Physicomechanical characterizations and in vitro release studies of electrospun ethyl cellulose fibers, solvent cast carboxymethyl cellulose films, and their composites.

Frequent change of wound dressings introduces wound inflammation and infections. In this study, we electrospun phenytoin (PHT) loaded ethyl cellulose (EC) microfibers and solvent cast tetracycline hydrochloride (TCH) loaded carboxymethyl cellulose (CMC) films with the aim to demonstrate tailorable in vitro drug release behaviors suitable for long-term use of wound dressings. Results from tensile testing showed a significant decrease in average elastic moduli from 8.8 ± 0.6 to 3.3 ± 0.3 MPa after incorporating PHT into EC fibers. PHT-loaded EC fibers displayed a slow and zero-ordered release up to 80 % of the total drug at 48 h, while TCH-loaded CMC films demonstrated a rapid and complete release within 30 min. Furthermore, drug-loaded EC/CMC composites were fabricated into fiber-in-film and fiber-on-film composites. Fiber-in-film composites showed stage release of TCH and PHT at 8 h, while fiber-on-film composites demonstrated simultaneous release of PHT and TCH with a prolonged release of TCH from CMC films. In general, electrospun PHT-loaded EC microfibers, solvent cast TCH-loaded CMC films, and their composites were studied to provide a fundamental scientific understanding on the novelty of the ability to modulate drug release characteristics based on the composite designs.

Effects of Poloxamers as Excipients on the Physicomechanical Properties, Cellular Biocompatibility, and In Vitro Drug Release of Electrospun Polycaprolactone (PCL) Fibers.

Electrospun microfibers are emerging as one of the advanced wound dressing materials for acute and/or chronic wounds, especially with their ability to carry drugs and excipients at a high loading while being able to deliver them in a controlled manner. Various attempts were made to include excipients in electrospun microfibers as wound dressing materials, and one of them is poloxamer, an amphiphilic polymer that exhibits wound debridement characteristics. In this study, we formulated two types of poloxamers (i.e., P188 and P338) at 30% (w/w) loading into electrospun polycaprolactone (PCL) fibers to evaluate their physicomechanical properties, biocompatibility, and in vitro drug release of a model drug. Our findings showed that the incorporation of poloxamers in the PCL solutions during electrospinning resulted in a greater "whipping" process for a larger fiber deposition area. These fibers were mechanically stiffer and stronger, but less ductile as compared to the PCL control fibers. The incorporation of poloxamers into electrospun PCL fibers reduced the surface hydrophobicity of fibers according to our water contact angle studies and in vitro degradation studies. The fibers' mechanical properties returned to those of the PCL control groups after "dumping" the poloxamers. Moreover, poloxamer-loaded PCL fibers accelerated the in vitro release of the model drug due to surface wettability. These poloxamer-loaded PCL fibers were biocompatible, as validated by MTT assays using A549 cells. Overall, we demonstrated the ability to achieve a high loading of poloxamers in electrospun fibers for wound dressing applications. This work provided the basic scientific understanding of materials science and bioengineering with an emphasis on the engineering applications of advanced wound dressings.

Physicomechanical properties and in vitro release behaviors of electrospun ibuprofen-loaded blend PEO/EC fibers.

Electrospinning is a fiber manufacturing technique with the possibility of encapsulating high levels of small molecule drugs while providing controlled release rates. In this study, electrospun blend fibers were produced from polyethylene oxide (PEO) and ethyl cellulose (EC) at various compositions to encapsulate a poorly water-soluble drug of ibuprofen (IBP) at 30% loading. Microscopic evaluation showed smooth and defect-free fiber morphologies for blank and IBP-loaded PEO/EC fibers. The average fiber diameters and fiber yields suggested a potential optimization on the blend fiber composition for the electrospun drug-eluting PEO/EC fibers, where the highest average fiber diameter and fiber yield occurred at 50PEO/50EC fiber composition. Surface wettability studies demonstrated the effects on surface hydrophobicity from blend fibers of water-soluble PEO and hydrophobic EC as well as the incorporation of IBP. In addition, blend fibers containing more PEO promoted the water absorption rates through dissolution of the polymer matrix. Furthermore, results from mechanical testing of the blend fibers showed the highest fiber elastic modulus and tensile strength at fiber compositions in between 75PEO/25EC and 50PEO/50EC, corresponding to the average fiber diameter measurements. The in vitro IBP release rates demonstrated a dependence on the EC compositions supported by the surface wettability and water absorption rate studies. In general, our work demonstrated the ability to electrospin blank and IBP-loaded PEO/EC fibers with the scientific understandings of EC compositions on modulations of fiber physicomechanical properties and in vitro drug release rates. The findings from the work indicated the potential engineering and pharmaceutical applications of electrospun drug-eluting fibers for topical drug delivery.

Electrospun Chitosan-based Fibers for Wound Healing Applications.

Chitosan, a natural-occurring biopolymer, is biocompatible to tissues with excellent antibacterial and hemostatic properties, which makes it a great candidate among wound dressing materials. In this paper, electrospun fiber-based wound dressings from blend chitosan and/or polyethylene oxide (PEO) and/or polyvinyl alcohol (PVA) fibers were reviewed. The incorporation of these water-soluble copolymers allows the entanglement of the rigid chitosan molecular chains during electrospinning leading to the production of continuous nonwoven fibers having average diameters ranging from several tenths to hundredths of nanometers. Increasing chitosan composition in the fibers improves the bulk mechanical strength of the fiber mats due to the rigid molecular structure of chitosan. The nano-sized pores within the fiber mats promote permeability of the fiber dressings, which further enhances the exchange of oxygen and nutrients with outside environment. In addition, the porous fiber mat structure facilitates the absorption of wound exudates while reducing the possibility of bacterial infections. Several studies in antibacterial and anti-inflammatory responses of chitosan-based electrospun fibers were discussed in this short review. More importantly, inclusions of small molecule drugs and/or biological agents are possible in chitosan-based electrospun fibers, which provide a multi-purpose treatment capability for wound healing applications.

Inhibition of Platelet Adhesion from Surface Modified Polyurethane Membranes.

Coronary thrombosis is one of the leading causes of mortality and morbidity in cardiovascular diseases, and patients who received vascular stent treatments are likely to suffer from restenosis due to tissue damage from stenting procedures (extrinsic pathway) and/or presence of unregulated factor XII (intrinsic pathway). Regardless of the pathway, coagulation factors and exposed collagen activate the G-protein-coupled receptors located at the plasma membrane of the resting platelets resulting in the change of their shapes with protrusions of filopodia and lamellipodia for surface adhesion. In this mini review, we discussed the mechanisms involved in platelet activation, adhesion, and aggregation. More importantly, we reviewed the use of polyurethane membranes with modified surface functional groups to down-regulate platelet adhesion and aggregation activities. Polyurethane membranes with hydrophilic and negatively charged surface properties showed a reduced αIIb-ß3 signaling from the activated platelets, resulting in the decrease of platelet adhesion and aggregation. The use of polyurethane membranes with modified surface properties as coatings on vascular stents provides an engineering approach to mitigate blood clotting associated with restenosis.

Thermal and Physico-Mechanical Characterizations of Thromboresistant Polyurethane Films.

Hemocompatibility remains a challenge for injectable and/or implantable medical devices, and thromboresistant coatings appear to be one of the most attractive methods to down-regulate the unwanted enzymatic reactions that promote the formation of blood clots. Among all polymeric materials, polyurethanes (PUs) are a class of biomaterials with excellent biocompatibility and bioinertness that are suitable for the use of thromboresistant coatings. In this work, we investigated the thermal and physico-mechanical behaviors of ester-based and ether-based PU films for potential uses in thromboresistant coatings. Our results show that poly(ester urethane) and poly(ether urethane) films exhibited characteristic peaks corresponding to their molecular configurations. Thermal characterizations suggest a two-step decomposition process for the poly(ether urethane) films. Physico-mechanical characterizations show that the surfaces of the PU films were hydrophobic with minimal weight changes in physiological conditions over 14 days. All PU films exhibited high tensile strength and large elongation to failure, attributed to their semi-crystalline structure. Finally, the in vitro clotting assays confirmed their thromboresistance with approximately 1000-fold increase in contact time with human blood plasma as compared to the glass control. Our work correlates the structure-property relationships of PU films with their excellent thromboresistant ability.

The Role of Electrospun Fiber Scaffolds in Stem Cell Therapy for Skin Tissue Regeneration.

Stem cell therapy has emerged as one of the topics in tissue engineering where undifferentiated and multipotent cells are strategically placed/ injected in tissue structure for cell regeneration. Over the years, stem cells have shown promising results in skin repairs for non-healing and/or chronic wounds. The addition of the stem cells around the wound site promotes signaling pathways for growth factors that regulate tissue reconstruction. However, injecting stem cells around the wound site has its drawbacks, including cell death due to lack of microenvironment cues. This particular issue is resolved when biomaterial scaffolds are involved in the cultivation and mechanical support of the stem cells. In this review, we describe the current models of stem cell therapy by injections and those that are done through cell cultures using electrospun fiber scaffolds. Electrospun fibers are considered as an ideal candidate for cell cultures due to their surface properties. Through the control of fiber morphology and fiber structure, cells are able to proliferate and differentiate into keratinocytes for skin tissue regeneration. Furthermore, we provide another perspective of using electrospun fibers and stem cells in a layer-by-layer structure for skin substitutes (dressing). Finally, electrospun fibers have the potential to incorporate bioactive agents to achieve controlled release properties, which is beneficial to the survival of the delivered stem cells or the recruitment of the cells. Overall, our work illustrates that electrospun fibers are ideal for stem cell cultures while serving as cell carriers for wound dressing materials.

Engineering Approaches to Prevent Blood Clotting from Medical Implants.

Injectable and/or Implantable medical devices are widely used in the treatment of diseases. Among them, vascular stents provide the medical solution to treat blood clotting. However, traditional metallic stents, even with current improvements in anticoagulation properties, have potential drawbacks in local inflammation when first implanted into the body and undesirable protein adsorption and cell adhesion after a prolonged period of time in the body. In this perspective, we discuss several engineering approaches, including drug-eluting materials, polymeric and non-polymeric coatings, and surface modifications to coating materials that can be applied to the surface of medical implants to significantly improve the hemocompatibility. These coatings are expected to have a slow degradation rate with the ability to either load drugs or attach biomacromolecules to form an architecture that mimics the surrounding cells. In general, our perspective provides a current view on the achievements of hemo-compatible coatings and future trends in coating materials that will extend the life of the medical implants.

Nanostructure-Enabled and Macromolecule-Grafted Surfaces for Biomedical Applications.

Advances in nanotechnology and nanomaterials have enabled the development of functional biomaterials with surface properties that reduce the rate of the device rejection in injectable and implantable biomaterials. In addition, the surface of biomaterials can be functionalized with macromolecules for stimuli-responsive purposes to improve the efficacy and effectiveness in drug release applications. Furthermore, macromolecule-grafted surfaces exhibit a hierarchical nanostructure that mimics nanotextured surfaces for the promotion of cellular responses in tissue engineering. Owing to these unique properties, this review focuses on the grafting of macromolecules on the surfaces of various biomaterials (e.g., films, fibers, hydrogels, and etc.) to create nanostructure-enabled and macromolecule-grafted surfaces for biomedical applications, such as thrombosis prevention and wound healing. The macromolecule-modified surfaces can be treated as a functional device that either passively inhibits adverse effects from injectable and implantable devices or actively delivers biological agents that are locally based on proper stimulation. In this review, several methods are discussed to enable the surface of biomaterials to be used for further grafting of macromolecules. In addition, we review surface-modified films (coatings) and fibers with respect to several biomedical applications. Our review provides a scientific update on the current achievements and future trends of nanostructure-enabled and macromolecule-grafted surfaces in biomedical applications.

Electrospun Fibers as a Dressing Material for Drug and Biological Agent Delivery in Wound Healing Applications.

Wound healing is a complex tissue regeneration process that promotes the growth of new tissue to provide the body with the necessary barrier from the outside environment. In the class of non-healing wounds, diabetic wounds, and ulcers, dressing materials that are available clinically (e.g., gels and creams) have demonstrated only a slow improvement with current available technologies. Among all available current technologies, electrospun fibers exhibit several characteristics that may provide novel replacement dressing materials for the above-mentioned wounds. Therefore, in this review, we focus on recent achievements in electrospun drug-eluting fibers for wound healing applications. In particular, we review drug release, including small molecule drugs, proteins and peptides, and gene vectors from electrospun fibers with respect to wound healing. Furthermore, we provide an overview on multifunctional dressing materials based on electrospun fibers, including those that are capable of achieving wound debridement and wound healing simultaneously as well as multi-drugs loading/types suitable for various stages of the healing process. Our review provides important and sufficient information to inform the field in development of fiber-based dressing materials for clinical treatment of non-healing wounds.

Bioengineered keratocyte spheroids fabricated on chitosan coatings enhance tissue repair in a rabbit corneal stromal defect model.

Cultivated cell spheroid transplantation is widely studied as a means of facilitating tissue regeneration. Chitosan biomaterial has been shown to promote keratocyte aggregation and multicellular spheroid formation. This study provides further evidence on application of bioengineered keratocyte spheroids for corneal stromal tissue engineering. In an allogeneic rabbit model of stromal destruction caused by bacterial keratitis, the corneas were intrastromally injected with isolated keratocyte suspensions or aggregated spheroid grafts at same cell number. Results of clinical observations and histological examinations on postoperative day 14 showed that when an antibiotic eye drop is only medication for inhibiting bacterial growth, permanent damage to stroma occurs, leading to disorganization of collagen lamellae and tissue structure as well as loss of corneal transparency and visual function. Intrastromal grafting of keratocytes provided additional benefits to overcome drawbacks of limited disease treatment performance associated with topically applied antibiotics. In particular, as compared to their cell suspension counterparts, bioengineered keratocyte spheroids had higher ability to preserve cellular phenotype, secrete collagen matrix, and enhance graft retention, suggesting excellent repair capability for managing stromal tissue defect and alleviating corneal haze/oedema. In summary, the findings emphasize the role of keratocyte configuration (i.e., two-dimensional monolayer or three-dimensional spheroid) in determining therapeutic potency of cellular allografts for stromal tissue reconstruction. Transplantation of keratocyte spheroids cultured on chitosan substrates may represent a promising strategy for corneal stromal repair.

Development of gelatin/ascorbic acid cryogels for potential use in corneal stromal tissue engineering.

To offer an ideal hospitable environment for corneal keratocyte growth, the carrier materials can be functionalized with incorporation of signaling molecules to regulate cell biological events. This study reports, for the first time, the development of gelatin/ascorbic acid (AA) cryogels for keratocyte carriers in vitro and in vivo. The cryogel samples were fabricated by blending of gelatin with varying amounts of AA (0-300 mg) and carbodiimide cross-linking via cryogelation technique. Hydrophilic AA content in the carriers was found to significantly affect cross-linking degree and pore dimension of cryogels, thereby dictating their mechanical and biological stability and AA release profile. The cryogel carriers with low-to-moderate AA loadings were well tolerated by rabbit keratocyte cultures and anterior segment eye tissues, demonstrating good ocular biocompatibility. Although higher incorporated AA level contributed to enhanced metabolic activity and biosynthetic capacity of keratocytes grown on cryogel matrices, the presence of excessive amounts of AA molecules could lead to toxic effect and limit cell proliferation and matrix production. The cytoprotective activity against oxidative stress was shown to be strongly dependent on AA release, which further determined cell culture performance and tissue reconstruction efficiency. With the optimum AA content in carrier materials, intrastromally implanted cell/cryogel constructs exhibited better capability to enhance tissue matrix regeneration and transparency maintenance as well as to mitigate corneal damage in an alkali burn-induced animal model. It is concluded that understanding of antioxidant molecule-mediated structure-property-function interrelationships in gelatin/AA cryogels is critical to designing carrier materials for potential use in corneal stromal tissue engineering. STATEMENT OF SIGNIFICANCE: Multifunctional cryogel material can offer an ideal hospitable environment for cell-mediated tissue reconstruction. To our knowledge, this is the first report describing the use of gelatin/ascorbic acid (AA) cryogels as keratocyte carriers for corneal stromal tissue engineering. The AA loading during cryogel fabrication is found to have a significant effect on cross-linking degree and pore dimension, mechanical and biological stability, ocular biocompatibility, cell culture performance, and cytoprotective activity, giving comprehensive insight into fine-tuning the structure-property-function interrelationships of keratocyte carrier material. Using an alkali burn-induced animal model, we present evidence that with the optimum AA loading into cryogel materials, intrastromally implanted cell/carrier constructs exhibited better capability to enhance tissue matrix regeneration and transparency maintenance as well as to mitigate corneal damage.

In vivo Pharmacological Evaluations of Pilocarpine-Loaded Antioxidant-Functionalized Biodegradable Thermogels in Glaucomatous Rabbits.

To alleviate oxidative stress-induced ocular hypertension, grafting of antioxidant molecules to drug carriers enables a dual-function mechanism to effectively treat glaucomatous intraocular pressure (IOP) dysregulation. Providing potential application for intracameral administration of antiglaucoma medications, this study, for the first time, aims to examine in vivo pharmacological efficacy of pilocarpine-loaded antioxidant-functionalized biodegradable thermogels in glaucomatous rabbits. A series of gallic acid (GA)-grafted gelatin-g-poly(N-isopropylacrylamide) (GN) polymers were synthesized via redox reactions at 20-50 °C. Our results showed that raising redox radical initiation reaction temperature maximizes GA grafting level, antioxidant activity, and water content at 40 °C. Meanwhile, increase in overall hydrophilicity of GNGA carriers leads to fast polymer degradation and early pilocarpine depletion in vivo, which is disadvantageous to offer necessary pharmacological performance at prolonged time. By contrast, sustained therapeutic drug concentrations in aqueous humor can be achieved for long-term (i.e., 28 days) protection against corneal aberration and retinal injury after pilocarpine delivery using dual-function optimized carriers synthesized at 30 °C. The GA-functionalized injectable hydrogels are also found to contribute significantly to enhancement of retinal antioxidant defense system and preservation of histological structure and electrophysiological function, thereby supporting the benefits of drug-containing antioxidant biodegradable thermogels to prevent glaucoma development.

Relationships between mechanical properties and drug release from electrospun fibers of PCL and PLGA blends.

Electrospun nanofibers have the potential to achieve high drug loading and the ability to sustain drug release. Mechanical properties of the drug-incorporated fibers suggest the importance of drug-polymer interactions. In this study, we investigated the mechanical properties of electrospun polycaprolactone (PCL) and poly (D,L-lactic-co-glycolic) acid (PLGA) fibers at various blend ratios in the presence and absence of a small molecule hydrophilic drug, tenofovir (TFV). Young׳s modulus of the blend fibers showed dependence on PLGA content and the addition of the drug. At a PCL/PLGA (20/80) composition, Young׳s modulus and tensile strength were independent of drug loading up to 40wt% due to offsetting effects from drug-polymer interactions. In vitro drug release studies suggested that release of TFV significantly decreased fiber mechanical properties. In addition, mechanically stretched fibers displayed a faster release rate as compared to the non-stretched fibers. Finally, drug partition in the blend fibers was estimated using a mechanical model and then experimentally confirmed with a composite of individually stacked fiber meshes. This work provides scientific understanding on the dependence of drug release and drug loading on the mechanical properties of drug-eluting fibers.

Role of solvent-mediated carbodiimide cross-linking in fabrication of electrospun gelatin nanofibrous membranes as ophthalmic biomaterials.

Due to their ability to mimic the structure of extracellular matrix, electrospun gelatin nanofibers are promising cell scaffolding materials for tissue engineering applications. However, the hydrophilic gelatin molecules usually need stabilization before use in aqueous physiological environment. Considering that biomaterials cross-linked via film immersion technique may have a more homogeneous cross-linked structure than vapor phase cross-linking, this work aims to investigate the chemical modification of electrospun gelatin nanofibrous membranes by liquid phase carbodiimide in the presence of ethanol/water co-solvents with varying ethanol concentrations ranging from 80 to 99.5vol%. The results of characterization showed that increasing water content in the binary reaction solvent system increases the extent of cross-linking of gelatin nanofibers, but simultaneously promotes the effect of biopolymer swelling and distortion in fiber mat structure. As compared to non-cross-linked counterparts, carbodiimide treated gelatin nanofibrous mats exhibited better thermal and biological stability where the shrinkage temperature and resistance to enzymatic degradation varied in response to ethanol/water solvent composition-mediated generation of cross-links. Irrespective of their cross-linking density, all studied membrane samples did not induce any responses in ocular epithelial cell cultures derived from cornea, lens, and retina. Unlike many other cross-linking agents and/or methods (e.g., excessive vapor phase cross-linking) that may pose a risk of toxicity, our study demonstrated that these nanofibrous materials are well tolerated by anterior segment tissues. These findings also indicate the safety of using ethanol/water co-solvents for chemical cross-linking of gelatin to engineer nanofibrous materials with negligible biological effects. In summary, the present results suggest the importance of solvent-mediated carbodiimide cross-linking in modulating structure-property relationship without compromising in vitro and in vivo biocompatibility of electrospun gelatin nanofibers for future ophthalmic applications.

On the importance of Bloom number of gelatin to the development of biodegradable in situ gelling copolymers for intracameral drug delivery.

To overcome the drawbacks associated with conventional antiglaucoma eye drops, this work demonstrated the feasibility of an effective alternative strategy to administer pilocarpine directly via intracameral injections of drug-containing biodegradable in situ gelling GN copolymers composed of gelatin and poly(N-isopropylacrylamide). Specifically, this study aims to understand the importance of Bloom number of gelatin, a physicochemical parameter, to the development of GN carriers for intracameral drug delivery in glaucoma therapy. Our results showed that both imino acid and triple-helix contents increased with increasing Bloom index from 75-100 to 300. The drug encapsulation efficiency in response to temperature-triggered phase transition in GN copolymers was affected by the Bloom index of gelatin. In addition, the differences in protein secondary structure significantly influenced the degradation rates of GN carriers, which were highly correlated with drug release profiles. The increase in released pilocarpine concentration led to a high intracellular calcium level in rabbit ciliary smooth muscle cell cultures, indicating a beneficial pharmacological response to a drug. Irrespective of Bloom number of gelatin, all carrier materials exhibited excellent in vitro and in vivo biocompatibility with corneal endothelium. In a glaucomatous rabbit model, intracameral injections of pilocarpine-containing GN synthesized from gelatins with various Bloom numbers had different abilities to improve ocular hypertension and induce pupillary constriction, indicating distinct antiglaucoma efficacies due to in vivo drug release. It is concluded that the effects on pharmacological treatment using GN carriers for intracameral pilocarpine administration demonstrate a strong dependence on the Bloom number of gelatin.

Gallic acid grafting effect on delivery performance and antiglaucoma efficacy of antioxidant-functionalized intracameral pilocarpine carriers.

UNLABELLED: Functionalization of therapeutic carrier biomaterials can potentially provide additional benefits in drug delivery for disease treatment. Given that this modification determines final therapeutic efficacy of drug carriers, here, we investigate systematically the role of grafting amount of antioxidant gallic acid (GA) onto GN in situ gelling copolymers made of biodegradable gelatin and thermo-responsive poly(N-isopropylacrylamide) for intracameral delivery of pilocarpine in antiglaucoma treatment. As expected, increasing redox reaction time increased total antioxidant activities and free radical scavenging abilities of synthesized carrier biomaterials. The hydrophilic nature of antioxidant molecules strongly affected physicochemical properties of carrier materials with varying GA grafting amounts, thereby dictating in vitro release behaviors and mechanisms of pilocarpine. In vitro oxidative stress challenges revealed that biocompatible carriers with high GA content alleviated lens epithelial cell damage and reduced reactive oxygen species. Intraocular pressure and pupil diameter in glaucomatous rabbits showed correlations with GA-mediated release of pilocarpine. Additionally, enhanced pharmacological treatment effects prevented corneal endothelial cell loss during disease progression. Increasing GA content increased total antioxidant level and decreased nitrite level in the aqueous humor, suggesting a much improved antioxidant status in glaucomatous eyes. This work significantly highlights the dependence of physicochemical properties, drug release behaviors, and bioactivities on intrinsic antioxidant capacities of therapeutic carrier biomaterials for glaucoma treatment. STATEMENT OF SIGNIFICANCE: Development of injectable biodegradable polymer depots and functionalization of carrier biomaterials with antioxidant can potentially provide benefits such as improved bioavailability, controlled release pattern, and increased therapeutic effect in intracameral pilocarpine administration for glaucoma treatment. For the first time, this study demonstrated that the biodegradable in situ gelling copolymers can incorporate different levels of antioxidant gallic acid to tailor the structure-property-function relationship of the intracameral drug delivery system. The systematic evaluation fully verified the dependence of phase transition, degradation behavior, drug release mechanism, and antiglaucoma efficacy on intrinsic antioxidant capacities of carrier biomaterials. The report highlights the significant role of grafting amount of gallic acid in optimizing performance of antioxidant-functionalized polymer therapeutics as new drug delivery platforms in disease treatment.

Coaxially electrospun fiber-based microbicides facilitate broadly tunable release of maraviroc.

Electrospun fibers show potential as a topical delivery system for vaginal microbicides. Previous reports have demonstrated delivery of anti-HIV and anti-STI (sexually transmitted infection) agents from fibers formulated using hydrophilic, hydrophobic, or pH-responsive polymers that result in rapid, prolonged, or stimuli-responsive release, respectively. However, coaxial electrospun fibers have yet to be evaluated as a highly tunable microbicide delivery vehicle. In this research, we explored the opportunities and limitations of a model coaxial electrospun fiber system to provide broad and tunable release rates for the HIV entry inhibitor maraviroc. Specifically, we prepared ethyl cellulose (EC)-shell and polyvinylpyrrolidone (PVP)-core fibers that were capable of releasing actives over a range of hours to several days. We further demonstrated simple and effective methods for combining core-shell fibers with rapid-release formulations to provide combined instantaneous and sustained maraviroc release. In addition, we investigated the effect of varying release media on maraviroc release from core-shell fibers, and found that release was strongly influenced by media surface tension and drug ionization. Finally, in vitro cell culture studies show that our fiber formulations were not cytotoxic and that electrospun maraviroc maintained similar antiviral activity compared to neat maraviroc.

Dependence of corneal keratocyte adhesion, spreading, and integrin ß1 expression on deacetylated chitosan coating.

This study reports, for the first time, the regulation of corneal keratocyte adhesion, spreading, morphology, and integrin gene expression on chitosan coating due to the effects of deacetylation. The degree of deacetylation (DD) in chitosan materials was confirmed by elemental analysis, gel permeation chromatography, and Fourier transform infrared spectroscopy. In this study, chitosan samples with the same molecular weight level but varying DD (74.1 ± 0.5%, 84.4 ± 0.7%, and 94.2 ± 0.5%) were obtained by heat-alkaline treatment under a nitrogen atmosphere. For higher DD groups, the biopolymer carried abundant amino groups since the deacetylation process removed larger amount of acetyl groups from the chitosan molecules. Results showed that the mechanical stability and crystallinity of the chitosan coatings significantly increased with increasing DD value. Fibronectin adsorption, keratocyte adhesion, and cell spreading exhibited a positive correlation with DD due to the chemical functionality of polysaccharides (bearing acetyl and amino groups) and increase of substrate stiffness and crystallinity. In particular, when adhered to chitosan coatings with a DD value of 74.1%, the keratocytes appeared to be fibroblastic, elongated, and spindle shape, indicating a loss of their characteristic dendritic morphology. Furthermore, the gene expression of integrin ß1 (i.e., a cell-matrix adhesion molecule) was significantly up-regulated on the chitosan coatings with higher DD, which supports favorable attachment of corneal keratocytes. Our findings suggest that DD-mediated physicochemical properties of chitosan coatings greatly affect cell-substrate crosstalk during corneal keratocyte cultivation.

Relationships between surface roughness/stiffness of chitosan coatings and fabrication of corneal keratocyte spheroids: Effect of degree of deacetylation.

Fabrication of the cell spheroids from corneal keratocytes has important implications to the advance in tissue engineering while stimulation from the interface of a biopolymer coating has the ability to modulate this event. This study aims to investigate the dependence of keratocyte migration, proliferation, and differentiation on the surface roughness/stiffness of the chitosan coatings through modifications by degree of deacetylation (DD). After a series of deacetylation process, chitosan coatings with increasing DD exhibited significantly decreased surface roughness and increased surface stiffness. Relationships between the behaviors of rabbit corneal keratocytes (RCKs) and biopolymer coatings with varying DDs (between 75% and 96%) were also found during in vitro cultivation. Both the surface roughness increase and stiffness decrease could lead to enhanced cell migration, which is the main driving force for the early stage spheroid formation on chitosan substrates (e.g., within 8h). With these stimulations from the substrate interfaces, the size and morphology of RCK spheroids were greatly affected by the DD of chitosan. When fabricated on a lowered DD of chitosan material, the spheroids had a larger size with abundant extracellular matrix produced around the cells. At a later stage of spheroid cultivation (e.g., 5 days), significantly higher amount of RCKs on chitosan coatings was noted with increasing DD, indicating the substrate interface effects on cell proliferation. The keratocan expression of RCK spheroids grown on a lowered DD of chitosan was up-regulated, suggesting that both the surface roughness increase and stiffness decrease may facilitate the microenvironment for preservation of cellular phenotype. Overall, our work contributes to the scientific understanding of the keratocyte behaviors and spheroid fabrications in response to DD-mediated surface roughness/stiffness of chitosan coatings.