Albrecht Berg

Release of metronidazole from electrospun poly(l-lactide-co-d/l-lactide) fibers for local periodontitis treatment

OBJECTIVES We aimed to achieve detailed biomaterials characterization of a drug delivery system for local periodontitis treatment based on electrospun metronidazole-loaded resorbable polylactide (PLA) fibers. METHODS PLA fibers loaded with 0.1-40% (w/w) MNA were electrospun and were characterized by SEM and DSC. HPLC techniques were used to analyze the release profiles of metronidazole (MNA) from these fibers. The antibacterial efficacy was determined by measuring inhibition zones of drug-containing aliquots from the same electrospun fiber mats in an agar diffusion test. Three pathogenic periodontal bacterial strains: Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis were studied. Cytotoxicity testing was performed with human gingival fibroblasts by: (i) counting viable cells via live/dead staining methods and (ii) by exposing cells directly onto the surface of MNA-loaded fibers. RESULTS MNA concentration influenced fiber diameters and thus w/w surface areas: diameter being minimal and area maximal at 20% MNA. HPLC showed that these 20% MNA fibers had the fastest initial MNA release. From the third day, MNA release was slower and nearly linear with time. All fiber mats released 32-48% of their total drug content within the first 7 days. Aliquots of media taken from the fiber mats inhibited the growth of all three bacterial strains. MNA released up to the 28th day from fiber mats containing 40% MNA significantly decreased the viability of F. nucleatum and P. gingivalis and up to the 2nd day also for the resistant A. actinomycetemcomitans. All of the investigated fibers and aliquots showed excellent cytocompatibility. SIGNIFICANCE This study shows that MNA-loaded electrospun fiber mats represent an interesting class of resorbable drug delivery systems. Sustained drug release properties and cytocompatibility suggest their potential clinical applicability for the treatment of periodontal diseases.

Biocompatible polysaccharide-based cryogels.

This study focuses on the development of novel biocompatible macroporous cryogels by electron-beam assisted free-radical crosslinking reaction of polymerizable dextran and hyaluronan derivatives. As a main advantage this straightforward approach provides highly pure materials of high porosity without using additional crosslinkers or initiators. The cryogels were characterized with regard to their morphology and their basic properties including thermal and mechanical characteristics, and swellability. It was found that the applied irradiation dose and the chemical composition strongly influence the material properties of the resulting cryogels. Preliminary cytotoxicity tests illustrate the excellent in vitro-cytocompatibility of the fabricated cryogels making them especially attractive as matrices in tissue regeneration procedures.

Two-photon techniques in tissue engineering

PURPOSE NIR radiation in the range of about 800 nm is less absorbed by biological tissues and is suited for triggering photonic effects using femtosecond pulsed Ti:Sa lasers. Especially in the life sciences, two-photon techniques are gaining greater importance. We introduce two laser applications for tissue engineering: the autofluorescent visualization of cells seeded on 3D scaffolds after two-photon excitation; and the manufacturing of 3D-structured hydrogel-like scaffolds by triggering free-radical polymerization processes within polymerizable precursors. METHODS Primary bovine chondrocytes were cultivated on collagen I/III scaffolds using a flow chamber system coupled with a two-photon laser scanning microscope (2PLSM). During the incubation the cell population was hydrostatically stimulated. The selective visualization of unlabeled cells and scaffolds was achieved by spectral autofluorescence imaging. To gain some insight into scaffold-mediated effects on cell growth and cell differentiation, hydrogel-like scaffolds with well defined 3D structures were generated by two-photon polymerization (2PP) using methacrylated urethane and polyethyleneglycol diacrylate. RESULTS We were able to show that spectral autofluorescence imaging provides spatially resolved data for the non-invasive online control of the tissue engineering process as well as the quantification of cell distribution within the scaffold. The fabrication of 3D 2PP scaffolds made from hydrogel-forming monomers and their effect on cell attachment and cell growth were also shown. CONCLUSIONS Two-photon techniques provide powerful tools for both the non-invasive online visualization of 3D cell-scaffold constructs and the structuring of 3D cultivation environments. The application of these techniques is also suitable for integration into micro-systems technology (e.g. BioMEMS, Cells-on-Chip, Lab-on-a Chip).

Development of novel electrospun dual-drug fiber mats loaded with a combination of ampicillin and metronidazole

OBJECTIVE Our study was performed with the aim of preparing electrospun polylactide fibers with a combination of ampicillin (AMP) and metronidazole (MNZ) and investigating their drug release behavior and the antibacterial effect on Aggregatibacter actinomycetemcomitans and other oral pathogens. METHODS AMP and MNZ were integrated as a combination in two separate fibers (dual fiber mats - DFW mix) of electrospun PLA fiber mats by means of multijet electrospinning and in a single fiber (single fiber mats - SFW mix). HPLC (high-performance liquid chromatography) was used to measure the released drug quantities. Agar diffusion tests were used to determine the antibacterial effect of the eluates on A. actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Enterococcus faecalis. The neutral red test was made to examine the cytocompatibility of the eluates with human gingival fibroblasts (hGFs). RESULTS The release of the active agents varied with the antibiotic concentrations initially used in the fiber mats, but also with the distribution of the active agents in one or two fibers. Of the total quantity of MNZ (AMP), the SFW mix fiber mats released >60% (>70%) within a span of 5min, and 76% (71%) after 96h. With these drug concentrations released by the fiber mats (≥5m%), an antibacterial effect was achieved on A. actinomycetemcomitans and on all other species tested. Fiber mats and their eluates have no cytotoxic influence on human gingival fibroblasts (hGFs). SIGNIFICANCE Electrospun AMP/MNZ-loaded polymer fibers are a potential drug delivery system for use in periodontal and endodontic infections.

Cytocompatibility of polymer-based periodontal bone substitutes in gingival fibroblast and MC3T3 osteoblast cell cultures

OBJECTIVES Inflammatory periodontal diseases are accompanied by destruction of periodontal tissue and alveolar bone. Infrabony lesions can be regenerated with adequate bone substitutes, which require high biocompatibility of the material. METHODS To rate the biocompatibility of nine polymeric periodontal bone substitutes (Bio 1-Bio 9), cell viability and cytotoxicity assays were performed. For viability, human gingival fibroblasts (HGFs) and MC3T3 osteoblasts were cultured on the bone substitutes. For cytotoxicity, biomaterial extracts were prepared by incubation with culture medium for maximally 28days, and cells were exposed to the extracts for 1day. Polymers Bio 1 to Bio 5 were prepared by solvent casting, Bio 6 to Bio 9 by photopolymerization of the monomers at wavelengths of 400-500nm in the presence of a suitable photoinitiation system. RESULTS Bio 1, Bio 3, Bio 4, Bio 5, and Bio 7 showed moderate to excellent cytocompatibility for both HGFs and osteoblasts in viability tests. Together with the results of the cytotoxicity assays, four of the nine tested polymers were considered cytocompatible: Bio 1 (poly(vinyl butyral-co-vinyl alcohol-co-vinyl acetate; PVB)), Bio 4 and Bio 5 (functionalized oligolactones), and, to a limited degree, Bio 7 (urethane methacrylate). Except for Bio 7, the cytocompatible polymers showed intermediate water contact angles (74-85°) and therefore moderate to low hydrophilicity. SIGNIFICANCE The non-cross-linked polymers Bio 1, Bio 4, or Bio 5, and the photopolymerized polymeric network Bio 7 display good/excellent cytocompatibility and are therefore potential candidates for tissue engineering in alveolar bone substitution.

Formation of new, cytocompatible hydrogels based on photochemically crosslinkable levan methacrylates

Levan is a high molecular weight fructose-based biotechnologically available polysaccharide with a range of interesting properties qualifying this molecule for applications in biomedicine. In this study, new levan derivatives containing methacrylate groups attached either via ester or urethane linkages to the fructan backbone could be synthesized and structurally characterized by conventional analytical techniques. The photochemical crosslinking of these substances applying different photoinitiator systems and reaction conditions resulted in hydrogels of diverse properties which were investigated with regard to mechanical behaviour, hydrolytic degradability, and cytocompatibility. It was found that crosslinkable levan derivatives represent a new class of promising biopolymer-based macromers broadening the spectrum of available biomaterials to facilitate the adaption to the requirements of specific applications.

Antimicrobial photodynamic active biomaterials for periodontal regeneration

OBJECTIVE Biomaterials for periodontal regeneration may have insufficient mechanical and antimicrobial properties or are difficult to apply under clinical conditions. The aim of the present study was to develop a polymeric bone grafting material of suitable physical appearance and antimicrobial photodynamic activity. METHODS Two light curable biomaterials based on urethane dimethacrylate (BioM1) and a tri-armed oligoester-urethane methacrylate (BioM2) that additionally contained a mixture of β-tricalcium phosphate microparticles and 20wt% photosensitizer mTHPC (PS) were fabricated and analyzed by their compressive strength, flexural strength and modulus of elasticity. Cytotoxicity was observed by incubating eluates and in direct-contact to MC3T3-E1 cells. Antimicrobial activity was ascertained on Porphyromonas gingivalis and Enterococcus faecalis upon illumination with laser light (652nm, 1×100J/cm2, 2×100J/cm2). RESULTS The compressive strength, flexural strength and elastic modulus were, respectively, 311.73MPa, 22.81MPa and 318.85MPa for BioM1+PS and 742.37MPa, 7.58MPa and 406.23MPa for BioM2+PS. Both materials did not show any cytotoxic behavior. Single laser-illumination (652nm) caused total suppression of P. gingivalis (BioM2+PS), while repeated irradiation reduced E. faecalis by 3.7 (BioM1+PS) and 3.1 (BioM2+PS) log-counts. SIGNIFICANCE Both materials show excellent mechanical and cytocompatible properties. In addition, irradiation with 652nm induced significant bacterial suppression. The manufactured biomaterials might enable a more efficient cure of periodontal bone lesions. Due to the mechanical properties functional stability might be increased. Further, the materials are antimicrobial upon illumination with light that enables a trans-mucosal eradication of residual pathogens.

Biodegradable (Meth)acrylate-based Adhesives for Surgical Applications

The use of adhesives in surgery is an old but mostly unfulfilled dream (Donkerwolcke et al., 1998). Compared to conventional bonding techniques employed in surgery today like stitching, fixing with screws, pins, and plates, gluing has several advantages because it represents a fast and uncomplicated technique that causes no or only slight injuries of surrounding tissue and enables a homogenous load distribution between bonded materials (Rimpler, 1996). If such an adhesive would be gradually self-degrading in the body, newly formed tissue could replace the adhesive during the healing process and a complete regeneration of the damaged tissue would be possible. A gradual degradation of the adhesive would also maintain the necessary bonding strength within the tissue repair period and finally no foreign material would remain in the body.