Shyh-Dar Lee

Plasma-induced grafted polymerization of acrylic acid and subsequent grafting of collagen onto polymer film as biomaterials

Polyacrylic acid (pAA) was introduced onto Ar-plasma treatment silicone rubber (SR) membrane surfaces by plasma-induced grafted polymerization. Collagen (type III) was also linked with the carboxylic group of pAA grafted onto the SR surface via a carbodiimine agent to obtain a secondary structure of SR. The SR surface properties were characterized by ATR-FTIR, ESCA, contact angle, and SEM. The biocompatibility of the SR surface was evaluated by a culture of cornea epithelial (CE) cells. Subsequently, 75-450 micrograms cm-2 of pAA were obtained on the SR surfaces under different reactive conditions; 3-12 micrograms cm-2 of collagen were linked on modified surfaces of SR. Moreover, ATR-FTIR and ESCA were utilized to confirm the proceedings of these reactions. The hydrophility of the modified SR was measured by a contact angle meter. The values of contact angle for SR grafted with pAA were approximately 45-50 degrees; a 50-55 degrees contact angle on pAA-g-SR to be further linked with collagen was subsequently obtained. Moreover, the influence of surface properties toward migration, growth and attachment of CE cells on the modified surfaces was also examined. Here, untreated SR was used as a control. Experimental results indicated that the number of CE cells attached onto the controlled SR was negligible. The attachment of cells onto pAA-grafted surfaces was clearly observed and peusopoda occurred; however, cell growth was depressed. This depression may have been caused by the acid environment of the pAA-grafted membrane. Nevertheless, both cell attachment and growth onto collagen-linked surfaces were significant. In addition, the morphology of the cells attached onto this surface was considered normal for primary cells. Collagen introduced on the SR surface was not denatured, i.e the natural properties of collagen were maintained. The results obtained in this study will hopefully lead to the successful development of modified SR for clinical applications.

Artificial cornea: surface modification of silicone rubber membrane by graft polymerization of pHEMA via glow discharge.

A method for producing various surfaces of silicone rubber membrane (SR) was developed in this study by grafting various amounts of poly(2-hydroxy ethyl methacrylate) (pHEMA) onto SR by plasma-induced grafted polymerization (PIP) as a homobifunctional membrane. The elemental composition and different carbon bindings on the surface of SR were examined by electron spectroscopy for chemical analysis with the amount of O1s/C1s being approximately 0.7 at 1 min, 60 W, 200 mTorr of Ar-plasma treatment. The peroxide group introduced on SR was measured via 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the amount of 6.85 x 10(-8) mol cm-2 reached optimum value at 1 min of Ar-plasma treatment. After Ar-plasma treated SR, the peroxide group (33D peak) was introduced on the surface of SR by negative spectra of secondary ion mass spectroscopy analysis, whereas ester groups (72D peak) were observed for pHEMA-grafted SR. For the in vitro test, the influence of various surfaces of SR on attachment and growth of rabbit corneal epithelial cells (CEC) was studied by cell culture assay. These results indicated that 56-150 micrograms cm-2 of pHEMA grafted onto SR were suitable values for attachment and growth of CEC. On the contrary, the large grafted amounts (500-1650 micrograms cm-2) of pHEMA on SR were insufficient for attachment and growth of CEC. For the in vivo test, the migration of CEC from host cornea to implant was investigated by slit lamp microscopy. The experimental results indicated that SRs grafted with pHEMA were completely covered with CEC 3 weeks after implantation of the membranes into the host cornea. These results provide a valuable reference for developing an artificial cornea.

Surface characterization and biological properties study of silicone rubber membrane grafted with phospholipid as biomaterial via plasma induced graft copolymerization

Poly(2-methacryloyloxyethyl phosphorylcholine) (pMPC) was grafted onto the surface of a silicon rubber (SR) membrane (pMPC-SR) by plasma induced grafted copolymerization (PIP). Argon plasma was used to activate the SR surfaces. Determination was also made of the influences of grafted copolymerization reaction time, reaction temperature, and monomer concentration on polymerization yield. The surface properties of SR were characterized by ATR-FTIR, ESCA, and SEM. In those analyses the ATR-FTIR spectra indicated that the pMPC grafted onto the SR surface at 1720 and 3300 cm(-1). The elemental composition and different carbon bindings on the surface of the SR were examined by ESCA. An increasing P1s/C1s value g was obtained in the grafted polymerization yield with a concentration of 0.05-0.5M of MPC in the isolated ethanol solution. The surface morphologies of pMPC-SR differed more than those of control and Ar plasma treated surfaces. The difference could have been caused by the homogeneous graft polymerization of pMPC onto the SR membrane. In the biological analyses, protein adsorption on pMPC-SR surfaces was reduced. The reduced level increased with an increase in the pMPC grafted amount. The epithelial cell attachment and growth onto these samples were suppressed. The blood compatibility for a series of pMPC-SR surfaces was examined by platelet adhesion. Blood platelet morphologies in contact with the high ratio of pMPC-SR surfaces were maintained, meaning that in this case the release reaction for platelets never occurred. Consequently, the high amount of pMPC-SR surface had excellent blood compatibility, further suggesting that prevention of adhesion, activation of platelets, and adsorption of blood protein could be achieved.

Preparation and characterization of a homobifunctional silicone rubber membrane grafted with acrylic acid via plasma‐induced graft copolymerization

Polyacrylic acid (PAA) was grafted onto the surface of silicone rubber membrane (SR) by plasma-induced graft copolymerization (PIP). Ar-plasma was used to activate the surface of SR. Also, a determination was made of the influences of plasma treatment power, pressure, time, grafted copolymerization reaction time, and monomer concentration on polymerization yield. The surface properties of SR were measured by ATR-FTIR, ESCA, and SIMS. In those analyses, the elemental composition and different carbon bindings on the surface of SR were examined by ESCA with the amount of O1s/C1s being approximately 0.7 at 60 s by Ar-plasma treatment (60 W, 200 mtorr). The peroxide group introduced on SR was measured via 1,1-diphenyl-2-picryhydrazyl (DPPH). The optimum amount of peroxide groups was 6.85 × 10−8 mol/cm2 at 60 s of Ar-plasma treatment. The peroxide group (33D peak) was introduced onto the surface of SR by negative spectra of SIMS analysis after SR treatment by Ar-plasma. An increase was obtained in grafted polymerization yield with a region of 5 to 50% (v/v) of acrylic acid aqueous solution. Both sites of polyacrylic acid were detected after staining by toluidine blue O. That is, a homobifunctional membrane was developed via this surface modification method.

Heterobifunctional membranes by plasma induced graft polymerization as an artificial organ for penetration keratoprosthesis

Highly biocompatible polymer membrane was developed for an artificial cornea in this surface modification study. Heterobifunctional silicone rubber membranes (hetero-SR) were prepared by grafting different functional polymers on each side of a silicone rubber membrane (SR). A novel type of bifunctional membrane was developed with the upper-side favoring cell attachment and growth, and the lower-side suppressing cell adhesion. The preparation of heterobifunctional membranes, characterization of polymer membrane surface properties such as ATR-FTIR and ESCA and contact angle, and biological analysis (in vitro and in vivo studies) were investigated in this work. Based on the biological analysis, the heterobifunctional membrane displays promising potential for use as an artificial cornea.

The effect of plasma-induced graft copolymerization of PHEMA on silicone rubber towards improving corneal epithelial cells growth

A PHEMA grafted polymer film was prepared by plasma induced graft copolymerization onto an elastic material, silicone rubber. The control, Ar plasma-treated, and PHEMA-grafted silicone rubber surfaces were characterized by ESCA, FTIR-ATR, and SEM techniques. ESCA verified the respective chemical shift of control and Ar plasma-treated films. The presence of the grafted PHEMA was also verified by ESCA. The amounts of grafted PHEMA did not monotonously increase with the plasma exposure conditions, but decreased after passing a maximum. The introduction of PHEMA onto a hydrophobic support provided an adequate surface for rabbit corneal epithelium cell attachment and growth. Cell attachment and growth onto these surfaces were examined by light microscopy. Cell attachment onto the control and Ar plasma-treated surface was negligible, while improved attachment and growth of rabbit corneal epithelium cells was demonstrated on the PHEMA-grafted polymer surface. The PHEMA-grafted silicone rubber surface demonstrated a confluent cell layer after 72 h.

Platelet adhesion and cellular interaction with poly(ethylene oxide) immobilized onto silicone rubber membrane surfaces.

Cellular interaction and platelet adsorption were investigated on poly(ethylene oxide) (PEO) immobilized silicone rubber membrane (SR) which has polyacrylic acid grafts on the surfaces. Polyacrylic acid (PAA) had been introduced to the SR surface after Ar plasma treatment of SR surfaces to introduce peroxide groups. Surface characterizations were made using ATR-FTIR, ESCA, SEM, and contact angle measurements. Experimental results obtained by ESCA high resolution curve fitting spectra indicated that the amount of bisamino PEO of different molecular weights immobilized onto SR surfaces were similar, which showed that the influence of the length of molecular chains (-C-C-O-) on the reactivity of terminal amino group is negligible. The wettability of modified SR surfaces increased with an increase in PEO molecular weight. Biological studies such as corneal epithelial cell culture and blood platelet adhesion were performed to understand the biocompatibility of modified SR surfaces. Biological studies using corneal epithelial cells showed that cell migration, attachment and proliferation onto PEO-20000 immobilized SR surface were suppressed, whereas these biological activities on PEO-600 were enhanced. Another study on platelet adhesion revealed that many platelets attached to PEO-600 immobilized SR, while platelet deposition was rarely observed on SR grafted with PEO-3350. The effects of different PEO molecular chains on biological response were discussed.

Novel Biomaterials as Artificial Cornea via Plasma Induced Grafted Polymerization

The primary objective of this study is to prepare a highly biocompatible polymer membrane by surface modification and to further develop an artificial cornea. Novel heterobifunctional membranes prepared by grafting different functional polymers onto silicone rubber membranes were achieved. In this work, we report preparation and surface characterization of the heterobifunctional membranes, and their biological analysis (in vitro and in vivo studies). Based on the biological analysis, the heterobifunctional membrane exhibits high potential to be used as artificial cornea.