Corneal replacement using a synthetic hydrogel cornea, AlphaCor: device, preliminary outcomes and complications.

PURPOSE: Clinical assessment of outcome of corneal replacement with a synthetic cornea, AlphaCor, in patients considered at too high risk for conventional penetrating keratoplasty with donor tissue to be successful, but excluding indications such as end-stage dry eye that might be suited to traditional prosthokeratoplasty. METHODS: All patients in the multicentre clinical trial were managed according to an approved protocol, with Ethics Committee approval in each centre. Preoperative visual acuity ranged from perception of light (PL) to 6/60 (20/200). Implantation was by means of an intralamellar technique, with a conjunctival flap in most cases. Tissues anterior to the optic were removed as a secondary procedure. RESULTS: Up to 30 November 2001, 40 AlphaCor devices had been implanted in 38 patients, of mean age 60 years. Follow-up ranged from 0.5 months to 3 years. There had been one extrusion (2.5%) and four cases (10%) where a device had been removed due to melt-related complications. All five of these cases received a donor corneal graft after the device was removed, with these grafts remaining anatomically satisfactory and epithelialised to date. Corneal melts in AlphaCor recipients were found to be strongly associated with a history of ocular herpes simplex infection. Two further devices (5%) were removed owing to reduced optic clarity after presumed drug-related deposition, and have been successfully replaced with second devices. Mean preoperative best-corrected visual acuity was hand movements. Visual acuities after surgery ranged from PL to 6/6(-2) (20/20(-2)). CONCLUSIONS: Early results suggest that the AlphaCor, previously known as the Chirila keratoprosthesis (Chirila KPro), has a low incidence of the complications traditionally associated with keratoprostheses and can be effective in restoring vision in patients considered untreatable by conventional corneal transplantation. Importantly, the device can be replaced with a donor graft in the event of development of a significant complication. A history of ocular herpes simplex is a contraindication to AlphaCor implantation. Ongoing monitoring of clinical outcomes in all patients will allow the indications for AlphaCor, as opposed to donor grafts, to be determined.

Effect of a standard paired arcuate incision and augmentation sutures on postkeratoplasty astigmatism.

To analyze the efficacy of a standardized paired arcuate incision and augmentation suture technique in the treatment of various levels of post-penetrating keratoplasty (PKP) astigmatism. Lions Eye Institute, Royal Perth Hospital, Perth, Australia, and University of Dundee Department of Ophthalmology, Dundee, United Kingdom.A standardized paired arcuate incision and paired augmentation suture technique was used to treat 34 eyes with post-PKP astigmatism ranging from -3.50 to -20.00 diopters (D) at the spectacle plane. The technique consisted of paired arcuate incisions of 3 clock hours, 480 microm deep in the graft-host junction, and 2 pairs of augmentation 10-0 nylon sutures. The mean preoperative cylinder was -9.14 D +/- 4.38 (SD) and the mean postoperative cylinder, -3.59 +/- 1.92 D at the corneal plane after a mean follow-up of 50 +/- 43 weeks. This represents an empirical reduction in mean cylinder of 5. 55 D (60.7%). The Alpins correction index (surgically induced astigmatism [SIA] divided by target induced astigmatism) was calculated for each case, and the mean was 1.01 +/- 0.34, with a median of 0.91. Approximately 53.1% of cases achieved a correction index between 0.80 and 1.20, and the correction index correlated poorly with the initial magnitude of cylinder. A direct numerical relationship between SIA and the initial magnitude of cylinder was observed, although a standard surgical procedure was used in all cases.A simple standardized technique using paired arcuate incisions in the graft-host junction with paired augmentation sutures reduces the amount of cylinder in proportion to the magnitude of the preoperative cylinder and effectively reduces post-PKP astigmatism.

Development of a poly(2-hydroxyethyl methacrylate) orbital implant allowing direct muscle attachment and tissue ingrowth.

PURPOSE: To develop a poly(2-hydroxyethyl methacrylate) orbital implant that allows tissue ingrowth and direct muscle attachment to minimize the risk of extrusion and to enhance cosmesis. METHODS: Assessment of clinical outcomes and histologic findings after implantation of 18 prototype prostheses into rabbits. The implants were not wrapped with other tissues or materials. RESULTS: One case of infection was observed but there were no extrusions, with up to 21 months follow-up. Biocolonization was confirmed histologically. Good movement was observed when a cosmetic shell was fitted. CONCLUSIONS: The prototype prosthesis appears promising, with particular advantages being the direct attachment of extraocular muscles, good cosmesis and movement, and a low complication rate in this pilot study.

Correlation of histological findings with gadolinium enhanced MRI scans during healing of a PHEMA orbital implant in rabbits.

BACKGROUND/AIMS: To investigate a poly(2-hydroxyethyl methacrylate) (PHEMA) orbital implant with a spongy anterior hemisphere and a smooth gel posterior hemisphere, by histology correlated with magnetic resonance images. METHODS: Following enucleation, eight rabbits received PHEMA implants to which the muscles were directly sutured, and underwent gadolinium enhanced magnetic resonance imaging (MRI) from 3 to 52 weeks. After the rabbits were killed, the implants were removed, cut in a plane corresponding to the scan, and processed for light and electron microscopy. RESULTS: All eight rabbits retained their implant to the end of the study period without complications. The scans demonstrated muscle attachment to the anterior half of the implant, and enhancement was seen on injection of gadolinium chelate. Histology confirmed muscle attachment, and cellular and vascular ingrowth. Over time, a transformation from reactive inflammatory to relatively non-vascular scar tissue was seen within the implant. Calcium deposits in one implant were detected by imaging and histology. CONCLUSION: The implants are readily visualised on MRI. Muscle attachment and fibrovascular ingrowth into the anterior hemisphere are seen, while encapsulation of the posterior hemisphere is minimal. Histological findings confirm the progress of the healing response, with initial inflammation and marked vascularisation, developing later into quiescent scar tissue predominantly of fibroblasts.

Implantation of PHEMA keratoprostheses after alkali burns in rabbit eyes.

PURPOSE: We have previously examined histologically the healing of a PHEMA core-and-skirt keratoprosthesis (the Chirila KPro) as a full-thickness implant in healthy animal corneas. The present study was carried out to determine whether a diseased cornea could also generate biocolonization of the skirt region of a KPro. METHODS: Ten KPros were placed as full-thickness corneal implants under conjunctival flaps in 10 alkali-burned rabbit corneas. Histological findings at intervals from 2 weeks to 6 months postoperatively were compared with earlier findings in 10 rabbits that had received identical KPros without prior alkali injury. RESULTS: Despite severe corneal injury and the reduced keratocyte population present, there were no clinically detected complications in 60%. Histological findings established that, compared with healthy host tissue, skirt biocolonization and KPro-cornea healing after an alkali burn were impaired, with evidence of epithelial downgrowth in 40%. One animal required euthanasia earlier than the planned end point, but no KPro extrusions occurred. CONCLUSION: Biocolonization of a KPro skirt is reduced but not prevented in an alkali-induced corneal inflammation model. Although no extrusions occurred, close follow-up and anticollagenolytic medication would be required to minimize the complication rate.

Clinical results of implantation of the Chirila keratoprosthesis in rabbits.

AIMS/BACKGROUND: An ideal keratoprosthesis (KPro) would closely resemble a donor corneal button in terms of its surgical handling, optics, and capacity to heal with host tissue in order to avoid many of the complications associated with the KPros which are currently in clinical use. This study was carried out to assess the long term clinical outcomes on implantation of the core and skirt poly(2-hydroxyethyl methacrylate) KPro in animals. METHODS: 20 KPros were made and implanted as full thickness corneal replacements into rabbits and followed for up to 21 months to date. RESULTS: 80% of the prostheses have been retained, with a low incidence of complications such as cataract, glaucoma, and retroprosthetic membrane formation which are frequently associated with KPro surgery. CONCLUSIONS: KPros of this type may offer promise in the treatment of patients for whom penetrating keratoplasty with donor material carries a poor prognosis. Refinement of the KPro and further animal trials, including implantation into abnormal corneas, are however mandatory before human implantation could be planned.

Assessment of anticollagenase treatments after insertion of a keratoprosthetic material in the rabbit cornea.

PURPOSE: This study was performed to evaluate the enzyme production in response to implantation of the hydrogel material used in the experimental Chirila keratoprosthesis (KPro) and to assess the effects of five topical drugs on enzyme production and activity. KPros may be extruded from the cornea as a result of tissue melting, a process that involves excessive enzyme activity. To reduce the possibility of implant loss for the hydrogel Chirila KPro, a number of antiinflammatory drugs that have been used to treat other corneal melting conditions were investigated for their effect on initial collagenase activity after the implantation of KPro material into the rabbit cornea. METHODS: Poly(2-hydroxyethyl methacrylate) sponge pieces were implanted into rabbit corneas. Prednisolone, tetracycline, medroxyprogesterone, acetylcysteine, and sodium citrate were assessed for effects on gelatinolytic activity and stromal collagenase [matrix metalloprotease-1 (MMP-1)] production in vivo and in vitro by using zymography and Western blotting techniques. RESULTS: Whereas all five anticollagenase drugs were effective in reducing gelatinolytic activity in vitro, many were ineffective in vivo. However, medroxyprogesterone caused a reduction of gelatinolytic activity in vivo. The amount of MMP-1, as measured by immunoblotting, also was reduced by medroxyprogesterone treatment when compared with untreated controls. An increase in the apparent molecular weight of MMP-1 in operated corneas appears to be the result of the association of MMP-1 with collagen fragments resulting from the surgical trauma. CONCLUSION: This study indicates that topical medroxyprogesterone may be a useful adjunctive therapy after prosthokeratoplasty.

Cell viability and inflammatory response in hydrogel sponges implanted in the rabbit cornea.

In the quest for the development of a functional keratoprosthesis, the biocompatibility of the porous skirt material in the Chirila keratoprosthesis (KPro) was investigated. The population of live and dead cells within, and the inflammatory response to, a tissue-integrating poly(2-hydroxyethyl methacrylate) (PHEMA) sponge were studied. Samples of the hydrogel sponge were implanted in rabbit corneas and explanted at predetermined time points up to 12 weeks. The explanted sponges were subjected to cell viability assay using two types of fluoroprobes, 5-chloromethylfluorescein diacetate and ethidium homodimer-1. A semiquantitative analysis was performed to assess the number of dead cells within the sponge and in the area of corneal stroma proximal to the sponge. Five rabbits were used for each end point (2, 4 and 12 weeks). To investigate the inflammatory response to the sponge, immunocytochemistry, using specific antibodies to rabbit macrophages, enzyme histochemistry of chloroacetate esterase (to detect neutrophils) and transmission electron microscopy (TEM) were also employed at 24 h, 2, 4 and 12 weeks after implantation. Four weeks after implantation, fewer viable cells were observed in the sponge when compared to the 2-week implant. However, the proportion of viable cells increased dramatically by 12 weeks. The proportion of nonviable cells decreased gradually with time; central sponge contained 34+/-11 % dead cells after 2 weeks, and 15+/-4.3% after 12 weeks. The staining of inflammatory cells demonstrated the presence of macrophages and neutrophils up to 12 weeks after implantation. TEM confirmed the presence of these cell types and others. including eosinophils and myofibroblasts, as well as blood capillaries. The presence of a significant number of viable cells at each time point and the uniform reduction of the nonviable cell proportion with time suggests that the sponge is a conducive environment supporting a prolific, viable cellular colonization. Dead cells observed in the first instance indicate a normal injury pattern. However, the presence of a small but significant proportion of invading inflammatory cells 12 weeks after implantation confirms a characteristic pattern of wound healing within the sponges.

Keratoprostheses: advancing toward a true artificial cornea.

Keratoprosthesis surgery is carried out in very few centers. Elaborate surgical techniques and high complication rates limit the application of currently available keratoprostheses (KPros). However, the clinical need for an alternative to donor tissue has sparked considerable research interest in the development of new KPros. This paper charts the evolution of KPros from the earliest devices to those currently used, describes their drawbacks and discusses the specifications of an ideal device. Recent research focuses upon the use of porous polymers as the skirt component of core-and-skirt KPros in order to obtain improved biological integration of the prosthetic material. Developments in biomaterials technology make a KPro analogous to a donor corneal button an increasingly realistic goal. However, two particular problems still need to be addressed. First, it must be demonstrated that secure long-term fixation that is able to withstand trauma is achievable in a full-thickness artificial cornea. Second, an ideal artificial cornea for a wet eye requires an epithelialized surface, and this has yet to be achieved.

Scedosporium prolificans corneoscleritis: a successful outcome.

BACKGROUND: A case of Scedosporium prolificans corneoscleritis is reported in a patient who had developed scleral necrosis following pterygium surgery, with adjunctive beta-irradiation. This fungus has been reported to be the causative organism in only two previous cases of corneoscleritis. METHODS: The patient presented with signs and symptoms typical of corneoscleritis. When the fungus was isolated from a biopsy specimen, aggressive scleral debridement was carried out and intensive antifungal therapy was instigated. RESULTS: After a prolonged course, the eye was rendered sterile. CONCLUSION: Early conjunctival recession and aggressive scleral debridement combined with intensive instillation of antifungals are crucial to the successful management of fungal corneoscleritis.

Keratoprosthesis results in animals: an update.

BACKGROUND: The report presented is an update on continuing development work on modified PHEMA core-and-shirt KPros in animals. METHODS: Two variations (improved wet-eye, and dry-eye) of a prototype core-and-skirt Chirila KPro are described. The clinical success rate on implantation of these versions of the Chirila KPro was assessed. RESULTS: It was found that a significant improvement in retention rate was shown in the improved model but that the dry-eye model failed early in two of the three implanted. CONCLUSIONS: The significance of the improved strength and the reasons for disappointing results with the early dry-eye KPros are discussed. Ongoing work is briefly outlined.

Histologic evaluation during healing of hydrogel core-and-skirt keratoprostheses in the rabbit eye.

PURPOSE: We developed two models that are modifications of our original poly(2-hydroxyethyl methacrylate) (PHEMA) core-and-skirt keratoprosthesis. In these keratoprostheses, the mechanical strength of the skirt has been considerably increased with divinyl glycol (DVG) as a cross-linking agent during polymerization. In one (KPro I), methyl methacrylate (MMA) was added as comonomer to increase cell adhesion, and in the other (KPro II), HEMA was polymerized with DVG without comonomer. The aim of this study was to evaluate the process of healing and biocolonization and to ascertain whether KPro I demonstrates better ingrowth than the mechanically stronger KPro II, after implantation in rabbit eyes. METHODS: Ten rabbits were used for each model and studied at five predetermined end points up to 26 weeks. The device was implanted as a full-thickness keratoprosthesis covered with a conjunctival flap. RESULTS: Neither prosthesis demonstrated extrusion or retroprosthetic membrane formation. There was no significant difference between the two types of prosthesis with respect to tissue ingrowth and surrounding tissue melting. Histologically, inflammation was not severe, but calcification was seen in most specimens. Evidence of biodegradation of the prosthesis also was seen. CONCLUSION: In our original keratoprosthesis, fibrovascular invasion had occurred into the prosthetic skirt, but wound dehiscence and low mechanical strength resulted in an unfavorable outcome. In this series, the mechanical properties were improved, and KPro II was stronger than KPro I. Therefore KPro II would be the preferred polymer combination for surgical manipulation. However, biodegradation and calcification require further investigation into the degree and significance of these adverse reactions.

Keratoprosthesis: preliminary results of an artificial corneal button as a full-thickness implant in the rabbit model.

PURPOSE: To develop a prototype artificial cornea and evaluate it in the rabbit model. METHODS: Hydrogel core-and-skirt keratoprostheses were made and were inserted as full-thickness implants covered with conjunctival flaps in the right eyes of eight rabbits. RESULTS: Peroperative complications related to inadequate mechanical strength led to failure in the early postoperative period in three animals, one was euthanased for an unrelated reason and the remaining four have been successful for up to 16 weeks' follow-up. CONCLUSIONS: Full-thickness implantation of an artificial cornea, analogous to penetrating keratoplasty, has been achieved in the rabbit model. Histological findings confirm that integration of the prosthesis with host tissue occurs. The main complications encountered in this preliminary series were related to inadequate strength of the sponge skirt of this prototype device. Work in our laboratories is now concentrated upon improving the mechanical qualities of the hydrogel skirt and on the enhancement of biointegration.

Preliminary evaluation of a hydrogel core-and-skirt keratoprosthesis in the rabbit cornea.

BACKGROUND: We developed a core-and-skirt keratoprosthesis, with both components made from poly(2-hydroxyethyl methacrylate) (PHEMA) hydrogels. The identical chemical nature of both spongy skirt and transparent core assures a permanent union between them. We have previously shown that PHEMA sponges, within a certain range of pore size, can support cellular invasion and neovascularization when implanted into the rabbit cornea. The present study is the first to evaluate the behavior of the whole prosthesis after implantation into the rabbit cornea. METHODS: Hydrogel keratoprostheses were inserted intrastromally into the corneas of seven rabbits and histologically examined by light microscopy in five eyes enucleated at 8, 12, and 14 weeks. RESULTS: None of the implants extruded over this period. Both clinical and histopathologic examination showed that the keratoprostheses were well tolerated by the host tissue. The porous skirt was fully integrated into the stroma by fibrovascular invasion, and no capsule formed around the implants. Stromal melting anterior to the implant occurred in two cases, but this did not affect the fixation of the keratoprostheses. CONCLUSIONS: This study indicates that our keratoprosthesis can prevent extrusion in the short term when inserted into an intrastromal pocket of the rabbit eye.

Production of neocollagen by cells invading hydrogel sponges implanted in the rabbit cornea.

BACKGROUND: Poly(2-hydroxyethyl methacrylate) sponges are artificial tissue-equivalent matrices with potential value as materials for the peripheral zone of artificial corneas. A keratoprosthetic device was developed incorporating a poly(HEMA) spongy skirt which allowed cellular invasion. The present in vivo study investigated the biosynthetic activity of stromal fibroblasts growing within a poly(HEMA) sponge implanted into the rabbit cornea. METHODS: A porous poly(HEMA) hydrogel was synthesized by polymerization in a large excess of water. Specimens with a pore size larger than 10 microns were impregnated with collagen type I and then implanted into the limbal region of cornea in four rabbits. The animals were followed clinically for 28 days, when they were anaesthetized and new sponge specimens were implanted in their second eye. After 2 h, both eyes were enucleated. The 28-day and 2-h explants were subjected to autoradiographic analysis following labelling with tritiated proline and to an immunostaining technique using antibodies to collagen types I-VI. RESULTS: The autoradiographic analysis showed that the fibroblasts within the 28-day explants continued to be synthetically active and deposited proteins. Using the immunostaining technique, the deposition was most clearly demonstrated by the localization of collagen type III in the tissue invading the sponge. Both techniques failed to indicate any cellular activity in the short-time implants. CONCLUSIONS: The presence of collagen type III is consistent with a normal healing response of the stromal fibroblasts and indicates that poly(-HEMA) sponges are able to function as tissue-equivalent matrices.

Enhancement of neovascularization in regenerating skeletal muscle by the sustained release of erucamide from a polymer matrix.

The angiogenic agent erucamide (cis-13-docosenamide), incorporated into a polymeric biomaterial (Elvax 40P, a copolymer of ethylene and vinyl acetate), was used to determine whether angiogenesis can be increased in the regenerating skeletal muscle, and whether the enhanced revascularization improves the new muscle formation. The angiogenic nature of this lipid was confirmed in a rat cornea-micropocket assay, prior to insertion of small strips of the polymer containing either 3 micrograms, 300 micrograms erucamide or only polymer as a control into the mid-region of crush-injured tibialis anterior (TA) muscles of forty-five adult male BALB/c mice. All TA muscles were sampled ten days after injury and analyzed morphometrically. Statistical analyses of the mean blood vessel area density in lesions from twelve perfused TA muscles (three from each of the erucamide-treated or control group), revealed a dose-dependent angiogenic effect of erucamide: a dosage of 3 micrograms increased mean blood vessel area density to 5.1% compared to 2.0% in controls, due to numerous large caliber, thin-walled vessels, whereas the mean vessel area density in both the 30-micrograms (3.5%) and 300-micrograms (1.5%) doses were similar to controls. However, at all three doses tested, erucamide did not significantly alter the degree of new muscle formation, connective tissue deposition, or removal of necrotic debris.

Severe microbial keratitis in temperate and tropical Western Australia.

To determine the patterns of severe microbial keratitis in Western Australia, all acute admissions over a 2 year period to the Department of Ophthalmology, Royal Perth Hospital were assessed. Fifty-three cases (n = 53) of severe, presumed microbial keratitis were identified. Seventy per cent of these eyes had a visual acuity of 6/60 or less on admission and only 38.8% had 6/12 or better corrected acuity following resolution of the keratitis. The most commonly identified predisposing factors were: prior ocular surgery with or without exposed monofilament sutures (43.4%); contact lens wear (22.6%); lid malposition (17.0%); history of ocular trauma (15.1%); and history of previous herpes simplex keratitis (13.2%). It is notable that 26.4% of the subjects had been applying topical ophthalmic corticosteroids prior to admission. Following corneal scrape or biopsy a positive microbial diagnosis was made in 71% of samples, with Gram-negative and Gram-positive bacterial isolates being equally frequent. Five cases of Acanthamoeba keratitis were identified following corneal biopsy. Where antibiotic sensitivities were available, it was noted that 61.5% of Gram-positive and 46.1% of Gram-negative bacteria were susceptible to chloramphenicol, with 84.6% of Gram-negative bacteria being sensitive to gentamicin. Many of these severe cases of microbial keratitis might have been avoided, or their severity lessened, by earlier identification of predisposing risk factors, more intensive and appropriate antibiotic administration, and improved patient education following ocular surgery.