Deposition of PLA/CDHA composite coating via electrospraying

Composite coatings composed of carbonated calcium deficient hydroxyapatite (CDHA) and polylactic acid (PLA) were deposited on a PLA substrate surface via electrospraying. The operation parameters, structural properties, bioactivity, cell adhesion, and growth capability of as-fabricated PLA/CDHA coatings were investigated. The composite coating showed good biocompatibility and bioactivity. The deposited coating was also applied as a carrier to assist alendronate sodium (AS) local release. AS, an approved bisphosphonate drug used for the treatment of osteoporosis, was incorporated into a composite coating matrix via coelectrospraying. Its release behavior showed a long-term sustained release. This approach can be a potential coating technique for the surface modification of biopolymer implants.     

Controlled release of sirolimus from a multilayered PLGA stent matrix

The release of sirolimus from a bi-layer biodegradable polymeric film is reported in this study. Approved drug-eluting metal stents use a thin polymer coating to control drug release, but the degree of control is limited. In a fully polymeric stent, the use of multilayers allows a range of release kinetics. A bi-layer system, with PLLA as the supporting layer and PLGA as the drug-eluting layer, was used in this study to simulate release of sirolimus from a stent. The results show that the release of sirolimus is diffusion and degradation-controlled, and that the amount of sirolimus loading does not affect its release kinetics. The release of sirolimus is, however, accelerated by the addition of a plasticizer, such as PEG, as water uptake is increased. An increased water uptake increases polymer degradation, and changes the dominant mode of release to degradation-control. The release of sirolimus can, on the other hand, be retarded by using a coating of a biodegradable polyester with a lauryl ester end group. Therefore, multilayered systems offer many options for controlling sirolimus release over months.     

A mathematical model for predicting drug release from a biodurable drug-eluting stent coating

Drug-eluting stents (DESs) are drug-device combination products that have been commercialized and demonstrated to be safe and efficacious in treating coronary artery disease. They have been very effective in reducing the extent of neointimal hyperplasia and therefore in preventing or minimizing the occurrence of in-stent restenosis. In order to develop a successful DES, it is imperative that the coating be designed so as to deliver, after stent implantation, a therapeutic dose of the drug for the desired time duration at the site of the arterial blockage. Mathematical models are very valuable tools that can be used to study the effect of different coating parameters on drug delivery and can therefore help in coating design. We have developed a bimodal lumped-parameter mass transport model to describe the release of the drug everolimus from a biodurable fluoropolymer-based DES coating. We assume that the dispersed drug phase contributes to two discrete modes of drug transport through the coating. These are the fast mode (mode I) which is the release of the drug from a highly percolated structure of drug phase within the polymer, and the slow mode (mode II) which is the release of the drug from a nonpercolated, polymer-encapsulated phase of the drug within the coating. The three coefficients in the governing equations describing the model, i.e. the two effective diffusivities corresponding to each of the two modes and the fraction of the drug in one of the two modes, were determined by fitting with available DES release data. The predictive power of the model is demonstrated by comparing the release rate from different coating configurations (thickness and drug to polymer ratios) with experimental data. Also, it is demonstrated that if limited experimental data are available at early time points, the model can be used to predict drug release at subsequent time points.     

Chitin/PLGA blend microspheres as a biodegradable drug delivery system: a new delivery system for protein

Novel chitin/PLGAs and chitin/PLA based microspheres were developed for the delivery of protein. These biodegradable microspheres were prepared by polymers blending and wet phase-inversion methods. The parameters such as selected non-solvents, temperature of water and ratio of polylactide to polyglycolide were adjusted to improve thermodynamic compatibility of individual polymer (chitin and PLGAs or chitin/PLA), which affects the hydration and degradation properties of the blend microspheres. Triphasic pattern of drug release model is observed from the release of protein from the chitin/PLGAs and chitin/PLA microspheres: the initially fast release (the first phase), the following slow release (the second phase) and the second burst release (the third phase). Formulations of the blends, which are based on the balance among the hydration rate of the chitin phase and degradation of chitin/PLA and PLGA phase, can lead to a controllable release of bovine serum albumin (BSA). In conclusion, such a chitin/PLGA 50/50 microsphere is novel and interesting, and may be used as a protein delivery system.     

Computational Investigation of the Delamination of Polymer Coatings During Stent Deployment

Recent advances in angioplasty have involved the application of polymer coatings to stent surfaces for purposes of drug delivery. Given the high levels of deformation developed in the plastic hinge of a stent during deployment, the achievement of an intact bond between the coating and the stent presents a significant mechanical challenge. Problems with coating delamination have been reported in recent experimental studies. In this paper, a cohesive zone model of the stent–coating interface is implemented in order to investigate coating debonding during stent deployment. Simulations reveal that coatings debond from the stent surface in tensile regions of the plastic hinge during deployment. The critical parameters governing the initiation of delamination include the coating thickness and stiffness, the interface strength between the coating and stent surface, and the curvature of the plastic hinge. The coating is also computed to debond from the stent surface in compressive regions of the plastic hinge by a buckling mechanism. Computed patterns of coating delamination correlate very closely with experimental images. This study provides insight into the critical factors governing coating delamination during stent deployment and offers a predictive framework that can be used to improve the design of coated stents.     

Drug loading and elution from a phosphorylcholine polymer-coated coronary stent does not affect long-term stability of the coating in vivo

A drug eluting coronary stent was developed for use in preclinical and clinical trial evaluation. The stent was coated with a phosphorylcholine (PC)-based polymer coating containing the cell migration inhibitor batimastat. A pharmacokinetic study was conducted in a rabbit iliac model using (14)C-radiolabeled version of the drug; this showed the drug release to be first order with 94% of it being released within 28 days. Unloaded and drug-loaded stents were implanted in a porcine coronary artery model; a number were explanted at 5 days and scanning electron microscopy was used to show that the presence of the drug did not affect the rate of stent endothelialization. The remainder of the stents were removed after 6 months and the stents carefully removed from the arterial tissue. Fourier-transform infrared (FT-IR) spectroscopy (both attenuated total reflectance and microscopic imaging) was used to show the presence of the PC coating on control unloaded, drug-loaded and explanted stents, providing evidence that the coating was still present. This was further confirmed by use of atomic force microscopy (AFM) amplitude-phase, distance (a-p,d) curves which generated the characteristic traces of the PC coating. Further AFM depth-profiling techniques found that the thicknesses of the PC coatings on an control unloaded stent was 252+/-19 nm, on an control batimastat-loaded stent 906+/-224 nm and on an explanted stent 405+/-224 nm. The increase in thickness after the drug-loading process was a consequence of drug incorporation in the film, and the return to the unloaded dimensions for the explanted sample indicative of elution of the drug from the coating. The drug delivery PC coating was therefore found to be stable following elution of the drug and after 6 months implantation in vivo.     

The pre-clinical assessment of rapamycin-eluting, durable polymer-free stent coating concepts

All four currently FDA-approved drug-eluting stents (DESs) contain a durable polymeric coating which can negatively impact vascular healing processes and eventually lead to adverse cardiac events. Aim of this study was the pre-clinical assessment of two novel rapamycin-eluting stent (RES) coating technologies that abstain from use of a durable polymer. Two distinctive RES coating technologies were evaluated in vitro and in the porcine coronary artery stent model. The R-poly(S) stent platform elutes rapamycin from a biodegradable polymer that is top coated with the resin shellac to minimize the amount of polymer. The R-pro(S) stent platform allows dual drug release of rapamycin and probucol, blended by shellac. HPLC-based determination of pharmacokinetics indicated drug release for more than 28 days. At 30 days, neointimal formation was found to be significantly decreased for both DESs compared to bare-metal stents. Assessment of vascular healing revealed absence of increased inflammation in both DESs, which is commonly observed in DES with non-erodible polymeric coating. In conclusion, the pre-clinical assessment of RESs with resin-based or dual drug coating indicated an adequate efficacy profile as well as a beneficial effect for vascular healing processes. These results encourage the transfer of these technologies to clinical evaluation.     

Reduced thrombogenicity of nitinol stents--in vitro evaluation of different surface modifications and coatings

The material and the surface patterns of intravascular stents play a pivotal role in activating platelets and triggering adherence of inflammatory cells that consecutively leads to renarrowing caused by neointimal hyperplasia. To improve these features, besides mechanical and chemical modifications, ways of masking the stent by covering have been developed. In addition, polymer-coated stents are used as vehicle for local drug delivery. But as substances used for this application are described to possess an inflammatory potential, this aspect has to be evaluated. In the present study we compared different approaches to surface alterations applied to a nitinol stent design. Besides commonly used techniques like passivation and electropolishing, we evaluated coatings with heparin, aluminium and a polyurethane polymer regarding their thrombogenic and inflammatory characteristics. By weaving thin elastomer fibres a graft was generated. The previously described Chandler loop was used to simulate arterial flow conditions ex vivo using rotating PVC tubings filled with human blood. All stents received 120 min of blood contact. To determine thrombocyte activation and inflammatory reaction, the platelet count and levels of beta-TG, TAT and PMN-elastase were assessed. Scanning electron microscopy was used to visualize the reactions. Mechanical polishing and passivation did not improve the stent surface characteristics while sandblasting, electropolishing and aluminium covering decreased activation of the coagulation cascade. In terms of thrombogenicity, the heparin coating had no beneficial effect. The lowest thrombogenic potential was found in the Polyurethane-coated stent group. All stents showed similar levels of polymorph nuclear granulocyte elastase except for the membrane design. While mechanical and chemical modifications are able to reduce thrombogenicity, coating with this particular polyurethane polymer seems to be superior to these approaches regarding the parameters assessed in this experimental setting. The Chandler loop is a valuable tool to test polymeric coatings ex vivo since these modifications may reduce drug performance by inducing inflammatory reaction themselves.     

Preparation and characterization of zwitterionic phospholipid polymer-coated poly(lactic acid) nanoparticles

Poly(lactic acid) (PLA) nanoparticles (NPs) are the most promising polymer NPs for drug delivery and targeting. However, they are easily recognized as a foreign body and rapidly cleared from the body by the mononuclear phagocyte system. Cell membrane mimetic random copolymers, bearing both zwitterionic phosphorylcholine groups and hydrophobic butyl side chains (PMB) and additional cross-linkable trimethoxysilylpropyl side chains (PMBT), were synthesized and coated on PLA NPs. Effects of the zwitterionic copolymer coatings on the NP size distribution, dispersion stability, and drug release behavior were investigated. Furthermore, the effect of the coatings on phagocytosis was also investigated. Compared with conventional polyvinyl alcohol coating, the cell membrane mimetic copolymer coatings decreased the size and increased the stability of the PLA NPs aqueous dispersions. More importantly, doxorubicin (DOX) release was well controlled and NPs phagocytosis by mouse peritoneal macrophage was decreased to one-third when the nanoparticles were coated with PMBT. This simple and effective zwitterionic polymer coating strategy may serve as a new route to design and optimize long-circulating intravenously injectable nanoparticle drug carriers.     

Paclitaxel release from single and double-layered poly(DL-lactide-co-glycolide)/poly(L-lactide) film for biodegradable coronary stent application

Our laboratory has been developing a completely biodegradable coronary stent which is made of bilayers of biodegradable polyesters. This article presents the preliminary work done to exploit the drug delivery potential of such a polymeric stent. An antiproliferative drug (paclitaxel) was added either only to the top layer or to both layers and the in vitro release profiles were monitored for up to 90 days. Within 90 days, the measured paclitaxel release was almost entirely from the P(DL)LGA layer. In general, the release profiles show three distinct stages: (a) extremely slow diffusional release, (b) accelerated diffusion-degradation release, and (c) saturation. Separate degradation studies (water absorption, molecular weight reduction, weight loss, and surface topography) were also conducted to better understand the observed release behavior.     

Physical characterization of controlled release of paclitaxel from the TAXUS Express2 drug-eluting stent

The polymer carrier technology in the TAXUS drug-eluting stent consists of a thermoplastic elastomer poly(styrene-b-isobutylene-b-styrene) (SIBS) with microphase-separated morphology resulting in optimal properties for a drug-delivery stent coating. Comprehensive physical characterization of the stent coatings and cast film formulations showed that paclitaxel (PTx) exists primarily as discrete nanoparticles embedded in the SIBS matrix. Thermal and chemical analysis did not show any evidence of solubility of PTx in SIBS or of any molecular miscibility between PTx and SIBS. Atomic force microscope data images revealed for the first time three-dimensional stent coating surfaces at high spatial resolutions in air and in situ under phosphate-buffered saline as drug was released. PTx release involves the initial dissolution of drug particles from the PTx/SIBS coating surface. Morphological examination of the stent coatings in vitro supported an early burst release in most formulations because of surface PTx followed by a sustained slower release of PTx from the bulk coating. The in vitro PTx release kinetics were dependent on the formulation and correlated to the drug-to-polymer ratio. Atomic force microscopy analysis confirmed this correlation and further supported the concept of a matrix-based drug-release coating.     

Hydroxyapatite/poly(epsilon-caprolactone) composite coatings on hydroxyapatite porous bone scaffold for drug delivery

Hydroxyapatite (HA) porous scaffold was coated with HA and polycaprolactone (PCL) composites, and antibiotic drug tetracycline hydrochloride was entrapped within the coating layer. The HA scaffold obtained by a polymeric reticulate method, possessed high porosity ( approximately 87%) and controlled pore size (150-200 microm). Such a well-developed porous structure facilitated usage in a drug delivery system due to its high surface area and blood circulation efficiency. The PCL polymer, as a coating component, was used to improve the brittleness and low strength of the HA scaffold, as well to effectively entrap the drug. To improve the osteoconductivity and bioactivity of the coating layer, HA powder was hybridized with PCL solution to make the HA-PCL composite coating. With alteration in the coating concentration and HA/PCL ratio, the morphology, mechanical properties, and biodegradation behavior were investigated. Increasing the concentration rendered the stems thicker and some pores to be clogged; as well increasing the HA/PCL ratio made the coating surface be rough due to the large amount of HA particles. However, for all concentrations and compositions, uniform coatings were formed, i.e., with the HA particles being dispersed homogeneously in the PCL sheet. With the composite coating, the mechanical properties, such as compressive strength and elastic modulus were improved by several orders of magnitude. These improvements were more significant with thicker coatings, while little difference was observed with the HA/PCL ratio. The in vitro biodegradation of the composite coatings in the phosphate buffered saline solution increased linearly with incubation time and the rate differed with the coating concentration and the HA/PCL ratio; the higher concentration and HA amount caused the increased biodegradation. At short period (<2 h), about 20-30% drug was released especially due to free drug at the coating surface. However, the release rate was sustained for prolonged periods and was highly dependent on the degree of coating dissolution, suggesting the possibility of a controlled drug release in the porous scaffold with HA+PCL coating.     

Porous polysulfone coatings for enhanced drug delivery

The synthesis of a porous polysulfone (PSU) coating for use in drug delivery applications is presented. PSU can serve as a functional surface coating for drug delivery vehicles, such as intraocular biomicrorobots. The coatings can be applied using spin coating or dip coating. The porosity is introduced by selectively dissolving calcium carbonate nanoparticles embedded in the bulk polymer. The network of pores thus formed increases by a factor of thirty the amount of Rhodamine B (model drug) that can be loaded and by a factor of fifteen the amount that can be released. The films do not affect cell viability and exhibit poor cell adhesion. The straightforward synthesis and predictability of porosity enables the tuning of the amount of drug that can be loaded.     

Semi-interpenetrating network of poly(ethylene glycol) and poly(D,L-lactide) for the controlled delivery of protein drugs

We have prepared a semi-interpenetrating network (IPN) of poly(ethylene glycol) dimethacrylate (PEGDMA) with entrapped poly(D,L-lactide) (PLA) using photochemical techniques. These IPNs were developed for the controlled delivery of protein drugs such as growth factors. The PEG component draws water into the network, forming a hydrogel within the PLA matrix, controlling and facilitating release of the protein drug, while the PLA component both strengthens the PEG hydrogel and enhances the degradation and elimination of the network after the protein drug is released. The rate and extent of swelling and the resultant protein release kinetics could be controlled by varying the PEG/PLA ratio and total PLA content. These IPNs were prepared using a biocompatible benzyl benzoate/benzyl alcohol solvent system that yields a uniform, fine dispersion of the protein throughout the PEG/PLA IPN matrix. IPNs composed of high molecular mass PLA and lower PEG/PLA ratios exhibited lower equilibrium swelling ratios. The release of bovine serum albumin (BSA), a model protein, from these IPNs was characterized by a large initial burst, regardless of the PEG/PLA ratio, due to the entrapment of residual solvent within the network. Microparticles of the PEG/PLA IPNs were also prepared using a modified Prolease strategy. Residual solvent removal was significantly enhanced using this process. The microparticles also exhibited a significant reduction in the initial burst release of protein. Mixtures of different compositions of PEG/PLA microparticles should be useful for the delivery of a variety of protein drugs with different release kinetics from any tissue-engineering matrix.     

Self-assembled molecular structures as ultrasonically-responsive barrier membranes for pulsatile drug delivery

Noninvasive ultrasound has been shown to increase the release rate on demand from drug delivery systems; however, such systems generally suffer from background drug leaching. To address this issue, a drug-containing polymeric monolith coated with a novel ultrasound-responsive coating was developed. A self-assembled molecular structure coating based on relatively impermeable, ordered methylene chains forms an ultrasound-activated on-off switch in controlling drug release on demand, while keeping the drug inside the polymer carrier in the absence of ultrasound. The orderly structure and molecular orientation of these C12 n-alkyl methylene chains on polymeric surfaces resemble self-assembled monolayers on gold. Their preparation and characterization have been published recently (Kwok et al. [Biomacromolecules 2000;1(1):139-148]). Ultrasound release studies showed that a copolymer of 2-hydroxyethyl methacrylate and ethylene glycol dimethacrylate (MW 400) coated with such an ultrasound-responsive membrane maintained sufficient insulin for multiple insulin delivery, compared with a substantial burst release during the first 2 h from uncoated samples. With appropriate surface coating coverage, the background leach rate can be precisely controlled. The biological activity of the insulin releasate was tested by assessing its ability to regulate [C14]-deoxyglucose uptake in 3T3-L1 adipocyte cells in a controlled cell culture environment. Uptake triggered by released insulin was comparable to that of the positive insulin control. The data demonstrate that the released insulin remains active even after the insulin had been exposed to matrix synthesis and the methylene chain coating process.     

Metallic wires with an adherent lubricious and blood-compatible polymeric coating and their use in the manufacture of novel slippery-when-wet guidewires: possible applications related to controlled local drug delivery

A new procedure was developed for the controlled application of adherent hydrophilic and biocompatible coatings onto the surface of "endless" metallic wires. Use of copolymers of 1-vinyl-2-pyrrolidinone and alkylmethacrylates provided coatings with excellent adherence and lubricity, and markedly low thrombogenicity. Coated wires could be spiralized without damaging the coating; the resulting coils are potentially useful as lubricious guidewires for use in, for example, interventional cardiology or urology. This study demonstrates that the lubricity of the coating is dependent on the composition (hydrophilicity) of the coating biomaterial, as well as on the thickness of the coating. Furthermore, the results imply that the adherence of the hydrophilic coating is essentially due to entanglement of the binder polymer chains and the hydrophilic copolymer chains. Moreover, the idea to use the hydrophilic coating on the wire as a temporary depot for controlled local drug delivery was explored. The coating was loaded with the dye rhodamine, and release of the dye upon immersion of the coated wire in water was studied. This work revealed that release of the drug is dependent on the composition of the coating. The potential utility of such wires with a drug-charged coating for controlled local drug delivery is discussed briefly.     

Drug release from ion-exchange microspheres: mathematical modeling and experimental verification

This paper presents for the first time a mathematical model for a mechanism of controlled drug release involving both ion exchange and transient counter diffusion of a drug and counterions. Numerical analysis was conducted to study the effect of different factors on drug release kinetics including environmental condition, material properties, and design parameters. The concentration profiles of counterions and drug species, the moving front of ion exchange, and three distinct regions inside a microsphere, namely unextracted region, ion-exchange region and drug diffusion region, were revealed by model prediction. The numerical results indicated that the rate of drug release increased with an increase in the initial drug concentration in the microspheres, the salt concentration in the external solution, or the valence of the counterions, whereas it decreased with increasing Langmuir isotherm constant. The mathematical and experimental procedures for determination of the equilibrium constant and the usefulness of the model were demonstrated using verapamil hydrochloride and sulfopropyl dextran microsphere system as an example. This work has provided a very useful mathematical tool for predicting kinetics and equilibrium of drug release and for optimizing the design of ion-exchange drug delivery systems.     

Silk coatings on PLGA and alginate microspheres for protein delivery

Bombyx mori silk fibroin self-assembles on surfaces to form ultrathin nanoscale coatings based on our prior studies using layer-by-layer deposition techniques driven by hydrophobic interactions between silk fibroin protein molecules. In the present study, poly(lactic-co-glycolic acid) (PLGA) and alginate microspheres were used as substrates and coated with silk fibroin. The coatings were visualized by confocal laser scanning microscopy using fluorescein-labeled silk fibroin. On PLGA microspheres, the coating was approximately 1microm and discontinuous, reflecting the porous surface of these microspheres determined by SEM. In contrast, on alginate microspheres the coating was approximately 10microm thick and continuous. The silk fibroin penetrated into the alginate gel matrix. The silk coating on the PLGA microspheres delayed PLGA degradation. The silk coating on the alginate microspheres survived ethylenediamine tetraacetic acid (EDTA) treatment used to remove the Ca(2+)-cross-links in the alginate gels to solubilize the alginate. This suggests that alginate microspheres can be used as templates to form silk microcapsules. Horseradish peroxidase (HRP) and tetramethylrhodamine-conjugated bovine serum albumin (Rh-BSA) as model protein drugs were encapsulated in the PLGA and alginate microspheres with and without the silk fibroin coatings. Drug release was significantly retarded by the silk coatings when compared to uncoated microsphere controls, and was retarded further by methanol-treated silk coating when compared to silk water-based coatings on alginate microspheres. Silk coatings on PLGA and alginate microspheres provide mechanically stable shells as well as a diffusion barrier to the encapsulated protein drugs. This coating technique has potential for biosensor and drug delivery applications due to the aqueous process employed, the ability to control coating thickness and crystalline content, and the biocompatibility of the silk fibroin protein used in the process.     

Mechanical properties, drug eluting characteristics and in vivo performance of a genipin-crosslinked chitosan polymeric stent

A limitation with the use of polymers as stent matrices is their inherent mechanical weakness. In this study, a polymeric stent, made from chitosan-based films fixed by genipin which has a cyclic molecular structure, was developed (the genipin stent). The mechanical properties of the genipin stent were investigated; its counterpart fixed by a linear epoxy compound (the epoxy stent) and a commercially available metallic stent were used as controls. The results indicated that the cyclic crosslinking structures formed within the genipin stent matrix were beneficiary to the improvement of its mechanical property. Additionally, the tolerable compression load of the genipin stent was superior to that of the control metallic stent. The cytotoxicity of the genipin stent was significantly lower than the epoxy stent. The deployment of the genipin stent in rabbit infrarenal abdominal aortas was performed using a French sheath. At 3 months postoperatively, the retrieved arteries remained patent; no thrombosis was observed. A nearly intact layer of endothelial cells was seen on the stent-implanted vessel wall. To evaluate its possibility as a drug delivery vehicle, sirolimus (an anti-proliferative drug) was loaded in the genipin stent. It was found that the genipin stent with heparin coating exhibited a linear sustained-release profile and the released sirolimus still possessed its original activity in inhibiting smooth muscle cell proliferation. These findings suggest that the genipin stent with enhanced mechanical strength can be used as an attractive stent platform for local drug delivery.     

Development of rationally designed affinity-based drug delivery systems

Many drug delivery systems have been developed to provide sustained release of proteins in vivo. However, the ability to predict and control the rate of release from delivery systems is still a challenge. Toward this goal, we screened a random drug-binding peptide library (12 amino acids) to identify peptides of varying (i.e. low, moderate, and high) affinity for a model polysaccharide drug (heparin). Peptide domains of varying affinity for heparin identified from the library were synthesized using standard solid phase chemistry. A mathematical model of drug release from a biomaterial scaffold containing drug-binding peptide domains identified from the library was developed. This model describes the binding kinetics of drugs to the peptides, the diffusion of free drug, and the kinetics of enzymatic matrix degradation. The effect of the ratio of binding sites to drug, the effect of varying the binding kinetics and the rate of enzymatic matrix degradation on the rate of drug release was examined. The in vitro release of the model drug from scaffold containing the peptide drug-binding domains was measured. The ability of this system to deliver and modulate the biological activity of protein drugs was also assessed using nerve growth factor (NGF) in a chick dorsal root ganglia (DRG) neurite extension model. These studies demonstrate that our rational approach to drug delivery system design can be used to control drug release from tissue-engineered scaffolds and may be useful for promoting tissue regeneration in vivo.