Objective To observe the biocompatibility of the acellular corneal stroma materials prepared by three different methods. Methods Three different serial digestion methods were used to produce the acellular corneal stroma materials. The biocompatibility of the materials was investigated by the cell seeding and the materials were implanted into the rabbit corneal stroma layer. Results The cells in the materials 1 and 2 were not decellularized completely. The rabbit corneal fibroblasts died on the materials 1 and 2 after the cell seeding for 3-4 days. An obvious rejection could be observed after the implantation. The cells in material 3 were decellularized completely and the collagen fibers or elastic fibers were reserved integrally，showing a typical three-dimensional net work. The rabbit corneal fibroblasts could expand on the materials in vitro. No obvious rejection could be observed and the materials were gradually absorbed. Conclusion The acellular porcine cornea stroma materials prepared by trypsin-Dnase-Rnase are suitable for reconstruction of the tissue engineered cornea.
The aim of this article is to study the effect of tumor necrosis factor alpha (TNF-α) on the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) in keratoconus fibroblasts in vitro. Normal cornea and keratoconus fibroblasts were extracted using enzyme digestion method and were cultured in the medium containing TNF-α (0, 10 and 100 ng/mL). The expression of MMPs proteins in the supernatant of corneal fibroblasts and the expression of TIMPs in the normal cornea and keratoconus fibroblasts were detected by Western blot and real-time quantitative polymerase chain reaction respectively. The active form of MMP1 could be detected in the supernatant of keratoconus fibroblasts and upregulated by TNF-α. TNF-α could increase the protein expression of MMP2, MMP3, MMP9 in the supernatant of keratoconus fibroblasts and decrease the gene expression of TIMP1, TIMP2 in keratoconus fibroblasts. The increased MMPs and the decreased TIMPs can increase the degradation of the extracellular matrix. TNF-α may play an important role in the occurrence and development of keratoconus by regulating the expression of MMPs/TIMPs.
Objective To compare the effects of the denudedfreeze-dried-amniotic-membrane and the denuded freeze-dried bovine corneal stroma when they were explanted to repair the corneal defect of rabbits. Methods The amnia from healthy human placentae were prepared with the method reported by LUO Jingcong, which were freeze-dried and sterilized. The bovine cornea was also denuded by typsin, rinsed, freeze-dried, and sterilized. Twenty Japan rabbits weredivided into group A(the amniontic group) and group B(the bovine-corneal-stroma group) at random. The defect was made, which was 7.5 mm in diameter and 1/3 ofthe thickness of the cornea, and the two kinds of materials were explanted to repair the defect. The vascularization and the changes of the operated eye were observed. The samples were taken at 2, 4 and 8 weeks for histologicalexamination. Results The explanted materials were not melted or excluded. There were visible neovessels in both groups, yet there was no significant difference between them. According to the histological observation, there was severe inflammation in both groups 2 weeks after operation, the fibroblasts were proliferated, and the collagen fibers were disorganized; however,the reactions became milder from 4 weeks after operations,andthe neovessles could be seen in groups A and B; at 8 weeks, the collagen fibers were more organized in groups A and B; however,there was still a small area of disorganized fibers left. Conclusion The two materials can lead to rejection to some extent, and so they need to be improved.
Study of the mechanical properties of in vivo corneal materials is an important basis for further study of corneal physiological and pathological phenomena by means of finite element method. In this paper, the elastic coefficient (E) and viscous coefficient (η) of normal cornea and keratoconus under pulse pressure are calculated by using standard linear solid model with the data provided by corneal visualization scheimpflug technology. The results showed that there was a significant difference of E and η between normal cornea and keratoconus cornea (P < 0.05). Receiver operating characteristic curve analysis showed that the area under curve (AUC) for E, η and their combined indicators were 0.776, 0.895 and 0.948, respectively, which indicated that keratoconus could be predicted by E and η. The results of this study may provide a reference for the early diagnosis of keratoconus and avoid the occurrence of keratoconus after operation, so it has a certain clinical value.
Objective To analyze the risk factors for persistent corneal epithelial defects (PCED) after pars plana vitrectomy (PPV) in patients with proliferative diabetic retinopathy (PDR). Methods A total of 201 PDR patients (201 eyes) who received PPV were enrolled in this retrospective study. There were 86 males (86 eyes) and 115 females (115 eyes). The patients aged from 30 to 81 years, with the mean age of (57.94±9.65) years. Among them, 159 patients were ≥50 years of age, and 42 patients were ＜50 years of age. There were 36 patients with HbA1c ＜7.0%, 165 patients with HbA1c ≥7.0%. There were 93 right eyes and 108 left eyes. There were 93 right eyes and 108 left eyes. The diabetic retinopathy stages were as follows: stage Ⅳ in 24 eyes, stage Ⅴ in 78 eyes and stage Ⅵ in 99 eyes. The operation time was ranged from 1 to 4 hours, with an average of 2 hours. Among the 201 eyes, corneal epidermis were scraped on 25 eyes; 70 eyes were combined with cataract surgery; a laser photocoagulation count ＜1000 points was performed in 78 eyes, and ＞1000 points were performed in 123 eyes. Sixty-one eyes involved intravitreal silicone oil tamponade, 18 eyes involved intravitreal tamponade with C3F8, and 122 eyes were not involved with intraocular tamponade. Postoperative persistent intraocular hypertension was defined as an intraocular pressure (IOP) ≥21 mmHg (1 mmHg=0.133 kPa) after PPV with necessary treatment using IOP-lowering medications for ≥2 weeks. The diagnostic criteria for corneal epithelial defects were taken from the Expert Consensus on Clinical Diagnosis and Treatment of Corneal Epithelial Defect in China (2016). The corneal epithelial defect was diagnosed as PCED if it was treated with common methods such as a lacrimal substitute or corneal contact lens, but showed no improvement and no signs of healing for ≥2 weeks. The incidence of PCED after eye surgery was recorded and its related risk factors were analyzed. A multivariate logistic regression was used to analyze the risk factors for PCED, which were expressed as a odds ratio (OR) and a 95% confidence interval (CI). Results Of 201 eyes, 16 eyes (7.96%) presented with PCED after surgery and 185 eyes (92.04%) with no PCED. There was no significant difference in the age, sex, and eyes between the patients with or without PCED (χ2=6.548, 0.927, 0.044; P=0.011, 0.336, 0.833). A multivariate logistic regression showed that intraoperative epithelial debridement (OR=13.239, 95%CI 2.999−58.442, P=0.001), intraoperative treatment in combination with cataract surgery (OR=7.448, 95%CI 1.975−28.091, P=0.003), intravitreal tamponade with C3F8 (OR=11.344, 95%CI 2.169−59.324, P=0.004), and postoperative persistent intraocular hypertension (OR=10.462, 95%CI 2.464−44.414, P=0.001) were risk factors for PCED after PPV. Conclusion Intraoperative epithelial debridement, intraoperative treatment in combination with cataract surgery, intravitreal tamponade with C3F8, and postoperative persistent intraocular hypertension are risk factors for PCED in patients with PDR after PPV.
As the most important refraction part in the optical system, cornea possesses characteristics which are important parameters in ophthalmology clinical surgery. During the measurement of the cornea in our study, we acquired the corneal data of Orbscan Ⅱ corneal topographer in real time using the Hook technology under Windows, and then took the data into the corneal analysis software. We then further analyzed and calculated the data to obtain individual Q-value of overall corneal 360 semi-meridian. The corneal analysis software took Visual C++6.0 as development environment, used OpenGL graphics technology to draw three-dimensional individual corneal morphological map and the distribution curve of the Q-value, and achieved real-time corneal data query. It could be concluded that the analysis would further extend the function of the corneal topography system, and provide a solid foundation for the further study of automatic screening of corneal diseases.
In the evaluation of tear film stability based on corneal topography, a pretreatment algorithm for tear film video was proposed for eye movement, eyelash reflection and background interference. First, Sobel operator was used to detect the blur image. Next, the target image with highlighted ring pattern was obtained by the morphological open operation performed on the grayscale image. Then the ring pattern frequency of the target image was extracted through the Hough circle detection and fast Fourier transform, and a band-pass filter was applied to the target image according to the ring pattern frequency. Finally, binarization and morphological closed operation were used for the localization of the ring pattern. Ten tear film videos were randomly selected from the database and processed frame by frame through the above algorithm. The experimental results showed that the proposed algorithm was effective in removing the invalid images in the video sequence and positioning the ring pattern, which laid a foundation for the subsequent evaluation of tear film stability.
ObjectiveTo compare the changes of corneal astigmatism after long-term spherical and toric orthokeratology wearing, and to investigate the effects of different orthokeratology design on corneal astigmatism.MethodsThe medical records of myopic adolescent patients who have been prescribed spherical and toric orthokeratology in the contact lens clinic of West China Hospital, Sichuan University between January 2019 and December 2021 were analyzed retrospectively.The differences of corneal astigmatism changes after wearing spherical and toric orthokeratology for a long time and one month discontinuation were compared. The influencing factors of corneal astigmatism changes were analyzed. ResultsA total of 156 patients were included. There were 76 cases (76 eyes) in spherical orthokeratology group and 80 cases (80 eyes) in toric orthokeratology group. There was no significant difference between the two groups in age, gender, baseline myopia diopter and total lens wearing time (P>0.05). There were statistically significant differences between the spherical orthokeratology group and the toric orthokeratology group in the baseline corneal flat K value [42.1 (41.3, 43.3) vs. 43.1 (42.0, 44.1) D], baseline corneal steep K value [(43.4±1.3) vs. (44.6±1.5) D], baseline corneal astigmatism [(1.1±0.5) vs. (1.6±0.6) D], and baseline total eye astigmatism [−0.6 (−1.2, 0.0) vs. −1.4 (−1.8, −1.0) D] before wearing the orthokeratology (P<0.05). Compared with the baseline value, 1 month after the two groups stopped wearing the orthokeratology, the corneal flat K values became flatter [spherical orthokeratology group: 42.09 (41.28, 43.34) vs. 41.73 (40.98, 43.16) D, toric orthokeratology group: 43.09 (41.95, 44.10) vs. 42.61 (41.52, 43.56) D; P<0.05], the changes of corneal steep K values were not statistically significant (P>0.05), but the corneal astigmatism values increased [spherical orthokeratology group: (1.05±0.49) vs. (1.37±0.56) D, toric orthokeratology group: (1.62±0.57) vs. (1.99±0.63) D; P<0.05]. There was no significant difference in the changes of corneal flat K value, corneal steep K value and corneal astigmatism between the two groups (P>0.05). Multivariate analysis showed that age (P=0.011) and the total duration of orthokeratology wearing (P=0.004) were the main factors affecting the changes of corneal astigmatism. ConclusionAfter 1 month of non-wearing, the flat K value of the cornea becomes flat, the steep K value remains unchanged, and the corneal astigmatism increases. There is no difference in the effect of the spherical and toric orthokeratology on corneal astigmatism. The change of corneal astigmatism is related to the patient’s age and the total duration of wearing the orthokeratology. The younger the age, the longer the duration of orthokeratology wearing, the more significant the increase of corneal astigmatism after stopping wearing the orthokeratology.
Due to lack of the practical technique to measure the biomechanical properties of the ocular cornea in vivo, clinical ophthalmologists have some difficulties in understanding the deformation mechanism of the cornea under the action of physiological intraocular pressures. Using Young's theory analysis of the corneal deformation during applanation tonometry, the relation between the elasticity moduli of the cornea and the applanated corneal area and the measured and true intraocular pressures can be obtained. A new applanation technique has been developed for measuring the biomechanical properties of the ocular cornea tissue in vivo, which can simultaneously acquire the data of the applanation area and displacement of the corneal deformation as well as the exerted applanation force on the cornea. Experimental results on a rabbit's eyeball demonstrated that the present technique could be used to measure the elasticity moduli and creep properties of the ocular cornea nondestructively in vivo.
The decrease of corneal stiffness is the key factor leading to keratoconus, and the corneal collagen fiber stiffness and fiber dispersion are closely related to the corneal biomechanical properties. In this paper, a finite element model of human cornea based on corneal microstructure, namely collagen fiber, was established before and after laser assisted in situ keratomileusis (LASIK). By simulating the Corvis ST process and comparing with the actual clinical results, the hyperelastic constitutive parameters and corneal collagen fiber stiffness modulus of the corneal material were determined before and after refractive surgery. After LASIK, the corneal collagen fiber stiffness modulus increased significantly, and was highly correlated with central corneal thickness (CCT). The predictive relationship between the corneal collagen fiber stiffness modulus and the corresponding CCT before and after surgery was: k1 before = exp(9.14 − 0.009CCTbefore), k1 after = exp(8.82 − 0.008CCTafter). According to the results of this study, the central corneal thickness of the patient can be used to estimate the preoperative and postoperative collagen fiber stiffness modulus, and then a personalized corneal model that is more consistent with the actual situation of the patient can be established, providing a theoretical reference for more accurately predicting the safe surgical cutting amount of the cornea.