The ability to estimate the diffusion coefficient of a solute within hydrogels has important application in the design and analysis of hydrogels used in drug delivery, tissue engineering, and ...regenerative medicine. A number of mathematical models have been derived for this purpose; however, they often rely on fitted parameters and so have limited predictive capability. Herein we assess the ability of the obstruction-scaling model to provide reasonable estimates of solute diffusion coefficients within hydrogels, as well as the assumption that a hydrogel can be represented as an entangled polymer solution of an equivalent concentration. Fluorescein isothiocyanate dextran solutes were loaded into sodium alginate solutions as well as hydrogels of different polymer volume fractions formed from photoinitiated cross-linking of methacrylate sodium alginate. The tracer diffusion coefficients of these solutes were measured using fluorescence recovery after photobleaching (FRAP). The measured diffusion coefficients were then compared to the values predicted by the obstruction-scaling model. The model predictions were within ±15% of the measured values, suggesting that the model can provide useful estimates of solute diffusion coefficients within hydrogels and solutions. Moreover, solutes diffusing in both sodium alginate solutions and hydrogels were demonstrated to experience the same degree of solute mobility restriction given the same effective polymer concentration, supporting the assumption that a hydrogel can be represented as an entangled polymer solution of equivalent concentration.
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Degenerative intervertebral disc disease is a common source of chronic pain and reduced quality of life in people over the age of 40. While degeneration occurs throughout the disc, it most often ...initiates in the nucleus pulposus (NP). Minimally invasive delivery of NP cells within hydrogels that can restore and maintain the disc height while regenerating the damaged NP tissue is a promising treatment strategy for this condition. Towards this goal, a biohybrid ABA dimethacrylate triblock copolymer was synthesized, possessing a lower critical solution temperature below 37 °C and which contained as its central block an MMP-degradable peptide flanked by poly(trimethylene carbonate) blocks bearing pendant oligoethylene glycol groups. This triblock prepolymer was used to form macroporous NP cell-laden hydrogels via redox initiated (ammonium persulfate/sodium bisulfite) crosslinking, with or without the inclusion of thiolated chondroitin sulfate. The resulting macroporous hydrogels had water and mechanical properties similar to those of human NP tissue and were mechanically resilient. The hydrogels supported NP cell attachment and growth over 28 days in hypoxic culture. In hydrogels prepared with the triblock copolymer but without the chondroitin sulfate the NP cells were distributed homogeneously throughout in clusters and deposited collagen type II and sulfated glycosaminoglycans but not collagen type I. This hydrogel formulation warrants further investigation as a cell delivery vehicle to regenerate degenerated NP tissue.
The intervertebral disc between the vertebral bones of the spine consists of three regions: a gel-like central nucleus pulposus (NP) within the annulus fibrosis, and bony endplates. Degeneration of the intervertebral disc is a source of chronic pain in the elderly and most commonly initiates in the NP. Replacement of degenerated NP tissue with a NP cell-laden hydrogel is a promising treatment strategy. Herein we demonstrate that a crosslinkable polymer with a lower critical solution temperature below 37 °C can be used to form macroporous hydrogels for this purpose. The hydrogels are capable of supporting NP cells, which deposit collagen II and sulfated glycosaminoglycans, while also possessing mechanical properties matching those of human NP tissue.
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Specialized pro-resolving mediators (SPMs) are being considered for the treatment of chronic inflammatory diseases. However, these polyunsaturated fatty acids are prone to oxidation ...and as a result have a short biological half-life. It was reasoned that a micelle formulation would provide sustained delivery of SPMs while providing protection from oxidation. Thus, micelle formulations were prepared with poly(ethylene glycol) (PEG) as the hydrophilic block and poly(trimethylene carbonate) (PT) containing unsaturated pendant groups, specifically benzyloxy (BT) and sorbate (ST) groups, as the hydrophobic block. The potential of these micelles was assessed using linoleic acid as a model SPM. Loading into a micelle core reduced the extent of oxidation of the model SPM and a sustained release of non-oxidized model drug was achieved for up to 20 days in vitro from the PEG-P(T-BT) micelles. These micelles were also non-cytotoxic over a wide concentration range, demonstrating the potential of this formulation for effective SPM release in vivo.
Abstract Decellularized adipose tissue (DAT) has shown promise as an adipogenic bioscaffold for soft tissue augmentation and reconstruction. The objective of the current study was to investigate the ...effects of allogeneic adipose-derived stem/stromal cells (ASCs) on in vivo fat regeneration in DAT bioscaffolds using an immunocompetent rat model. ASC seeding significantly enhanced angiogenesis and adipogenesis, with cell tracking studies indicating that the newly-forming tissues were host-derived. Incorporating ASCs also mediated the inflammatory response and promoted a more constructive macrophage phenotype. A fraction of the CD163+ macrophages in the implants expressed adipogenic markers, with higher levels of this “adipocyte-like” phenotype in proximity to the developing adipose tissues. Our results indicate that the combination of ASCs and adipose extracellular matrix (ECM) provides an inductive microenvironment for adipose regeneration mediated by infiltrating host cell populations. The DAT scaffolds are a useful tissue-specific model system for investigating the mechanisms of in vivo adipogenesis that may help to develop a better understanding of this complex process in the context of both regeneration and disease. Overall, combining adipose-derived matrices with ASCs is a highly promising approach for the in situ regeneration of host-derived adipose tissue.
Melt electrospinning writing (MEW) is an emerging additive manufacturing technique that enables the design and fabrication of micrometer-thin fibrous scaffolds made of biocompatible and biodegradable ...polymers. By using a computer-aided deposition process, a unique control over pore size and interconnectivity of the resulting scaffolds is achieved, features highly interesting for tissue engineering applications. However, MEW has been mainly used to process low melting point thermoplastics such as poly(ε-caprolactone). Since this polymer exhibits creep and a reduction in modulus upon hydration, we manufactured scaffolds of poly(l-lactide-co-ε-caprolactone-co-acryloyl carbonate) (poly(LLA-ε-CL-AC)), a photo-cross-linkable and biodegradable polymer, for the first time. We show that the stiffness of the scaffolds increases significantly (up to ∼10-fold) after cross-linking by UV irradiation at room temperature, compared with un-cross-linked microfiber scaffolds. The preservation of stiffness and high average fiber modulus (370 ± 166 MPa) within the cross-linked hydrated scaffolds upon repetitive loading (10% strain at 1 Hz up to 200,000 cycles) suggests that the prepared scaffolds may be of potential interest for soft connective tissue engineering applications. Moreover, the approach can be readily adapted through manipulation of polymer properties and scaffold geometry to prepare structures with mechanical properties suitable for other tissue engineering applications.
A mathematical model that can provide good predictions of the solute diffusion coefficient in hydrogels would be highly beneficial in designing hydrogels for biomedical and industrial applications, ...and a number of such models have been derived. Mesh size plays a prominent role in determining the solute diffusion coefficient within a hydrogel. However, in assessing the predictive ability of models derived for this purpose, we have employed various values of the mesh size, i.e., the correlation length or the mesh radius. Herein, a systematic examination of the use of the correlation length or the mesh radius as the mesh size was performed in assessing the predictive quality of four recent models: a semiempirical Cukier hydrodynamic model, an obstruction model, an obstruction-exclusion model, and a combined free volume/obstruction model. The use of the correlation length as the mesh size along with the obstruction model yielded the most consistent agreement between experimental data and model predictions. In contrast, use of the mesh radius did not yield good agreement with the experimental data when used with any of the models.
Poly(5-hydroxy-trimethylene carbonate) (PHTMC) was synthesized through ring-opening polymerization of 5-benzyloxy trimethylene carbonate followed by debenzylation. The water solubility of the ...resulting homopolymer was measured as a function of its molecular weight, and its degradation in acidic (pH 2.0), and basic (pH 12.0) solutions of deionized water, as well as in phosphate buffered saline (PBS, pH 7.4) assessed. PHTMC exhibited notable hydrophilicity, even at molecular weights as high as 49 kDa. The pendant hydroxyl functional groups also allowed PHTMC to degrade rapidly (<1 h) under basic conditions via an intramolecular cyclization process along the polymer backbone. The ultimate degradation products were the biologically benign glycerol and carbon dioxide.
•Poly(5-hydroxy trimethylene carbonate) (PHTMC) exhibits appreciable water solubility.•Under alkaline aqueous conditions, PHTMC degrades rapidly through an intramolecular cyclization reaction.•The resulting degradation products are glycerol carbonate, carbon dioxide and glycerol.•The glycerol carbonate in turn gradually hydrolyses to glycerol and carbon dioxide.
Abstract Poly(trimethylene carbonate) (PTMC) with molecular weights greater than 100 kg/mol is known to degrade readily in vivo while PTMC of less than 70 kg/mol is resistant to degradation. The ...reason for the molecular weight dependent degradation rate of PTMC is unclear, and may be due to differences in macrophage behavior or enzyme adsorption or activity. Macrophage number and production of reactive oxygen species (ROS) and esterase were measured when cultured on 60 and 100 kg/mol PTMC. Cholesterol esterase and lipase were adsorbed to 60 and 100 kg/mol PTMC and mass and viscoelastic properties of the adsorbed enzyme layers were measured. No significant differences were observed in macrophage number or production of degradative species. Significant differences were measured in mass, shear modulus and viscosity of the adsorbed cholesterol esterase layer, suggesting that the cholesterol esterase is adsorbing in a different conformation on the 60 and 100 kg/mol PTMC. Despite similar bulk moduli, the surface modulus of 60 kg/mol PTMC was significantly lower than 100 kg/mol. It is proposed that the difference in surface stiffness and polymer chain flexibility affect the arrangement of water bound to and freed from the polymer chains during adsorption, thus affecting enzymatic adsorption, conformation, and activity.
Abstract An injectable tissue-engineered adipose substitute that could be used to deliver adipose-derived stem cells (ASCs), filling irregular defects and stimulating natural soft tissue ...regeneration, would have significant value in plastic and reconstructive surgery. With this focus, the primary aim of the current study was to characterize the response of human ASCs encapsulated within three-dimensional bioscaffolds incorporating decellularized adipose tissue (DAT) as a bioactive matrix within photo-cross-linkable methacrylated glycol chitosan (MGC) or methacrylated chondroitin sulphate (MCS) delivery vehicles. Stable MGC- and MCS-based composite scaffolds were fabricated containing up to 5 wt% cryomilled DAT through initiation with long-wavelength ultraviolet light. The encapsulation strategy allows for tuning of the 3-D microenvironment and provides an effective method of cell delivery with high seeding efficiency and uniformity, which could be adapted as a minimally-invasive in situ approach. Through in vitro cell culture studies, human ASCs were assessed over 14 days in terms of viability, glycerol-3-phosphate dehydrogenase (GPDH) enzyme activity, adipogenic gene expression and intracellular lipid accumulation. In all of the composites, the DAT functioned as a cell-supportive matrix that enhanced ASC viability, retention and adipogenesis within the gels. The choice of hydrogel also influenced the cell response, with significantly higher viability and adipogenic differentiation observed in the MCS composites containing 5 wt% DAT. In vivo analysis in a subcutaneous Wistar rat model at 1, 4 and 12 weeks showed superior implant integration and adipogenesis in the MCS-based composites, with allogenic ASCs promoting cell infiltration, angiogenesis and ultimately, fat formation.
Abstract Myocardial infarction (MI) is one of the leading causes of mortality worldwide and is associated with irreversible cardiomyocyte death and pathological remodeling of cardiac tissue. In the ...past 15 years, several animal models have been developed for pre-clinical testing to assess the potential of stem cells for functional tissue regeneration and the attenuation of left ventricular remodeling. The promising results obtained in terms of improved cardiac function, neo-angiogenesis and reduction in infarct size have motivated the initiation of clinical trials in humans. Despite the potential, the results of these studies have highlighted that the effective delivery and retention of viable cells within the heart remain significant challenges that have limited the therapeutic efficacy of cell-based therapies for treating the ischemic myocardium. In this review, we discuss key elements for designing clinically translatable cell-delivery approaches to promote myocardial regeneration. Key topics addressed include cell selection, with a focus on mesenchymal stem cells derived from the bone marrow (bMSCs) and adipose tissue (ASCs), including a discussion of their potential mechanisms of action. Natural and synthetic biomaterials that have been investigated as injectable cell delivery vehicles for cardiac applications are critically reviewed, including an analysis of the role of the biomaterials themselves in the therapeutic scheme.
