Biomedical subjects

H Chaouk

Publications and source records attributed to H Chaouk.

4 recordsLinked to original sources

PubMed2000

Epithelialization of a synthetic polymer in the feline cornea: a preliminary study.

PURPOSE: This study examined the potential of a synthetic polymer to support stable epithelial growth when implanted in the feline cornea. METHODS: A perfluoropolyether-based polymer was cast into lenticules that were coated with collagen I and implanted in four feline corneas. Epithelial growth onto the lenticules was monitored clinically for 6 weeks, after which time the animals were killed, and three corneas were evaluated histologically. Immunohistochemistry was used to identify proteins associated with the formation of a basement membrane (laminin) and adhesion complexes (bullous pemphigoid antigen and collagen VII). Electron microscopy was used to examine the tissue-polymer interface for evidence of the assembly of these adhesive structures. RESULTS: Postoperative epithelial growth began on days 2 to 3, and lenticules were fully epithelialized by days 5 to 9. Lenticules were clinically well tolerated and histology showed epithelium consisting of multiple layers adherent to the lenticule's surface. Laminin, bullous pemphigoid antigen and collagen VII were identified at the tissue-polymer interface using immunohistochemistry. Ultrastructural examination showed evidence of assembly of these proteins into a recognizable basement membrane and hemidesmosomal plaques. CONCLUSIONS: A perfluoropolyether-based polymer coated with collagen I was implanted in the feline cornea and supported epithelial growth that showed signs of persistent adhesion, both clinically and histologically. This polymer shows potential for ophthalmic applications that require sustained epithelialization.

Animals↗

PubMed1999

New ligand, N-(2-pyridylmethyl)aminoacetate, for use in the immobilised metal ion affinity chromatographic separation of proteins.

A new chelating compound has been developed for use in the immobilised metal ion affinity chromatographic separation of proteins. The tridentate ligand, sodium N-(2-pyridylmethyl)aminoacetate (carbpyr), 1, was prepared via a one-step synthesis from 2-picolylamine, 3 and then immobilised onto Sepharose CL-4B through the epoxide coupling procedure. The binding behaviour of the resulting IMAC sorbent, following chelation with Cu2+ ions to a density of 152 micromol Cu2+ ions/g gel was characterised by frontal analysis experiments using horse heart myoglobin (HMYO) at pH 7.0 and pH 9.0. From the derived isotherms, the adsorption capacity, q(m), for the binding of HMYO to immobilised Cu2+-N-(2-pyridylmethyl)aminoacetate (im-Cu2+-carbpyr)-Sepharose CL-4B at these pH values was found to be 1.92 and 1.91 micromol/g sorbent, respectively, whilst the dissociation constants K(D) were 0.0092 x 10(-6) M and 0.0062 x 10(-6) M at pH 7.0 and pH 9.0, respectively, indicating that the HMYO-im-Cu2+-N-(2-pyridylmethyl)aminoacetate complex was more stable under alkaline conditions, although the binding capacity in terms of micromol protein/g gel remained essentially unchanged. The selectivity features of the im-Cu2+-carbpyr-Sepharose CL-4B sorbent were further characterised in terms of the binding properties with several human serum proteins at pH 5.0, pH 7.0 and pH 9.0.

Adsorption↗

PubMed1999

Examination of the protein binding behaviour of immobilised copper (II)-2,6-diaminomethylpyridine and its application in the immobilised metal ion affinity chromatographic separation of several human serum proteins.

A new metal ion chelator has been developed for use in the immobilised metal ion affinity chromatography (IMAC) of proteins. The aromatic tridentate ligand 2,6-diaminomethylpyridine (bisampyr), 1, was prepared as the dihydrochloride salt, via a two step synthesis from 2,6-pyridinedimethanol, 2, and immobilised onto Sepharose CL-4B through an epoxide coupling procedure. The resulting sorbent was chelated with Cu2+ ions to a density of 420 micromol Cu2+ ions per g gel and then characterised by frontal analysis using the protein, horse heart myoglobin (HMYO), at pH 7.0 and 9.0. From the resulting adsorption isotherms, the adsorption capacity, qm, for HMYO at pH 7.0 and pH 9.0 with the immobilised Cu2+-bisampyr Sepharose sorbent was found to be 1.27 micromol protein/g gel and 1.43 micromol protein/g gel, whilst the corresponding dissociation constants, K(D)s, were 18.0 x 10(-6) M and 16.0 x 10(-6) M respectively. The results confirm that the HMYO-Cu2+-bisampyr complex had similar stability at these pH values. This finding is in contrast with the situation observed with some other commonly used IMAC chelating ligates such as Cu2+-iminodiacetic acid (Cu2+-IDA) or Cu2+-nitrilotriacetic acid (Cu2+-NTA). Using human serum proteins, the interactive properties of the immobilised Cu2+-bisampyr Sepharose sorbent were further characterised at pH 5.0, 7.0 and 9.0 with specific reference to the binding behaviour of albumin, transferrin, and alpha2-macroglobulin.

Adsorption↗

PubMed1999

Cell interactions with perfluoropolyether-based network copolymers.

We have investigated the potential of several polymers based on perfluoropolyether (PFPE) macromonomers for use in biomaterial applications. Polymer networks were synthesised from the PFPE macromonomers of increasing chain length and the adhesion and proliferation of corneal, vascular and bone cells was evaluated on these polymers. The polymer surfaces were quite hydrophobic, having sessile air-water contact angles ranging between 96 and 125 degrees. However, these polymers supported the attachment and growth of bovine corneal epithelial and endothelial cells and fibroblasts at 60-100% of the rate of cell growth on the culture substratum, TCPS. Furthermore, the PFPE polymers supported the attachment and growth of vascular endothelial cells (from human umbilical artery) and human bone-derived cells over a 7 day period at an equal level to TCPS. The relationship between the macromonomer chain length (n = 1 to 4) and the ability of the resulting PFPE homopolymer to support the overgrowth of corneal epithelial tissue was also evaluated. The PFPE-containing polymers supported corneal epithelial tissue overgrowth, with the most effective having a performance equivalent to that of TCPS. In addition to these homopolymers, copolymers comprising of PFPE and N,N-dimethylaminoethyl methacrylate (DMAEMA) were also synthesised. Surprisingly, the addition of DMAEMA to the PFPE polymer network lead to a reduction in the growth and attachment of corneal epithelial cells and fibroblasts. These results indicate that PFPE-based materials show a potential for use in the development of biomaterials in the ocular, vascular and orthopaedic areas.

Animals↗