- Email: [email protected]
49 Biocompatibility evaluation of residual hydrogen peroxide solution using in vitro cytotoxicity assays
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Abstracts of the 2011 BCLA Annual Clinical Conference / Contact Lens & Anterior Eye 34, Supplement 1 (2011) S1–S43
47 Differences in time-matched profiles of corneal staining severity and extent with three lens disinfecting solution chemistries
Conclusions: CR was found to be less cytotoxic than OFR or OFE in MPSCLs biocompatibility, as defined by lower in vitro cytotoxicity scores. Cytotoxicity scores with CR did not correlate with preservative uptake, but a weak correlation with alexidine dihydrochloride release was observed.
Eugenia Y Kao 1, *, Ling C Huang 1 , Linda Tsai 1 , Nicholas Tarantino 1 , Marc Odrich 2 Abbott Medical Optics Inc., Santa Ana, USA; 2 Riverdale Laser Center, New York, USA
1
*E-mail address: [email protected] Purpose: Corneal staining associated with contact lens wear and multipurpose solutions (MPS) may indicate ocular surface toxicity. However, corneal staining varies with the time of evaluation. These studies examined corneal staining over time of three MPSs with balafilcon A lenses. Method: Three MPSs were evaluated in two studies: Solution A (COMPLETE® RevitaLens Multi-Purpose Disinfecting Solution [MPDS], Abbott Medical Optics Inc.); Solution B (Opti-Free® RepleniSH® MPDS, Alcon, Inc.] and Solution C (Biotrue™ MPS, Bausch & Lomb Incorporated). These studies were double-masked, contralateral eye, with 29 to 30 subjects. Subjects were randomized to wear lenses soaked in different MPSs in each eye. After baseline evaluations, follow-up visits occurred after one, two, four and six+ hours (end-of-day) of lens wear. Corneal staining was graded on severity and extent by a five-point scale, while comfort was assessed by a 10-point scale. Results: Significant differences in corneal staining extent were found between Solutions A and B at two hours (p=0.037) and Solutions A and C at every follow-up time point (p<0.002). Significant differences in corneal staining severity were found between Solutions A and C at two and four hours (p<0.001). Corneal staining incidence and area were highest with Solution C. No significant difference in comfort was found between any of the solutions at any time point. Conclusions: The clinical performance of different MPSs can be distinctly different between different solution chemistries. Comparative conclusions on corneal staining should factor in the time of evaluation and nature of the kinetics curve.
49 Biocompatibility evaluation of residual hydrogen peroxide solution using in vitro cytotoxicity assays Ann M Wright 1, *, Charlon Tolliver 1 , Leroy Muya 1 , Colleen Toole 2 1
CIBA VISION, Duluth, GA, USA; 2 CEE TOX, Kalamazoo, MI, USA
*E-mail address: [email protected] Purpose: Evaluate residual hydrogen peroxide (H2O2) and silicone hydrogel (SiHy) contact lenses (CL) cycled in experimental hydrogen peroxide soft contact lens care solutions (EH2O2LCS) using in vitro models. Method: Lotrafilcon B CLwas cycled in a EH2O2LCPsystem for the 6-hour regimen time. L929 cells were exposed to cycled SiHy CL and evaluated using the Direct Contact and Agar Overlay Tests for a 24-hour exposure. Additionally, the EH2O2LCS neutralized solution at 6 hours was measured for residual H2O2 and tested using the modified Elution and Agar Overlay Tests for a 48-hour exposure. Finally, 3D human corneal Epithelia (HCE) model (SkinEthicTM) was exposed to 100 ppm HP for 6 hours and evaluated for viability (MTT) and cytokine release (IL-1a). Results: The residual H2O2 levels for EH2O2LCS at 6 hours were approximately 50 ppm. SiHy CL cycled/neutralized for 6 hours with EH2O2LCS were non-cytotoxic in the Direct Contact and the Agar Overlay Tests. HCE 3-D model exposed to 100 ppm residual H2O2 levels for 6 hours were viable and did NOT exhibit significant IL1-α cytokines. The residual peroxide, when tested as a liquid, was cytotoxic in the modified Elution and Agar Overlay Tests. Conclusions: Use of Direct Contact and Agar Overlay tests for cycled lenses and use of 3D human corneal model for lens care solutions are appropriate for simulating biocompatibility on eye, and for determining the relative cytotoxicity of lens care systems. Human 3D models using corneal epithelial cells support literature for the non cytotoxicity of low level HP on the eye.
48 Comparative biocompatibility of contact lens multi-purpose disinfecting solutions with soft contact lenses – potential correlations with lens preservative uptake and release profiles Ling C Huang 1, *, Lauren Crawford 1 , Charles H Powell 1 , Lisa Hoong 1 , Amit R Agarwal 2 Abbott Medical Optics Inc., Santa Ana, CA, USA; 2 University of Southern California, Los Angeles, CA, USA
1
*E-mail address: [email protected] Purpose: To evaluate effects of contact lens multi-purpose disinfecting solutions (MPDS) by in vitro biocompatibility assessment using mouse fibroblast (L929) cells and in relationship to preservative uptake and release profiles with various soft contact lenses (CLs). Method: Complete RevitaLens MPDS (CR) was evaluated against Opti-Free RepleniSH MPDS (OFR) and Opti-Free Express MPS (OFE). Five CL brands were examined: polymacon (Soflens38), galyfilcon A (Acuvue Advance), lotrafilcon A (O2 Optix), balafilcon A (Pure Vision) and comfilcon A (Biofinity) lenses. MPS-treated CLs (100ml, 4 days, n=3) were placed onto L929 cells (24hrs). Cells were scored for reactivity according to USP criteria. Comparative antimicrobial uptake of lenses soaked in CR, and subsequent release kinetics in buffered saline (1ml/lens, n=3–5), were determined out to equilibrium (28 days max). Results: In vitro cytotoxicity showed that CR-treated Pure Vision, Acuvue Advance, and O2 Optix lenses (grade 1–2) were less cytotoxic compared with OFR and OFE (grade 3–4). All MPS-treated Soflens38 CLs demonstrated low cytotoxicity scores (grade 2). MPS-treated Biofinity CLs displayed moderate cytotoxicity (grade 3–4). Lens uptake of alexidine dihydrochloride (0.4–4.1 ug/mg dry lens) showed Soflens38 < O2 Optix < Acuvue Advance < Biofinity < Pure Vision. Lens release of alexidine dihydrochloride (6.6–30.9 ng/mg dry lens) showed O2 Optix < Pure Vision < Acuvue Advance < Biofinity < Soflens 38. No quantifiable uptake or release of polyquaternium-1 from lenses was found.
50 Use of a bovine lens primary organ cultureto evaluate the toxicity of contact lens caresolution products David J McCanna*, Elizabeth Kao, Heather Marriott, Jacob G Sivak University of Waterloo, Waterloo, Canada *E-mail address: [email protected] Purpose: We evaluated the toxicity of contact lenses that had been soaked in various contact lens solution products on a primary lens epithelial culture. This test model is proposed to be used in the in vitro test battery to assess contact lens care solution toxicity due to its ability to evaluate epithelial cells from a primary organ culture that has not been immortalized. Method: Etafilcon A contact lenses (AcuVue 2 lenses) were soaked in various concentrations of benzalkonium chloride (BAK) and the contact lens care solutions Solocare, Optifree Express and ReNu MultiPlus. The contact lenses were then placed on the epithelial cells of intact whole cultured bovine lenses. After incubation the epithelial cells were assessed for viability using the alamarBlue assay and for mitochondrial integrity using rhodamine. Results: It was shown that there was a dose dependent significant increase in toxicity (p<0.05) as the concentration of the BAK soaking solution was increased from 0.0001% to 0.01% with severe toxicity seen in the lens epithelial exposed to 1% BAK. The contact lenses soaked in the currently marketed contact lens solutions did not cause a reduction in viability or in the mitochondria integrity of the epithelium. Conclusions: This experiment demonstrated that the bovine lens epithelium can be utilized as a primary cell culture model to assess the toxicity contact lens care products. The sensitivity of the assay to a known ocular irritant (BAK) was demonstrated and the assay confirmed the safety of currently marketed contact lens solution products.
Abstracts of the 2011 BCLA Annual Clinical Conference / Contact Lens & Anterior Eye 34, Supplement 1 (2011) S1–S43
47 Differences in time-matched profiles of corneal staining severity and extent with three lens disinfecting solution chemistries
Conclusions: CR was found to be less cytotoxic than OFR or OFE in MPSCLs biocompatibility, as defined by lower in vitro cytotoxicity scores. Cytotoxicity scores with CR did not correlate with preservative uptake, but a weak correlation with alexidine dihydrochloride release was observed.
Eugenia Y Kao 1, *, Ling C Huang 1 , Linda Tsai 1 , Nicholas Tarantino 1 , Marc Odrich 2 Abbott Medical Optics Inc., Santa Ana, USA; 2 Riverdale Laser Center, New York, USA
1
*E-mail address: [email protected] Purpose: Corneal staining associated with contact lens wear and multipurpose solutions (MPS) may indicate ocular surface toxicity. However, corneal staining varies with the time of evaluation. These studies examined corneal staining over time of three MPSs with balafilcon A lenses. Method: Three MPSs were evaluated in two studies: Solution A (COMPLETE® RevitaLens Multi-Purpose Disinfecting Solution [MPDS], Abbott Medical Optics Inc.); Solution B (Opti-Free® RepleniSH® MPDS, Alcon, Inc.] and Solution C (Biotrue™ MPS, Bausch & Lomb Incorporated). These studies were double-masked, contralateral eye, with 29 to 30 subjects. Subjects were randomized to wear lenses soaked in different MPSs in each eye. After baseline evaluations, follow-up visits occurred after one, two, four and six+ hours (end-of-day) of lens wear. Corneal staining was graded on severity and extent by a five-point scale, while comfort was assessed by a 10-point scale. Results: Significant differences in corneal staining extent were found between Solutions A and B at two hours (p=0.037) and Solutions A and C at every follow-up time point (p<0.002). Significant differences in corneal staining severity were found between Solutions A and C at two and four hours (p<0.001). Corneal staining incidence and area were highest with Solution C. No significant difference in comfort was found between any of the solutions at any time point. Conclusions: The clinical performance of different MPSs can be distinctly different between different solution chemistries. Comparative conclusions on corneal staining should factor in the time of evaluation and nature of the kinetics curve.
49 Biocompatibility evaluation of residual hydrogen peroxide solution using in vitro cytotoxicity assays Ann M Wright 1, *, Charlon Tolliver 1 , Leroy Muya 1 , Colleen Toole 2 1
CIBA VISION, Duluth, GA, USA; 2 CEE TOX, Kalamazoo, MI, USA
*E-mail address: [email protected] Purpose: Evaluate residual hydrogen peroxide (H2O2) and silicone hydrogel (SiHy) contact lenses (CL) cycled in experimental hydrogen peroxide soft contact lens care solutions (EH2O2LCS) using in vitro models. Method: Lotrafilcon B CLwas cycled in a EH2O2LCPsystem for the 6-hour regimen time. L929 cells were exposed to cycled SiHy CL and evaluated using the Direct Contact and Agar Overlay Tests for a 24-hour exposure. Additionally, the EH2O2LCS neutralized solution at 6 hours was measured for residual H2O2 and tested using the modified Elution and Agar Overlay Tests for a 48-hour exposure. Finally, 3D human corneal Epithelia (HCE) model (SkinEthicTM) was exposed to 100 ppm HP for 6 hours and evaluated for viability (MTT) and cytokine release (IL-1a). Results: The residual H2O2 levels for EH2O2LCS at 6 hours were approximately 50 ppm. SiHy CL cycled/neutralized for 6 hours with EH2O2LCS were non-cytotoxic in the Direct Contact and the Agar Overlay Tests. HCE 3-D model exposed to 100 ppm residual H2O2 levels for 6 hours were viable and did NOT exhibit significant IL1-α cytokines. The residual peroxide, when tested as a liquid, was cytotoxic in the modified Elution and Agar Overlay Tests. Conclusions: Use of Direct Contact and Agar Overlay tests for cycled lenses and use of 3D human corneal model for lens care solutions are appropriate for simulating biocompatibility on eye, and for determining the relative cytotoxicity of lens care systems. Human 3D models using corneal epithelial cells support literature for the non cytotoxicity of low level HP on the eye.
48 Comparative biocompatibility of contact lens multi-purpose disinfecting solutions with soft contact lenses – potential correlations with lens preservative uptake and release profiles Ling C Huang 1, *, Lauren Crawford 1 , Charles H Powell 1 , Lisa Hoong 1 , Amit R Agarwal 2 Abbott Medical Optics Inc., Santa Ana, CA, USA; 2 University of Southern California, Los Angeles, CA, USA
1
*E-mail address: [email protected] Purpose: To evaluate effects of contact lens multi-purpose disinfecting solutions (MPDS) by in vitro biocompatibility assessment using mouse fibroblast (L929) cells and in relationship to preservative uptake and release profiles with various soft contact lenses (CLs). Method: Complete RevitaLens MPDS (CR) was evaluated against Opti-Free RepleniSH MPDS (OFR) and Opti-Free Express MPS (OFE). Five CL brands were examined: polymacon (Soflens38), galyfilcon A (Acuvue Advance), lotrafilcon A (O2 Optix), balafilcon A (Pure Vision) and comfilcon A (Biofinity) lenses. MPS-treated CLs (100ml, 4 days, n=3) were placed onto L929 cells (24hrs). Cells were scored for reactivity according to USP criteria. Comparative antimicrobial uptake of lenses soaked in CR, and subsequent release kinetics in buffered saline (1ml/lens, n=3–5), were determined out to equilibrium (28 days max). Results: In vitro cytotoxicity showed that CR-treated Pure Vision, Acuvue Advance, and O2 Optix lenses (grade 1–2) were less cytotoxic compared with OFR and OFE (grade 3–4). All MPS-treated Soflens38 CLs demonstrated low cytotoxicity scores (grade 2). MPS-treated Biofinity CLs displayed moderate cytotoxicity (grade 3–4). Lens uptake of alexidine dihydrochloride (0.4–4.1 ug/mg dry lens) showed Soflens38 < O2 Optix < Acuvue Advance < Biofinity < Pure Vision. Lens release of alexidine dihydrochloride (6.6–30.9 ng/mg dry lens) showed O2 Optix < Pure Vision < Acuvue Advance < Biofinity < Soflens 38. No quantifiable uptake or release of polyquaternium-1 from lenses was found.
50 Use of a bovine lens primary organ cultureto evaluate the toxicity of contact lens caresolution products David J McCanna*, Elizabeth Kao, Heather Marriott, Jacob G Sivak University of Waterloo, Waterloo, Canada *E-mail address: [email protected] Purpose: We evaluated the toxicity of contact lenses that had been soaked in various contact lens solution products on a primary lens epithelial culture. This test model is proposed to be used in the in vitro test battery to assess contact lens care solution toxicity due to its ability to evaluate epithelial cells from a primary organ culture that has not been immortalized. Method: Etafilcon A contact lenses (AcuVue 2 lenses) were soaked in various concentrations of benzalkonium chloride (BAK) and the contact lens care solutions Solocare, Optifree Express and ReNu MultiPlus. The contact lenses were then placed on the epithelial cells of intact whole cultured bovine lenses. After incubation the epithelial cells were assessed for viability using the alamarBlue assay and for mitochondrial integrity using rhodamine. Results: It was shown that there was a dose dependent significant increase in toxicity (p<0.05) as the concentration of the BAK soaking solution was increased from 0.0001% to 0.01% with severe toxicity seen in the lens epithelial exposed to 1% BAK. The contact lenses soaked in the currently marketed contact lens solutions did not cause a reduction in viability or in the mitochondria integrity of the epithelium. Conclusions: This experiment demonstrated that the bovine lens epithelium can be utilized as a primary cell culture model to assess the toxicity contact lens care products. The sensitivity of the assay to a known ocular irritant (BAK) was demonstrated and the assay confirmed the safety of currently marketed contact lens solution products.