Direct and Indirect Cationization of Cellulose Nanocrystals: Structure-Properties Relationship and Virus Capture Activity.

Due to increasing public concern over hygiene, there have been many studies investigating antimicrobial and antiviral agents recently. With the aim of developing biobased virucidal/virus capture agents, we report a chemical modification of the cellulose nanocrystals (CNCs) surface with poly(2-dimethylamino) ethyl acrylate) methyl chloride quaternary salt (Q-PDMAEA) to introduce the positively charged functional groups. The surface of CNCs was modified through direct and indirect graft polymerization. Subsequently, the direct and indirect cationization effect on the degree of functionalization, thermal stability, crystallinity, and antiviral activity of CNCs was investigated. Indirect cationization produced the highest degree of polymer grafting, increasing particle size and thermal stability. Further, the modified CNCs were tested for their ability to capture nonenveloped bacteriophages PhiX174 (ΦX174) and MS2. We observed a significant (>4.19 log10) reduction in total viral load by specific functionalized CNCs. However, the activity depended on the structure of functional groups, surface charge density, and the type of virus under study. Overall, the direct and indirect cationization of CNC leads to biobased agents with immobilized cationic charge, with good virus capture activity. Such agents can be used for various applications including textiles, packaging, wastewater treatment, etc.

Strontium-Substituted Nanohydroxyapatite-Incorporated Poly(lactic acid) Composites for Orthopedic Applications: Bioactive, Machinable, and High-Strength Properties.

Traditional metal-alloy bone fixation devices provide structural support for bone repair but have limitations in actively promoting bone healing and often require additional surgeries for implant removal. In this study, we focused on addressing these challenges by fabricating biodegradable composites using poly(lactic acid) (PLA) and strontium-substituted nanohydroxyapatite (SrHAP) via melt compounding and injection molding. Various percentages of SrHAP (5, 10, 20, and 30% w/w) were incorporated into the PLA matrix. We systematically investigated the structural, morphological, thermal, mechanical, rheological, and dynamic mechanical properties of the prepared composites. Notably, the tensile modulus, a critical parameter for orthopedic implants, significantly improved from 2.77 GPa in pristine PLA to 3.73 GPa in the composite containing 10% w/w SrHAP. The incorporation of SrHAP (10% w/w) into the PLA matrix led to an increased storage modulus, indicating a uniform dispersion of SrHAP within the PLA and good compatibility between the polymer and nanoparticles. Moreover, we successfully fabricated screws using PLA composites with 10% (w/w) SrHAP, demonstrating their formability at room temperature and radiopacity when observed under X-ray microtomography (micro-CT). Furthermore, the water contact angle decreased from 93 ± 2° for pristine PLA to 75 ± 3° for the composite containing SrHAP, indicating better surface wettability. To assess the biological behavior of the composites, we conducted in vitro cell-material tests, which confirmed their osteoconductive and osteoinductive properties. These findings highlight the potential of our developed PLA/SrHAP10 (10% w/w) composites as machinable implant materials for orthopedic applications. In conclusion, our study presents the fabrication and comprehensive characterization of biodegradable composites comprising PLA and strontium-substituted nanohydroxyapatite (SrHAP). These composites exhibit improved mechanical properties, formability, and radiopacity while also demonstrating desirable biological behavior. Our results suggest that these PLA/SrHAP10 composites hold promise as machinable implant materials for orthopedic applications.

Additive Manufacturing of Bioactive Poly(trimethylene carbonate)/ß-Tricalcium Phosphate Composites for Bone Regeneration.

Implants of bioresorbable materials combined with osteoconductive calcium phosphate ceramics show promising results to replace and repair damaged bone tissue. Here we present additive manufacturing of patient-specific porous scaffolds of poly(trimethylene carbonate) (PTMC) including high amounts of ß-tricalcium phosphate (ß-TCP). Tensile testing of composite networks showed that addition of ß-tricalcium phosphate reinforces the composites significantly. Three-dimensional structures containing up to 60 wt % ß-TCP could be built by stereolithography. By lowering the content to 51 wt %, manufacturing of a large-sized patient-specific prototype was possible at high resolution. Closer examination revealed that the created scaffolds contained more ß-TCP on the surface of the builds. Stereolithography therefore provides a manufacturing technique where the bioactive agent is directly available for creating an enhanced microenvironment for cell growth. The biocompatibility and bioresorption of PTMC coupled with the osteoconductivity of ß-TCP are an important candidate to consider in additive manufacturing of bone regeneration implants.

Native Structure of the Plant Cell Wall Utilized for Top-Down Assembly of Aligned Cellulose Nanocrystals into Micrometer-Sized Nanoporous Particles.

Despite their sustainable appeal, biomass components are currently undervalued in nanotechnology because means to control the assembly of bio-based nanoparticles are lagging behind the synthetic counterparts. Here, micrometer-sized particles consisting of aligned cellulose nanocrystals (CNCs) are prepared by crosslinking cellulose in cotton linter fibers that are prehydrolyzed with gaseous HCl, resulting in chemical cleavage necessary for CNC formation but retaining the morphology of the native fibers. That way, the intrinsic alignment of cellulose microfibrils within the fiber cell wall can be retained and utilized for top-down CNC alignment. Subsequent crosslinking with citric acid cements the alignment and preserves it, following the dispersion of CNCs trapped end-to-end, connected, and crosslinked within the colloidally stable micrometer-sized particles. Furthermore, thermoporosimetry and cryogenic transmission electron microscopy (Cryo TEM) shows that the particles possess mainly nanoporous (<2 nm) character in water. The approach challenges the current paradigm of predominantly bottom-up methods for nanoparticle assembly.

Ectopic bone formation in and soft-tissue response to P(CL/DLLA)/bioactive glass composite scaffolds.

OBJECTIVES: To characterize biological response to subcutaneously implanted macroporous poly(ε-caprolactone/D,L-lactide)-based scaffolds, and to evaluate the effect of bioactive glass (BAG) filler and osteogenic cells to the tissue response and ectopic bone formation. MATERIAL AND METHODS: In the first part of this study, six different scaffold types were screened in a rat subcutaneous implantation model. The polymer scaffolds with 70/30 caprolactone/lactide ratio and corresponding composites with < 45 µm BAG filler size were chosen for the further ectopic bone formation assay. The scaffolds were loaded with differentiating bone marrow stromal cells and implanted subcutaneously in syngeneic rats. RESULTS: With plain scaffolds, only mild foreign body reaction with no signs of gross inflammation was observed after 4 weeks of implantation. Furthermore, the scaffolds were fully invaded by well-vascularized soft connective tissue. Overall, all the tested scaffold types showed an appropriate host response. With cell-seeded scaffolds, several loci of immature mineralizing tissue and small amounts of mature bone were observed after 4 weeks. The incidence of mature bone formation was two and four in polymer scaffolds and composites, respectively (n = 8). After twelve weeks, mature bone was observed in only one polymer scaffold but in seven composites (n = 8). Excluding bone formation, the host response was considered similar to that with cell-free scaffolds. CONCLUSIONS: Plain scaffolds supported the ingrowth of well-vascularized fibroconnective tissue. Furthermore, cell seeded composites with BAG filler showed enhanced ectopic bone formation in comparison with corresponding neat polymer scaffolds.

Preparation and enzymatic degradation of porous crosslinked polylactides of biomass origin.

To understand the enzymatic degradation behavior of crosslinked polylactide (PLA), the preparation and enzymatic degradation of both thermoplastic (linear) and crosslinked PLAs that have pore structures with different dimensions were carried out. The porous structures of the linear PLA samples were of micro and nanoporous nature, and prepared by batch foaming with supercritical CO2 and compared with the porous structures of crosslinked PLA (Lait-X) created by the salt leaching method. The surface and cross-sectional morphologies of the porous structures were investigated by using scanning electron microscopy. The morphological analysis of porous Lait-X showed a rapid loss of physical features within 120 h of exposure to proteinase-K enzymatic degradation at 37 °C. Due to the higher affinity for water, enhanced enzymatic activity as compared to the linear PLA porous structures in the micro and nanoporous range was observed.

In Vitro and In Vivo Degradation of Photo-Crosslinked Poly(Trimethylene Carbonate-co-ε-Caprolactone) Networks.

Three-armed poly(trimethylene carbonate) (PTMC) and poly(trimethylene carbonate-co-Ɛ-caprolactone) (P(TMC-co-ε-CL)) macromers with molecular weights of approximately 30 kg mol-1 are synthesized by ring-opening polymerization and subsequent functionalization with methacrylic anhydride. Networks are then prepared by photo-crosslinking. To investigate the in vitro and in vivo degradation properties of these photo-crosslinked networks and assess the effect of ε-caprolactone content on the degradation properties, PTMC networks, and copolymer networks with two different TMC:ε-CL ratios are prepared. PTMC networks degraded slowly, via an enzymatic surface erosion process, both in vitro and in vivo. Networks prepared from P(TMC-co-ε-CL) macromers with a 74:26 ratio are found to degrade slowly as well, via a surface erosion process, albeit at a higher rate compared to PTMC networks. Increasing the ε-CL content to a ratio of 52:48, resulted in a faster degradation. These networks lost their mechanical properties much sooner than the other networks. Thus, PTMC and P(TMC-co-ε-CL) networks are interesting networks for tissue engineering purposes and the exact degradation properties can be tuned by varying the TMC:ε-CL ratio, providing researchers with a tool to obtain copolymer networks with the desired degradation rate depending on the intended application.

Heat-Induced Actuator Fibers: Starch-Containing Biopolyamide Composites for Functional Textiles.

This study introduces the development of a thermally responsive shape-morphing fabric using low-melting-point polyamide shape memory actuators. To facilitate the blending of biomaterials, we report the synthesis and characterization of a biopolyamide with a relatively low melting point. Additionally, we present a straightforward and solvent-free method for the compatibilization of starch particles with the synthesized biopolyamide, aiming to enhance the sustainability of polyamide and customize the actuation temperature. Subsequently, homogeneous dispersion of up to 70 wt % compatibilized starch particles into the matrix is achieved. The resulting composites exhibit excellent mechanical properties comparable to those reported for soft and tough materials, making them well suited for textile integration. Furthermore, cyclic thermomechanical tests were conducted to evaluate the shape memory and shape recovery of both plain polyamide and composites. The results confirmed their remarkable shape recovery properties. To demonstrate the potential application of biocomposites in textiles, a heat-responsive fabric was created using thermoresponsive shape memory polymer actuators composed of a biocomposite containing 50 wt % compatibilized starch. This fabric demonstrates the ability to repeatedly undergo significant heat-induced deformations by opening and closing pores, thereby exposing hidden functionalities through heat stimulation. This innovative approach provides a convenient pathway for designing heat-responsive textiles, adding value to state-of-the-art smart textiles.

Adsorption of PEO-PPO-PEO triblock copolymers with end-capped cationic chains of poly(2-dimethylaminoethyl methacrylate).

We study the adsorption of a symmetric triblock copolymer of ethylene oxide, EO, and propylene oxide, PO, end-capped with quarternized poly(2-dimethylaminoethyl methacrylate), DMAEMA (DMAEMA(24)-EO(132)PO(50)EO(132)-DMAEMA(24)). Light scattering and tensiometry are used to measure the relative size of the associated structures and surface excess at the air-liquid interface. The adsorbed amount, the amount of coupled water, and the viscoelasticity of the adsorbed polymer layer are measured on hydrophobic and hydrophilic surfaces (polypropylene, cellulose, and silica) by using quartz crystal microgravimetry (QCM) and surface plasmon resonance (SPR) at different ionic strengths and temperatures. The results of the experiments are compared with those obtained after adsorption of the uncharged precursor copolymer, without the cationic end-caps (EO(132)PO(50)EO(132)). DMAEMA(24)-EO(132)PO(50)EO(132)-DMAEMA(24) possesses higher affinity with the negatively charged silica and cellulose surfaces while the uncharged copolymer adsorbs to a larger extent on polypropylene surfaces. In this latter case, adsorption increases with increasing solution ionic strength and temperature. Adsorption of EO(132)PO(50)EO(132) on silica surfaces has little effect on the water contact angle (WCA), while adsorption of DMAEMA(24)-EO(132)PO(50)EO(132)-DMAEMA(24) increases the WCA of silica to 32°, indicating a large density of exposed PPO blocks upon adsorption. After adsorption of EO(132)PO(50)EO(132) and DMAEMA(24)-EO(132)PO(50)EO(132)-DMAEMA(24) on PP, the WCA is reduced by ≈14° and ≈28°, respectively, due to the exposed hydrophilic EO and highly water-soluble DMAEMA segments on the surfaces. The extent of surface coverage at saturation at the polypropylene/liquid interfaces (≈31 and 40 nm(2)/molecule obtained by QCM and SPR, respectively) is much lower, as expected, when compared with results obtained at the air/liquid interface, where a tighter packing is observed. The percentage of water coupled to the adsorbed cationic polymer decreases with solution ionic strength. Overall, these observations are ascribed to the effects of electrostatic screening, polymer hydrodynamic size, and solvency.

Photo-cross-linked biodegradable poly(ester anhydride) networks prepared from alkenylsuccinic anhydride functionalized poly(ε-caprolactone) precursors.

Biodegradable poly(ester anhydride) networks based on linear and star-shaped poly(ε-caprolactone)-based precursors were synthesized with the aim of obtaining matrixes suitable for release of macromolecular active agents. The ring-opening polymerization yielded hydroxyl telechelic oligomers, which were end-functionalized with succinic anhydride or with alkenylsuccinic anhydrides containing 8, 12, or 18 carbons in their alkenyl chains. Before cross-linking, the acid-terminated oligomers were reacted with methacrylic anhydride to obtain methacrylated precursors containing labile anhydride bonds. The degrees of substitution for the acid functionalization and methacrylation were >93%. Cross-linking of the precursors was carried out with visible light at room temperature. Gel contents and cross-linking densities were higher for networks cross-linked from the star-shaped precursors than for networks prepared from the linear precursors. In in vitro erosion tests, the presence of the alkenyl chain slowed down the erosion rate. The networks exhibited characteristic surface erosion: the mass loss was linear, whereas the dimensions of the specimens decreased steadily. A macromolecular release study showed the release of the model compound to be linear and in proportion to the mass loss.

Effect of moisture on electrospun nanofiber composites of poly(vinyl alcohol) and cellulose nanocrystals.

The effect of humidity on the morphological and thermomechanical properties of electrospun poly(vinyl alcohol) (PVA) fiber mats reinforced with cellulose nanocrystals (CNs) was investigated. Scanning electron microscopy (SEM) images revealed that the incorporation of CNs improved the morphological stability of the composite fibers even in high humidity environments. Thermal and mechanical properties of the electrospun fiber mats were studied by using differential scanning calorimetry (DSC), dynamic mechanical analysis (DMA), and large deformation tensile tests under controlled humidity and temperatures. The balance between the moisture-induced plasticization and the reinforcing effect of rigid CN particles was critical in determining the thermomechanical behaviors of the electrospun fiber mats. Results indicated that the stabilizing effect of the CNs in the PVA matrix might be compromised by water absorption, disrupting the hydrogen bonding within the structure. The amount of this disruption depended on the surrounding humidity and the CN loading. The reduction in tensile strength of neat PVA fiber mats as they were conditioned from low relative humidity (10% RH) to high relative humidity (70% RH) was found to be about 80%, from 1.5 to 0.4 MPa. When the structure was reinforced with CNs, the reduction in strength was limited to 40%, from 2 to 0.8 MPa over the same range in relative humidity. More importantly, the CN-loaded PVA fiber mats showed a reversible recovery in mechanical strength after cycling the relative humidity. Finally, humidity treatments of the composite PVA fiber mats induced significant enhancement of their strength as a result of the adhesion between the continuous matrix and the CNs.

Synthesis and hydrolysis behaviour of poly(ester anhydrides) from polylactone precursors containing alkenyl moieties.

Hydroxyl-group functional polylactones were prepared and converted to acid- terminated polyesters in a reaction with a series of alkenylsuccinic anhydrides containing 8, 12, or 18 carbons in their alkenyl chains. These polyester precursors were then linked into higher molecular weight poly(ester anhydrides) containing alkenyl moieties in their polyester blocks. The hydrolysis behaviour of the poly(ester anhydrides) was found to depend on the thermal properties of the polyester precursors. For poly(ester anhydrides) prepared from low molecular weight prepolymers with thermal transitions below 37 degrees C, the presence of hydrophobic alkenyl chains in the polyester precursors slowed the rate of weight loss. Poly(ester anhydrides) prepared from higher molecular weight prepolymers showed the opposite weight-loss behaviour; i.e., the crystallinity and thermal transitions of the alkenyl chain-containing poly(ester anhydrides) were low, and the weight loss was faster than for poly(ester anhydrides) without the alkenyl chains. The differences in length of the alkenyl chain, as such, had little effect on the hydrolysis behaviour and thermal properties of the poly(ester anhydrides).

Three-dimensional fabrication of cell-laden biodegradable poly(ethylene glycol-co-depsipeptide) hydrogels by visible light stereolithography.

Stereolithography (SLA) holds great promise in fabrication of cell-laden hydrogels with biomimetic complexity for use in tissue engineering and pharmaceutics. However, the availability of biodegradable photocrosslinkable hydrogel polymers for SLA is very limited. In this study, a water-soluble methacrylated poly(ethylene glycol-co-depsipeptide) was synthesized to yield a biodegradable photocrosslinkable macromer for SLA. Structural analysis confirmed the inclusion of biodegradable peptide and ester groups and photocrosslinkable methacrylate groups into the polymer backbone. The new macromer combined with RGDS peptide was used for SLA fabrication of hydrogels in absence and presence of cells. With the increasing light exposure time in SLA, mechanical stiffness of the hydrogels increased from 3 ± 1 kPa to 38 ± 13 kPa. Total mass loss of the samples within 7 days in PBS was 13%-21% and within 24 days 35%-66%. Due to degradation, the mechanical stiffness decreased by one order magnitude within 7-day incubation in PBS. Encapsulated endothelial cells proliferated in the hydrogels during 10-day in vitro cell culturing study. The macromer was further used in SLA to fabricate bifurcating tubular structures as preliminary vessel grafts. The new biodegradable, photocrosslinkable polymer is a significant addition to the very limited material selection currently available for SLA-based fabrication of cell-laden tissue engineering constructs.

Degradable polyesters through chain linking for packaging and biomedical applications.

The major route to convert lactic acid to high-molecular-weight polymers is ring-opening polymerization of lactide. We have investigated alternative synthesis routes based on oligomerization and chain linking to produce high-molecular-weight thermoplastic degradable polymers cost-effectively. Chain linking also offers new possibilities to prepare degradable polyesters for biomedical applications by extending the range of polymer properties achievable. In this paper, we briefly review different chain linking techniques used in our laboratory. Typically, lactic acid prepolymers with molecular weights of around 3,000-15,000 g x mol(-1) have been prepared by direct polycondensation. Hydroxyl terminated oligomers have been chain linked by using diisocyanate coupling agents, preferably 1,4-butane diisocyanate, forming poly(ester-urethanes). Poly(ester-amides) have been prepared by using 2,2'-bis(2-oxazoline) as coupling agent for carboxylic acid telechelic oligomers. Chain linking by end functionalization has been used in the preparation of poly(ester-anhydrides). In addition, a variety of crosslinked degradable polymers and copolymers have been synthesized through different crosslinking routes, by using methacrylic, itaconic or maleic double bonds or triethoxysilane moieties. A biodegradation test and ecotoxicological evaluation of the degradation products were carried out in addition to hydrolysis tests. Lactic acid based chain linked polymers were biodegradable and the degradation products were harmless. In hydrolysis tests, enzymatic degradation was pronounced in the chain linked poly(epsilon-caprolactone).

Enhanced osteogenicity of bioactive composites with biomimetic treatment.

PURPOSE: This study aimed to explore if initiation of biomimetic apatite nucleation can be used to enhance osteoblast response to biodegradable tissue regeneration composite membranes. MATERIALS AND METHODS: Bioactive thermoplastic composites consisting of poly(ε-caprolactone/DL-lactide) and bioactive glass (BAG) were prepared at different stages of biomimetic calcium phosphate deposition by immersion in simulated body fluid (SBF). The modulation of the BAG dissolution and the osteogenic response of rat mesenchymal stem cells (MSCs) were analyzed. RESULTS: SBF treatment resulted in a gradual calcium phosphate deposition on the composites and decreased BAG reactivity in the subsequent cell cultures. Untreated composites and composites covered by thick calcium phosphate layer (14 days in SBF) expedited MSC mineralization in comparison to neat polymers without BAG, whereas other osteogenic markers--alkaline phosphatase activity, bone sialoprotein, and osteocalcin expression--were initially decreased. In contrast, surfaces with only small calcium phosphate aggregates (five days in SBF) had similar early response than neat polymers but still demonstrated enhanced mineralization. CONCLUSION: A short biomimetic treatment enhances osteoblast response to bioactive composite membranes.

Photocrosslinked poly(ester anhydride)s for peptide delivery: Effect of oligomer hydrophobicity on PYY3-36 delivery.

The treatment for many diseases can be improved by developing more efficient peptide delivery technologies, for example, biodegradable polymers. In this work, photocrosslinked poly(ester anhydride)s based on functionalized poly(ε-caprolactone) oligomers were investigated for their abilities to achieve controlled peptide delivery. The effect of oligomer hydrophobicity on erosion and peptide release from poly(ester anhydride)s was evaluated by developing a sustained subcutaneous delivery system for an antiobesity drug candidate, peptide YY3-36 (PYY3-36). Oligomer hydrophobicity was modified with alkenylsuccinic anhydrides containing a 12-carbon alkenyl chain. PYY3-36 was mixed as a solid powder with methacrylated poly(ester anhydride) precursors, and this mixture was photocrosslinked at room temperature to form an implant for subcutaneous administration in rats. The oligomer hydrophobicity controlled the polymer erosion and PYY3-36 release as the increased hydrophobicity via the alkenyl chain prolonged polymer erosion in vitro and sustained in vivo release of PYY3-36. In addition, photocrosslinked poly(ester anhydride)s increased the bioavailability of PYY3-36 by up to 20-fold in comparison with subcutaneous administration of solution, evidence of remarkably improved delivery. In conclusion, this work demonstrates the suitability of photocrosslinked poly(ester anhydride)s for use in peptide delivery.

Characterization of internal structure, polymer erosion and drug release mechanisms of biodegradable poly(ester anhydride)s by X-ray microtomography.

Surface-eroding biodegradable polymers can provide many advantages in drug delivery, such as controllable and zero-order drug release. Photocrosslinkable poly(ester anhydride)s are a recently developed family of surface-eroding polymers with readily modifiable oligomer chemistry allowing tailoring of polymer properties. For example, in vivo release rate of peptide from photocrosslinked poly(ester anhydride)s can be controlled by oligomer hydrophobicity. In this study, X-ray microtomography (micro-CT) was used to gain a deeper understanding on internal structure, polymer erosion and drug release mechanisms of photocrosslinked poly(ester anhydride)s. Micro-CT is non-destructive and able to provide quantitative and qualitative information on the 3D structure of the sample in micrometer resolution. Photocrosslinked poly(ester anhydride) samples with varying drug loading degrees (propranolol HCl 0%, 10% and 60% w/w) and hydrophobicity (with and without 12-carbon alkenyl chain) were prepared. The samples, both freshly prepared and exposed to buffer solution for varying durations were characterized by micro-CT. The results showed that drug release from photocrosslinked poly(ester anhydride)s was primarily controlled by the surface erosion. However, drug diffusion had also a significant role in drug release from less hydrophobic samples with very high (60% w/w) drug loading degrees. In conclusion, micro-CT is a valuable tool in the characterization of surface-eroding polymers.

Preparation of poly(ε-caprolactone)-based tissue engineering scaffolds by stereolithography.

A photocrosslinkable poly(ε-caprolactone) (PCL)-based resin was developed and applied using stereolithography. No additional solvents were required during the structure preparation process. Three-armed PCL oligomers of varying molecular weights were synthesized, functionalized with methacrylic anhydride, and photocrosslinked, resulting in high gel content networks. Stereolithography was used to build designed porous scaffolds using the resin containing PCL macromer, Irgacure 369 photoinitiator, inhibitor and dye. A suitable resin viscosity was obtained by heating the resin during the curing process. The scaffolds precisely matched the computer-aided designs, with no observable material shrinkage. The average porosity was 70.5 ± 0.8%, and the average pore size was 465 µm. The pore network was highly interconnected. The photocrosslinkable, biodegradable PCL resin is well suited for the solvent-free fabrication of tissue engineering scaffolds by stereolithography.

Osteoblast response to continuous phase macroporous scaffolds under static and dynamic culture conditions.

Average scaffold pore sizes in the order of several hundred microns are generally required for efficient bone tissue ingrowth in vivo, whereas the culture of large bone engineering constructs in vitro can require bioreactor cultures to decrease diffusional constraints on the cells. In this study, we prepared poly(epsilon-caprolactone/D,L-lactide)-based scaffolds with continuous phase macroporosity using a novel CaCl(2) . 6H(2)O porogen agent. Osteogenic differentiation and scaffold colonization in rat bone marrow stromal cell cultures were compared in such polymer scaffolds, and in composites with 30 wt % bioactive glass filler. The effect of a rotating wall bioreactor culture on the cell response was also evaluated. Bioactive filler enhanced proliferation, early osteogenic differentiation, and mineralization of the cultured cells under static conditions. Dynamic cultures, in turn, resulted in decreased cell numbers and inhibition of the differentiation process irrespective of the scaffold type. This effect was ascribed to the harsh mechanical stresses caused by constant collisions of the scaffolds in the bioreactor vessels. However, cells were able to penetrate into the scaffold interior only under dynamic culture conditions. Thus, interconnected macroporosity is an essential, but not sufficient, condition to allow for full colonization of millimeter scale tissue engineering scaffolds in vitro.

Adhesion of respiratory-infection-associated microorganisms on degradable thermoplastic composites.

The purpose of this study was to evaluate bacterial adhesion and early colonization on a composite consisting of bioactive glass (BAG) particles and copolymer of epsilon-caprolactone/D,L-lactide. Materials were incubated with suspensions of both type strains and clinical isolates of Streptococcus pneumoniae, Haemophilus influenzae, and Pseudomonas aeruginosa for 30 minutes (adhesion) and 4 hours (colonization). Clear differences exist in the microorganisms' ability to adhere on the experimental materials. However, the presence of BAG particles does not inhibit bacterial adhesion, but early colonization of the materials with P. aeruginosa was inhibited by the addition of 90-315 mum BAG particles.