited double-bond conversion. Additionally, the networks degrade via a surface erosion mechanism by which the degradation rate can be controlled through varying the degree of oligomerization of the multifunctional monomer backbone, varying the copolymer precursor composition and changing the monomer backbone chemistry. Finally, the copolymer was found to have improved mechanical properties over homopolymerized MSA. Compressive strengths as high as 78 +/- 5 MPa were attained with a 70/30 wt% MSA/methacrylic anhydride copolymer, which are comparable to measured (91 +/- 7 MPa) and literature (approx. 100 MPa) values for conventional poly(methyl methacrylate) bone cement.

Biocompatible Materials↗

PubMed2002

Delivery of osteoinductive growth factors from degradable PEG hydrogels influences osteoblast differentiation and mineralization.

Degradable poly(ethylene glycol) (PEG) hydrogels with varying mass loss profiles were investigated to assess their applicability as delivery vehicles for osteoinductive growth factors in bone tissue engineering. Protein release is readily controlled by changes in both the structure (i.e., macromer concentration) and chemistry (i.e., number of degradable units) of the starting macromer. In vitro studies indicate an increase in total protein levels, alkaline phosphatase, and mineralization by osteoblasts cultured in the presence of osteoinductive growth factors compared to cells cultured with standard media. When growth factors are delivered from a 25 wt% hydrogel, a significant increase in both alkaline phosphatase and mineralization was seen after 3 weeks of culture over growth factor delivery in a bolus fashion. Additionally, gene expression levels of both osteocalcin and type I collagen were higher at early timepoints when growth factors were released from hydrogels. These results indicate that growth factors remain active after photoencapsulation, that the sustained delivery of growth factors alters various markers of osteoblastic differentiation, and that these networks could be useful as delivery vehicles for growth factors in bone tissue engineering. Finally, ectopic bone formation was present in subcutaneous rat tissue after implantation of hydrogel networks loaded with osteoinductive growth factors.

Animals↗

PubMed2002

In situ forming degradable networks and their application in tissue engineering and drug delivery.

Multifunctional macromers based on poly(ethylene glycol) and poly(vinyl alcohol) were photopolymerized to form degradable hydrogel networks. The degradation behavior of the highly swollen gels was characterized by monitoring changes in their mass loss, degree of swelling, and compressive modulus. Experimental results show that the modulus decreases exponentially with time, while the volumetric swelling ratio increases exponentially. A degradation mechanism assuming pseudo first-order hydrolysis kinetics and accounting for the structure of the crosslinked networks successfully predicted the experimentally observed trends in these properties with degradation. Once verified, the proposed degradation mechanism was extended to correlate network degradation kinetics, and subsequent changes in network structure, with release behavior of bioactive molecules from these dynamic systems. A theoretical model utilizing a statistical approach to predict the cleavage of crosslinks within the network was used to predict the complex erosion profiles produced by these hydrogels. Finally, the application of these macromers as in situ forming hydrogel constructs for cartilage tissue engineering is demonstrated.

Animals↗

PubMed2002

An investigation of the cytotoxicity and histocompatibility of in situ forming lactic acid based orthopedic biomaterials.

The cytotoxicity and biocompatibility of polymer networks prefabricated from multifunctional lactic acid based oligomers that are being developed for orthopedic applications were assessed through in vitro cytotoxicity analysis and subcutaneous implantation. After 7 and 14 days, no significant difference was observed in the relative viability or alkaline phosphatase activity of primary rat calvarial osteoblasts cultured in the presence or absence of degrading polymer networks, indicating that the degradation products had no detrimental effect on the function or activity of the cultured cells. The tissue response to preformed lactic acid networks implanted in rats consisted of a mild inflammatory response with an increase in fibrous capsule thickness and inflammation correlating with faster degrading polymer compositions. This relatively neutral response is indicative of a biocompatible, degradable polymer that has potential medical applications. Finally, porous scaffolds were implanted subcutaneously in rats, and vascularized fibrous tissue infiltration was highly dependent on the scaffold porosity and architecture. This finding indicates that an in situ forming porous scaffold of this composition may support the infiltration of surrounding vascularized tissue, and thus be applicable to orthopedic treatments of large bone defects.

Alkaline Phosphatase↗

PubMed2002

Hydrogel properties influence ECM production by chondrocytes photoencapsulated in poly(ethylene glycol) hydrogels.

When using hydrogel scaffolds for cartilage tissue engineering, two gel properties are particularly important: the equilibrium water content (q, equilibrium swelling ratio) and the compressive modulus, K. In this work, chondrocytes were photoencapsulated in degrading and nondegrading poly(ethylene glycol)-based hydrogels to assess extracellular matrix (ECM) formation as a function of these gel properties. In nondegrading gels, the glycosaminoglycan (GAG) content was not significantly different in gels when q was varied from 4.2 to 9.3 after 2 and 4 weeks in vitro. However, gels with a q of 9.3 allowed GAGs to diffuse throughout the gels homogenously, but a q < or = 5.2 resulted in localization of GAGs pericellularly. Interestingly, in the moderately crosslinked gels with a K of 360 kPa, an increase in type II collagen synthesis was observed compared with gels with a higher (960 kPa) and lower (30 kPa) K after 4 weeks. With the incorporation of degradable linkages into the network, gel properties with an initially high K (350 kPa) and final high q (7.9) were obtained, which allowed for increased type II collagen synthesis coupled with a homogenous distribution of GAGs. Thus, a critical balance exists between gel swelling, mechanics, and degradation in forming a functional ECM.

Animals↗

PubMed2002

Synthesis and characterization of photocrosslinkable, degradable poly(vinyl alcohol)-based tissue engineering scaffolds.

Hydrogels have many advantages that make them prime candidates for tissue engineering applications: high water content, tissue-like elasticity, and relative biocompatibility. We aim to tissue engineer heart valves using a hydrogel scaffold based on poly(vinyl alcohol) (PVA), and the design parameters for a suitable tissue engineering scaffold are quite stringent. In this research, we develop degradable and photocrosslinkable poly(lactic acid)-g-PVA multifunctional macromers that can be reacted in solution to form degradable networks. The mass loss profiles and bulk properties of the resulting scaffolds are easily tailored by modifying the structure of the starting macromers. Specifically, altering the number of lactide repeat units per crosslinking side chain, percent substitution, molecular weight of PVA backbone, and macromer solution concentration, the rate of mass loss from these degradable networks is controlled. In addition, by increasing the network's hydrophobicity, valve interstitial cell adhesion is improved.

Animals↗

PubMed2002

Photoencapsulation of osteoblasts in injectable RGD-modified PEG hydrogels for bone tissue engineering.

Poly(ethylene glycol) (PEG) hydrogels were investigated as encapsulation matrices for osteoblasts to assess their applicability in promoting bone tissue engineering. Non-adhesive hydrogels were modified with adhesive Arg-Gly-Asp (RGD) peptide sequences to facilitate the adhesion, spreading, and, consequently, cytoskeletal organization of rat calvarial osteoblasts. When attached to hydrogel surfaces, the density and area of osteoblasts attached were dramatically different between modified and unmodified hydrogels. A concentration dependence of RGD groups was observed, with increased osteoblast attachment and spreading with higher RGD concentrations, and cytoskeleton organization was seen with only the highest peptide density. A majority of the osteoblasts survived the photoencapsulation process when gels were formed with 10% macromer, but a decrease in osteoblast viability of approximately 25% and 38% was seen after 1 day of in vitro culture when the macromer concentration was increased to 20 and 30wt%, respectively. There was no statistical difference in cell viability when peptides were added to the network. Finally, mineral deposits were seen in all hydrogels after 4 weeks of in vitro culture, but a significant increase in mineralization was observed upon introduction of adhesive peptides throughout the network.

Animals↗

PubMed2002

Application of a kinetic gelation simulation to the characterization of in situ cross-linking biomaterials.

Multifunctional monomers that are polymerized in situ to form highly cross-linked biomaterials have been an area of recent interest for medical applications. From a biomaterial application standpoint, the relationship between the reaction conditions, polymer structure, and final physical properties is important, particularly for in situ formed materials, as they must have optimum network properties immediately after Formation. However, multifunctional monomer reaction mechanisms are complicated by mobility restrictions on the reacting species, cyclization of pendant groups, unequal reactivity of functional groups, and structural heterogeneities. Also, experimental characterization of these complexities is difficult and limited by the insolubility of the resulting cross-linked structure. Thus, in this contribution, a kinetic gelation simulation was used to understand and characterize the evolution of in situ forming, three-dimensional polymer structures. Specifically, the reaction of tetrafunctional monomers (i.e. divinyl monomers) to form high strength networks with degradable cross-links was modeled. This work focuses on using a lattice-based model to characterize network properties important for biomaterial applications and compare them with an experimental system (i.e. cross-linked polyanhydrides) where appropriate. Simulated results for pendant group reactivity, kinetic chain lengths, and radical concentrations (trapped and free) are presented herein.

Biocompatible Materials↗

PubMed2002

Controlled release from crosslinked degradable networks.

This article reviews controlled release from crosslinked degradable networks. Network formulations include those derived from wholly synthetic components, natural components, and combinations thereof. This includes, but is not limited to, poly(orthoesters), poly(anhydrides), poly(ethylene glycol) (PEG) derivatives, and dextran functional macromonomers. In addition, we present a discussion of the chemistry behind novel degradable networks with potential use in the controlled release realm.

Biodegradation, Environmental↗

PubMedDate not supplied

Synthesis of immunoisolation barriers that provide localized immunosuppression for encapsulated pancreatic islets.

Pancreatic islet encapsulation into synthetic, passive material matrixes can provide protection for transplanted islets from destruction via cell-contacted mediated interactions with autoreactive immune cells for treatment of Type I diabetes mellitus. However, one of the fundamental deficiencies with current encapsulation technology is that passive material barriers cannot protect islets from exposure to cytokines and other small, diffusible cytotoxic molecules produced by activated immune cells, subsequently leading to beta-cell destruction. Preparation of material matrixes that can actively provide localized immunosuppression of autoreactive immune cells may prolong the viability, and hence function, of encapsulated islet grafts. We have demonstrated the ability to conjugate apoptosis-inducing anti-Fas monoclonal antibodies (MAbs) to the surfaces of poly(ethylene glycol)-modified hydrogels, providing a surface that actively attempts to locally down-regulate the autoimmune response by destroying autoreactive T cells against pancreatic islet cells. We have conjugated anti-Fas MAbs to a high degree to the surface of these hydrogels, with retention of anti-Fas recognition of the Fas antigen as shown by ELISA testing. Apoptosis induction of Fas-sensitive Jurkat T cells was enhanced in the presence of anti-Fas conjugated hydrogels. In addition, this apoptosis induction was specific to anti-Fas MAbs, with no apoptosis induction with control antibodies or with Fas-insensitive T cells. These experiments promote the concept that surface-conjugated hydrogel constructs can provide localized immunosuppression for encapsulated grafted tissue.

Antibodies, Monoclonal↗

PubMedDate not supplied

Tailoring the degradation of hydrogels formed from multivinyl poly(ethylene glycol) and poly(vinyl alcohol) macromers for cartilage tissue engineering.

Tuning the degradation profiles of polymer cell carriers to match cell and tissue growth is an important design parameter for (cartilage) tissue engineering. In this study, degradable hydrogels were fabricated from divinyl, tetrafunctional poly(ethylene glycol) (PEG) and multivinyl, multifunctional poly(vinyl alcohol) (PVA) macromers to form homopolymer and copolymer gels. These gels were characterized by their volumetric swelling ratio and mass loss profiles as a function of degradation time. By variation of the macromer chemistry and functionality, the degradation time changed from less than 1 day for homopolymer PVA gels to 34 days for pure PEG gels. Furthermore, the degrading medium influenced mass loss, and a marked decrease in degradation time, from 34 to 12 days, was observed with the PEG gels when a chondrocyte-specific medium containing fetal bovine serum was employed. Interestingly, when copolymer gels of PEG and PVA were formed, PVA was released throughout the degradation (as determined by gel permeation chromatography) suggesting that covalent cross-linking of the PVA in the network was facilitated by copolymerizing with the PEG macromer. To assess these novel gels for cartilage tissue engineering applications, chondrocytes were photoencapsulated in the copolymer networks and cultured in vitro for up to 6 weeks. DNA, glycosaminoglycan (GAG), and total collagen contents increased with culture time, and the resulting neocartilaginous tissue at 6 weeks was homogeneously distributed as seen histologically. Biochemical analysis revealed that the constructs were comprised of 0.66 +/- 0.04 microg of DNA/mg wet weight (ww), 1.0 +/- 0.05% GAG/ww, and 0.29 +/- 0.07% total collagen/ww at 6 weeks. Furthermore, the compressive modulus increased during culture from 7 to 97 kPa as the neocartilaginous tissue evolved and the gel degraded. In summary, fabricating hydrogels through the copolymerization of PEG and PVA macromers is an effective tool for encapsulating chondrocytes, controlling gel degradation profiles, and generating cartilaginous tissue.

Cartilage↗