A composite hydrogel-3D printed thermoplast osteochondral anchor as example for a zonal approach to cartilage repair: in vivo performance in a long-term equine model.

Recent research has been focusing on the generation of living personalized osteochondral constructs for joint repair. Native articular cartilage has a zonal structure, which is not reflected in current constructs and which may be a cause of the frequent failure of these repair attempts. Therefore, we investigated the performance of a composite implant that further reflects the zonal distribution of cellular component both in vitro and in vivo in a long-term equine model. Constructs constituted of a 3D-printed poly(ϵ-caprolactone) (PCL) bone anchor from which reinforcing fibers protruded into the chondral part of the construct over which two layers of a thiol-ene cross-linkable hyaluronic acid/poly(glycidol) hybrid hydrogel (HA-SH/P(AGE-co-G)) were fabricated. The top layer contained Articular Cartilage Progenitor Cells (ACPCs) derived from the superficial layer of native cartilage tissue, the bottom layer contained mesenchymal stromal cells (MSCs). The chondral part of control constructs were homogeneously filled with MSCs. After six months in vivo, microtomography revealed significant bone growth into the anchor. Histologically, there was only limited production of cartilage-like tissue (despite persistency of hydrogel) both in zonal and non-zonal constructs. There were no differences in histological scoring; however, the repair tissue was significantly stiffer in defects repaired with zonal constructs. The sub-optimal quality of the repair tissue may be related to several factors, including early loss of implanted cells, or inappropriate degradation rate of the hydrogel. Nonetheless, this approach may be promising and research into further tailoring of biomaterials and of construct characteristics seems warranted.

Biodistribution of quantum dots in the kidney after intravenous injection.

The biodistribution of nanoparticles is a major subject of current nanomedical research. To date, however, the exact investigation of nanoparticle fate in the microenvironment of a main excretory organ, the kidney has largely been neglected. In this study, the biodistribution of polyethylene glycol-coated quantum dots (Qdots) with special focus on their interaction with the kidney is investigated. Upon intravenous injection, nanoparticles showed effective blood circulation in mice and significant renal accumulation after two hours. Histological analysis of the kidney revealed that Qdots were strongly associated to the intraglomerular mesangial cells. This preferential deposition of nanoparticles in the kidney mesangium is highly promising, since it could be of utmost value for site-specific treatment of severe kidney diseases like diabetic nephropathy in the future.

Enhanced bone morphogenetic protein-2 performance on hydroxyapatite ceramic surfaces.

The immobilization of biomolecules on biomaterial surfaces allows for the control of their localization and retention. In numerous studies, proteins have been simply adsorbed to enhance the biological performance of various materials in vivo. We investigated the potential of surface modification techniques on hydroxyapatite (HA) ceramic discs in an in vitro approach. A novel method for protein immobilization was evaluated using the aminobisphosphonates pamidronate and alendronate, which are strong Ca chelating agents, and was compared with the established silanization technique. Lysozyme and bone morphogenetic protein-2 (BMP-2) were used to assess the suitability of the two surface modification methods with regard to the enzymatic activity of lysozyme and to the capacity of BMP-2 to stimulate the osteoblastic differentiation of C2C12 mouse myoblasts. After immobilization, a 2.5-fold increase in enzymatic activity of lysozyme was observed compared with the control. The alkaline phosphatase activity per cell stimulated by immobilized BMP-2 was 2.5-fold higher [9 x 10(-6) I.U.] than the growth factor on unmodified surfaces [2-4 x 10(-6) I.U.]. With regard to the increase in protein activity, both procedures lead to equivalent results. Thus, the bisphosphonate-based surface modification represents a safe and easy alternative for the attachment of proteins to HA surfaces.

Polymer coating of quantum dots--a powerful tool toward diagnostics and sensorics.

The use of quantum dots for biological and biomedical applications is one of the fastest moving fields of nanotechnology today. The unique optical properties of these nanometer-sized semiconductor crystals make them an exciting fluorescent tool for in-vivo and in-vitro imaging as well as for sensoric applications. To apply them in biological fluids or aqueous environment it is essential to modulate the chemical nature of quantum dot surfaces to alter their solubility and add additional chemical functionalities. By employing different coating technologies they cannot only be rendered water soluble but also functionalized to fulfill different tasks, like receptor targeting or sensing of low molecular weight substances. To achieve this goal different polymeric coatings are applied to provide solubility in water and additional functional groups for attachment. Taken together the versatile modifications described in this review make quantum dots a promising alternative to conventional fluorescent dyes and may offer possibilities for new future developments.

Influence of wettability and surface activity on release behavior of hydrophilic substances from lipid matrices.

The aim of this study was to investigate the role of matrix and drug properties on controlled release from triglyceride matrices. Mini-cylinders of 2 mm diameter, 2.2 mm height and 7 mg weight were produced by compression of lipid powder obtained by using a polyethylene glycol (PEG) co-lyophilization method for the model substances lysozyme and FITC-dextran (Mw 4000 Da). Lysozyme was released with decreasing velocity from glyceryl trilaurate, -myristate, -palmitate and -stearate for more than 14 months. Release correlated well with triglyceride lipophilicity defined by the chain length of the fatty acids. Contact angle measurements and the analysis of buffer penetration visualized by confocal microscopy emphasized the role of matrix wettability as a prerequisite for release. A comparison with FITC-dextran revealed that the protein itself enhances matrix wettability and hence its release due to its surface active properties. FITC-dextran remained trapped within the matrix only to be released at lower compression force or after the addition of surfactant. Protein added externally to the release buffer at 0.1% (w/v) was not efficient in lowering the contact angle and increasing the release rate of FITC-dextran. Tween 20 and 81 could be used in different concentrations (0.1, 0.01 and 0.001% (w/v)) to alter lysozyme and FITC-dextran release profiles: resulting release rates showed a close dependence on the contact angle of the respective release medium and triglyceride matrix material. However, both Tweens seem to act not only by reducing the release medium contact angle but also by moderately affecting interparticulate adhesion of the matrix material.

Mediating specific cell adhesion to low-adhesive diblock copolymers by instant modification with cyclic RGD peptides.

One promising strategy to control the interactions between biomaterial surfaces and attaching cells involves the covalent grafting of adhesion peptides to polymers on which protein adsorption, which mediates unspecific cell adhesion, is essentially suppressed. This study demonstrates a surface modification concept for the covalent anchoring of RGD peptides to reactive diblock copolymers based on monoamine poly(ethylene glycol)-block-poly(D,L-lactic acid) (H(2)N-PEG-PLA). Films of both the amine-reactive (ST-NH-PEG(2)PLA(20)) and the thiol-reactive derivative (MP-NH-PEG(2)PLA(40)) were modified with cyclic alphavbeta3/alphavbeta5 integrin subtype specific RGD peptides simply by incubation of the films with buffered solutions of the peptides. Human osteoblasts known to express these integrins were used to determine cell-polymer interactions. The adhesion experiments revealed significantly increased cell numbers and cell spreading on the RGD-modified surfaces mediated by RGD-integrin-interactions.

Biomimetic polymers in pharmaceutical and biomedical sciences.

This review describes recent developments in the emerging field of biomimetic polymeric biomaterials, which signal to cells via biologically active entities. The described biological effects are, in contrast to many other known interactions, receptor mediated and therefore very specific for certain cell types. As an introduction into this field, first some biological principles are illustrated such as cell attachment, cytokine signaling and endocytosis, which are some of the mechanisms used to control cells with biomimetic polymers. The next topics are then the basic design rules for the creation of biomimetic materials. Here, the major emphasis is on polymers that are assembled in separate building blocks, meaning that the biologically active entity is attached to the polymer in a separate chemical reaction. In that respect, first individual chemical standard reactions that may be used for this step are briefly reviewed. In the following chapter, the emphasis is on polymer types that have been used for the development of several biomimetic materials. There is, thereby, a delineation made between materials that are processed to devices exceeding cellular dimensions and materials predominantly used for the assembly of nanostructures. Finally, we give a few current examples for applications in which biomimetic polymers have been applied to achieve a better biomaterial performance.

Poly(D,L-lactic acid)-poly(ethylene glycol)-monomethyl ether diblock copolymers control adhesion and osteoblastic differentiation of marrow stromal cells.

Biodegradable polymers, such as poly(lactic acid) (PLA) and poly(lactic-coglycolic acid) (PLGA), are attractive materials for tissue engineering because of their degradative and mechanical properties, which permit scaffolds to be tailored to the individual requirements of different tissues. Although these materials support tissue development, their chemical properties offer no control of cell adhesion or function because their surfaces become immediately masked by adsorbing serum proteins when the materials come into contact with body fluids. Furthermore, adhesion proteins undergo conformational changes and a decrease in bioactivity when adsorbed to hydrophobic materials, such as PLA. To overcome these limitations, we modified the properties of PLA by synthesizing a diblock copolymer with poly(ethylene glycol) (PEG), which is known to reduce the amount of adsorbed proteins and to modify their conformation. By altering the PEG content of these diblock copolymers we were able to control the adsorption of adhesion proteins and, because cell adhesion takes place only in the presence of serum proteins, to control cell adhesion and cell shape. Marrow stromal cell differentiation to the osteoblastic phenotype was strongly improved on PEG-PLA compared with PLA, PLGA and tissue culture polystyrene and led to a 2-fold increase in alkaline phosphatase activity and mineralization.

Polyanhydride degradation and erosion.

It was the intention of this paper to give a survey on the degradation and erosion of polyanhydrides. Due to the multitude of polymers that have been synthesized in this class of material in recent years, it was not possible to discuss all polyanhydrides that have gained in significance based on their application. It was rather the intention to provide a broad picture on polyanhydride degradation and erosion based on the knowledge that we have from those polymers that have been intensively investigated. To reach this goal this review contains several sections. First, the foundation for an understanding of the nomenclature are laid by defining degradation and erosion which was deemed necessary because many different definitions exist in the current literature. Next, the properties of major classes of anhydrides are reviewed and the impact of geometry on degradation and erosion is discussed. A complicated issue is the control of drug release from degradable polymers. Therefore, the aspect of erosion-controlled release and drug stability inside polyanhydrides are discussed. Towards the end of the paper models are briefly reviewed that describe the erosion of polyanhydrides. Empirical models as well as Monte-Carlo-based approaches are described. Finally it is outlined how theoretical models can help to answer the question why polyanhydrides are surface eroding. A look at the microstructure and the results from these models lead to the conclusion that polyanhydrides are surface eroding due to their fast degradation. However they switch to bulk erosion once the device dimensions drop below a critical limit.

Biodegradable poly(D,L-lactic acid)-poly(ethylene glycol)-monomethyl ether diblock copolymers: structures and surface properties relevant to their use as biomaterials.

To obtain biodegradable polymers with variable surface properties for tissue culture applications, poly(ethylene glycol) blocks were attached to poly(lactic acid) blocks in a variety of combinations. The resulting poly(D,L-lactic acid)-poly(ethylene glycol)-monomethyl ether (Me.PEG-PLA) diblock copolymers were subject to comprehensive investigations concerning their bulk microstructure and surface properties to evaluate their suitability for drug delivery applications as well as for the manufacture of scaffolds in tissue engineering. Results obtained from 1H-NMR, gel permeation chromatography, wide angle X-ray diffraction and modulated differential scanning calorimetry revealed that the polymer bulk microstructure contains poly(ethylene glycol)-monomethyl ether (Me.PEG) domains segregated from poly(D,L-lactic acid) (PLA) domains varying with the composition of the diblock copolymers. Analysis of the surface of polymer films with atomic force microscopy and X-ray photoelectron spectroscopy indicated that there is a variable amount of Me.PEG chains present on the polymer surface, depending on the polymer composition. It could be shown that the presence of Me.PEG chains in the polymer surface had a suppressive effect on the adsorption of two model peptides (salmon calcitonin and human atrial natriuretic peptide). The possibility to modify polymer bulk microstructure as well as surface properties by variation of the copolymer composition is a prerequisite for their efficient use in the fields of drug delivery and tissue engineering.

[Long-term sequelae of neonatal group B streptococcal septicemia/meningitis (author's transl)]. / Dauerschäden nach B-Streptokokken-Sepsis/Meningitis im Neugeborenenalter.

In recent years group B streptococci accounted for nearly 40% of all cases of neonatal septicemia in our intensive-care-unit. Nineteen of 38 babies did not survive the acute illness (mortality 50%). Six of 18 surviving children showed abnormalities related to the septicemia/meningitis (morbidity 33%). The severity of chronic complications ranged from minor neurological problems to marked retardation, deafness, blindness, and epilepsy. In our series there seemed to be a relation between the severity of complications during the acute illness (meningitis, convulsions), and later neurological sequelae. Early detection and early treatment were found to be most important for the final outcome.