Expansion of human bone marrow stromal cells on poly-(DL-lactide-co-glycolide) (PDL LGA) hollow fibres designed for use in skeletal tissue engineering.

Strategies to expand human bone marrow stromal cells (HBMSC) for bone tissue engineering are a key to revolutionising the processes involved in three-dimensional skeletal tissue reconstruction. To facilitate this process we believe the use of biodegradable porous poly(DL-lactide-co-glycolide) (PDL LGA) hollow fibres as a scaffold used in combination with HBMSC to initiate natural bone repair and regeneration offers a potential solution. In this study, the biocompatibility of 75:25 PDL LGA fibres with HBMSC and the capacity of a PDL LGA fibre-associated HBMSC-monolayer to establish an osteogenic phenotype in vivo was examined. A high proportion of HBMSC survived when expanded on PDL LGA fibres for 6 days, with only 10% of the propidium iodide (pI)-labelled population represented in the sub-G1 DNA peak on analysis by flow cytometry. Tracking carboxy-fluorescein diacetate, succinimidyl ester (CFSE)-labelled HBMSC by flow cytometry indicated that HBMSC attachment to the P(DL)LGA fibres does not interfere with their rate of proliferation. Furthermore, in response to osteogenic stimuli, HBMSC expanded on PDL LGA fibres can differentiate, as expected, along the osteogenic lineage with associated alkaline phosphatase activity. Following implantation into SCID mice, osteogenic-conditioned PDL LGA fibre-HBMSC graft resulted in type I collagen deposition and associated bone mineralisation and osteoid formation, as evidenced by immunohistochemistry and histology. These studies provide evidence that porous PDL LGA hollow fibre-HBMSC graft is an innovative biomaterial that offers new approaches to mesenchymal cell expansion, which could be utilised as a scaffold for skeletal tissue generation.

Effects of common sterilization methods on the structure and properties of poly(D,L lactic-co-glycolic acid) scaffolds.

While methods for the production of scaffolds with the appropriate mechanical properties and architecture for tissue engineering are attracting much attention, the effects of subsequent sterilization processes on the scaffold properties have often been overlooked. This study sought to determine the effects of sterilization with ethanol, peracetic acid, ultraviolet irradiation, and antibiotic solution on the structure of 50:50 (mol:mol) 65:35, and 85:15 poly(D,L-lactic-co-glycolic acid [PLGA]) flat-sheet and hollow-fiber scaffolds. All methods resulted in scaffold sterilization, but scanning electron microscopy revealed deformations to the scaffold surface for all treatments. The extent of surface damage increased with treatment duration. This was further investigated by measurement of pore sizes, water flux, breaking strain, and Young's modulus. External pore size and water flux was found to be increased by all treatments in the following order: ethanol (largest), antibiotics, ultraviolet light, and peracetic acid. Pore sizes were 0.25 to 0.17 microm and water flux ranged from 0.01 kg x m(-2) x s(-1) to 3.34 kg x m(-2) x s(-1). For all samples, the Young's modulus was 1.0 to 31.1 MPa and breaking strain was 1.2 to 2.4 MPa. The results of this study suggest that antibiotic treatment shows the most potential to sterilize PLGA hollow fibers for tissue engineering.

Comparison of drug release from poly(lactide-co-glycolide) microspheres and novel fibre formulations.

Intraperitoneal cisplatin delivery has recently been shown to benefit ovarian cancer patients. Cisplatin-containing poly(lactide-co-glycolide) (PLGA) microspheres have been proposed for cisplatin delivery. The drug loading of cisplatin containing microspheres produced elsewhere is 3-10%w. Similar microspheres are reported here with a mean diameter of 38.8 µm, and a drug loading of 11.7%w, but using ethyl acetate as a safer solvent. In addition, novel formulations of cisplatin-containing solid and hollow PLGA 65:35 (lactide:glycolide) fibres were prepared and are reported here for the first time. PLGA hollow fibres were produced by phase inversion with a high drug loading of 27%w. Mechanistic mathematical models were applied to the cisplatin release profiles to allow quantitative comparison of microsphere, solid fibre and hollow fibre formulations. The diffusion coefficient of cisplatin eluting from a typical batch of PLGA microspheres was 4.8 × 10(-13) cm(2) s(-1); this low diffusivity of cisplatin in microspheres was caused by the low porosity of the polymer matrix. The diffusion coefficients of cisplatin eluting from a batch of PLGA solid fibres and hollow fibres were 6.1 × 10(-10) and 3.3 × 10(-10) cm(2) s(-1), respectively. These fibres allowed the controlled release of high doses of cisplatin over four days and may represent an improvement in slow release technology for treatment of ovarian cancer.

The initial release of cisplatin from poly(lactide-co-glycolide) microspheres.

PLGA microspheres loaded with cisplatin were produced using a single emulsion method. A semi-empirical model, with bi-exponential terms, was found to give a better fit to the drug release profiles compared to a mono-exponential model. This model suggests that there are two separate fractions of drug present in the depot. A fraction of the drug is located near/at the surface of the depot, and is readily released during immersion in buffer. A second fraction of drug is entrapped deeper within the depot and is subsequently released. It was also found that the initial release of cisplatin from PLGA microsphere is highly diffusion-controlled and the classical Higuchi model provides a good fit. From studies of water diffusion using PFG-NMR, results suggested that 50:50 PLGA microsphere was most susceptible to swelling and this might have promoted the faster initial drug release. Results from NMR cryoporometry also indicated that the developed PLGA microspheres could have "ink-bottle" pores.