Cell Delivery Therapy Research Articles

Hydrogel-Bondable Asymmetric Planar Membranes with Hierarchical Pore Structures for Cell Scaffolding and Encapsulation.

Biomaterials for cell replacement therapy could facilitate the delivery, function, and retrieval of transplanted therapeutic cells. However, the limited capacity to accommodate a sufficient quantity of cells in biomedical devices has hindered the success of clinical application, resulting from the suboptimal spatial organization of cells and insufficient permeation of nutrients in the materials. Herein, through the immersion-precipitation phase transfer (IPPT) process from polyether sulfone (PES), we develop planar asymmetric membranes with a hierarchical pore architecture spanning from nanopores (∼20 nm) in the dense skin and open-ended microchannel arrays with gradient pore size increasing vertically from microns to ∼100 μm. The nanoporous skin would be an ultrathin diffusion barrier, while the microchannels could support high-density cell loading by acting as separate chambers allowing uniform distribution of cells in the scaffold. Alginate hydrogel could permeate into the channels and form a sealing layer after gelation, which could slow down the invasion of host immune cells into the scaffold. The hybrid thin-sheet encapsulation system (∼400 μm thick) could protect allogeneic cells over half-year after intraperitoneal (IP) implantation in immune-competent mice. Such structural membranes and plastic-hydrogel hybrids of thin dimensions could find important applications in cell delivery therapy.

Preparation and evaluation of a temperature-responsive methylcellulose/polyvinyl alcohol hydrogel for stem cell encapsulation

Cell delivery therapy holds great prospects in regenerative medicine. However, low cell survival and retention are still the main problems in current studies. To address these challenges, a temperature-sensitive hydrogel suitable for cell encapsulation was developed by mixing methylcellulose (MC) and polyvinyl alcohol (PVA) solution simply in this work. Based on the physical hydrogen bonds between MC and PVA chains, porous hydrogels with excellent water-retention ability were achieved at 37 °C. The hydrogels exhibited superior performances such as fast-gelling, stable modulus, great injectability, and high light transmittance. Additionally, cell viability analysis showed MC/PVA hydrogels were cytocompatible. More importantly, cell encapsulation experiments demonstrated that rabbit adipose-derived stem cells (rADSCs) cultured within the MC/PVA hydrogels remain in high viability. Taken together, these results indicated the potential of MC/PVA hydrogels as a clinically translatable delivery system for stem cells.

Injectable conductive micro-cryogel as a muscle stem cell carrier improves myogenic proliferation, differentiation and in situ skeletal muscle regeneration

Volumetric muscle loss (VML) results in the impediment of skeletal muscle function, and there were still great challenges in cell delivery approach with the minimally invasive operation to repair muscle defects. To deliver cells to the VML defects site efficiently, the injectable conductive porous nanocomposite microcryogels based on gelatin (GT) and reduced graphene oxide (rGO) were designed and prepared. The microcryogels were loaded with myoblasts to form an injectable cell delivery system and show the ability to protect cells during injection. Conductive microcryogel with 4 mg/mL rGO (GT/rGO4) enhanced C2C12 cell proliferation and myogenic differentiation during 3D culture compared with pure gelatin microcryogel. In a mice VML model, injection of microcryogel loaded with muscle-derived stem cells into the injury site significantly improved the generation of new muscle fibers and blood vessels, and anti-inflammatory properties. The results show that injectable biodegradable conductive microcryogel can be used as muscle stem cell carriers with the potential to maintain cell activity and deliver cells to defective sites, thereby in situ enhancing skeletal muscle regeneration. Statement of significanceVolumetric muscle loss overwhelms the regenerative capacity of skeletal muscle, which results in severe damage to muscle tissues. In the treatment of volumetric muscle loss, conductive niche and muscle stem cells are needed to alleviate excessive scar formation and inflammation to improve muscle regeneration. Injectable gelatin/reduced graphene oxide based nanocomposite microcryogel can enhance the differentiation of seeded muscle stem cells. The improved repair of volumetric muscle loss was achieved via reducing collagen deposition and inflammation in the injected region through the microcryogel cell-delivery therapy, suggesting great potential of the injectable microcryogel as a cell carrier in soft tissue repair.

Endothelial cell secreted VEGF-C enhances NSC VEGFR3 expression and promotes NSC survival

Although delivery of neural stem cell (NSC) as a therapeutic treatment for intracerebral hemorrhage (ICH) provides promise, NSC delivery typically has extremely low survival rates. Here, we investigate endothelial cell (EC) and pericyte (PC) interactions with NSC, where our results demonstrate that EC, and not PC, promote NSC cell proliferation and reduce cytotoxicity under glucose deprivation (GD). Additionally, NSC proliferation was increased upon treatment with EC conditioned media, inhibited with antagonism of VEGFR3. In an NSC + EC co-culture we detected elevated levels of VEGF-C, not seen for NSC cultured alone. Exogenous VEGF-C induced NSC upregulation of VEGFR3, promoted proliferation, and reduced cytotoxicity. Finally, we delivered microbeads containing NSC + EC into a murine ICH cavity, where VEGF-C was increasingly present in the injury site, not seen upon delivery NSC encapsulated alone. These studies demonstrate that EC-secreted VEGF-C may promote NSC survival during injury, enhancing the potential for cell delivery therapies for stroke.

Development and novel design of clustery graphene oxide formed Conductive Silk hydrogel cell vesicle to repair and routine care of myocardial infarction: Investigation of its biological activity for cell delivery applications

Cell delivery therapy is a talented and hopeful strategic approach to repair the damaged cardiac tissues after myocardial infarction. The biocompatible injectable hydrogel with bioactive nanomaterials with effective self-healing capability has been highly required for the cell delivery and regeneration therapy. In the present study, we have developed the biopolymeric and biocompatible self-healing silk protein hydrogel matrix with graphene oxide nanoformulations as cell delivery vehicles for the treatment of myocardial infarction regeneration. The materials combinations, structural interactions, thermal stability and morphology of the designed hydrogel were investigated and elaborated. To improve therapeutic potential of the hydrogel matrix, growth factor was loaded and investigated its biological activity such as cell survival and cell proliferations properties by using endothelial progenitor cells. Collected, these developed materials are excellent vesicle for cell therapy in myocardial infarction.

Dynamic Loading and Tendon Healing Affect Multiscale Tendon Properties and ECM Stress Transmission

The extracellular matrix (ECM) is the primary biomechanical environment that interacts with tendon cells (tenocytes). Stresses applied via muscle contraction during skeletal movement transfer across structural hierarchies to the tenocyte nucleus in native uninjured tendons. Alterations to ECM structural and mechanical properties due to mechanical loading and tissue healing may affect this multiscale strain transfer and stress transmission through the ECM. This study explores the interface between dynamic loading and tendon healing across multiple length scales using living tendon explants. Results show that macroscale mechanical and structural properties are inferior following high magnitude dynamic loading (fatigue) in uninjured living tendon and that these effects propagate to the microscale. Although similar macroscale mechanical effects of dynamic loading are present in healing tendon compared to uninjured tendon, the microscale properties differed greatly during early healing. Regression analysis identified several variables (collagen and nuclear disorganization, cellularity, and F-actin) that directly predict nuclear deformation under loading. Finite element modeling predicted deficits in ECM stress transmission following fatigue loading and during healing. Together, this work identifies the multiscale response of tendon to dynamic loading and healing, and provides new insight into microenvironmental features that tenocytes may experience following injury and after cell delivery therapies.

Tendon healing affects the multiscale mechanical, structural and compositional response of tendon to quasi-static tensile loading.

Tendon experiences a variety of multiscale changes to its extracellular matrix during mechanical loading at the fascicle, fibre and fibril levels. For example, tensile loading of tendon increases its stiffness, with organization of collagen fibres, and increases cell strain in the direction of loading. Although applied macroscale strains correlate to cell and nuclear strains in uninjured tendon, the multiscale response during tendon healing remains unknown and may affect cell mechanosensing and response. Therefore, this study evaluated multiscale structure-function mechanisms in response to quasi-static tensile loading in uninjured and healing tendons. We found that tendon healing affected the macroscale mechanical and structural response to mechanical loading, evidenced by decreases in strain stiffening and collagen fibre realignment. At the micro- and nanoscales, healing resulted in increased collagen fibre disorganization, nuclear disorganization, decreased change in nuclear aspect ratio with loading, and decreased indentation modulus compared to uninjured tendons. Taken together, this work supports a new concept of nuclear strain transfer attenuation during tendon healing and identifies several multiscale properties that may contribute. Our work also provides benchmarks for the biomechanical microenvironments that tendon cells may experience following cell delivery therapies.

Mouse model of endoscopically ablated enteric nervous system

BackgroundCurrent transgenic animal models of Hirschsprung disease are restricted by limited survival and need for special dietary care. We used small animal colonoscopy to produce chemically ablated enteric nervous system in the distal colon and rectum of normal mice. Materials and methodsAdult C57BL/6 mice underwent colonoscopy with submucosal injection of 75–100 μL of saline (n = 2) or 0.002% (n = 2), 0.02% (n = 15), or 0.2% (n = 2) benzalkonium chloride (BAC). Each mouse received 1–3 injections in the distal colon and rectum. Mice were sacrificed on postprocedure day 7 or 28. Injection sites were analyzed histologically and with immunostaining for β-tubulin III. ResultsSubmucosal injection of 0.02% BAC resulted in megacolon and obliteration of 82 ± 8.8% of myenteric ganglia at the injection site on postprocedure day 7 compared with normal colon. This effect was sustained until day 28. Injection of 0.002% BAC had little effect on the myenteric neuronal network at these time points. Multiple injections of 0.002% or 0.02% BAC (up to three injections per mouse) were well tolerated. Injection of 0.2% BAC caused acute toxicity or death. ConclusionsA novel model of chemically ablated enteric nervous system in the mouse colon and rectum is introduced. This model can be valuable in evaluating targeted cell delivery therapies for Hirschsprung disease.

Encapsulation and Survival of a Chondrocyte Cell Line within Xanthan Gum Derivative

A chemical derivative of xanthan gum polysaccharide is investigated as a new artificial matrix for the encapsulation of chondrocytic cells. Toward this goal, a novel micro-droplet generator is developed to produce microcapsules. Microcapsules with an average diameter of 500 µm, smooth surface, and homogeneous size distribution are obtained. ATDC5 cells encapsulated in carboxymethyl xanthan (CMX) microcapsules remain viable and are observed to proliferate for prolonged culture periods with enhanced metabolic activity. Furthermore, retention of the chondrogenic phenotype is exhibited by the cells within CMX, suggesting the ability of this material to be applied in cell-delivery therapies.

Modulating the Gelation Properties of Self-Assembling Peptide Amphiphiles

Peptide amphiphiles (PAs) are self-assembling molecules that form interwoven nanofiber gel networks. They have gained a lot of attention because of their excellent biocompatibility, adaptable peptide structure that allows for specific biochemical functionality, and nanofibrous assembly that mimics natural tissue formation. However, variations in molecule length, charge, and intermolecular bonding between different bioactive PAs cause contrasting mechanical properties. This potentially limits cell-delivery therapies because scaffold durability is needed to withstand the rigors of clinician handling and transport to wound implant sites. Additionally, the mechanical properties have critical influence on cellular behavior, as the elasticity and stiffness of biomaterials have been shown to affect cell spreading, migration, contraction, and differentiation. Several different PAs have been synthesized, each endowed with specific cellular adhesive ligands for directed biological response. We have investigated mechanical means for modulating and stabilizing the gelation properties of PA hydrogels in a controlled manner. A more stable, biologically inert PA (PA-S) was synthesized and combined with each of the bioactive PAs. Molar ratio (M(r) = PA/PA-S) combinations of 3:1, 1:1, and 1:3 were tested. All PA composites were characterized by observed nanostructure and rheological analysis measuring viscoelasticity. It was found that the PAs could be combined to successfully control and stabilize the gelation properties, allowing for a mechanically tunable scaffold with increased durability. Thus, the biological functionality and natural degradability of PAs can be provided in a more physiologically relevant microenvironment using our composite approach to modulate the mechanical properties, thereby improving the vast potential for cell encapsulation and other tissue engineering applications.