Effects of Qingguang'an containing serum on the expression levels of autophagy-related genes in human Tenon's fibroblasts induced by transforming growth factor beta 1.

OBJECTIVE: To explore the effects of Qingguang'an () containing serum on the expression levels of autophagy related genes in the transforming growth factor beta 1 (TGF-ß1)-activated human Tenon's fibroblasts (HTFs). METHODS: (a) Primary HTFs were stimulated by TGF-ß1 and underwent immunohistochemistry, which established a cell model after Glaucoma filtration surgery (GFS). (b) The cell models were divided into 4 group: normal group (normal cells), model group (+TGF-ß1),treatment group (+TGF-ß1+ medicated serum), and positive control group (TGF-ß1+ rapamycin). Then, Qingguang'an medicated serum with optimum concentration was added to the corresponding group. The autophagy positive cells were identified by the Cyto-ID autophagy detection kits under fluorescent microscope and Cytation 5 multifunctional instrument for cell imaging. And the mean fluorescence intensity of autophagy positive cells was determined by flow cytometry. The expression levels of autophagy related genes － Beclin-1, autophagy related gene 5 (ATG-5), and microtubule-associated protein 1 light chain 3 (LC-3Ⅱ were detected by quantitative reverse transcription-polymerase chain reaction and Western blot analysis. RESULTS: Compared with the normal group and the model group, the relative mRNA expression levels of autophagy-related genes (Beclin-1, ATG-5 and LC-3Ⅱ in the experimental group were notably increased (P < 0.05, P < 0.01), and with the extension of treatment time, it had an increasing trend (48 h was more obvious), which showed a certain time dependency; the protein expression levels of autophagy-related genes (Beclin-1, ATG-5, and LC-3Ⅱ were significantly increased in the experimental group (P < 0.05, P < 0.01). With the prolongation of treatment time, there was an increasing trend (48 h was relatively obvious), and it revealed a certain time dependency. CONCLUSION: The Qingguang'an medicated serum could up-regulate autophagy related genes (Beclin1, ATG5, and LC3Ⅱ in the TGF-ß1-activated HTFs.

A Tenon's capsule/bulbar conjunctiva interface biomimetic to model fibrosis and local drug delivery.

Glaucoma filtration surgery is one of the most effective methods for lowering intraocular pressure in glaucoma. The surgery efficiently reduces intra-ocular pressure but the most common cause of failure is scarring at the incision site. This occurs in the conjunctiva/Tenon's capsule layer overlying the scleral coat of the eye. Currently used antimetabolite treatments to prevent post-surgical scarring are non-selective and are associated with potentially blinding side effects. Developing new treatments to target scarring requires both a better understanding of wound healing and scarring in the conjunctiva, and new means of delivering anti-scarring drugs locally and sustainably. By combining plastic compression of collagen gels with a soft collagen-based layer, we have developed a physiologically relevant model of the sub-epithelial bulbar conjunctiva/Tenon's capsule interface, which allows a more holistic approach to the understanding of subconjunctival tissue behaviour and local drug delivery. The biomimetic tissue hosts both primary human conjunctival fibroblasts and an immune component in the form of macrophages, morphologically and structurally mimicking the mechanical proprieties and contraction kinetics of ex vivo porcine conjunctiva. We show that our model is suitable for the screening of drugs targeting scarring and/or inflammation, and amenable to the study of local drug delivery devices that can be inserted in between the two layers of the biomimetic. We propose that this multicellular-bilayer engineered tissue will be useful to study complex biological aspects of scarring and fibrosis, including the role of inflammation, with potentially significant implications for the management of scarring following glaucoma filtration surgery and other anterior ocular segment scarring conditions. Crucially, it uniquely allows the evaluation of new means of local drug delivery within a physiologically relevant tissue mimetic, mimicking intraoperative drug delivery in vivo.

Natural Plant Extract Berbamine Is a Potent Inhibitor of Cell Growth and Survival of Human Tenon's Fibroblasts.

INTRODUCTION: Post-trabeculectomy scarring due to excessive proliferation of human Tenon's fibroblasts (HTFs) often led to operation failure. Developing a new anti-fibrosis drug with high efficacy to inhibit HTF cell growth will greatly improve the effectiveness of trabeculectomy. OBJECTIVE: This study aims to investigate the effect of berbamine (BBM) treatment on the cell growth and survival of HTFs. METHODS: Cultured human fetal Tenon's fibroblasts (HFTFs) were treated with or without different concentrations of BBM. Cell morphology was observed with a phase contrast microscope. A CCK-8 method and Ki67 immunofluorescence were used to determine cell viability and cell proliferation. A scratch test was used to study cell migration. Flow cytometry and TUNEL staining were performed to detect cell apoptosis. The expression of BAX/BCL-2, ERK, and AKT/mTOR pathway components was determined by Western blotting. RESULTS: BBM treatment disrupted HFTF normal morphology and inhibited its cell growth in a dose-dependent manner. Ki67 immunofluorescence and scratch assay showed BBM suppressed HFTF cell proliferation and migration. Importantly, BBM dose-dependently increased the BAX/BCL-2 ratio and induced apoptosis in HFTF cells. Western blotting showed BBM significantly inhibited the ERK and AKT/mTOR pathway, and PTEN inhibition ameliorated the inhibitory effect of BBM on cell viability and survival in HFTFs. CONCLUSIONS: BBM potently inhibits the cell growth and survival of HTFs through AKT/mTOR and has the potential to serve as an anti-fibrosis drug after trabeculectomy.

Efficacy and Safety of a Novel Blunt Cannula Trans-Sub-Tenon's Retrobulbar Block for Vitreoretinal Surgery.

PURPOSE: To evaluate a novel trans-sub-Tenon's retrobulbar block (TSTRB) compared to sub-Tenon's block (STB) and peribulbar block (PBB) anesthesia for vitreoretinal surgery. METHODS: This study was a prospective evaluation of cases undergoing TSTRB, STB, or PBB. The Kallio scale and Brahma scales were used to score hemorrhage and extraocular motility, respectively. Pain was documented on a visual analog score graded (1-10) at induction, intraoperatively, and postoperatively, any confounding variables were noted. RESULTS: Seventy eyes have been used in this analysis, of which TSTRB was used in 37% (n = 26), PBB in 34% (n = 24), and STB in 29% (n = 20). Postoperative analgesia was required by 10% (n = 2) of STB and 8% (n = 2) of PBB; none of the TSTRB cases required analgesia (P = 0.003). The mean volume required with each technique was as follows: TSTRB, 4.8 ml; STB, 5.3 ml; and PBB, 10.4 ml (P = 0.030). The volume of anesthesia was correlated with the level of proptosis and even more important affected the ease of surgery most (P = 0.005). Akinesia was greatest with TSTRB > PBB > STB (P = 0.040). There were no complications such as brainstem anesthesia, globe perforation, or retrobulbar hemorrhage. CONCLUSION: Intentionally extending a STB into the retrobulbar space, via a TSTRB fenestration utilizes a familiar skill set. TSTRB produced the best levels of reduced kinesia during surgery and increased duration of postoperative analgesia. The technique uses a small-volume anesthesia.

Incidence and impact factors of intraoperative loss of light perception under sub-Tenon's anesthesia in patients with macular diseases.

PURPOSE: To investigate the incidence and impact factors of intraoperative loss of light perception (LP) under sub-Tenon's anesthesia in patients with macular diseases. METHODS: Eighty-five consecutive patients received standard phacoemulsification combined pars plana vitrectomy (PPV) under sub-Tenon's anesthesia. At several checkpoints during the surgery (the end of phacoemulsification, the end of vitrectomy, and the end of surgery), participants were interviewed about whether they had LP or not after removing the influence of contralateral eye and the photo-bleaching effect. In patients treated with retinal photocoagulation, visual experience on laser flashes was evaluated. RESULTS: Under routine draping, no patients reported loss of LP at all the checkpoints. When the contralateral eye was tightly covered, the rates of LP loss were 84.7%, 97.6%, and 87.1% at the end of phacoemulsification, the end of vitrectomy, and the end of surgery, respectively. When the photo-bleaching effect was also removed, the rates of LP loss were 61.2%, 82.4%, and 56.5% at each checkpoint, respectively, and there were 87.1% (74/85) of patients reporting visual loss in at least one checkpoint. In addition, 76.9% (50/65) of patients could not feel laser flashes during retinal photocoagulation. CONCLUSION: Intraoperative loss of LP under sub-Tenon's anesthesia was a relatively common and reversible event. The conduction block of optic nerve by anesthetic mainly contributed to the visual loss during surgery. Photo-bleaching effect also has some effect on the LP evaluation. Surgeons need to inform and counsel the patients about the intraoperative loss of LP, to prevent any sudden panic attacks in them.

Local Anesthesia With Blunt Sub-Tenon's Cannula Versus Sharp Retrobulbar Needle for Vitreoretinal Surgery: A Retrospective, Comparative Study.

BACKGROUND AND OBJECTIVE: To evaluate the safety and efficacy of a blunt sub-Tenon's cannula for local anesthesia before vitreoretinal surgery compared to a sharp retrobulbar needle. PATIENTS AND METHODS: Retrospective, comparative study of all patients undergoing vitreoretinal surgery at the Medical College of Wisconsin between August 2009 and November 2013. Institutional review board approval was obtained. RESULTS: Of 940 surgeries performed with a sub-Tenon's cannula, 99% (938 of 940) were completed. Of the 771 surgeries performed with a sharp retrobulbar needle, 99% (770 of 771) were completed. Factors associated with use of a sharp retrobulbar needle over sub-Tenon's cannula were presence of prior scleral buckle (P < .01) and inclusion of scleral buckle placement in the procedure (P < .01). No case of globe perforation, severe retrobulbar hemorrhage, or severe conjunctival chemosis was observed in either group. CONCLUSION: Blunt sub-Tenon's cannula appears as effective and safe as a sharp retrobulbar needle for local anesthesia during vitreoretinal surgery. Vitreoretinal surgeons may wish to consider a blunt sub-Tenon's cannula for local surgical anesthesia.

Effects of sub-Tenon's anaesthesia on ocular hemodynamics.

OBJECTIVE: To research the effects of sub-Tenon's anaesthesia (STA) on ocular hemodynamics in patients with cataract using colour Doppler imaging (CDI). DESIGN: Prospective clinical study. PARTICIPANTS: Thirty-four eyes of 34 patients with age-related cataract. METHODS: Peak systolic velocity (PSV), end-diastolic velocity (EDV), and resistive index (RI) values of the ophthalmic artery (OA), central retinal artery, and central retinal vein were measured by CDI before and immediately after performing sub-Tenon block using 2 mL of 2% lidocaine with adrenaline (1:200 000). RESULTS: Both PSV and EDV values were significantly decreased after sub-Tenon injection in all the studied vessels of the patients. However, no important changes were observed in the RI values of the vessels. CONCLUSIONS: STA markedly reduced ocular blood flow. The reduction may be more acceptable compared with other retrobulbar block methods. Sub-Tenon block should be performed carefully to senile patients with cataract with vascular occlusive disorder.

Staggered sub-Tenon's blocks: a novel anesthetic approach for bilateral strabismus surgery.

Bilateral strabismus surgery is usually performed under general anesthesia. However, sometimes general anesthesia is not in the best interest of patients presenting with advanced multiple comorbidities. These patients are best treated with local anesthesia instead. However, the bilateral nature of the surgery becomes an issue because bilateral akinetic needle blocks for ophthalmic surgery have not been described before. This article describes a novel approach for bilateral eye surgery in three patients who were at high risk from general anesthesia but needed to have the surgery performed in the same sitting. Staggered blunt needle sub-Tenon's blocks were chosen over sharp needle akinetic extraconal peribulbar or intraconal retrobulbar blocks on account of their predictability, relative safety, and efficacy. The potential systemic complications of bilateral injections were minimized by staggering the blocks. This technique may be an option for high-risk patients who are not candidates for general anesthesia but require bilateral ophthalmic surgery performed in a single surgical episode.

Two-quadrant high-volume sub-Tenon's anaesthesia for vitrectomy: a randomised controlled trial.

BACKGROUND: Total volume using a standard single inferonasal injection for sub-Tenon's anaesthesia is limited by an increase in intraocular pressure (IOP) and commonly requires the operating surgeon to top-up the block intraoperatively. This study compares the efficacy and safety of a two-quadrant technique that allows the use of a higher volume of local anaesthetic. METHODS: 54 patients undergoing vitrectomy were randomised into two groups. The control group (n=27) received a standard 5 ml single inferonasal sub-Tenon injection of a 50:50 mixture of 2% lidocaine and 0.5% bupivacaine with 150 IU hyaluronidase. The study group (n=27) received a 5 ml inferonasal and 5 ml superotemporal injection of the same mixture (10 ml total). The primary outcome measure was the number of intraoperative top-ups required. Secondary outcome measures were intraoperative and postoperative pain scores, IOP, block onset time, ocular akinesia, eyelid akinesia and chemosis. RESULTS: 24 patients required a top-up in the control group. No patients required a top-up in the study group (p<0.001). IOP measurements were similar in both groups. Block onset was shorter, eyelid akinesia was improved and pain scores were also reduced in the study group intraoperatively and at 0-2 h, 4-6 h, 10-14 h and 20-24 h postoperatively. CONCLUSIONS: Two-quadrant sub-Tenon's anaesthesia using 10 ml of a 50:50 mixture of 2% lidocaine and 0.5% bupivacaine with 150 IU hyaluronidase seems to be more effective than a single-quadrant technique at reducing intraoperative and postoperative pain during vitrectomy.