Simultaneous measurement of refractive index and thickness of multilayer systems using Fourier domain optical coherence tomography, part 2: implementation.

We introduce a theoretical method for simultaneous measurement of refractive index and thickness of multilayer systems using Fourier domain optical coherence tomography (FD-OCT) without any auxiliary arrangement. The input data to the formalism are the FD-OCT measured optical path lengths (OPLs) and properly selected spectral components of FD-OCT interference spectrum. The outputs of the formalism can be affected significantly by uncertainty in measuring the OPLs. An optimization method is introduced to deal with the relatively large amount of uncertainty in measured OPLs and enhance the final results. Simulation result shows that by using the optimization method, indices can be extracted with the absolute error ? 0.001 for transparent biological samples having indices < 1.55 .

Simultaneous measurement of refractive index and thickness of multilayer systems using Fourier domain optical coherence tomography, part 1: theory.

We introduce a theoretical framework for simultaneous refractive index and thickness measurement of multilayer systems using the Fourier domain optical coherence tomography (FD-OCT) system without any previous information about the item under investigation. The input data to the new formalism are the FD-OCT measured optical path lengths and properly selected spectral components of the FD-OCT interference spectrum. No additional arrangement, reference reflector, or mechanical scanning is needed in this approach. Simulation results show that the accuracy of the extracted parameters depends on the index contrast of the sample while it is insensitive to the sample's thickness profile. For transparent biological samples with smooth interfaces, when the object is in an aqueous medium and has indices < 1.55 , this method can extract indices and thicknesses with the absolute error ? 0.001 .

A focus on the optical properties of the regenerated newt lens.

Lens regeneration studies in the adult newt suggest that molecular aspects of lens regeneration are complete within 5 weeks of lentectomy. However, very little is known about the optical properties of the regenerated lens. In an aquatic environment, the lens accounts for almost all of the refractive power of the eye, and thus, a fully functional lens is critical. We compared the optical properties of 9- and 26-week regenerated lenses in the red spotted newt, Notophthalmus viridescens, with the original lenses removed from the same eyes. At 9 weeks, the regenerated lenses are smaller than the original lenses and are histologically immature, with a lower density of lens proteins. The 9 week lenses have greater light transmission, but significantly reduced focal length and refractive index than the original lenses. This suggests that following 9 weeks of regeneration, the lenses have not recovered the functionality of the original lens. By 26 weeks, the transmission of light in the more mature lens is reduced, but the optical parameters of the lens have recovered enough to allow functional vision.

Advanced corneal surface ablation efficacy in myopia: changes in higher order aberrations.

OBJECTIVE: To evaluate the change of ocular and corneal higher order aberrations (HOAs) after wavefront-guided advanced surface ablation (WF-ASA) for myopia using 4 different epithelial management techniques (AA-PRK, LASEK, Epi-PRK, and Epi-LASIK). DESIGN: Retrospective single centre excimer database analysis. PARTICIPANTS: Two hundred and forty eyes equally divided between the 4 WF-ASA techniques, matched for mean and range of required spherical correction. METHODS: Ocular wavefront aberrations were measured using the wavescan aberrometer and corneal aberrations were obtained from corneal topography elevation maps and calculated by ray-tracing. All data were collected preoperatively and 3 months following treatment. The aberrations were described as Zernike polynomials, and analysis focused on total HOAs and spherical aberration (SA). RESULTS: Three months postoperatively, there was a statistically significant surgically induced increase in total HOAs and SA both for ocular and corneal analysis (p < 0.001). There was no statistically significant difference in the induced ocular SA and HOAs between the groups, but the differences in induced corneal SA and HOAs were significant (p < 0.010). Ocular changes in SA were weakly correlated to preoperative SA (20.30, p < 0.001) but strongly correlated to applied spherical correction (20.68, p < 0.001). Surgically induced corneal SA was weakly correlated to preoperative corneal SA (20.34, p < 0.001) and applied spherical correction (20.46, p < 0.001). CONCLUSIONS: Three months postoperatively, all procedures resulted in an increase in ocular and cornealHOAs and SA. Induced aberrations were most strongly correlated to the applied spherical power correction. Modifying the existing ablation pattern to compensate for induced HOAs might improve the outcome.

Quantitative impact of small angle forward scatter on whole blood oximetry using a Beer-Lambert absorbance model.

It is well known that red blood cell scattering has an impact on whole blood oximetry as well as in vivo retinal oxygen saturation measurements. The goal of this study was to quantify the impact of small angle forward scatter on whole blood oximetry for scattering angles found in retinal oximetry light paths. Transmittance spectra of whole blood were measured in two different experimental setups: one that included small angle scatter in the transmitted signal and one that measured the transmitted signal only, at absorbance path lengths of 25, 50, 100, 250 and 500 µm. Oxygen saturation was determined by multiple linear regression in the 520-600 nm wavelength range and compared between path lengths and experimental setups. Mean calculated oxygen saturation differences between setups were greater than 10% at every absorbance path length. The deviations to the Beer-Lambert absorbance model had different spectral dependences between experimental setups, with the highest deviations found in the 520-540 nm range when scatter was added to the transmitted signal. These results are consistent with other models of forward scatter that predict different spectral dependences of the red blood cell scattering cross-section and haemoglobin extinction coefficients in this wavelength range.

Correlation between refractive error, corneal power, and thickness in a large population with a wide range of ametropia.

PURPOSE: To determine the correlations between mean keratometry (KM), central corneal thicknesses (CCT), and cycloplegic spherical equivalent (SE) in patients with a wide range of ametropia. METHODS: Retrospective analysis of the excimer laser surgery database at the University of Ottawa Eye Institute between 1993 and 2008 was performed. This study included 3395 eyes from 1858 subjects. The refractive error ranged from +6.75 to -14.00 D. CCT was obtained either by ultrasound pachymetry or anterior segment tomography. Keratometry was determined using an autokeratorefractometer. RESULTS: In the myopic group, the SE was observed to be inversely proportional to the KM (correlation coefficient, -0.18; P < 0.01). The KM and CCT were also inversely proportional (-0.11; P < 0.01). In hyperopes, a correlation between the cycloplegic SE and KM was also found (-0.25; P < 0.01), but the CCT did not correlate with either of these metrics. A direct correlation for the myopic group was found between KM and the difference in power of the principal meridians (keratometric astigmatism [KA]) (0.08; P < 0.01). This relationship was not observed for the hyperopic group. Within the myopic group the SE correlated with the refractive astigmatism (RA) (-0.04; P = 0 0.04). In all groups, a strong correlation was observed between RA and KA (0.78; P < 0.01). CONCLUSIONS: In the myopia group, the KM showed close correspondence with KA and an inverse relationship with SE and CCT. In hyperopes, an inverse correlation between the KM and SE was found, but no correlation with CCT was evident.

Effects of advanced surface ablations and intralase femtosecond LASIK on higher order aberrations and visual acuity outcome.

BACKGROUND/AIMS: To study the changes in wavefront (ocular) and corneal higher order aberrations (HOAs) and visual acuity (VA) outcome following wavefront-guided advanced surface ablation (ASA) techniques and intralase femtosecond LASIK (iLASIK) in myopia treatment. METHODS: Ocular aberration and corneal topography of 240 eyes in the ASA techniques (this was equally divided into a flap-on group where the epithelial flap was preserved and reapplied to the photoablated stromal bed and a flap-off group when the epithelial flap was discarded during the procedure), and 138 eyes in the iLASIK group were obtained before and 3 months following treatment. The correlation of aberrations with best spectacle-corrected visual acuity was analyzed. RESULTS: At 3 months, there was statistically significant (P < 0.001) surgically induced increase in spherical aberration (SA) in each of the techniques for both ocular and corneal analysis. iLASIK induced significantly less ocular and corneal HOAs (P < 0.001). The mean manifest refractive spherical equivalent was closer to attempted correction compared to other groups (P < 0.001). Eighty-three eyes (70%) of flap-on, 80 (67%) flap-off and 94 eyes (68%) in the iLASIK group achieved 20/20 uncorrected VA. Fifteen eyes (11%) accomplished 20/12.5 or better in iLASIK compared to 4 (3%) for flap-on and 7 (6%) for flap-off ASA group. Only the flap-off treatment showed a consistent correlation between the corrected aberrations and visual performance. CONCLUSION: At 3 months, all procedures resulted in a significant increase in HOAs and SA. All had comparable 20/20 VA and 11% of iLASIK patients achieved 20/12.5 or better level.

A biosynthetic alternative to human donor tissue for inducing corneal regeneration: 24-month follow-up of a phase 1 clinical study.

Corneas from human donors are used to replace damaged tissue and treat corneal blindness, but there is a severe worldwide shortage of donor corneas. We conducted a phase 1 clinical study in which biosynthetic mimics of corneal extracellular matrix were implanted to replace the pathologic anterior cornea of 10 patients who had significant vision loss, with the aim of facilitating endogenous tissue regeneration without the use of human donor tissue. The biosynthetic implants remained stably integrated and avascular for 24 months after surgery, without the need for long-term use of the steroid immunosuppression that is required for traditional allotransplantation. Corneal reepithelialization occurred in all patients, although a delay in epithelial closure as a result of the overlying retaining sutures led to early, localized implant thinning and fibrosis in some patients. The tear film was restored, and stromal cells were recruited into the implant in all patients. Nerve regeneration was also observed and touch sensitivity was restored, both to an equal or to a greater degree than is seen with human donor tissue. Vision at 24 months improved from preoperative values in six patients. With further optimization, biosynthetic corneal implants could offer a safe and effective alternative to the implantation of human tissue to help address the current donor cornea shortage.

Collagen-phosphorylcholine interpenetrating network hydrogels as corneal substitutes.

A biointeractive collagen-phospholipid corneal substitute was fabricated from interpenetrating polymeric networks comprising 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide and N-hydroxysuccinimide crosslinked porcine atelocollagen, and poly(ethylene glycol) diacrylate crosslinked 2-methacryloyloxyethyl phosphorylcholine (MPC). The resulting hydrogels showed an overall increase in mechanical strength beyond that of either original component and enhanced stability against enzymatic digestion (by collagenase) or UV degradation. More strikingly, these hydrogels retained the full biointeractive, cell friendly properties of collagen in promoting corneal cell and nerve in-growth and regeneration (despite MPC's known anti-adhesive properties). Measurements of refractive indices, white light transmission and backscatter showed the optical properties of collagen-MPC are comparable or superior to those of the human cornea. In addition, the glucose and albumin permeability were comparable to those of human corneas. Twelve-month post-implantation results of collagen-MPC hydrogels into mini-pigs showed regeneration of corneal tissue (epithelium, stroma) as well as the tear film and sensory nerves. We also show that porcine collagen can be substituted with recombinant human collagen, resulting in a fully-synthetic implant that is free from the potential risks of disease transmission (e.g. prions) present in animal source materials.

PEG-stabilized carbodiimide crosslinked collagen-chitosan hydrogels for corneal tissue engineering.

Implantable biomaterials that mimic the extracellular matrix (ECM) in key physical and physiological functions require components and microarchitectures that are carefully designed to maintain the correct balance between biofunctional and physical properties. Our goal was to develop hybrid polymer networks (HPN) that combine the bioactive features of natural materials and physical characteristics of synthetic ones to achieve synergy between the desirable mechanical properties of some components with the biological compatibility and physiological relevance of others. In this study, we developed collagen-chitosan composite hydrogels as corneal implants stabilized by either a simple carbodiimide cross-linker or a hybrid cross-linking system comprised of a long-range bi-functional cross-linker (e.g. poly(ethylene glycol) dibutyraldehyde (PEG-DBA)), and short-range amide-type cross-linkers (e.g. 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), and N-hydroxysuccinimide (NHS)). Optimum hybrid hydrogel demonstrated significantly enhanced mechanical strength and elasticity by 100 and 20%, respectively, compared to its non-hybrid counterpart. It demonstrated excellent optical properties, optimum mechanical properties and suturability, and good permeability to glucose and albumin. It had excellent biocompatibility and when implanted into pig corneas for 12 months, allowed seamless host-graft integration with successful regeneration of host corneal epithelium, stroma, and nerves.

Regeneration of corneal cells and nerves in an implanted collagen corneal substitute.

PURPOSE: Our objective was to evaluate promotion of tissue regeneration by extracellular matrix (ECM) mimics, by using corneal implantation as a model system. METHODS: Carbodiimide cross-linked porcine type I collagen was molded into appropriate corneal dimensions to serve as substitutes for natural corneal ECM. These were implanted into corneas of mini-pigs after removal of the host tissue, and tracked over 12 months, by clinical examination, slit-lamp biomicroscopy, in vivo confocal microscopy, topography, and esthesiometry. Histopathology and tensile strength testing were performed at the end of 12 months. Other samples were biotin labeled and implanted into mice to evaluate matrix remodeling. RESULTS: The implants promoted regeneration of corneal cells, nerves, and the tear film while retaining optical clarity. Mechanical testing data were consistent with stable, seamless host-graft integration in regenerated corneas, which were as robust as the untreated fellow corneas. Biotin conjugation is an effective method for tracking the implant within the host tissue. CONCLUSIONS: We show that a simple ECM mimetic can promote regeneration of corneal cells and nerves. Gradual turnover of matrix material as part of the natural remodeling process allowed for stable integration with host tissue and restoration of mechanical properties of the organ. The simplicity in fabrication and shown functionality shows potential for ECM substitutes in future clinical applications.

Tissue-engineered recombinant human collagen-based corneal substitutes for implantation: performance of type I versus type III collagen.

PURPOSE: To compare the efficacies of recombinant human collagens types I and III as corneal substitutes for implantation. METHODS: Recombinant human collagen (13.7%) type I or III was thoroughly mixed with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide. The final homogenous solution was either molded into sheets for in vitro studies or into implants with the appropriate corneal dimensions for transplantation into minipigs. Animals with implants were observed for up to 12 months after surgery. Clinical examinations of the cornea included detailed slit lamp biomicroscopy, in vivo confocal microscopy, and fundus examination. Histopathologic examinations were also performed on corneas harvested after 12 months. RESULTS: Both cross-linked recombinant collagens had refractive indices of 1.35, with optical clarity similar to that in human corneas. Their chemical and mechanical properties were similar, although RHC-III implants showed superior optical clarity. Implants into pig corneas over 12 months show comparably stable integration, with regeneration of corneal cells, tear film, and nerves. Optical clarity was also maintained in both implants, as evidenced by fundus examination. CONCLUSIONS: Both RHC-I and -III implants can be safely and stably integrated into host corneas. The simple cross-linking methodology and recombinant source of materials makes them potentially safe and effective future corneal matrix substitutes.

Innervation of tissue-engineered recombinant human collagen-based corneal substitutes: a comparative in vivo confocal microscopy study.

PURPOSE: To compare reinnervation in recombinant human collagen-based corneal substitutes with allografts during a 1-year postimplantation follow-up period in pigs. A retrospective comparison to innervation in porcine collagen-based biosynthetic grafts was also performed. METHODS: Pigs received a corneal allograft or a substitute made of either recombinant human type-I or -III collagen. In vivo confocal microscopic examination of the central cornea of surgical and untouched control eyes before surgery and at 2, 6, and 12 months after surgery was performed to quantify the number, density, and diameter of nerves at various corneal depths. RESULTS: By 12 months after surgery, the number and density of regenerated nerves in the anterior and deep anterior corneal stroma recovered to preoperative and control levels in both types of substitute grafts and in the allografts. In the subepithelial and subbasal regions, however, significantly fewer nerves were detected relative to those in control subjects at 12 months, regardless of graft type (P < 0.05), similar to the behavior of porcine collagen-based biosynthetic grafts. An absence of thick stromal nerve trunks (diameter, >10 mum) in all grafts, irrespective of material type, indicated that nerve regeneration in grafts was accompanied by persistent morphologic changes. CONCLUSIONS: Nerve regeneration in recombinant human collagen-based biosynthetic corneal grafts proceeded similarly to that in allograft tissue, demonstrating the suitability of recombinant human collagen constructs as nerve-friendly corneal substitutes. Furthermore, only minor differences were noted between type-I and -III collagen grafts, indicating an insensitivity of nerve regeneration to initial collagen type.

Recombinant human collagen for tissue engineered corneal substitutes.

We successfully fabricated transparent, robust hydrogels as corneal substitutes from concentrated recombinant human type I and type III collagen solutions crosslinked with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). White light transmission through these gels is comparable or superior to that of human corneas. Hydrogels from both type I and type III collagens supported in vitro epithelium and nerve over-growth. While both these biocompatible hydrogels have adequate tensile strength and elasticity for surgical manipulation, type III collagen hydrogels tended to be mechanically superior. Twelve-month post-implantation results of type I recombinant collagen-based corneal substitutes into mini-pigs showed retention of optical clarity, along with regeneration of corneal cells, nerves and tear film. For clinical use, implants based on fully characterized, recombinant human collagen eliminate the risk of pathogen transfer or xenogeneic immuno-responses posed by animal collagens.

Spectroscopic whole-blood indicators of end-stage renal disease and the hemodialysis treatment.

The diffuse reflection spectrum in the 500-1670 nm region for whole blood taken from healthy subjects and end-stage renal disease (ESRD) patients was measured to test the feasibility of optically monitoring ESRD and its treatment by hemodialysis. Spectral regions where optical absorption significantly differed between healthy subjects and ESRD patients were used to form a multiple linear discriminant classification model. With this model a total of 41 whole-blood samples were classified into healthy, pretreatment and posttreatment ESRD classes. 96.7% of original and cross-validated cases and 100% of independent validation cases were correctly classified, indicating ESRD and its treatment exhibit characteristic spectral features in whole blood. Upon comparison of the discriminant model variables with a few key clinical blood parameters, model variables were found to significantly correlate with hematocrit and plasma levels of urea and potassium (P<0.05). The results of this study suggest that the optical signature of whole blood conveys basic clinical status information, and provides a path for investigating improved indices of hemodialysis toxicity, adequacy and patient outcome.

Innervation of tissue-engineered corneal implants in a porcine model: a 1-year in vivo confocal microscopy study.

PURPOSE: To examine the pattern of nerve regeneration within tissue-engineered corneal substitutes grafted into host porcine corneas over a 1-year postoperative period. METHODS: Biodegradable corneal substitutes from cross-linked collagen were implanted into the left eyes of 12 pigs by deep lamellar keratoplasty. Regeneration of severed nerves into the central implant region was investigated with in vivo confocal microscopy. Both implant-recipient and control (right) eyes were examined before surgery and 2, 6, 10, and 12 months after surgery, to quantify the number, density, diameter, and branching of nerve fiber bundles at various corneal depths. Transmission electron microscopy was used to confirm the presence of nerve bundles. RESULTS: Two months after surgery, corneal nerve ingrowth was observed within the deep anterior stroma, with a number and density of regenerated nerves significantly higher than in nonsurgical control eyes (P < 0.01). Nerves within the superficial anterior stroma regenerated by 6 to 10 months after surgery, and the first subbasal epithelial nerves were seen 10 months after surgery. After 1 year, subbasal nerve density recovered to preoperative levels. Nerve fibers in the deep anterior stroma remained significantly thinner relative to control eyes after 1 year (P < 0.001), where both superficial anterior and subbasal nerve diameter did not change relative to control eyes. CONCLUSIONS: The pattern of reinnervation within tissue-engineered corneal substitutes has been quantified in vivo. Innervation proceeded rapidly in the deep anterior stroma, followed by repopulation of more superficial regions. One year after surgery, nerve density within the tissue-engineered cornea increased or remained unchanged relative to controls in all corneal regions examined.

Hemodialysis monitoring in whole blood using transmission and diffuse reflection spectroscopy: A pilot study.

Visible and near infrared transmission and diffuse reflection spectroscopy were used to monitor changes in whole blood resulting from hemodialysis treatment for end-stage renal disease. Blood samples from 8 patients on chronic hemodialysis therapy were measured in the 500- to 1700-nm wavelength range immediately before and after a single treatment. Principal component scores characteristic of each spectrum were derived, and mean pre- and posttreatment scores of the first principal component indicated a significant treatment-dependent change in both optical transmission (P = 0.004) and diffuse reflection (P < 0.001). Significant treatment-induced change persisted (P < 0.05) when the first four principal components were used to account for >97% of the treatment-dependent spectral variation. Some blood spectral changes expressed in terms of difference spectra (posttreatment - pretreatment) were consistent with standard clinical indicators of weight reduction, urea reduction, and potassium change, with probable origins at a molecular level. The results indicate the feasibility of using optical transmission and diffuse reflection spectroscopy to characterize clinically relevant blood changes for the future development of more comprehensive indicators of hemodialysis efficacy and long-term clinical outcomes. Moreover, the optical techniques employed are adaptable for potential online monitoring of blood changes during the hemodialysis treatment.

A simple, cross-linked collagen tissue substitute for corneal implantation.

PURPOSE: To develop a simple corneal substitute from cross-linked collagen. METHODS: Porcine type I collagen (10%; pH 5), was mixed with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The final homogenous solution was molded to corneal dimensions, cured, and then implanted into rabbits and minipigs by lamellar keratoplasty. The implants were followed for up to 6 months after surgery. Clinical examinations of the cornea included detailed slit lamp biomicroscopy, in vivo confocal microscopy, topography and esthesiometry for nerve function. Histopathologic examinations were also performed on rabbit corneas harvested after 6 months. RESULTS: Cross-linked collagen (refractive index, 1.35) had optical clarity superior to human corneas. Implanted into rabbit and porcine corneas, only 1 of 24 of the surgical corneas showed a slight haze at 6 months after surgery. All other implants showed no adverse reactions and remained optically clear. Topography showed a smooth surface and a profile similar to that of the contralateral nonsurgical eye. The implanted matrices promoted regeneration of corneal cells, tear film, and nerves. Touch sensitivity was restored, indicating some restoration of function. The corneas with implants showed no significant loss of thickness and demonstrated stable host-graft integration. CONCLUSIONS: Collagen can be adequately stabilized, using water soluble carbodiimides as protein cross-linking reagents, in the fabrication of corneal matrix substitutes for implantation. The simple cross-linking methodology would allow for easy fabrication of matrices for transplantation in centers where there is a shortage of corneas, or where there is need for temporary patches to repair perforations in emergency situations.

Wavefront-guided correction of ocular aberrations: are phase plate and refractive surgery solutions equal?

Wavefront-guided laser eye surgery has been recently introduced and holds the promise of correcting not only defocus and astigmatism in patients but also higher-order aberrations. Research is just beginning on the implementation of wavefront-guided methods in optical solutions, such as phase-plate-based spectacles, as alternatives to surgery. We investigate the theoretical differences between the implementation of wavefront-guided surgical and phase plate corrections. The residual aberrations of 43 model eyes are calculated after simulated refractive surgery and also after a phase plate is placed in front of the untreated eye. In each case, the current wavefront-guided paradigm that applies a direct map of the ocular aberrations to the correction zone is used. The simulation results demonstrate that an ablation map that is a Zernike fit of a direct transform of the ocular wavefront phase error is not as efficient in correcting refractive errors of sphere, cylinder, spherical aberration, and coma as when the same Zernike coefficients are applied to a phase plate, with statistically significant improvements from 2% to 6%.