Long-term sustained-released in situ gels of a water-insoluble drug amphotericin B for mycotic arthritis intra-articular administration: preparation,in vitro and in vivo evaluation

Amphotericin B (AMB) was often used in intra-articular injection administration for fungal arthritis, because it could often bring a satisfactory therapeutic efficacy and a minimum systemic toxic side effect. However, because of the multiple operations and the frequent injections, the compliance of the patients was bad. Therefore, to develop a long-term sustained-released preparation of AMB for mycotic arthritis intra-articular administration is of great significance. The purpose of present study was to develop a long-term sustained-released in situ gel of a water-insoluble drug AMB for mycotic arthritis intra-articular administration. Based on the evaluations of the in vitro properties of the formulations, the formulation containing 10% (w/w) ethanol, 15% (w/w) PG, 0.75% (w/w) HA, 5% (w/w) purified soybean oil, 0.03% (w/w) α-tocopherol, 15% (w/w) water and 55% (w/w) glyceryl monooleate was selected as a suitable intra-articular injectable in situ gel drug delivery system for water-insoluble drug AMB. Furthermore, the results of the in vivo study on rabbits showed that the selected formulation was a safe and effective long-term sustained-released intra-articular injectable AMB preparation. Therefore, the presented in situ AMB gel could reduce the frequency of the administration in the AMB treatment of fungal arthritis, and then would get a good patient compliance.     

Biodegradable hollow microfibres to produce bioactive scaffolds

Biodegradable hollow microfibres containing particles loaded with specific active agents can be potentially employed to produce a special kind of substrate for tissue engineering, able to function as a scaffold and at the same time to act as a drug‐releasing system. Biodegradable hollow microfibres based on poly(lactic acid) were produced by a dry–wet spinning procedure. Drug‐loaded microparticles were prepared by a simple oil‐in‐water emulsion and entrapped inside the fibres. The morphology of both fibres and particles was investigated by scanning electron microscopy. The mechanical and thermal properties of the fibres were investigated by tensile tests and differential scanning calorimetry. In vitro tests were performed to evaluate the release of the drug from the fibres loaded with the particles Copyright © 2004 Society of Chemical Industry     

A novel direct aerodynamically assisted threading methodology for generating biologically viable microthreads encapsulating living primary cells

In a recent discovery, coaxial electrospinning was explored to encapsulate living organisms within a continuous bio‐polymeric microthread from which active biological scaffolds were fabricated (Townsend‐Nicholson and Jayasinghe, Biomacromolecules 2006, 7, 3364). The cells were demonstrated to have gone through all expected cellular activity without their viability being compromised. These biologically active threads and scaffolds have direct and tremendous applicability from regenerative to therapeutic medicine. Currently these post‐processed cells as composite threads and scaffolds are being investigated in‐depth at a cellular level to establish if the processing methodology has any affect on the cellular make‐up. We now demonstrate a competing non‐electric field driven approach for fabricating composite threads and scaffolds influenced only by a differential pressure. We refer to this novel composite thread to scaffold fabrication methodology as coaxial aerodynamically assisted bio‐threading (CAABT). Our investigations firstly, demonstrate that this technique can process handle living organisms without biologically perturbing them in anyway. Secondly the process is elucidated as possessing the ability to form composite active threads from which biologically viable scaffolds are formed. Finally our study employs florescent activated cell sorting (FACScan), a method by which the cellular dynamics and viability are quantified on control and threaded cellular samples at two prescribed time points. In parallel with FACScan, optical comparison of cellular morphology at three time points within a period of three weeks is carried out to photographically observe any changes in the post‐processed cellular phenotype. Our developmental investigations into this novel aerodynamically assisted threading methodology has unearthed a unique biomicrofabrication approach, which joins cell electrospinning in the cell threading to scaffold fabrication endeavor. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Coaxial Alginate Hydrogels: From Self-Assembled 3D Cellular Constructs to Long-Term Storage

Alginate as a versatile naturally occurring biomaterial has found widespread use in the biomedical field due to its unique features such as biocompatibility and biodegradability. The ability of its semipermeable hydrogels to provide a favourable microenvironment for clinically relevant cells made alginate encapsulation a leading technology for immunoisolation, 3D culture, cryopreservation as well as cell and drug delivery. The aim of this work is the evaluation of structural properties and swelling behaviour of the core-shell capsules for the encapsulation of multipotent stromal cells (MSCs), their 3D culture and cryopreservation using slow freezing. The cells were encapsulated in core-shell capsules using coaxial electrospraying, cultured for 35 days and cryopreserved. Cell viability, metabolic activity and cell–cell interactions were analysed. Cryopreservation of MSCs-laden core-shell capsules was performed according to parameters pre-selected on cell-free capsules. The results suggest that core-shell capsules produced from the low viscosity high-G alginate are superior to high-M ones in terms of stability during in vitro culture, as well as to solid beads in terms of promoting formation of viable self-assembled cellular structures and maintenance of MSCs functionality on a long-term basis. The application of 0.3 M sucrose demonstrated a beneficial effect on the integrity of capsules and viability of formed 3D cell assemblies, as compared to 10% dimethyl sulfoxide (DMSO) alone. The proposed workflow from the preparation of core-shell capsules with self-assembled cellular structures to the cryopreservation appears to be a promising strategy for their off-the-shelf availability.     

Role of Scaffolds,Subchondral, Intra-Articular Injections of Fresh Autologous Bone Marrow Concentrate Regenerative Cells in Treating Human Knee Cartilage Lesions: Different Approaches and Different Results

The value of bone marrow aspirate concentrates for treatment of human knee cartilage lesions is unclear. Most of the studies were performed with intra-articular injections. However, subchondral bone plays an important role in the progression of osteoarthritis. We investigated by a literature review whether joint, subchondral bone, or/and scaffolds implantation of fresh autologous bone marrow aspirate concentrated (BMAC) containing mesenchymal stem cells (MSCs) would improve osteoarthritis (OA). There is in vivo evidence that suggests that all these different approaches (intra-articular injections, subchondral implantation, scaffolds loaded with BMAC) can improve the patient. This review analyzes the evidence for each different approach to treat OA. We found that the use of intra-articular injections resulted in a significant relief of pain symptoms in the short term and was maintained in 12 months. However, the clinical trials indicate that the application of autologous bone marrow concentrates in combination with scaffolds or in injection in the subchondral bone was superior to intra-articular injection for long-term results. The tendency of MSCs to differentiate into fibrocartilage affecting the outcome was a common issue faced by all the studies when biopsies were performed, except for scaffolds implantation in which some hyaline cartilage was found. The review suggests also that both implantation of subchondral BMAC and scaffolds loaded with BMAC could reduce the need for further surgery.     

Glass powders and reactive silicone binder: Interactions and application to additive manufacturing of bioactive glass-ceramic scaffolds

A novel concept for the additive manufacturing of three-dimensional glass-ceramic scaffolds, to be used for tissue engineering applications, was based on fine glass powders mixed with a reactive binder, in the form of a commercial silicone. The powders consisted of ‘silica-defective glass’ specifically designed to react, upon firing in air, with the amorphous silica yielded by the binder. By silica incorporation, the glass was intended to reach the composition of an already known CaONa₂OB₂O₃SiO₂ system. Silica from the binder provided up to 15 wt% of the total silica. With the same overall formulation, silicone-glass powder mixtures led to nearly the same phase assemblage formed by the reference system, crystallizing into wollastonite (CaSiO₃) and Ca-borate (CaB₂O₄). Samples from silicone-glass powder mixtures exhibited an excellent shape retention after firing, which was later exploited in highly porous reticulated scaffolds, obtained by means of direct ink writing (DIW).     

Specially elaborated thermally induced phase separation to fabricate poly(L‐lactic acid) scaffolds with ultra large pores and good interconnectivity

Poly(L ‐lactic acid) (PLLA) scaffolds with pore diameters from several micrometers to ～300 μm were fabricated by a specially elaborated thermally induced phase separation technique. Two different coarsening protocols, i.e., normal coarsening and multi‐step coarsening were compared in consideration of phase separation and domain growth. A normal coarsening route produced scaffolds with pore size from several micrometers to 150 μm depending on the coarsening time after phase separation, accompanying with the emergence of isolated pores at long time coarsening. Scaffolds with large pores with size up to ～300 μm were fabricated by the two‐step coarsening technique, e.g., the PLLA‐solvent (dioxane/water) system was coarsened at a temperature after phase separation for a period, followed by coarsening at a lower temperature for another period. In parallel with formation of the large pores, the interconnectivity between pores was also improved, which was evidenced by scanning electron microscopy, gelatin solution pervasion, and collagen entrapment. The present technique provides the ability to produce scaffolds with high purity, controllable microstructures, and ease of modification, and hence can be widely used in tissue engineering field. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 101: 3336–3342, 2006