Synthesis of nanosized 58S bioactive glass particles by a three-dimensional ordered macroporous carbon template.

Nanosized 58S bioactive glass (BG) particles were synthesized by using a three-dimensional ordered macroporous carbon template (OMC) with a pore size of 400nm. The obtained 58S BG particles possessed a diameter of 300nm, narrow size distribution and uniform spherical morphology. 58S/gelatin composites were prepared and showed much better mechanical properties than pure gelatin. The narrow size distribution of the 58S particles replicated from OMC was confirmed crucial to the mechanical properties of the 58S/gelatin composite, comparing to the contrast sample prepared with polydispersed particles. The outstanding bioactivity of the 58S BG particles was confirmed by inducing the formation of carbonated hydroxyapatite on the 58S/gelatin composite surface. This work showed a successful example that OMC template could be used to synthesize particles requiring a robust reaction condition, and a particle synthesis method that could well control particle size distribution was important for preparing materials with outstanding mechanical properties.

Functionalized poly(γ-Glutamic Acid) fibrous scaffolds for tissue engineering.

Poly(γ-glutamic acid) (γ-PGA) is a biocompatible, enzymatically-degradable, natural polymer with a higher resistance to hydrolysis than polyesters commonly used for tissue engineering scaffolds such as poly(L-lactide) (PLLA). Notably, γ-PGA's free carboxyl side groups allow for simple chemical functionalization, making it a versatile candidate for producing scaffolds. Here, a series of water-resistant fibrous scaffolds were engineered from ethyl (Et), propyl (Pr) and benzyl (Bn) esterifications of γ-PGA. All scaffolds were non-cytotoxic and γ-PGA-Bn showed an increase in cell adhesion of hMSCs compared to γ-PGA-Et and γ-PGA-Pr. Moreover, cells on γ-PGA-Bn showed three-fold higher viability at day 14 and significantly higher adhesion when compared with PLLA scaffolds, despite having a similar hydrophobicity. Cell attachment decreased by 40% when the polymer was only partially modified with benzyl groups (γ-PGA-Bn-77%), but was restored when integrin-binding RGD peptide was conjugated to the remaining free carboxylic groups, indicating the peptide was accessible and able to bind integrins. The mechanism behind the cell-material interactions on γ-PGA-Bn scaffolds was further investigated through protein adsorption and fibronectin conformation experiments. These results, in addition to the cell-adhesion studies, suggest an inherent effect of the benzyl modification in the mechanism of cell attachment to γ-PGA-Bn scaffolds. Finally, γ-PGA-Bn scaffolds cultured in osteogenic media were also efficient in supporting hMSCs differentiation towards an osteogenic lineage as determined by alkaline phosphatase and Runx2 gene expression. Taken together these data suggest that esterified γ-PGA polymer scaffolds are new and versatile candidates for tissue engineering applications and that, intriguingly, aromatic functionality plays a key role in the cell-scaffold interaction.

Template synthesis of ordered macroporous hydroxyapatite bioceramics.

Hydroxyapatite has found wide application in bone tissue engineering. Here we use a macroporous carbon template to generate highly ordered macroporous hydroxyapatite bioceramics composed of close-packed hollow spherical pores with interconnected channels. The template has advantages for the preparation of ordered materials.

Preparation and evaluation of a novel gastric mucoadhesive sustained-release acyclovir microsphere.

OBJECTIVE: The objective of this study was to prepare a novel gastric mucoadhesive sustained-release acyclovir (AV)-resinate microsphere. METHODS: First, AV absorption ratio was quantified in a rat gastrointestinal (GI) tract model. AV-resinate was prepared by bath method and used as cores to prepare microspheres by an emulsion solvent diffusion technique with carbopol 934 as coating material. GI transit test of the prepared microspheres was carried out in rats and beagle dogs, followed by the in vivo bioavailability evaluation of the microspheres in beagle dogs. RESULTS: The AV absorption ratio in different segments of rat's GI track for 3 hours was as following: stomach 9.46 +/- 0.62%, duodenum 20.22 +/- 1.50%, jejunum 15.7 +/- 1.33%, ileum 9.15 +/- 1.01%, and colon 4.59 +/- 0.48%. These results showed that AV was mainly absorbed in the stomach and upper intestine. The average diameter of the microspheres was 115.3 microm. The microspheres had a drug content of 33.3 +/- 0.7% (w/w) and a sustained-release profile for 12 hours in vitro. The mucoadhesive test in rats and beagle dogs showed that most of the microspheres were retained in the stomach 6 hours after oral administration. The in vivo pharmacokinetics test revealed that the microsphere and reference (AV tablets) preparations have no significant difference for C(max). The t(max) has increased from 2.33 hours (reference) to 5 hours (test). Meanwhile, the relative bioavailability of AV microspheres was 145%. CONCLUSION: A novel AV-resinate microsphere was prepared. The microspheres were proved to be gastric mucoadhesive and sustained-release with higher bioavailability.

Distinctive degradation behaviors of electrospun polyglycolide, poly(DL-lactide-co-glycolide), and poly(L-lactide-co-epsilon-caprolactone) nanofibers cultured with/without porcine smooth muscle cells.

Biodegradable nanofibers have become a popular candidate for tissue engineering scaffolds because of their biomimetic structure that physically resembles the extracellular matrix. For certain tissue regeneration applications, prolonged in vitro culture time for cellular reorganization and tissue remodeling may be required. Therefore, extensive understanding of cellular effects on scaffold degradation is needed. There are only few studies on the degradation of nanofibers, and also the studies on degradation throughout cell culture are rare. In this study, polyglycolide (PGA), poly(DL-lactide-co-glycolide) (PLGA) and poly(L-lactide-co-epsilon-caprolactone) [P(LLA-CL)] were electrospun into nanofibrous meshes. The nanofibers were cultured with porcine smooth muscle cells for up to 3 months to evaluate their degradation behavior and cellular response. The results showed that the degradation rates are in the order of PGA >> PLGA > P(LLA-CL). PGA nanofibers degraded in 3 weeks and supported cell growth only in the first few days. PLGA nanofiber scaffolds facilitated cell growth during the first 30 days after seeding, but cell growth was slow thereafter. P(LLA-CL) nanofibers facilitated long-term (1-3 months) cell growth. mRNA quantification using real-time polymerase chain reaction revealed that some smooth muscle cell markers (alpha-actinin and calponin) and extracellular matrix genes (collagen and integrin) seemed to be downregulated with increased cell culture time. Cell culture significantly increased the degradation rate of PGA nanofibers, whereas the effect on PLGA and P(LLA-CL) nanofibers was limited. We found that the molecular weight of P(LLA-CL) and PLGA nanofibers decreased linearly for up to 100 days. Half lives of PLGA and P(LLA-CL) nanofibers were shown to be 80 and 110 days, respectively. In summary, this is the first study to our knowledge to evaluate long-term polymeric nanofiber degradation in vitro with cell culture. Cell culture accelerated the nanofibrous scaffold degradation to a limited extent. P(LLA-CL) nanofibers could be a good choice as scaffolds for long-term smooth muscle cell culture.

Degradation behaviors of electrospun resorbable polyester nanofibers.

Biodegradable materials are widely used in the biomedical field because there is no postoperative surgery after implantation. Widely used synthetic biodegradable materials are polyesters, especially those used in tissue engineering. Advances in the tissue engineering field have brought much attention in terms of scaffold fabrication, such as with biodegradable polyester nanofibers. The rationale for using nanofibers for tissue engineering is that the nonwoven polymeric meshwork is a close representation of the nanoscale protein fiber meshwork in native extracellular matrix (ECM). Electrospinning technique is a promising way to fabricate controllable continuous nanofiber scaffold mimicking the ECM structure. Electrospun nanofibers provide high surface-to-volume ratio and high porosity as a promising scaffold for tissue engineering. Because the degradation behaviors of scaffolds significantly affect new tissue regeneration, the degradation of the material becomes one of the crucial factors when considering using polyester nanofibers as scaffolds in tissue engineering. In this review paper, we focus on the degradation studies of several bioresorbable polyester nanofibrous scaffolds used in tissue engineering. The degradable properties of nanofibers were compared with the corresponding degradable materials in macroscale. The factors that might affect the degradation behaviors were analyzed.

Long-term viability of coronary artery smooth muscle cells on poly(L-lactide-co-epsilon-caprolactone) nanofibrous scaffold indicates its potential for blood vessel tissue engineering.

Biodegradable polymer nanofibres have been extensively studied as cell culture scaffolds in tissue engineering. However, long-term in vitro studies of cell-nanofibre interactions were rarely reported and successful organ regeneration using tissue engineering techniques may take months (e.g. blood vessel tissue engineering). Understanding the long-term interaction between cells and nanofibrous scaffolds (NFS) is crucial in material selection, design and processing of the tissue engineering scaffolds. In this study, poly(L-lactide-co-epsilon-caprolactone) [P(LLA-CL)] (70:30) copolymer NFS were produced by electrospinning. Porcine coronary artery smooth muscle cells (PCASMCs) were seeded and cultured on the scaffold to evaluate cell-nanofibre interactions for up to 105 days. A favourable interaction between this scaffold and PCASMCs was demonstrated by cell viability assay, scanning electron microscopy, histological staining and extracellular matrix (ECM) secretion. Degradation behaviours of the scaffolds with or without PCASMC culture were determined by mechanical testing and gel permeation chromatography (GPC). The results showed that the PCASMCs attached and proliferated well on the P(LLA-CL) NFS. Large amount of ECM protein secretion was observed after 50 days of culture. Multilayers of aligned oriented PCASMCs were formed on the scaffold after two months of in vitro culture. In the degradation study, the PCASMCs were not shown to significantly increase the degradation rate of the scaffolds for up to 105 days of culture. The in vitro degradation time of the scaffold could be as long as eight months by extrapolating the results from GPC. These observations further supported the potential use of the P(LLA-CL) nanofibre in blood vessel tissue engineering.