Characterization of an encapsulated insulin secreting human pancreatic beta cell line in a modular microfluidic device.

Type I diabetes mellitus is characterised by the destruction of the insulin producing beta cells within the pancreas by the immune system. After the success of Edmonton protocol, islet transplantation has shown to be a promising therapy, but with the Achilles´ heel of the need of using immunosuppressive drugs. Currently, cell encapsulation technology represents a real alternative to protect transplanted islets from the host´s immune attack. Although preliminary in vitro studies with encapsulated cells have been traditionally performed under static conditions in terms of viability and efficiency, these static cultures do not represent a close approach to in vivo environments. We have developed and characterised different alginate-poly-l-lysine-alginate (APA) microcapsules loaded with the insulin producing 1.1B4 cell line. Static in vitro studies confirmed a constant insulin secretion and a boost of the secretion when the medium was enriched with glucose. Nevertheless, these results were not completely reproduced in a dynamic system by APA liquefied microcapsules containing 1.1B4 cells. The dynamic culture setting created by a microfluidic device, allowed the determination of the glucose response in APA liquefied microcapsules, showing that dynamic conditions can mimic better physiological in vivo environments.

Benznidazole Nanoformulates: A Chance to Improve Therapeutics for Chagas Disease.

This article describes the characterization of various encapsulated formulations of benznidazole, the current first-line drug for the treatment of Chagas disease. Given the adverse effects of benznidazole, safer formulations of this drug have a great interest. In fact, treatment of Chagas disease with benznidazole has to be discontinued in as much as 20% of cases due to side effects. Furthermore, modification of delivery and formulations could have potential effects on the emergence of drug resistance. The trypanocidal activity of new nanostructured formulations of benznidazole to eliminate Trypanosoma cruzi was studied in vitro as well as their toxicity in two cultured mammalian cell lines (HepG2 and Fibroblasts). Nanoparticles tested included nanostructured lipid carriers, solid lipid nanoparticles, liposomes, quatsomes, and cyclodextrins. The in vitro cytotoxicity of cyclodextrins-benznidazole complexes was significantly lower than that of free benznidazole, whereas their trypanocidal activity was not hampered. These results suggest that nanostructured particles may offer improved therapeutics for Chagas disease.

Hybrid Alginate-Protein-Coated Graphene Oxide Microcapsules Enhance the Functionality of Erythropoietin Secreting C2C12 Myoblasts.

The beneficial effect of combining alginate hydrogel with graphene oxide (GO) on microencapsulated C2C12-myoblast viability has recently been described. However, the commercially available GO lacks homogeneity in size, this parameter being of high relevance for the cell fate in two-dimensional studies. In three-dimensional applications the capacity of this material for binding different kinds of proteins can result in the reduction of de novo released protein that can effectively reach the vicinity of the microcapsules. Undoubtedly, this could be an important hurdle in its clinical use when combined with alginate-PLL microcapsules. Here, we demonstrate that the homogenization of GO nanoparticles is not a mandatory preparation step in order to get the best of this material upon cell microencapsulation. In fact, when the superficial area of these particles is increased, higher amounts of the therapeutic protein erythropoietin (EPO) are adsorbed on their surface. On the other hand, we have been able to improve even more the favorable effects of this graphene derivative on microencapsulated cell viability by forming a protein biocorona. These proteins block the potential binding sites of EPO and, therefore, enhance the amount of therapeutic drug that is released. Finally, we prove that these hybrid alginate-protein-coated GO-microcapsules are functional in vivo.

Multifunctional hydrogel-based scaffold for improving the functionality of encapsulated therapeutic cells and reducing inflammatory response.

Since the introduction of cell immunoisolation as an alternative to protect transplanted cells from host immune attack, much effort has been made to develop this technology into a realistic clinical proposal. Several promising approaches have been investigated to resolve the biotechnological and biosafety challenges related to cell microencapsulation. Here, a multifunctional hydrogel-based scaffold consisting of cell-loaded alginate-poly-l-lysine-alginate (APA) microcapsules and dexamethasone (DXM)-loaded poly(lactic-co-glycolic) acid (PLGA) microspheres embedded in alginate hydrogel is developed and evaluated. Initially, the feasibility of using an alginate hydrogel for enclosing APA microcapsules was studied in a xenogeneic approach. In addition, the performance of the local release of DXM was addressed. The in vitro studies confirmed the correct adaptation of the enclosed cells to the scaffolds in terms of metabolic activity and viability. The posterior implantation of the hydrogel-based scaffolds containing cell-loaded microcapsules revealed that the hematocrit levels were maintained high and constant, and the pericapsular overgrowth was reduced in the DXM-treated rats for at least 2months. This multifunctional scaffold might have a synergistic effect: (1) providing a physical support for APA microcapsules, facilitating administration, ensuring retention and recuperation and preventing dissemination; and (2) reducing post-transplantation inflammation and foreign body reaction, thus prolonging the lifetime of the implant by the continuous and localized release of DXM.

Therapeutic applications of encapsulated cells.

The synergy of some promising advances in the fields of cell therapy and biomaterials together with improvements in the fabrication of more refined and tailored microcapsules for drug delivery have triggered the progress of cell encapsulation technology. Cell microencapsulation involves immobilizing the transplanted cells within a biocompatible scaffold surrounded by a membrane in attempt to isolate the cells from the host immune attack and enhance or prolong their function in vivo. This technology represents one strategy which aims to overcome the present difficulties related to local and systemic controlled release of drugs and growth factors as well as to organ graft rejection and thus the requirements for use of immunomodulatory protocols or immunosuppressive drugs. This chapter gives an overview of the current situation of cell encapsulation technology as a controlled drug delivery system, and the essential requirements of the technology, some of the therapeutic applications, the challenges, and the future directions under investigation are highlighted.

Hydrogel-based scaffolds for enclosing encapsulated therapeutic cells.

Cell encapsulation technology holds promise for the sustained and controlled delivery of different therapeutic proteins. Alginate-poly-L-lysine-alginate (APA) microcapsules represent one of the most widely studied alginate-polycation microcapsules. On the basis of this technology, two types of hydrogel-based scaffolds have been developed and analyzed with the aim of improving the retention and the retrieval of erythropoietin (Epo) secreting cell-loaded microcapsules in the tissue where they are implanted. Furthermore, these hydrogels may help to reduce the post-transplant inflammation and pericapsular fibrotic overgrowth typically observed around capsules. The hydrogel-based scaffolds could be administered as implantable forms (preformed scaffolds) or injectable forms (in situ formed scaffolds). The in vitro studies confirmed the correct adaptation of the enclosed cells to the scaffolds in terms of viability and protein expression. The posterior implantation of the cell-loaded capsules containing hydrogel-based scaffolds in mice revealed that the hematocrit levels were maintained up to 80% for at least 2 months. The histological analysis of the explanted microcapsules performed at that point showed that pericapsular overgrowth was reduced when cell-loaded microcapsules were enclosed in the hydrogels scaffolds. Incorporating microencapsulated cells within hydrogel-based scaffolds may help to improve their administration protocol and retention while reducing post-transplantation inflammation.

A preliminary approach to the repair of myocardial infarction using adipose tissue-derived stem cells encapsulated in magnetic resonance-labelled alginate microspheres in a porcine model.

Adipose tissue-derived stem cells (ASCs) have properties of self-renewal, pluripotency and high proliferative capability that make them useful for the treatment of cardiac ventricular function following ischaemic injury. However, their therapeutic use is limited due to the low retention of the cells at the targeted site. To address this issue, we developed semipermeable membrane microcapsules labelled with Endorem (magnetocapsules) that provide mechanical and immunological immune protection to the cells while maintaining internal cell microenvironment. In addition, the particles allow tracking the presence and migration of injected cells in vivo by Magnetic Resonance Imaging (MRI). Results indicate that after 21 days in culture, the cells encapsulated in the magnetocapsules showed similar viabilities than cells encapsulated in conventional microcapsules. MRI confirmed a gradual loss of the intensity of the iron oxide label in the non-encapsulated Endorem labelled cells, while magnetocapsules were detected throughout the study period, suggesting that cell retention in the myocardium is improved when cells are enclosed within the magnetocapsules. To further evaluate treatment's effect on global cardiac function, MRI determination of infarct size and left ventricular ejection fraction (LVEF) was performed. In vivo results showed no statistically significant differences in heart rate and cardiac output between treatment groups. In conclusion, cells enclosed within magnetocapsules have shown suitable viability and have been detected in vivo throughout the study period. Further studies will evaluate whether increasing cell loading with the particles may help to improve the therapeutic results.

A perspective on bioactive cell microencapsulation.

Bioactive cell encapsulation has emerged as a promising tool for the treatment of patients with various disorders including diabetes mellitus, central nervous system diseases, and cardiovascular diseases. The implantation of encapsulated cells that secrete a therapeutic product (protein, peptide, or antibody) within a semipermeable membrane provides a physical barrier to mask the implant from immune surveillance at a local level without the need for systemic immunosuppression; this serves to achieve a successful therapeutic function following in vivo implantation. The aim of this review article is to provide an update on the progress in this field. The current state of cell encapsulation technology as a controlled drug delivery system will be covered in detail, and the essential requirements of the technology, the challenges, and the future directions under investigation will be highlighted. The technical and biological advances, together with the increasing experience in the field, may lead to the realization of the full potential of bioactive cell encapsulation in the coming years.