ABSTRACT
Temperature-responsive polymers are attractive candidates for applications related to injectable delivery of biologically active therapeutics, such as stem cells. In this study, we evaluate the potential of thermosensitive hydroxybutyl chitosan (HBC) as a biomaterial for the culture of human mesenchymal stem cells (hMSC) and cells derived from the intervertebral disk, with the eventual goal of using the HBC polymer as an injectable matrix/cell therapeutic. Conjugation of hydroxybutyl groups to chitosan renders the polymer water soluble and thermally responsive. Below its lower critical solution temperature, a solution of HBC can be maintained indefinitely in its solvated state. Upon exposure to a 37 degrees C environment, within 60 s, a 3.8 wt% HBC solution rapidly forms a gel that can be maneuvered with forceps. Upon cooling, the gel once again is able to revert to its solvated state. The gel exhibits a dramatic increase in both G' and G'' with increasing temperature, signifying a temperature-dependent enhancement of gel mechanical properties. Although a solid structure upon gelation, due to its physical nature of polymer interaction and gel formation, the gel exhibits a fluid-like viscoelastic behavior when exposed to shear stresses of up to 10% strain, with both G' and G'' approaching zero with increasing shear stress. Formulations of HBC gels presented in this study have gelation temperatures ranging from 13.0 to 34.6 degrees C and water contents of 67-95%. Minimal cytotoxicity in MSC and disk cell cultures was observed with these polymers up to a concentration of 5 wt%. Detection of metabolic activity, genetic analysis of synthesized mRNA, and histological staining of MSC and disk cell cultures in these gels collectively indicate cell proliferation without a loss in metabolic activity and extracellular matrix production. This study suggests the potential of HBC gel as an injectable carrier for future applications of delivering therapeutics to encourage a biologically relevant reconstruction of the degenerated disk.
Subject(s)
Butanols/chemistry , Chitosan/pharmacology , Intervertebral Disc/cytology , Mesenchymal Stem Cells/cytology , Tissue Culture Techniques/methods , Aggrecans , Cell Proliferation/drug effects , Cell Survival/drug effects , Chitosan/analogs & derivatives , Chitosan/chemistry , Chondrocytes/cytology , Chondrocytes/drug effects , Chondrocytes/metabolism , Collagen/chemistry , Collagen Type I/genetics , Extracellular Matrix Proteins/genetics , Gels , Gene Expression/drug effects , Gene Expression/genetics , Humans , Lectins, C-Type/genetics , Mesenchymal Stem Cells/drug effects , Molecular Weight , Proteoglycans/genetics , Rheology , Transition Temperature , Water/chemistryABSTRACT
This paper describes the pH and temperature effects on drug release from polyelectrolyte complex (PEC) films composed of a cationic polymer, chitosan, and an anionic polymer, polyalkyleneoxide-maleic acid copolymer (PAOMA). In this study, we prepared and investigated PEC films in terms of the drug release properties as pH- and temperature-sensitive drug carriers. Drug release rates were tested at pH 3.8 and 7.2, and at 25 and 50 degrees C. Salicylic acid and phenol were selected as model drugs. An increase in pH from 3.8 to 7.2 resulted in an increase in the rate of drug release because of the repulsive forces between carboxyl groups in PAOMA and anionic groups in model drugs. When the hydrophobic PAOMA was used as a polyanion, the drug release rate increased at 50 degrees C. This is attributed to the increase of release area due to the phase transition of PAOMA and the increase of repulsive forces between carboxyl groups in PAOMA and anionic groups in model drugs.
Subject(s)
Chitosan/pharmacokinetics , Maleates/pharmacokinetics , Polymers/pharmacokinetics , Temperature , Chitosan/chemistry , Electrochemistry , Hydrogen-Ion Concentration , Maleates/chemistry , Microscopy, Electron, Scanning , Polyelectrolytes , Polymers/chemistry , Polysaccharides/chemistry , Polysaccharides/pharmacokinetics , Polysaccharides/ultrastructureABSTRACT
Polyelectrolyte complex films were prepared with polyethyleneoxide-maleic acid copolymer and chitosan using a casting/solvent evaporation method. The films were examined in terms of their IR spectra, surface and cross-section morphologies, cytotoxicity, and swelling behavior at different pH levels. To assess the potential of these films as a biomedical device, the profiles of the release of model drug from the CS/PEOMA films were examined at pH 4.8. The surface morphology of the films was quite smooth and uniform, and the cross-sectional morphology was dense and homogeneous. The swelling behaviors of CS/PEOMA films were found to depend on the pH of the solution as well as on the CS/PEOMA composition. Drug release from different CS/PEOMA films at pH 4.8 was found to be dependent on film composition. The results showed the potential applicability of CS/PEOMA film as a drug delivery vehicle.
