Biomedical subjects

E H Schacht

Publications and source records attributed to E H Schacht.

15 recordsLinked to original sources

PubMed2006

Osteoblast behaviour on in situ photopolymerizable three-dimensional scaffolds based on D, L-lactide, epsilon-caprolactone and trimethylene carbonate.

Polymer networks formed by photocrosslinking of multifunctional oligomers have great potential as injectable and in situ forming materials for bone tissue engineering. Porous scaffolds varying in polyester type and crosslinking density were prepared from methacrylate-endcapped oligomers based on D,L-lactide, epsilon -caprolactone and trimethylene carbonate: LA/CL-hexanediol, LA/CL-dipentaerythritol and LA/TMC-HXD. The biocompatibility and bone formation were related with the degradation time and mechanical properties. The viability of fibroblasts was evaluated after incubation with extraction medium by MTT-assay. All scaffolds showed a good biocompatibility. Rat bone marrow cells were cultured on the scaffolds for 21 days and were able to attach and differentiate on the scaffolds. The cells expressed high alkaline phosphatase activity, have formed a mineralized extracellular matrix and secreted osteocalcin. TEM of the polymer interface revealed osteoblasts which secreted an extracellular matrix containing matrix vesicles loaded with apatite crystals.LA/TMC-HXD, LA/CL-HXD and LA/CL-DPENT had a 50% mass loss at 3,5 months respectively 6 and 7, 5 months. The mechanical properties improve by increasing the branching of the precursor methacrylates (by replacing HXD by DPENT) but do not depend on their chemical composition. Hence, scaffolds with high elastic properties and variable degradation time can be obtained, which are promising for bone tissue engineering.

Alkaline Phosphatase↗

PubMed2002

Flow-through cell for on-line amperometric determination of Ce(lV) during polymerization reactions.

In this paper, a method is described for the chronoamperometric determination of Ce(IV) in acidic aqueous solutions in order to study the kinetics and the mechanism of polymerization reactions with Ce(IV) as the initiator. The method is based on the FIA principle, in which small samples are injected in a continuous 1.0 mol L(-1) H2SO4 flow, and Ce(IV) is detected chronoamperometrically by reduction at the surface of a gold CDtrode. Such an electrode is made from a commercial CD that has a gold coating acting as a reflecting layer for the laser beam. It was found that a detection limit of 1.5 10(-7) mol L(-1) could be obtained with this method, that high concentrations (order of 10(-2) mol L(-1)) can be detected without IR drop and that the system can be used over several days without renewal or recalibration of the gold CDtrode.

Journal Article↗

PubMed2000

Poly-L-glutamic acid derivatives as vectors for gene therapy.

This paper describes the synthesis and evaluation of biodegradable derivatives of poly-L-glutamic acid as suitable vectors for gene therapy. When mixed with DNA the new polymers self assemble and form polyelectrolyte complexes. The formation of the complexes and determination of their stability towards disruption by serum albumin was monitored by Ethidium bromide (EtBr) fluorescence spectroscopy. All polymers were able to form complexes and their size, determined by photon correlation spectroscopy, was between 84.5+/-2 nm and 96. 7+/-1.6 nm, depending on the type of polymer and the charge ratio. All complexes were stable towards serum albumin. The results from the biodegradability tests, using tritosomes, show that the polymers are biodegradable and the rate of degradation is influenced by the number of charged groups in the side chains. Haemolysis and red blood cell (RBC) agglutination were assessed and compared to poly(L-lysine) (pLL) and polyethyleneimine (pEI). RBC agglutination was monitored with optical microscopy. Results show that the new polymers are less toxic than pLL and pEI. Preliminary transfection studies show that the polymers are suitable vectors for gene delivery.

Absorptiometry, Photon↗

PubMed2000

Physicochemical and biological characterisation of an antisense oligonucleotide targeted against the bcl-2 mRNA complexed with cationic-hydrophilic copolymers.

The aim of this study was to evaluate the use of cationic-hydrophilic copolymers for self-assembly with antisense oligonucleotides targeted to the bcl-2 mRNA in order to improve their biocompatibility and modulation of their pharmacokinetics for greater therapeutic usefulness. Examination of the ability of poly(trimethylammonioethyl methacrylate chloride)-poly[N-(2-hydroxypropyl)methacrylamide] (pHPMA-b-pTMAEM) block copolymers to condense the oligonucleotide by fluorescence and electrophoresis techniques showed that complexes were formed more efficiently than with copolymers containing poly(ethylene glycol) blocks grafted onto the backbone of poly(L-lysine) (pLL-g-pEG). In addition, the copolymer pTMAEM-b-pHPMA produced oligonucleotide complexes with the most favourable physicochemical properties appropriate for in vivo applications. The complexes were small (approximately 36 nm in diameter), with low surface charge as measured by zeta potential, relatively stable to physiological salt conditions and could be formed at a DNA concentration of 500 microg/ml. Complex formation with the copolymer pTMAEM-b-pHPMA or pLL-g-pEG reduced the urinary clearance of the oligonucleotide after intravenous injection into mice. However after 30 min, the oligonucleotide complexes were cleared from the bloodstream. These results indicate that for the systemic delivery of oligonucleotides the polymer-derived complexes are not stable enough for prolonged circulation. Instead, these complexes may be more suitable for localised in vivo applications.

Animals↗

PubMed2000

Structural and rheological properties of methacrylamide modified gelatin hydrogels.

Dynamic shear oscillation measurements at small strain were used to characterize the viscoelastic properties and related differences in the molecular structure of hydrogels based on gelatin methacrylamide. Gelatin was derivatized with methacrylamide side groups and was subsequently cross-linked by radical polymerization via photoinitiation. The light treatment of methacrylamide gelatin solutions resulted in the production of hydrogel films with high storage modulus (G'). Mechanical spectra and thermal scanning rheology of the obtained hydrogels are described. The temperature scan of the network below and above melting point of gelatin allowed us to identify the respective contributions of chemical and physical cross-linkage to the hydrogel elastic modulus. The results indicate that the rheological properties of the gelatin-based hydrogels can be controlled by the degree of substitution, polymer concentration, initiator concentration, and UV irradiation conditions.

Acrylamides↗

PubMed1998

Novel vectors for gene delivery formed by self-assembly of DNA with poly(L-lysine) grafted with hydrophilic polymers.

Complexes formed between DNA and cationic polymers are attracting increasing attention as novel synthetic vectors for delivery of genes. We are trying to improve biological properties of such complexes by oriented self-assembly of DNA with cationic-hydrophilic block copolymers, designed to enshroud the complex within a protective hydrophilic polymer corona. Poly(L-lysine) (pLL) grafted with range of hydrophilic polymer blocks, including poly(ethylene glycol) (pEG), dextran and poly[N-(2-hydroxypropyl)methacrylamide] (pHPMA), shows efficient binding to DNA and mediates particle self-assembly and inhibition of ethidium bromide/DNA fluorescence. The complexes formed are discrete and typically about 100 nm diameter, viewed by atomic force microscopy. Surface charges are slightly shielded by the presence of the hydrophilic polymer, and complexes generally show decreased cytotoxicity compared with simple pLL/DNA complexes. pEG-containing complexes show increased transfection activity against cells in vitro. Complexes formed with all polymer conjugates showed greater aqueous solubility than simple pLL/DNA complexes, particularly at charge neutrality. These materials appear to have the ability to regulate the physicochemical and biological properties of polycation/DNA complexes, and should find important applications in packaging of nucleic acids for specific biological applications.

Amino Acid Sequence↗

PubMed1997

The GyneFix implant systems for interval, postabortal and postpartum contraception: a significant advance in long-term reversible contraception. International Study Group on Intrauterine Drug Delivery.

Intrauterine contraception has a number of important advantages over other forms of contraception and remains, therefore, an important method of birth control. However, side-effects and other drawbacks have reduced its overall acceptance. Also misconceptions and lack of updated scientific knowledge among the potential users and providers are major obstacles to the widespread use of intrauterine contraception. Ideally, an intrauterine device (IUD) should prevent pregnancy effectively, be well tolerated, not become displaced or expelled over time, cause a minimum of side-effects, be long-lasting, have a strictly local effect, and be easy to insert and remove. A group at the University of Ghent, Belgium, the International Study Group on Intrauterine Drug Delivery, has developed, since 1985, a totally new concept in order to improve current intrauterine contraceptive efficacy and enhance tolerance, by creating a harmonious relationship between the uterine cavity and the contraceptive 'foreign body'. The new concept (GyneFix) consists of a non-biodegradable suture thread made of surgical 00 monofilament polypropylene on which six copper tubes are threaded, providing a total surface area of 330 mm2. The upper and lower tubes are crimped onto the thread to keep the tubes in place. The upper extremity of the thread is provided with a knot which serves as an anchor. The knot is implanted in the myometrium of the uterine fundus with a specially designed insertion instrument, thereby permanently securing the device in the uterine cavity. Since the initial clinical investigations, over 10,000 woman years of experience and up to 10 years' follow-up in international multicenter, non-comparative and comparative clinical trials have been collected. The clinical material also included a large number of nulligravid and nulliparous women. Due to the design characteristics of the GyneFix and its anchoring in the uterine fundus, an optimal tolerance and almost complete absence of expulsion were obtained. The constant release of copper ions in the upper part of the uterine cavity results in the high effectiveness of the anchored device. The effectiveness is higher than in the high-load conventional copper IUDs which have a risk of becoming displaced, partially or totally expelled in 10% or more (nulliparous women), resulting in a significant number of accidental pregnancies. The absence of frame and, as a consequence, its flexibility, explain the low incidence of side-effects and the very low incidence of complications, such as pelvic inflammatory disease and ectopic pregnancies. This new concept could be a major step forward in the acceptance of intrauterine contraception worldwide and increase its popularity. This article reviews the experience with the new concept for interval, postabortal and postpartum contraception.

Abortion, Induced↗

PubMed1996

Characterization of vectors for gene therapy formed by self-assembly of DNA with synthetic block co-polymers.

Cationic polymers can self-assemble with DNA to form polyelectrolyte complexes capable of gene delivery, although biocompatibility of the complexes is generally limited. Here we have used A-B type cationic-hydrophilic block co-polymers to introduce a protective surface hydrophilic shielding following oriented self-assembly with DNA. Block co-polymers of poly(ethylene glycol)-poly-L-lysine (pEG-pLL) and poly-N-(2-hydroxypropyl)methacrylamide-poly(trimethylammonioethyl methacrylate chloride) (pHPMA-pTMAEM) both show spontaneous formation of complexes with DNA. Surface charge measured by zeta potential is decreased compared with equivalent polycation-DNA complexes in each case. Atomic force microscopy shows that pHPMA-pTMAEM/DNA complexes are discrete spheres similar to those formed between DNA and simple polycations, whereas pEG-pLL/DNA complexes adopt an extended structure. Biological properties depend on the charge ratio of formation. At optimal charge ratio, pEG-pLL/DNA complexes show efficient transfection of 293 cells in vitro, while pHPMA-pTMAEM/DNA complexes are more inert. Both block co-polymer-DNA complexes show only limited cytotoxicity. Careful selection of block co-polymer structure can influence the physicochemical and biological properties of the complexes and should permit design of materials for specific applications, including targeted delivery of genes in vivo.

Cell Survival↗

PubMed1996

Polymeric prodrugs of mitomycin C designed for tumour tropism and sustained activation.

Prodrugs of mitomycin C (MMC) based on soluble poly-[N-(2-hydroxyethyl)-L-glutamine] (pHEG) polymers have been evaluated as tumour-targeted drugs. These materials are designed to exploit the enhanced permeability of tumour vasculature, combining a passive tumour tropism with decreased systemic liberation of free MMC. A tri- or tetrapeptide linkage (e.g. Gly-Phe-Ala-Leu) between pHEG and the aziridine nitrogen of MMC can combine good hydrolytic stability with rapid cleavage by lysosomal enzymes, releasing free MMC. The conjugates showed decreased systemic toxicity and could be administered to mice at a total MMC dose of 15 mg/kg i.v., compared with just 6 mg/kg for free MMC. Conjugates also showed better activity against animal models of established tumours, achieving up to 77% increased life span (ILS) against solid P388 leukaemia, compared with only 23% for free MMC, and up to 121% ILS against solid C26 colorectal carcinoma, compared with no activity for the free drug. Improving the therapeutic index of anticancer drugs by combining tumour tropism with decreased systemic toxicity is a versatile approach that should produce a new generation of improved anticancer agents.

Amino Acid Sequence↗

PubMed1992

Biodegradable polymers. II. Degradation characteristics of hydrolysis-sensitive poly[(organo)phosphazenes].

Polyphosphazenes with hydrolytic labile substituents have potential as biodegradable materials. By proper choice of the substituents, polymers can be prepared which can degrade to harmless products. The rate of biodegradation and the nature of the degradation products can be widely varied by changing the chemical composition of the polymers. The degradation properties of a series of new polyphosphazene derivatives are discussed. The synthesis of phosphazene polymers with variable amounts of ethyl 2-(O-glycyl)lactate or ethyl 2-(O-alanyl)lactate as cosubstituents was described previously. These polymers were prepared by partial reaction of poly[(dichloro)phosphazene] with the corresponding amine compound. Total halogen replacement was achieved by subsequent introduction of glycine ethyl ester cosubstituents. The degradation characteristics of these polymers in organic solution or in vitro was investigated. It was demonstrated that the introduction of hydrolysis-sensitive side-groups along the polymer chain results in an increased degradability of the poly[(amino acid ester)phosphazenes]. A plausible mechanism for the hydrolysis of these materials is proposed. The main hydrolysis pathway of poly[(amino acid ester)phosphazene] devices in vitro involves release of the amino acid ester side-group followed by hydrolysis of the ester with formation of the amino acid and ethanol. Initial hydrolysis of the ester bond with subsequent release of glycine cannot be excluded but is probably predominant.

Biocompatible Materials↗

PubMed1992

Biodegradable polymers. I. Synthesis of hydrolysis-sensitive poly[(organo)phosphazenes].

Polyphosphazenes with hydrolytic labile substituents have potential as biodegradable materials. By proper choice of the substituents, polymers can be prepared which degrade in vivo to form harmless products. The rate of biodegradation and the nature of the degradation products can be varied by changing the chemical composition of the polymers. A series of new degradable polyphosphazene derivatives are described. The synthesis of phosphazene polymers with various amounts of ethyl 2-(O-glycyl)lactate or ethyl 2-(O-alanyl)lactate as cosubstituents is described. These polymers are prepared by reaction of poly[(dichloro)phosphazene) with the corresponding amine compound. Total halogen replacement was achieved by subsequent introduction of glycine ethyl ester cosubstituents. Introduction of these hydrolysis-sensitive side-groups along the polymer chain increases the degradability of poly[(amino acid ester]phosphazenes).

Biocompatible Materials↗

PubMed1992

Di-iodo-L-tyrosine-labelled dextrans as molecular size markers of nasal absorption in the rat.

A series of fractionated di-iodo-L-tyrosine-labelled dextrans (DIT dextrans), with a narrow range of number average molecular weights from 1260 to 45,500 Da, was administered intranasally and intravenously to anaesthetized rats. The nasal absorption of these compounds ranged from 0.6 to 52.7%. There was an inverse relationship between molecular size and the proportion of an intranasal dose absorbed. The study demonstrated the usefulness of DIT dextrans as molecular weight markers and confirmed the relationship between molecular size and nasal absorption for highly water soluble compounds. These results also supported the proposition that there is a continuous range of aqueous pores in the nasal mucosa.

Administration, Intranasal↗

PubMedDate not supplied

Phosphonylation of purified human, canine and porcine cholinesterase by soman. Stereoselective aspects.

Cholinesterases (EC 3.1.1.8, acylcholine acylhydrolase) from the sera of man, dog and pig were purified 400-600-fold using a combination of ion-exchange and affinity chromatography. In a first approach, phosphonylation by soman was studied by using the half-resolved epimers C(+)P(+/-)-soman and C(-)P(+/-)-soman. The degradation of soman at the nanomolar level was followed in time by determining the remaining soman by capillary gas chromatography with NP detection. In the three sera investigated the P-(-)-epimer phosphonylates at a higher rate than its corresponding P(+)-counterpart and the stereoselectivity is greater for the C(+)-epimers than for the C(-)-epimers. Individual soman isomers were isolated from C(+)- and C(-)-epimers and quantified by gas chromatography. Second-order rate constants were determined for the phosphonylation of purified cholinesterase by isolated soman isomers. The C(+)P(-)-isomer has the highest phosphonylation rate for the three species; the other toxic isomer, C(-)P(-), has a five to ten-fold lower rate. The overall stereoselectivity is more marked in human cholinesterase than in canine. Porcine serum cholinesterase is phosphonylated by the P(-)-isomers at a slightly higher rate than the human enzyme.

Animals↗

PubMedDate not supplied

Poly(ethylene glycol) multiblock copolymer as a carrier of anti-cancer drug doxorubicin.

The synthesis of a novel water-soluble polymer drug carrier system based on biodegradable poly(ethylene glycol) block copolymer is described in this paper. The copolymer consisting of PEG blocks of molecular weight 2000 linked by means of an oligopeptide with amino end groups was prepared by interfacial polycondensation of the diamine and PEG bis(succinimidyl carbonate). The structure of the oligopeptide diamine consisting of glutamic acid and lysine residues was designed as a substrate for cathepsin B, a lysosomal enzyme, which was assumed to be one of the enzymes responsible for the degradation of the polymer carrier in vivo. Each of the oligopeptide blocks incorporated in the carrier contained three carboxylic groups of which some were used for attachment of an anti-cancer drug, doxorubicin (Dox), via a tetrapeptide spacer Gly-Phe-Leu-Gly. This tetrapeptide spacer is susceptible to enzymatic hydrolysis. In vitro release of Dox and the degradation of the polymer chain by cathepsin B as well as preliminary evaluation of in vivo anti-cancer activity of the conjugate are also demonstrated.

Animals↗

PubMedDate not supplied

Using biodegradable polymers in advanced drug delivery systems.

Research currently is under way to design novel drug delivery systems that enhance the duration of activity and that provide site-specific release of the drug. Biodegradable polymers represent an important class of materials used in the manufacture of these systems. This article discusses some major types of biodegradable polymers and reviews their applications in advanced drug delivery.

Biodegradation, Environmental↗