Evaluation of the Therapeutic Efficacy of Lamellar Keratoconjunctival Transplantation in Treating Corneal Ulcers in Rabbits

Evaluation of the Therapeutic Efficacy of Lamellar Keratoconjunctival Transplantation in Treating Corneal Ulcers in Rabbits | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Evaluation of the Therapeutic Efficacy of Lamellar Keratoconjunctival Transplantation in Treating Corneal Ulcers in Rabbits Zhuangzhuang Wang, Haoyang Miao, Tiantian Li, Mingru Cui, Qun Yang, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7307393/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Corneal ulcer exhibits an extremely high incidence in the field of rabbit ophthalmic diseases and stands as one of the crucial blinding eye conditions leading to blindness in rabbits. The occurrence of severe complications, such as corneal perforation, can inflict severe damage on the visual function of rabbits, resulting in a rapid decline in their vision and even causing blindness.For deep-seated corneal ulcers or refractory corneal ulcers that persist and cannot be effectively controlled despite standardized drug therapy, surgical intervention has become an indispensable treatment approach. Lamellar keratoconjunctival transplantation, as a highly significant ocular surface reconstruction surgery, plays an irreplaceable role in restoring the normal structure and function of the rabbit's eye.This study aimed to systematically evaluate the therapeutic efficacy of lamellar keratoconjunctival transplantation in treating corneal ulcers in rabbits. Twenty-four rabbits underwent corneal burns to establish ulcer models. After three days, rabbits were randomly divided into three treatment groups (n = 8 each): Group A received conservative medical treatment, Group B underwent conjunctival flap transplantation, and Group C underwent lamellar keratoconjunctival transplantation. Postoperatively, corneal transparency, edema, and epithelial healing at the ulcer sites were regularly evaluated. Histopathological observations of corneal tissues and measurements of serum interleukin-6 (IL-6) and transforming growth factor-beta 1 (TGF-β1) were also conducted. Results indicated that the lamellar keratoconjunctival transplantation group (Group C) showed superior outcomes compared to the other two groups in terms of improved corneal transparency, reduced edema, enhanced epithelial healing, and favorable histopathological features. No significant differences were observed among the three groups in serum levels of IL-6 and TGF-β1. These findings suggest that lamellar keratoconjunctival transplantation is effective in supporting damaged corneas and promoting tissue regeneration, providing a promising method and alternative approach for the clinical management of corneal ulcers in rabbits and potentially in veterinary ophthalmology practice. Health sciences/Diseases Health sciences/Medical research Rabbit corneal ulcer Keratoconjunctival lamellar transplantation Figures Figure 1 Figure 2 Figure 3 1. Introduction Corneal ulcers represent a common and serious ophthalmic condition frequently encountered in rabbit farming and ophthalmological research. Any loss of corneal epithelium, irrespective of its cause, can be defined as a corneal ulcer 1 . In rabbits, corneal ulcers may result from multiple etiological factors, including trauma, infections (bacterial, viral, or fungal), and secondary complications related to ocular diseases 2 – 4 . Once affected by corneal ulcers, rabbits typically exhibit ocular pain, tearing, blepharospasm, and corneal opacity, significantly impacting their vision and overall quality of life. Without timely and effective treatment, corneal ulcers may deteriorate rapidly, potentially leading to severe complications such as corneal perforation, endophthalmitis, and even blindness 5 . Currently, various therapeutic strategies are employed for managing corneal ulcers in rabbits, yet each has certain limitations. Traditional pharmaceutical therapies, such as topical antibiotic eye drops or ointments, have demonstrated effectiveness mainly in mild corneal ulcers, as reported by Jun-Yi and Sha X-Y 6 , 7 . However, in cases of moderate to severe ulcers, particularly those involving significant corneal tissue defects, conventional treatments often fail to achieve satisfactory outcomes. Surgical methods, such as corneal perforation repair, have been described by researchers including Nishi E N and Tsao W-S as technically challenging procedures demanding high surgical expertise and frequently associated with postoperative complications, including infection and excessive corneal scarring. Furthermore, these methods typically yield suboptimal tissue repair, often failing to fully restore the cornea’s normal structure and function 8 , 9 . Lamellar keratoconjunctival transplantation, an emerging surgical technique, has achieved notable advancements within human ophthalmology. By transplanting healthy lamellar keratoconjunctival tissue, this procedure provides structural support and bioactive factors to damaged corneas, thus promoting epithelial repair and regeneration, enhancing corneal transparency and refractive status, and reducing immunological rejection 10 – 14 . Nevertheless, research evaluating the therapeutic effects of lamellar keratoconjunctival transplantation on corneal ulcers in rabbits remains insufficient. 2. Materials and Methods 2.1 Ethics statement (Animals) All procedures involving animals were reviewed and approved by the Henan Institute of Science and Technology Institutional Animal Care and Use Committee (Approval No. LLSC2022044). All methods were carried out in accordance with relevant guidelines and regulations, including the Regulations for the Administration of Affairs Concerning Experimental Animals (P. R. China), the Guidelines for the Ethical Review of Laboratory Animal Welfare (GB/T 35892 − 2018), the General Principles of Laboratory Animal Welfare (GB/T 42011 − 2022), and Laboratory Animals—Environment and Facilities (GB 14925 − 2023), as well as institutional policies. This study is reported in accordance with the ARRIVE guidelines (ARRIVE 2.0). Appropriate anesthesia and postoperative analgesia were provided, predefined humane endpoints were applied, and euthanasia was performed using an approved method to minimize pain and distress. 2.2 Establishment of the Rabbit Corneal Ulcer Model Twenty-four rabbits were randomly divided into three groups (n = 8 per group): Group A received conservative medical treatment, Group B underwent conjunctival flap transplantation, and Group C underwent lamellar keratoconjunctival transplantation. Three days before surgery, fur surrounding the right eyes was clipped, and the eye sockets were cleaned. Rabbits were anesthetized using a 1:1 mixture of Zoletil 50 and Sumianxin II (0.1 mL/kg). After placing rabbits in the left lateral recumbent position, the right eyes were disinfected and topically anesthetized using proparacaine hydrochloride eye drops. Corneal ulcer models were induced by placing filter paper saturated with 1 mol/L NaOH solution on the central cornea for 1 min, followed by thorough rinsing with saline. Postoperatively, all rabbits received subcutaneous injections of meloxicam (0.5 mg/kg) and enrofloxacin (10 mg/kg), chloramphenicol eye drops four times daily, and protective Elizabethan collars. 2.3 Conservative Medical Therapy, Conjunctival Flap Transplantation, and Lamellar Keratoconjunctival Transplantation for the Treatment of Corneal Ulcers The conservative medical treatment consisted of chloramphenicol eye drops applied three times daily, followed by recombinant bovine basic fibroblast growth factor (rb-bFGF) eye drops (Beifushu) 10 minutes later, continued for 60 days. For conjunctival flap transplantation, the corneal lesion was excised, extending 0.5-1 mm beyond the ulcer margin. An appropriately sized conjunctival flap was harvested from the dorsal bulbar conjunctiva, placed to cover the corneal defect without tension, and secured with 10 − 0 ophthalmic sutures. The lateral eyelids were partially closed with 5 − 0 absorbable sutures, which were removed after 7 days. In the lamellar keratoconjunctival transplantation group, lamellar keratoconjunctival grafts were harvested from the dorsal region of the globe. After excision of the diseased corneal tissue, a graft slightly larger (approximately 1 mm beyond defect margins) than the defect was prepared and secured tension-free onto the corneal wound using 10 − 0 ophthalmic sutures. The lateral eyelids were partially closed with 5 − 0 absorbable sutures, which were similarly removed after 7 days. 2.4 Clinical Assessment Following the methods of Holland and Shaopan Wang 15 – 17 . each animal was examined weekly, and quantitative assessments were performed based on scores ranging from 0 to 4 for corneal transparency, corneal edema, and corneal epithelial defects (assessed by fluorescein staining), as detailed in Table 1 . Table 1 Grade evaluation of clinical recovery of corneal ulcers in rabbits Grade Corneal Transparency Edema Severity Corneal Epithelial Defects (Fluorescein Staining) 0 Completely transparent cornea None Untainted 1 Mild opacity at ulcer site Slight Mild staining with fewer than 5 scattered staining spots 2 Moderate opacity at ulcer site, anterior chamber still visible Moderate Moderate staining with more than 5 staining spots 3 Marked opacity at ulcer site, anterior chamber barely visible Severe Staining spots merging into lines or patches 4 Complete opacity at ulcer site, anterior chamber not visible Extreme Extensive patchy staining covering a large area 2.5 Cytokine and Histological Evaluation Blood samples were collected intravenously from rabbits at 1 day prior to corneal injury, 3 days post-injury, and 3, 7, 14, and 28 days after transplantation. Serum IL-6 and TGF-β1 levels were measured using ELISA kits. All recipient animals were euthanized by intravenous injection of a lethal dose of sodium pentobarbital at 9 weeks postoperatively, and excised corneas were subjected to hematoxylin-eosin (HE) staining for histological evaluation. 2.6 Data Analysis Statistical analysis was conducted using SPSS 26.0, and experimental data were expressed as mean ± standard deviation (X ± SD). Differences were considered significant at P < 0.05 and highly significant at P < 0.01. GraphPad Prism 9.0.0 software was used for plotting data. 3. Result 3.1 Conservative Medical Treatment Group A total of 8 rabbits underwent conservative drug treatment. At the beginning of the treatment, the area of corneal sodium hydroxide burn showed significant edema (grade 0–2). By the end of the first week, the congestion of the conjunctival capillaries decreased. In the second week, obvious edema also appeared around the burned area of the healthy cornea (grade 2–3). The edema subsided after 3 weeks of treatment. At 5 weeks of treatment, new blood vessels on the cornea approached the ulcer site and reached the central area of the cornea. By 6 weeks, the new blood vessels on the cornea showed signs of regression. By the 9th week of treatment, the new capillaries on the cornea almost disappeared (Fig. 1 A), but the condition of the corneal burn site was not significantly different from that at the beginning of treatment. No deterioration of the condition such as corneal perforation was observed. The HE staining of the corneas at 9 weeks after the surgery (Fig. 1 B) showed that the epithelial layer became thinner, the stroma layer thickened, the fibrin arrangement was disordered, and there were a moderate number of fibroblasts; the posterior elastic layer and the endothelial layer detached. 3.2 Conjunctival Flap Transplantation Group In the conjunctival flap transplantation group, edema markedly decreased by week two (grade 0–2), and good vascularization of the conjunctival flap was observed immediately after surgery, completely covering the ulcer site. Throughout treatment, all eight rabbits maintained good conjunctival flap viability. By day 60, the conjunctival flap gradually thinned and became increasingly transparent at its margins (grade 0–2, Fig. 1 C). No flap detachment, corneal perforation, or other complications occurred during the 60-day observation period. Histological evaluation via HE staining at 9 weeks postoperatively (Fig. 1 D) revealed a continuous epithelial layer completely covering the ulcerated area, characterized by increased cell density, thickened epithelium with dense cellular arrangement, and an intact Descemet’s membrane and endothelial layer closely adhering to the stromal layer. 3.3 Lamellar Keratoconjunctival Transplantation Group After completion of the lamellar keratoconjunctival transplantation, the ulcerated region was fully covered by corneal tissue and partially by conjunctival tissue, with a strip-like white area indicating the limbus. At 7 days post-transplantation, abundant vascularization was observed in the conjunctival portion of the graft, and extensive new capillary formation was also evident within the corneal portion. Beginning at 21 days postoperatively and continuing up to day 60, the graft gradually became thinner and more transparent, with an increasingly larger area of the anterior chamber becoming visible through the previous ulcer site (Fig. 1 E). During the entire 60-day observation period, no adverse events such as graft detachment or corneal perforation were observed. Histological evaluation by HE staining at 9 weeks postoperatively (Fig. 1 F) showed a continuous epithelial layer completely covering the ulcer site, with mild epithelial thickening at the junctional area. The stromal layer demonstrated well-organized fibrillar arrangement and contained a low density of fibroblasts. Additionally, Descemet’s membrane and the endothelial layer were intact and closely adhered to the stroma. 3.4 Assessment of Corneal Ulcer Treatment Efficacy During the 60-day treatment period, corneal transparency at the ulcer site in the conservative medical treatment group exhibited little improvement (Fig. 2 ). The anterior chamber remained completely obscured by the ulcer throughout treatment, preventing any light from penetrating through the lesion. Although no severe complications such as further corneal damage or perforation occurred, the persistent opacity significantly impaired subsequent visual function. Corneal transparency in the conjunctival flap transplantation group was comparable to that of the conservative treatment group until day 28. Transparency gradually improved from day 35 to day 60, but this improvement was not statistically significant compared with conservative treatment. In contrast, the lamellar keratoconjunctival transplantation group showed significantly superior transparency compared to both other groups from day 7 to day 60 postoperatively (P < 0.01). In the first 14 days postoperatively, the clinical scores for corneal healing in the lamellar keratoconjunctival transplantation group were significantly better than those in the conjunctival flap transplantation group (P < 0.01). From day 14 to day 35, the differences between these two groups gradually diminished, and no significant differences were observed from day 45 to day 60. Compared with the conservative medical treatment group, both the conjunctival flap transplantation and lamellar keratoconjunctival transplantation groups exhibited statistically significant improvements at all time points, except on day 7 postoperatively when the difference between conjunctival flap transplantation and conservative treatment was not significant. Toward the end of the treatment period (days 45–60), some rabbits in both surgical groups still exhibited positive fluorescein staining, possibly due to eyelid friction against unremoved sutures, causing secondary epithelial damage (Fig. 3 ). Measurements of serum IL-6 and TGF-β1 levels showed no significant differences among the three groups. IL-6 has been associated with corneal injury repair, and its increased serum levels at day 28 postoperatively may be indicative of ongoing corneal recovery. Growth factors and cytokines play a crucial role in corneal healing; activation of the TGF-β system promotes differentiation of stromal cells into myofibroblasts, aiding stromal repair. The observed initial rise and subsequent decline of TGF-β1 and β2 levels may correlate with stromal healing progression during ulcer repair. 4. Discussion Corneal ulcers are a common ophthalmic disease that severely threatens vision. Without timely or proper intervention, corneal ulcers can lead to corneal perforation, intraocular infections, and even blindness 18 . This study established a rabbit model of corneal ulcers to comprehensively evaluate the therapeutic efficacy of lamellar keratoconjunctival transplantation. Compared to other existing treatments, lamellar keratoconjunctival transplantation demonstrated superior therapeutic outcomes and significant potential for clinical application in treating rabbit corneal ulcers 19 , 20 . By transplanting healthy lamellar keratoconjunctival tissue onto the ulcerated areas of recipient corneas, the procedure effectively replaces diseased tissue and provides an optimal physiological environment for corneal repair 21 , 22 . Following transplantation, the ulcerated areas of rabbit corneas exhibited marked improvements within a short period, characterized by reduced ulcer size, decreased opacity, and diminished anterior chamber reactions. These improvements were not limited to corneal appearance but also facilitated functional restoration, significantly enhancing visual quality in rabbits 23 – 27 . Furthermore, lamellar keratoconjunctival transplantation exhibited low rejection rates and high success. Since the procedure involves only superficial corneal layers and preserves the endothelial layer of the recipient cornea, it minimizes the risk of immune rejection. In the experiments, transplanted tissues integrated well with recipient corneas, without evident rejection responses or transplant failures. This advantage underscores the reliability and safety of lamellar keratoconjunctival transplantation for treating rabbit corneal ulcers. Histologically, lamellar keratoconjunctival transplantation promoted the proliferation of corneal squamous epithelial cells and facilitated ulcer healing. Confocal microscopy and histological staining revealed abundant regeneration of epithelial cells in the rabbit corneas after transplantation, with the ulcerated areas progressively replaced by healthy tissue. This process not only restored corneal transparency but also improved physiological functions, bringing the structural organization of the corneal tissue closer to normal conditions, characterized by orderly arrangement of stromal cells and notably reduced inflammatory cell infiltration. These findings further confirm the positive role of this surgical technique in promoting corneal tissue repair, suppressing inflammatory responses, and laying a foundation for vision recovery in rabbits. The successful implementation of lamellar keratoconjunctival transplantation also depends on meticulous surgical techniques and rigorous postoperative care. The thickness and size of the transplanted tissues must be precisely controlled during surgery to ensure compatibility with the recipient corneal bed 28 , 29 . Moreover, postoperative care requires careful monitoring of the rabbits' ocular condition, timely administration of antibiotic eye drops to prevent infections, and the use of ophthalmic ointments to protect the cornea 30 . These practices help to enhance surgical success rates and reduce the risk of postoperative complications. 5. Conclusion This study demonstrates that lamellar keratoconjunctival transplantation promotes more effective healing of corneal ulcers in rabbits compared with conservative medical treatment and conjunctival flap transplantation. The procedure accelerates epithelial repair, enhances corneal transparency and stromal structure, suppresses inflammatory cell infiltration and neovascularization, and significantly improves therapeutic efficacy and safety, representing a promising new therapeutic approach for managing corneal ulcers. Declarations Competing interests The authors declare no competing interests. Funding This work was supported by the Henan Science and Technology Attack Project (252102110047). Author Contribution Conceived and designed the experiments: Z.Z.W., H.Y.M. and T.T.L. Performed the experiments: Z.Z.W., M.R.C., Q.Y., H.Z., J.P.W. and Z.M.Z. Analyzed the data: Z.Z.W., X.F.Y., Q.G.H., M.Y., Z.X.A. and X.H.L. Wrote the paper: Z.Z.W. and H.Y.M. and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. Data availability Data will be made available on request. References Dhamayanti, F. A., Himayani, R. & Ismunandar, H. 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04:18:31","extension":"png","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":167559,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7307393/v1/f3fe9c8262154e96e337e238.png"},{"id":94622859,"identity":"cc9321e2-6136-4163-89d1-9b26921f43be","added_by":"auto","created_at":"2025-10-29 04:18:36","extension":"png","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":247441,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7307393/v1/697c4ca68769eb86561a8cba.png"},{"id":94640306,"identity":"a2544c8d-9e41-4b6e-8a12-be3ccc34bfb5","added_by":"auto","created_at":"2025-10-29 07:49:03","extension":"png","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":161979,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7307393/v1/4a445ebfe1a43ac9ef7b318e.png"},{"id":94622572,"identity":"842560bd-82ca-4fd0-b8eb-69a5ef0b280c","added_by":"auto","created_at":"2025-10-29 04:18:23","extension":"xml","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":66145,"visible":true,"origin":"","legend":"","description":"","filename":"2ee3364f1198429e82824d347f8156d51structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7307393/v1/54ae9d871ae426b8fc1ef156.xml"},{"id":94623019,"identity":"66e89327-114c-46c4-9bf6-f1104f4035c5","added_by":"auto","created_at":"2025-10-29 04:18:44","extension":"html","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":74346,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7307393/v1/945d533a46e4a50aaed0d136.html"},{"id":94622850,"identity":"c1a59977-928e-4030-9694-c9ecb3a48376","added_by":"auto","created_at":"2025-10-29 04:18:35","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1925519,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe effect of keratoconjunctival lamellar transplantation group and HE staining. \u003c/strong\u003e(A): After 9 weeks of conservative medical treatment, the anterior chamber remained completely obscured at the ulcer site, and newly formed corneal capillaries had almost entirely regressed. (B): HE staining at 9 weeks following conservative medical treatment revealed detachment of Descemet's membrane and endothelial layer. (C): At 9 weeks post conjunctival flap transplantation, the flap demonstrated good vascularization and completely covered the ulcerated region. (D): At 9 weeks following conjunctival flap transplantation, HE staining showed a loose stromal structure, with intact Descemet's membrane and endothelial layers closely adherent to the stroma. (E): At 9 weeks after lamellar keratoconjunctival transplantation, the graft had become transparent, and the visibility of the anterior chamber had increased. (F): HE staining at 9 weeks post lamellar keratoconjunctival transplantation showed well-organized stromal fibrillar arrangement with minimal fibroblast infiltration, intact Descemet's membrane, and endothelial layers tightly connected to the stroma.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7307393/v1/afdac37bad63cdf122ca4daf.png"},{"id":94623035,"identity":"ed30dc4f-bb63-43e8-9552-943f66716196","added_by":"auto","created_at":"2025-10-29 04:18:46","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1609091,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eTransparency score of ulcer site.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7307393/v1/b0c8c3a4656819bdf3a1d094.png"},{"id":94622950,"identity":"e39feaf0-7cfd-43b8-9770-f809b7c0c80f","added_by":"auto","created_at":"2025-10-29 04:18:40","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1020004,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGrading of fluorescein staining.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7307393/v1/c2865277979a3274441083ec.png"},{"id":100363285,"identity":"62eb3640-f792-4475-96eb-3d650ceb979c","added_by":"auto","created_at":"2026-01-16 07:49:19","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6980287,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7307393/v1/34ee03d5-3374-4017-a0c2-ab28b36f7535.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Evaluation of the Therapeutic Efficacy of Lamellar Keratoconjunctival Transplantation in Treating Corneal Ulcers in Rabbits","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eCorneal ulcers represent a common and serious ophthalmic condition frequently encountered in rabbit farming and ophthalmological research. Any loss of corneal epithelium, irrespective of its cause, can be defined as a corneal ulcer\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. In rabbits, corneal ulcers may result from multiple etiological factors, including trauma, infections (bacterial, viral, or fungal), and secondary complications related to ocular diseases\u003csup\u003e\u003cspan additionalcitationids=\"CR3\" citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. Once affected by corneal ulcers, rabbits typically exhibit ocular pain, tearing, blepharospasm, and corneal opacity, significantly impacting their vision and overall quality of life. Without timely and effective treatment, corneal ulcers may deteriorate rapidly, potentially leading to severe complications such as corneal perforation, endophthalmitis, and even blindness\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eCurrently, various therapeutic strategies are employed for managing corneal ulcers in rabbits, yet each has certain limitations. Traditional pharmaceutical therapies, such as topical antibiotic eye drops or ointments, have demonstrated effectiveness mainly in mild corneal ulcers, as reported by Jun-Yi and Sha X-Y\u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e,\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. However, in cases of moderate to severe ulcers, particularly those involving significant corneal tissue defects, conventional treatments often fail to achieve satisfactory outcomes. Surgical methods, such as corneal perforation repair, have been described by researchers including Nishi E N and Tsao W-S as technically challenging procedures demanding high surgical expertise and frequently associated with postoperative complications, including infection and excessive corneal scarring. Furthermore, these methods typically yield suboptimal tissue repair, often failing to fully restore the cornea\u0026rsquo;s normal structure and function\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e,\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. Lamellar keratoconjunctival transplantation, an emerging surgical technique, has achieved notable advancements within human ophthalmology. By transplanting healthy lamellar keratoconjunctival tissue, this procedure provides structural support and bioactive factors to damaged corneas, thus promoting epithelial repair and regeneration, enhancing corneal transparency and refractive status, and reducing immunological rejection\u003csup\u003e\u003cspan additionalcitationids=\"CR11 CR12 CR13\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. Nevertheless, research evaluating the therapeutic effects of lamellar keratoconjunctival transplantation on corneal ulcers in rabbits remains insufficient.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1 Ethics statement (Animals)\u003c/h2\u003e\u003cp\u003e All procedures involving animals were reviewed and approved by the Henan Institute of Science and Technology Institutional Animal Care and Use Committee (Approval No. LLSC2022044). All methods were carried out in accordance with relevant guidelines and regulations, including the Regulations for the Administration of Affairs Concerning Experimental Animals (P. R. China), the Guidelines for the Ethical Review of Laboratory Animal Welfare (GB/T 35892\u0026thinsp;\u0026minus;\u0026thinsp;2018), the General Principles of Laboratory Animal Welfare (GB/T 42011\u0026thinsp;\u0026minus;\u0026thinsp;2022), and Laboratory Animals\u0026mdash;Environment and Facilities (GB 14925\u0026thinsp;\u0026minus;\u0026thinsp;2023), as well as institutional policies. This study is reported in accordance with the ARRIVE guidelines (ARRIVE 2.0). Appropriate anesthesia and postoperative analgesia were provided, predefined humane endpoints were applied, and euthanasia was performed using an approved method to minimize pain and distress.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2 Establishment of the Rabbit Corneal Ulcer Model\u003c/h2\u003e\u003cp\u003eTwenty-four rabbits were randomly divided into three groups (n\u0026thinsp;=\u0026thinsp;8 per group): Group A received conservative medical treatment, Group B underwent conjunctival flap transplantation, and Group C underwent lamellar keratoconjunctival transplantation. Three days before surgery, fur surrounding the right eyes was clipped, and the eye sockets were cleaned. Rabbits were anesthetized using a 1:1 mixture of Zoletil 50 and Sumianxin II (0.1 mL/kg). After placing rabbits in the left lateral recumbent position, the right eyes were disinfected and topically anesthetized using proparacaine hydrochloride eye drops. Corneal ulcer models were induced by placing filter paper saturated with 1 mol/L NaOH solution on the central cornea for 1 min, followed by thorough rinsing with saline. Postoperatively, all rabbits received subcutaneous injections of meloxicam (0.5 mg/kg) and enrofloxacin (10 mg/kg), chloramphenicol eye drops four times daily, and protective Elizabethan collars.\u003c/p\u003e\u003cp\u003e\u003cb\u003e2.3 Conservative Medical Therapy, Conjunctival Flap Transplantation, and Lamellar Keratoconjunctival Transplantation for the Treatment of Corneal Ulcers\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe conservative medical treatment consisted of chloramphenicol eye drops applied three times daily, followed by recombinant bovine basic fibroblast growth factor (rb-bFGF) eye drops (Beifushu) 10 minutes later, continued for 60 days. For conjunctival flap transplantation, the corneal lesion was excised, extending 0.5-1 mm beyond the ulcer margin. An appropriately sized conjunctival flap was harvested from the dorsal bulbar conjunctiva, placed to cover the corneal defect without tension, and secured with 10\u0026thinsp;\u0026minus;\u0026thinsp;0 ophthalmic sutures. The lateral eyelids were partially closed with 5\u0026thinsp;\u0026minus;\u0026thinsp;0 absorbable sutures, which were removed after 7 days. In the lamellar keratoconjunctival transplantation group, lamellar keratoconjunctival grafts were harvested from the dorsal region of the globe. After excision of the diseased corneal tissue, a graft slightly larger (approximately 1 mm beyond defect margins) than the defect was prepared and secured tension-free onto the corneal wound using 10\u0026thinsp;\u0026minus;\u0026thinsp;0 ophthalmic sutures. The lateral eyelids were partially closed with 5\u0026thinsp;\u0026minus;\u0026thinsp;0 absorbable sutures, which were similarly removed after 7 days.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.4 Clinical Assessment\u003c/h2\u003e\u003cp\u003eFollowing the methods of Holland and Shaopan Wang\u003csup\u003e\u003cspan additionalcitationids=\"CR16\" citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. each animal was examined weekly, and quantitative assessments were performed based on scores ranging from 0 to 4 for corneal transparency, corneal edema, and corneal epithelial defects (assessed by fluorescein staining), as detailed in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eGrade evaluation of clinical recovery of corneal ulcers in rabbits\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrade\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCorneal Transparency\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eEdema Severity\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eCorneal Epithelial Defects (Fluorescein Staining)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCompletely transparent cornea\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eUntainted\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMild opacity at ulcer site\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSlight\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eMild staining with fewer than 5 scattered staining spots\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eModerate opacity at ulcer site, anterior chamber still visible\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eModerate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eModerate staining with more than 5 staining spots\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMarked opacity at ulcer site, anterior chamber barely visible\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSevere\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eStaining spots merging into lines or patches\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eComplete opacity at ulcer site, anterior chamber not visible\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eExtreme\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eExtensive patchy staining covering a large area\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e2.5 Cytokine and Histological Evaluation\u003c/h2\u003e\u003cp\u003eBlood samples were collected intravenously from rabbits at 1 day prior to corneal injury, 3 days post-injury, and 3, 7, 14, and 28 days after transplantation. Serum IL-6 and TGF-β1 levels were measured using ELISA kits. All recipient animals were euthanized by intravenous injection of a lethal dose of sodium pentobarbital at 9 weeks postoperatively, and excised corneas were subjected to hematoxylin-eosin (HE) staining for histological evaluation.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e2.6 Data Analysis\u003c/h2\u003e\u003cp\u003eStatistical analysis was conducted using SPSS 26.0, and experimental data were expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (X\u0026thinsp;\u0026plusmn;\u0026thinsp;SD). Differences were considered significant at P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 and highly significant at P\u0026thinsp;\u0026lt;\u0026thinsp;0.01. GraphPad Prism 9.0.0 software was used for plotting data.\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Result","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n \u003ch2\u003e3.1 Conservative Medical Treatment Group\u003c/h2\u003e\n \u003cp\u003eA total of 8 rabbits underwent conservative drug treatment. At the beginning of the treatment, the area of corneal sodium hydroxide burn showed significant edema (grade 0\u0026ndash;2). By the end of the first week, the congestion of the conjunctival capillaries decreased. In the second week, obvious edema also appeared around the burned area of the healthy cornea (grade 2\u0026ndash;3). The edema subsided after 3 weeks of treatment. At 5 weeks of treatment, new blood vessels on the cornea approached the ulcer site and reached the central area of the cornea. By 6 weeks, the new blood vessels on the cornea showed signs of regression. By the 9th week of treatment, the new capillaries on the cornea almost disappeared (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eA), but the condition of the corneal burn site was not significantly different from that at the beginning of treatment. No deterioration of the condition such as corneal perforation was observed. The HE staining of the corneas at 9 weeks after the surgery (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eB) showed that the epithelial layer became thinner, the stroma layer thickened, the fibrin arrangement was disordered, and there were a moderate number of fibroblasts; the posterior elastic layer and the endothelial layer detached.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n \u003ch2\u003e3.2 Conjunctival Flap Transplantation Group\u003c/h2\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003cp\u003eIn the conjunctival flap transplantation group, edema markedly decreased by week two (grade 0\u0026ndash;2), and good vascularization of the conjunctival flap was observed immediately after surgery, completely covering the ulcer site. Throughout treatment, all eight rabbits maintained good conjunctival flap viability. By day 60, the conjunctival flap gradually thinned and became increasingly transparent at its margins (grade 0\u0026ndash;2, Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eC). No flap detachment, corneal perforation, or other complications occurred during the 60-day observation period. Histological evaluation via HE staining at 9 weeks postoperatively (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eD) revealed a continuous epithelial layer completely covering the ulcerated area, characterized by increased cell density, thickened epithelium with dense cellular arrangement, and an intact Descemet\u0026rsquo;s membrane and endothelial layer closely adhering to the stromal layer.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003e3.3 Lamellar Keratoconjunctival Transplantation Group\u003c/h2\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003cp\u003eAfter completion of the lamellar keratoconjunctival transplantation, the ulcerated region was fully covered by corneal tissue and partially by conjunctival tissue, with a strip-like white area indicating the limbus. At 7 days post-transplantation, abundant vascularization was observed in the conjunctival portion of the graft, and extensive new capillary formation was also evident within the corneal portion. Beginning at 21 days postoperatively and continuing up to day 60, the graft gradually became thinner and more transparent, with an increasingly larger area of the anterior chamber becoming visible through the previous ulcer site (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eE). During the entire 60-day observation period, no adverse events such as graft detachment or corneal perforation were observed. Histological evaluation by HE staining at 9 weeks postoperatively (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eF) showed a continuous epithelial layer completely covering the ulcer site, with mild epithelial thickening at the junctional area. The stromal layer demonstrated well-organized fibrillar arrangement and contained a low density of fibroblasts. Additionally, Descemet\u0026rsquo;s membrane and the endothelial layer were intact and closely adhered to the stroma.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003e3.4 Assessment of Corneal Ulcer Treatment Efficacy\u003c/h2\u003e\n \u003cp\u003eDuring the 60-day treatment period, corneal transparency at the ulcer site in the conservative medical treatment group exhibited little improvement (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). The anterior chamber remained completely obscured by the ulcer throughout treatment, preventing any light from penetrating through the lesion. Although no severe complications such as further corneal damage or perforation occurred, the persistent opacity significantly impaired subsequent visual function. Corneal transparency in the conjunctival flap transplantation group was comparable to that of the conservative treatment group until day 28. Transparency gradually improved from day 35 to day 60, but this improvement was not statistically significant compared with conservative treatment. In contrast, the lamellar keratoconjunctival transplantation group showed significantly superior transparency compared to both other groups from day 7 to day 60 postoperatively (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01).\u003c/p\u003e\n \u003cp\u003eIn the first 14 days postoperatively, the clinical scores for corneal healing in the lamellar keratoconjunctival transplantation group were significantly better than those in the conjunctival flap transplantation group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01). From day 14 to day 35, the differences between these two groups gradually diminished, and no significant differences were observed from day 45 to day 60. Compared with the conservative medical treatment group, both the conjunctival flap transplantation and lamellar keratoconjunctival transplantation groups exhibited statistically significant improvements at all time points, except on day 7 postoperatively when the difference between conjunctival flap transplantation and conservative treatment was not significant. Toward the end of the treatment period (days 45\u0026ndash;60), some rabbits in both surgical groups still exhibited positive fluorescein staining, possibly due to eyelid friction against unremoved sutures, causing secondary epithelial damage (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). Measurements of serum IL-6 and TGF-\u0026beta;1 levels showed no significant differences among the three groups. IL-6 has been associated with corneal injury repair, and its increased serum levels at day 28 postoperatively may be indicative of ongoing corneal recovery. Growth factors and cytokines play a crucial role in corneal healing; activation of the TGF-\u0026beta; system promotes differentiation of stromal cells into myofibroblasts, aiding stromal repair. The observed initial rise and subsequent decline of TGF-\u0026beta;1 and \u0026beta;2 levels may correlate with stromal healing progression during ulcer repair.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eCorneal ulcers are a common ophthalmic disease that severely threatens vision. Without timely or proper intervention, corneal ulcers can lead to corneal perforation, intraocular infections, and even blindness\u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e. This study established a rabbit model of corneal ulcers to comprehensively evaluate the therapeutic efficacy of lamellar keratoconjunctival transplantation. Compared to other existing treatments, lamellar keratoconjunctival transplantation demonstrated superior therapeutic outcomes and significant potential for clinical application in treating rabbit corneal ulcers\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e,\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. By transplanting healthy lamellar keratoconjunctival tissue onto the ulcerated areas of recipient corneas, the procedure effectively replaces diseased tissue and provides an optimal physiological environment for corneal repair\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e,\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. Following transplantation, the ulcerated areas of rabbit corneas exhibited marked improvements within a short period, characterized by reduced ulcer size, decreased opacity, and diminished anterior chamber reactions. These improvements were not limited to corneal appearance but also facilitated functional restoration, significantly enhancing visual quality in rabbits\u003csup\u003e\u003cspan additionalcitationids=\"CR24 CR25 CR26\" citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. Furthermore, lamellar keratoconjunctival transplantation exhibited low rejection rates and high success. Since the procedure involves only superficial corneal layers and preserves the endothelial layer of the recipient cornea, it minimizes the risk of immune rejection. In the experiments, transplanted tissues integrated well with recipient corneas, without evident rejection responses or transplant failures. This advantage underscores the reliability and safety of lamellar keratoconjunctival transplantation for treating rabbit corneal ulcers.\u003c/p\u003e\u003cp\u003eHistologically, lamellar keratoconjunctival transplantation promoted the proliferation of corneal squamous epithelial cells and facilitated ulcer healing. Confocal microscopy and histological staining revealed abundant regeneration of epithelial cells in the rabbit corneas after transplantation, with the ulcerated areas progressively replaced by healthy tissue. This process not only restored corneal transparency but also improved physiological functions, bringing the structural organization of the corneal tissue closer to normal conditions, characterized by orderly arrangement of stromal cells and notably reduced inflammatory cell infiltration. These findings further confirm the positive role of this surgical technique in promoting corneal tissue repair, suppressing inflammatory responses, and laying a foundation for vision recovery in rabbits. The successful implementation of lamellar keratoconjunctival transplantation also depends on meticulous surgical techniques and rigorous postoperative care. The thickness and size of the transplanted tissues must be precisely controlled during surgery to ensure compatibility with the recipient corneal bed\u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e,\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e. Moreover, postoperative care requires careful monitoring of the rabbits' ocular condition, timely administration of antibiotic eye drops to prevent infections, and the use of ophthalmic ointments to protect the cornea\u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e. These practices help to enhance surgical success rates and reduce the risk of postoperative complications.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eThis study demonstrates that lamellar keratoconjunctival transplantation promotes more effective healing of corneal ulcers in rabbits compared with conservative medical treatment and conjunctival flap transplantation. The procedure accelerates epithelial repair, enhances corneal transparency and stromal structure, suppresses inflammatory cell infiltration and neovascularization, and significantly improves therapeutic efficacy and safety, representing a promising new therapeutic approach for managing corneal ulcers.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eCompeting interests\u003c/h2\u003e\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e\u003cp\u003eThis work was supported by the Henan Science and Technology Attack Project (252102110047).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eConceived and designed the experiments: Z.Z.W., H.Y.M. and T.T.L. Performed the experiments: Z.Z.W., M.R.C., Q.Y., H.Z., J.P.W. and Z.M.Z. Analyzed the data: Z.Z.W., X.F.Y., Q.G.H., M.Y., Z.X.A. and X.H.L. Wrote the paper: Z.Z.W. and H.Y.M. and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eData availability\u003c/h2\u003e\u003cp\u003eData will be made available on request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eDhamayanti, F. A., Himayani, R. \u0026amp; Ismunandar, H. Perforated Corneal Ulcer and Iris Oculi Sinistra Prolapse [Z]. (2020-01-01).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDuran-Ospina, J. P. et al. Global research on keratomycosis: New insights from latent Dirichlet allocation and HJ-Biplot-driven knowledge mapping Study [J]. \u003cem\u003eDiagn. Microbiol. Infect. Dis.\u003c/em\u003e \u003cb\u003e110\u003c/b\u003e (1), 116442 (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eThajudeen, B. \u003cem\u003eCorneal Ulcers and Contact Lens Keratitis [M]. 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Repeat corneal transplantation in Southern China: Indications, surgical technique, outcomes, and risk factors for repeat keratoplasty Failure [J]. \u003cem\u003eIndian J. Ophthalmol.\u003c/em\u003e \u003cb\u003e72\u003c/b\u003e (Suppl 3), S482\u0026ndash;S487 (2024).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Rabbit, corneal ulcer, Keratoconjunctival lamellar transplantation","lastPublishedDoi":"10.21203/rs.3.rs-7307393/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7307393/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCorneal ulcer exhibits an extremely high incidence in the field of rabbit ophthalmic diseases and stands as one of the crucial blinding eye conditions leading to blindness in rabbits. The occurrence of severe complications, such as corneal perforation, can inflict severe damage on the visual function of rabbits, resulting in a rapid decline in their vision and even causing blindness.For deep-seated corneal ulcers or refractory corneal ulcers that persist and cannot be effectively controlled despite standardized drug therapy, surgical intervention has become an indispensable treatment approach. Lamellar keratoconjunctival transplantation, as a highly significant ocular surface reconstruction surgery, plays an irreplaceable role in restoring the normal structure and function of the rabbit's eye.This study aimed to systematically evaluate the therapeutic efficacy of lamellar keratoconjunctival transplantation in treating corneal ulcers in rabbits. Twenty-four rabbits underwent corneal burns to establish ulcer models. After three days, rabbits were randomly divided into three treatment groups (n\u0026thinsp;=\u0026thinsp;8 each): Group A received conservative medical treatment, Group B underwent conjunctival flap transplantation, and Group C underwent lamellar keratoconjunctival transplantation. Postoperatively, corneal transparency, edema, and epithelial healing at the ulcer sites were regularly evaluated. Histopathological observations of corneal tissues and measurements of serum interleukin-6 (IL-6) and transforming growth factor-beta 1 (TGF-β1) were also conducted. Results indicated that the lamellar keratoconjunctival transplantation group (Group C) showed superior outcomes compared to the other two groups in terms of improved corneal transparency, reduced edema, enhanced epithelial healing, and favorable histopathological features. No significant differences were observed among the three groups in serum levels of IL-6 and TGF-β1. These findings suggest that lamellar keratoconjunctival transplantation is effective in supporting damaged corneas and promoting tissue regeneration, providing a promising method and alternative approach for the clinical management of corneal ulcers in rabbits and potentially in veterinary ophthalmology practice.\u003c/p\u003e","manuscriptTitle":"Evaluation of the Therapeutic Efficacy of Lamellar Keratoconjunctival Transplantation in Treating Corneal Ulcers in Rabbits","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-29 04:06:13","doi":"10.21203/rs.3.rs-7307393/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"cd7c782f-209c-416b-b9c3-6fc10d1b1f84","owner":[],"postedDate":"October 29th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":56880434,"name":"Health sciences/Diseases"},{"id":56880435,"name":"Health sciences/Medical research"}],"tags":[],"updatedAt":"2026-01-12T08:43:33+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-29 04:06:13","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7307393","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7307393","identity":"rs-7307393","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}