曲古抑菌素A对人Tenon囊成纤维细胞作用的研究

目的：研究曲古抑菌素A( trichostatin A,TSA)对体外培养的人Tenon囊成纤维细胞( human Tenon capsule fibroblast, HTF)增殖能力以及组蛋白去乙酰化酶1( histone deacetylase 1,HDAC1)和HDAC2蛋白表达的影响。
　　方法：取青光眼滤过术中Tenon囊组织进行HTF体外培养。选取第3~6代细胞进行实验。设置空白对照组及TSA组(600nmol/L TSA加入培养液中),分别于培养后1,2,3 d应用 MTT法检测细胞活力变化;以600 nmol/L TSA处理 HTF 2 d 后, Western blot 法检测细胞中 HDAC1和HDAC2蛋白的表达。
　　结果：与对照组比较,TSA作用1d后HTF活力下降,呈时间依赖性,具有统计学差异( P<0.05)。 TSA处理HTF后2d,Western blot法检测HDAC1和HDAC2蛋白表达受到明显抑制。
　　结论：TSA可以通过抑制HDAC1和HDAC2的表达量,抑制人Tenon囊成纤维细胞增殖,从而减少术后结膜瘢痕形成可能。     

小檗胺抑制兔Tenon囊和滤过道内成纤维细胞增殖的实验研究

目的:研究钙调素拮抗剂小檗胺(berbamine,BER)对兔Tenon囊成纤维细胞和兔小梁切除术后滤过道内成纤维细胞增殖的抑制作用。方法:体外培养兔Tenon囊成纤维细胞,经不同浓度BER处理不同时间后,细胞计数Kit8(CCK8)法检测BER对兔Tenon囊成纤维细胞增殖的抑制作用,流式细胞仪检测其凋亡率及细胞周期的变化。HE染色检测BER对兔小梁切除术后滤过道内成纤维细胞增殖的抑制作用。结果:兔Tenon囊成纤维细胞经不同浓度BER处理不同时间后细胞增殖受到抑制,并呈时间剂量依赖性。流式细胞仪检测发现BER处理后兔Tenon囊成纤维细胞呈现G1期阻滞,细胞凋亡率明显增加,20mg/LBER处理9h,细胞凋亡率从0.64%增加到31.86%。HE染色显示BER显著抑制兔小梁切除术后滤过道内成纤维细胞的增殖。结论:小檗胺在体内外均能抑制成纤维细胞的增殖,其可能是通过诱导细胞凋亡方式抑制兔Tenon囊和滤过道内成纤维细胞的增殖。     

汉防己甲素对体外人眼Tenon囊成纤维细胞的抑制效应

目的:研究汉防己甲素(tetrandrine,Tet)对人眼Tenon囊成纤维细胞(Tenon’s capsule fibroblasts,TCFs)的抑制效应。方法:用组织块和混合消化液培养法体外培养人眼Tenon囊成纤维细胞,并对培养的传代细胞进行形态学的鉴定。采用MTT法检测不同浓度的汉防己甲素(10-4,10-5,10-6,10-7mol/L)及0.2g/L丝裂霉素(MMC)对人眼Tenon囊成纤维细胞的抑制作用。结果:人眼Tenon成纤维细胞体外生长良好,经光镜和免疫组化观察证明该细胞为成纤维细胞。MTT法证实,随着Tet药物浓度的增大,TCFs增生活性降低,对应的TCFs抑制率升高。结论:Tet对人眼Tenon囊成纤维细胞的增殖具有明显的抑制作用。     

自体眼球筋膜囊充填修补术在外伤性角膜缺损手术中的应用分析

目的：调查研究自体眼球筋膜囊充填修补术在外伤性角膜缺损手术中的应用效果。
　　方法：采用随机数字表法将100例外伤性角膜缺损患者分成治疗组和对照组各50例,给予治疗组患者自体眼球筋膜囊充填修补手术治疗,给予对照组患者常规性的手术治疗。
　　结果：治疗组患者的临床治疗总有效率为88%,对照组患者的临床治疗总有效率为68%,两者相比具有显著性的差异(P<0.05),所有患者接受治疗过程中均未出现异常现象。
　　结论：采用自体眼球筋膜囊充填修补手术治疗外伤性角膜缺损,有较好的临床治疗效果,且手术安全性较高,值得在临床上进一步推广运用。     

The efficacy and safety of intraoperative and/or postoperative 5-fluorouracil in trabeculectomy and phacotrabeculectomy

Purpose : First, to assess the safety and efficacy of using 5‐fluorouracil (5‐FU) to improve trabeculectomy and phacotrabeculectomy success rates, and second, to assess the efficacy of intraoperative and reduced postoperative 5‐FU following trabeculectomy compared with a more intensive course of postoperative 5‐FU alone. Methods : In a retrospective, unmatched, non‐randomized consecutive series study, 186 eyes of 186 patients who had filtration surgery were followed for 2 years in four groups: 51 patients had undergone trabeculectomy surgery with postoperative 5‐FU, 51 had phacotrabeculectomy with postoperative 5‐FU, 56 had trabeculectomy with both intraoperative and postoperative 5‐FU, and 28 patients had trabeculectomy without antifibrotics. Results : At all times mean intraocular pressure (IOP) was reduced in all groups (P < 0.001 for each group). Success was defined as IOP < 16 mmHg and > 30% IOP reduction at the 2‐year follow up. It was achieved in 71% of the trabeculectomy patients with intraoperative and postoperative 5‐FU, 76% of the trabeculectomy group with only postoperative 5‐FU, 55% of the phacotrabeculectomy/ 5‐FU group, and in 29% of the trabeculectomy‐only eyes (between‐group differences P < 0.01). Success rates were not significantly different for the intraoperative and postoperative 5‐FU trabeculectomy versus the postoperative 5‐FU‐only eyes, but the former had fewer postoperative 5‐FU injections and corneal ulceration (P < 0.01 for both). Conclusions : 5‐Fluorouracil was safe and improved trabeculectomy survival. Intraoperative 5‐FU allowed fewer postoperative 5‐FU injections and fewer side‐effects without compromising success rates. Phacotrabeculectomy with postoperative 5‐FU had a lower surgical success rate than did trabeculectomy with 5‐FU and this was not statistically different from trabeculectomy without 5‐FU.     

阿昔洛韦对体外培养人眼Tenon囊成纤维细胞增殖迁移的影响

目的:探索阿昔洛韦(ACV)对体外培养人眼Tenon囊成纤维细胞(HTFs)的增殖和凋亡的影响以及作用机制。方法:将HTFs分为ACV处理组和空白组;CCK8检测不同浓度梯度下的细胞增殖速率,划痕实验检测HTFs迁移能力,流式细胞术检测HTFs凋亡以及细胞周期。结果:与空白组相比,ACV处理组(终浓度分别为1.125、2.25、3.375、4.5mmol/L)HTFs增殖速率显著降低(P<0.05),且呈浓度依赖性。4.5mmol/L ACV处理组划痕细胞迁移率显著降低(P<0.05),细胞凋亡率显著增加(P=0.0005),细胞周期G0/G1期峰值升高(P=0.0011),S期下降(P=0.0006),细胞周期被阻滞在G0/G1期。结论:阿昔洛韦可以通过阻滞HTFs周期促进细胞凋亡,抑制HTFs的增殖和迁移。     

Postoperative 5-fluorouracil versus intraoperative mitomycin C in high-risk glaucoma filtering surgery: extended follow up

Purpose: To compare the long‐term efficacy and safety of postoperative subconjunctival 5‐fluorouracil (5‐FU) injections with that of intraoperative mitomycin C (MMC) in eyes at high risk for failure of trabeculectomy. Methods: In a retrospective, non‐randomized comparative trial, 36 eyes of 36 consecutive patients at high risk for failure of trabeculectomy underwent glaucoma filtering surgery with either postoperative subconjunctival 5‐FU injections (19 eyes) or intraoperative application of MMC (17 eyes). Intraocular pressure, number of postoperative antiglaucoma medications, postoperative visual acuity, interventions, and complications were evaluated. Results: Overall success (intraocular pressure ≤21 mmHg) at 1 year was 73.6% in the 5‐FU group and 82.3% in the MMC group. The cumulative 4‐year success was 52.6% in the 5‐FU group and 60.5% in the MMC group (P = 0.6). At 4‐year follow up, mean ± SD intraocular pressures were 17.58 ± 4.01 mmHg in the 5‐FU group and 13.33 ± 3.36 mmHg in the MMC group (P = 0.01). There was no significant difference in the number of post­operative medications (P = 0.84), appearance of blebs (P = 0.20), final visual acuity (P = 1.00), and complications (P > 0.05) between the groups. Conclusions: These results suggest that both postoperative 5‐FU injections and intraoperative MMC application have long‐term success in high‐risk patients. However, MMC results in a greater decrease in intraocular pressure than 5‐FU.     

Five Years’ Experience with Tenon-Conjunctival Flaps in Phthisical Eyes

Background: To evaluate the efficacy of total Tenon-conjunctival flaps (TCFs) for coverage of phthisical eyes with normal corneal sensitivity. Methods: A retrospective chart review was performed on 142 patients with mild monocular phthisis who underwent the TCF procedure to allow monocular scleral shell fitting at the King Khaled Eye Specialist Hospital, Riyadh, Saudi Arabia. All patients had normal corneal sensation in their painless phthisical eye and were intolerant to cosmetic scleral shell fitting preoperatively. Consecutive patients over a five-year period were enrolled. Data were collected on functional success, duration of follow-up, and complications. The Tenon’s and conjunctival layers were closed separately in 103 eyes and as a single layer in the remaining 39 eyes. Main outcome measures were functional success, defined as tolerance to the prosthetic scleral shell, and rate of postoperative complications such as wound retraction, epithelial cysts, and blepharoptosis. Fisher’s exact test was used to analyze the association between the main outcome measures and type of flap closure. Results: The mean duration of follow-up was 23.7±17.43 months. Functional success was achieved in 98.6% of eyes. Minor complications included epithelial inclusion cysts in 10 (7.04%) eyes and mild blepharoptosis in 9 (6.34%) eyes. Overall flap retraction was noted in 14 (9.86%) eyes. This complication occurred in 13 of 103 (33.3%) eyes with double layer closure and in 1 of 39 (0.97%) eyes with a single layer flap closure (p=0.11). Conclusions: TCF is an effective procedure to increase corneal thickness for coverage in phthisical eyes with positive corneal sensation. The incorporation of Tenon’s capsule in the flap did not increase complications with a frequency similar to the classic Gundersen procedure.     

Periocular Accumulation of Timolol and Betaxolol in Glaucoma Patients Under Long-term Therapy

Purpose: To determine if betaxolol or timolol is present in measurable concentration in the Tenon capsule in patients under long-term topical therapy. Methods: Small (1-cc) specimens of Tenon capsule were removed at the time of filtering surgery from 15 glaucoma patients under long-term preoperative topical therapy, nine of whom had been treated with timolol and six of whom had been receiving betaxolol. Methanol extracts of these tissue samples were analyzed quantitatively for the presence of either β-adrenergic antagonist by high-performance liquid chromatography. Results: Drug was detected in every specimen. A mean total of 2.6 (range, 0.1–30.0) μg of betaxolol was detected per 1-cc specimen. Conclusion: Timolol and betaxolol penetrate the conjunctiva and accumulate in the Tenon capsule. In patients under long-term therapy, the periocular tissue can accumulate a greater quantity of beta-antagonist than is present in a daily dosage of applied eyedrops, manyfold higher than the maximal intraocular concentration.     

Effect of cryopreserved amniotic membrane on the development of adhesions and fibrosis after extraocular muscle surgery in rabbits

Purpose: To histopathologically evaluate the effect of cryopreserved human amniotic membrane (AM) transplant on preventing the development of postoperative adhesions after extraocular muscle surgery. Methods: Ten albino rabbits were used. The superior rectus muscles were bilaterally resected. In right eyes, the muscle was wrapped with cryopreserved human AM (group AM). In left eyes, the muscle was not wrapped with AM and served as a control group (group C). The rabbits were killed, and the eyes were enucleated 6 weeks after surgery to perform histopathological examination. Results: On histopathological examination, the AM was present in eight eyes, surrounded by periamniotic inflammation, with no adhesions detected between rectus muscle and sclera, conjunctiva and Tenon’s capsule in the segment where the AM was present, but detected elsewhere. Adhesions were detected in the other two eyes of group AM, in which the AM was absent, and in all group C eyes. When comparing eye pairs of each rabbit, AM eyes showed significantly less adhesions between the muscle and sclera (p = 0.009) and between the muscle and Tenon’s capsule and conjunctiva (p = 0.008), in the region of AM application, and significantly more foreign body inflammation (p = 0.031), than C eyes. The differences between AM and C eye pairs, in terms of conjunctival inflammation and vascularity and muscle fibrosis, were insignificant (p > 0.05). Conclusions: Cryopreserved AM is effective in reducing postoperative extraocular muscle adhesions. Its application is, therefore, recommended during strabismus reoperations.     

改良小梁切除术联合生物羊膜植入治疗闭角型青光眼的临床观察

目的探讨Tenon’s筋膜切除联合生物羊膜植入的小梁切除术治疗闭角型青光眼临床疗效。方法对26例(30只眼)闭角型青光眼患者行Tenon’s筋膜切除联合生物羊膜植入的小梁切除术,术后随访1年,观察降眼压效果及其手术并发症。结果术后1周眼压(11.80±2.23)mmHg,较术前眼压(38.53±7.17)mmHg明显降低(P<0.01)。术后随访1年,眼压(17.80±4.11)mmHg,保持功能性滤泡27只眼(90%)。结论 Tenon’s筋膜切除联合生物羊膜植入的小梁切除术可作为治疗闭角型青光眼的一种有效方法,手术成功率高,长期眼压控制好,可有效地减少瘢痕组织形成。     

TGF-β2特异性siRNA载体干扰人结膜囊成纤维细胞表达的研究

目的:探讨人转化生长因子TGF-β2特异性siRNA真核表达载体转染人结膜囊成纤维细胞后对其TGF-β2mRNA表达的影响。方法:体外分离并培养人结膜囊成纤维细胞,在培养后传代3次的细胞以TGF-β2特异性siRNA真核表达载体进行转染,并以未转染细胞作为对照。转染后分别于24,48和72h收集细胞,采用RT-PCR技术检测TGF-β2特异性siRNA真核表达载体对TGF-β2mRNA表达的影响。结果:分离的人结膜囊成纤维细胞于接种约4h左右开始贴壁,同时细胞变长成为梭形,表现出明显的成纤维细胞特性,约36h后达到融合状态;RT-PCR结果示:与对照组相比,转染24,48和72h后的细胞TGF-β2表达抑制率分别为17.40%,52.80%和79.20%,抑制效率呈现随时间延长有所加强的趋势。结论:TGF-β2特异性siRNA真核表达载体能抑制人结膜囊成纤维细胞TGF-β2mRNA的表达。     

Effect of Limited Tenon Capsule and Intermuscular Membranes Dissection on the Outcome of Surgery in Patients with Horizontal Strabismus

Purpose: The aim of the study was to evaluate the effect of limited dissection of Tenon capsule on the outcome of strabismus surgery.

Methods: Patients between the ages of 2 and 50 years with pure horizontal strabismus were enrolled in a prospective study. Patients were divided into two groups: case and control as a non-randomized study. The method of operation was similar in both groups except for the amount of sheath Tenon dissection. In the control group the intermuscular connections and Tenon capsule were cut as much as possible. In the study group, Tenon capsule, 3-4 mm posterior to the location of the sutures over the muscle (recessed or resected), were cut and intermuscular connections remained intact.

Results: The study enrolled 54 patients with 27 patients in each group. In both groups, after operation, regardless of the type of surgery, the angle of strabismus was reduced (P< 0.05). For patients who had undergone bimedial recession, the angle of strabismus was corrected to 2.6 ± 0.4 prism diopters per mm (PD/mm) of muscle recession in the case group, and 2.2 ± 0.9 in the control group (P=0.2). For patients who had undergone monocular recession and resection (R&R) for esotropia, the angle of strabismus was corrected 3.4 ± 0.3 PD/mm of muscle recession or resection in the case group, and 3.2 ± 0.2 in the control group (P=0.05). For patients who had undergone bilateral recession, the angle of strabismus was corrected 2.3 ± 0.2 PD/mm of muscle recession in the case group, and 2.2±0.2 in the control group (P=0.03). For patients who had undergone R&R for exotropia, the angle of strabismus was corrected 3.1 ± 0.5 PD/mm of muscle recession or resection in the case group, and 2.7 ± 0.3 in the control group (P=0.02).

Conclusion: Connective tissue ensheathments, whether disturbed or removed, do not ultimately affect the success of the strabismus surgery.     

Bilateral cataract induced by unilateral UVR‐B exposure – evidence for an inflammatory response

Purpose: To investigate whether unilateral in vivo UVR‐B exposure of one eye affects the fellow eye in a co‐cataractogenic, sympathetic reaction and to determine whether an inflammatory response could be involved in the pathogenesis. Methods: C57BL/6 mice were unilaterally exposed in vivo to UVR‐B for 15 min. In the group of 24 animals each received 0×/2×/3×/or 4× cataract threshold equivalent dose. Following 48‐hr UVR‐B exposure, cataract morphology was documented in dark‐field illumination photography, and light scattering was quantified, in both lenses in vitro. Serum levels of pro‐inflammatory cytokines IL‐1ß, IL‐6 and TNF‐α were analysed with ELISA. Immunohistochemistry was performed for inflammatory infiltration in exposed and contralateral eyes. Results: UVR‐B exposure induced cataract in all exposed lenses. There was additionally a significant UVR dose–dependent increase in light scattering in the lenses of the non‐exposed fellow eye. Inflammatory infiltration was detected immunohistochemically in the anterior segment of both eyes. IL‐1β serum concentration increased with increasing UVR‐B exposure dose. There was a similar trend for serum IL‐6 but not for TNF‐α. Conclusion: Unilateral UVR‐B exposure to one eye is associated with intraocular inflammation and an increase in lens light scattering also in the unexposed, fellow eye. A resulting systemic inflammatory response might be mediated by IL‐1β and possibly IL‐6. The finding that an inflammatory response may play a role in UVR‐B‐induced cataract development might initiate new strategies in the prevention of the disease.     

Lenses from Brown‐Norway pigmented rats are more tolerant to in vitro ultraviolet irradiation than lenses from Fischer‐344 albino rats

Purpose: To investigate if the previously shown difference in in vivo‐induced ultraviolet B radiation (UVR‐B) cataractogenesis between pigmented and albino rats can be seen also with in vitro irradiation. The shielding effect of the iris and UVR absorption in the anterior segment is nullified, and inherent differences in lenticular UVR‐B sensitivity between the strains may be revealed. Methods: Lenses from albino (Fischer‐344) and pigmented (Brown‐Norway) rats were irradiated in vitro with 1.8 kJ/m² UVR‐B. The lenses were cultured in standard environment in a culture incubator. Cataract was quantified daily by measuring the amount of lens forward light scattering over a period of 1 week. All lenses were photographed during the week. Results: Two days after exposure, both strains developed significant cataract compared to control lenses, and the light scattering increased exponentially to the last day. From day 4, exposed Fischer lenses scattered more light than Brown‐Norway lenses. This difference increased towards the end of the week. The type of cataract (anterior subcapsular, equatorial, and posterior cortical cataract) was similar in both strains. No anterior polar or nuclear cataract was observed. Conclusions: Lenses from albino Fischer rats are more sensitive to in vitro UVR‐B than lenses from pigmented Brown‐Norway rats. Ultraviolet B radiation cataract type induced in vitro differs from in vivo cataract in pigmented rats, but not from albino rats. In vitro UVR‐B exposure induces more cataract than corresponding lenticular UVR‐B in vivo exposures, for both albino and pigmented rat.     

Topical exposure of mitomycin C reduces opacification of the residual anterior lens capsule and lenticular regeneration after vitrectomy and lensectomy in rabbits

Purpose To examine the effect of topical administration of mitomycin C (MMC) on the opacification of residual anterior lens capsule in a vitrectomized/lensectomized eye.Methods Adult Japanese albino rabbits (n=8) received two-port vitrectomy and lensectomy with or without topical administration of MMC (0.4 mg/ml) to the residual anterior lens capsule in one eye under both general and topical anesthesia. Four eyes after vitrectomy and lensectomy were exposed to MMC for 1 min and washed out with balanced salt solution. After 1 month, the anterior segment of each globe was observed with EAS-1000 in vivo under general anesthesia and then the enucleated globes were observed through the Miyake view. Histological examination was performed.Results During healing intervals in eyes without MMC exposure, regenerated lens structure of Soemmerings ring or fibrous tissue was formed in peripheral or central areas of the residual capsule, respectively. Both the anterior lens capsule opacification and lens fiber regeneration were statistically significantly reduced by MMC topical exposure.Conclusion Lens epithelial cells produce regenerated lenticular structure and fibrous tissue on the residual capsule following vitrectomy and lensectomy in rabbits. Exposure of the residual anterior capsule to MMC may be effective in reducing both its opacification and lens fiber regeneration after vitrectomy/lensectomy.Commercial relationship policy for all authors: none     

Enduragen Patch Grafts for Exposed Orbital Implants

Purpose: To present a new technique using the recently introduced Enduragen^® material (Tissue Science Laboratories) as a patch graft for exposed ocular implants.

Methods: A retrospective, interventional, non-comparative case series of 3 patients who had Enduragen patch grafts for the closure of Tenon’s capsule and conjunctiva over exposed ocular implants. Medical records were reviewed and the following parameters were collected: age, gender, indication for surgery, type of surgery, laterality, type of orbital implant, complications after repair and length of follow-up.

Results: Three patients were identified, 2 males and 1 female. One patient had a secondary quad-motility implant with supertemporal exposure. The second patient had a secondary implant with a fistula at the lateral aspect of the socket. The third patient had a centrally exposed primary hydroxyapatite implant. All patients received Enduragen patch grafts to cover the implant. Follow up ranged from 40 to 43 months (mean, 41.3 months; SD, ± 1.5). None of the 3 patients had any signs of implant re-exposure at the time of the last post-operative visit. There were no intra-operative or early complications observed.

Conclusions: This consecutive case series suggests that Enduragen could be used as a safe and effective patch graft for exposed ocular implants. However, a larger prospective study with longer follow-up would be useful in further defining the indications and limitations of the Enduragen patch graft for the treatment of exposed orbital implants.     

A Conjunctival Retractor during Strabismus Surgery

The use of a conjunctival retractor that expands a small incision in the conjunctiva/Tenon’s capsule facilitated the performance of strabismus surgery through a fornix incision. A wide field of exposure revealed the entire area of the muscle insertion. While there is a risk of conjunctival tearing in elderly patients, the use of the retractor is valuable.     

Effect of Cauterization on the Subconjunctival Fibrovascular Reaction in Rabbit Eyes

ABSTRACT

Objective: The purpose of our study is to investigate the effect of episcleral cauterization during conjunctival autograft surgery on subconjunctival fibrovascular reaction in rabbit eyes. Methods: Twenty-four healthy male New Zealand white rabbits were divided into Control Group (n?=?12) and Cautery Group (n?=?12). In the Control Group, a 4?×?4?mm free conjunctival graft was elevated, the underlying Tenon’s capsule was excised, and the free graft was sutured back to its original place with 10-0 nylon sutures. In the Cautery Group, in addition to the same surgical steps, the episclera was cauterized with six gentle touches before the conjunctival graft was sutured back to its original place. The scleroconjunctival block of the operated area was excised at the first month. The sections were stained with hematoxylin and eosine and Masson trichrome and were evaluated using light microscopy. Results: Subconjunctival collagen fibers were increased, and the fibroblast density increased more significantly in the Cautery Group than in the Control Group. The groups were similar regarding vascularization, lymphocytic, and plasmocytic infiltration. Macrophage density was normal in all animals in both groups. Conclusion: Our histopathological examinations indicate that cauterization seems to lead to an increase in subconjunctival fibrosis at the first postoperative month in rabbit eyes.