Air pressure changes in the creation and bursting of the type-1 big bubble in deep anterior lamellar keratoplasty: an ex vivo study

Purpose: To measure the pressure and volume of air required to create a big bubble (BB) in simulated deep anterior lamellar keratoplasty (DALK) in donor eyes and ascertain the bursting pressure of the BB.Patients and methods: Twenty-two human sclera-corneal discs were used. Air was injected into the corneal stroma to create a BB and the pressure measured by means of a pressure converter attached to the system via a side port. A special clamp was designed to prevent air leak from the periphery of the discs. The pressure at which air emerged in the corneal tissue; the bursting pressure measured after advancing the needle into the bubble cavity and injecting more air; the volume of air required to create a BB and the volume of the BB were ascertained.Results: Type-1 BB were achieved in 19 and type-2 BB in 3 eyes. The maximum pressure reached to create a BB was 96.25+/- 21.61 kpa; the mean type-1 intrabubble pressure was 10.16 +/- 3.65 kpa. The mean bursting pressure of a type-1 BB was 66.65 +/- 18.65 kpa, while that of a type-2 BB was 14.77 +/- 2.44 kpa. The volume of air required to create a type-1 BB was 0.54 ml and the volume of a type-1 BB was consistently 0.1 ml.Conclusions: Dua's layer baring DALK can withstand high intraoperative pressures compared to Descemet's membrane baring DALK. The study suggests that it could be safe to undertake procedures such as DALK-triple with a type-1 BB but not with a type-2 BB

In vitro evaluation of electrospun blends of gelatin and PCL for application as a partial thickness corneal graft

The advent of innovative surgical procedures utilizing partial thickness corneal grafts has created a need for the development of synthetic implants to recreate corneal stromal tissue. This work evaluates electrospun gelatin and polycaprolactone (PCL) scaffolds as a potential biomaterial suitable for use in regeneration of corneal stromal tissue. Electrospun gelatin has been used for many years in tissue engineering, however, post‐production modification, such as crosslinking, is usually required to mechanically strengthen such scaffolds. This paper aims therefore to compare glutaraldehyde (GA) cross‐linked electrospun gelatin scaffolds with electrospun blends of gelatin and PCL at different ratios. Scaffolds were fabricated using electrospinning and characterized by scanning electron microscopy, Attenuated Total Reflectance‐Fourier Transform Infrared Spectroscopy (ATR‐FTIR), and tensile testing. To evaluate biocompatibility, primary human corneal stromal cells (hCSC) were seeded upon the scaffolds to assess adherence, proliferation and phenotype. Results demonstrated that scaffolds fabricated from mixtures of gelatin and PCL showed increased mechanical strength and plasticity compared to scaffolds fabricated from GA cross‐linked gelatin alone. In addition, scaffolds fabricated from PCL and gelatin showed comparable support of hCSC adhesion and proliferation. In conclusion, blended mixtures of gelatin and PCL can be considered as an option in the selection of corneal repair materials in the future

Gamma-irradiated human amniotic membrane decellularised with sodium dodecyl sulfate is a more efficient substrate for the ex vivo expansion of limbal stem cells

yesThe gold standard substrate for the ex vivo expansion of human limbal stem cells (LSCs) remains the human amniotic membrane (HAM) but this is not a defined substrate and is subject to biological variabil-ity and the potential to transmit disease. To better define HAM and mitigate the risk of disease transmis-sion, we sought to determine if decellularisation and/or c-irradiation have an adverse effect on culture growth and LSC phenotype. Ex vivo limbal explant cultures were set up on fresh HAM, HAM decellularised with 0.5 M NaOH, and 0.5% (w/v) sodium dodecyl sulfate (SDS) with or without c-irradiation. Explant growth rate was measured and LSC phenotype was characterised by histology, immunostaining and qRT-PCR (ABCG2, DNp63, Ki67, CK12, and CK13). Ƴ-irradiation marginally stiffened HAM, as measured by Brillouin spectromicroscopy. HAM stiffness and c-irradiation did not significantly affect the LSC phe-notype, however LSCs expanded significantly faster on Ƴ-irradiated SDS decellularised HAM (p < 0.05) which was also corroborated by the highest expression of Ki67 and putative LSC marker, ABCG2. Colony forming efficiency assays showed a greater yield and proportion of holoclones in cells cultured on Ƴ-irradiated SDS decellularised HAM. Together our data indicate that SDS decellularised HAM may be a more efficacious substrate for the expansion of LSCs and the use of a c-irradiated HAM allows the user to start the manufacturing process with a sterile substrate, potentially making it safer

Immunomodulation of experimental autoimmune uveitis

SIGLEAvailable from British Library Document Supply Centre- DSC:DX174399 / BLDSC - British Library Document Supply CentreGBUnited Kingdo

Corneal Allograft Rejection:Risk Factors, Diagnosis, Prevention, and Treatment

Recent advances in corneal graft technology, including donor tissue retrieval, storage and surgical techniques, have greatly improved the clinical outcome of corneal grafts. Despite these advances, immune mediated corneal graft rejection remains the single most important cause of corneal graft failure. Several host factors have been identified as conferring a "high risk" status to the host. These include: more than two quadrant vascularisation, with associated lymphatics, which augment the afferent and efferent arc of the immune response; herpes simplex keratitis; uveitis; silicone oil keratopathy; previous failed (rejected) grafts; "hot eyes"; young recipient age; and multiple surgical procedures at the time of grafting. Large grafts, by virtue of being closer to the host limbus, with its complement of vessels and antigen-presenting Langerhans cells, also are more susceptible to rejection. The diagnosis of graft rejection is entirely clinical and in its early stages the clinical signs could be subtle. Graft rejection is largely mediated by the major histocompatibility antigens, minor antigens and perhaps blood group ABO antigens and some cornea-specific antigens. Just as rejection is mediated by active immune mediated events, the lack of rejection (tolerance) is also sustained by active immune regulatory mechanisms. The anterior chamber associated immune deviation (ACAID) and probably, conjunctiva associated lymphoid tissue (CALT) induced mucosal tolerance, besides others, play an important role. Although graft rejection can lead to graft failure, most rejections can be readily controlled if appropriate management is commenced at the proper time. Topical steroids are the mainstay of graft rejection management. In the high-risk situations however, systemic steroids, and other immunosuppressive drugs such as cyclosporin and tacrolimus (FK506) are of proven benefit, both for treatment and prevention of rejection

Fine needle diathermy occlusion of corneal vessels

Purpose: To develop a novel technique, fine needle diathermy (FND), for the occlusion of corneal vessels and to evaluate its safety and efficacy in a series of patients. Methods: Fourteen patients were treated with FND to occlude corneal vessels. Patients were categorized into four groups: group 1 (n = 4), high risk patients with stromal vascularization before keratoplasty; group 2 (n = 2), patients with progressive lipid keratopathy; group 3 (n = 4), post keratoplasty patients with active rejection episodes associated with vessels; and group 4 (n = 4), patients with disciform vascularized scars with recurrent inflammation. The success of the treatment in terms of vessel occlusion and the clinical outcome were monitored. Results: All patients in group 1 had successful corneal transplantation, and the grafts remained clear without graft rejection. Patients in group 2 with lipid keratopathy had 100% obliteration of vessels with stabilization of corneal scar. All four patients in group 3 had complete regression of vessels with reversal of graft rejection. Patients with vascularized disciform scar had resolution of the inflammation without recurrence. Average follow-up was 10.3 months (minimum, 6 months; maximum, 24 months). No serious complications were observed with FND. Conclusions: FND is a useful and inexpensive technique that can serve as an adjunct or alternative to laser occlusion for the treatment of established corneal vessels. It is fairly safe and effective, although complications such as intrastromal bleeding and crystalline deposits can occur and at times it may have to be repeated once or twice to achieve the desired result. <br/