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  • 1.
    Koulikovska, Marina
    et al.
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för neuro- och inflammationsvetenskap. Linköpings universitet, Medicinska fakulteten. Region Östergötland, Sinnescentrum, Ögonkliniken US/LiM.
    Szymanowski, Olena
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för neuro- och inflammationsvetenskap. Linköpings universitet, Medicinska fakulteten. Region Östergötland, Sinnescentrum, Ögonkliniken US/LiM.
    Lagali, Neil
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för neuro- och inflammationsvetenskap. Linköpings universitet, Medicinska fakulteten. Region Östergötland, Sinnescentrum, Ögonkliniken US/LiM.
    Fagerholm, Per
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för neuro- och inflammationsvetenskap. Linköpings universitet, Medicinska fakulteten. Region Östergötland, Sinnescentrum, Ögonkliniken US/LiM.
    Platelet Rich Plasma Prolongs Myofibroblast Accumulation in Corneal Stroma with Incisional Wound2015Ingår i: Current Eye Research, ISSN 0271-3683, E-ISSN 1460-2202, Vol. 40, nr 11, s. 1102-1110Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose: The purpose of this study was to determine whether platelet rich plasma (PRP) has an effect on corneal stromal cells in a rat model of wound healing following corneal incision. Materials and Methods: The effect of PRP on corneal wound healing in vivo was investigated in a corneal incision wound model in rats. 40 rats were wounded by deep corneal incision, and treated with either topically administered PRP (20 rats) or sodium chloride (20 rats). At 4 hours and 1, 3, and 5 days after incision, α-smooth muscle actin (α SMA), SMAD2 and SMAD3 expression and apoptosis in stromal cells were evaluated by immunohistochemistry, and IL-1β mRNA expression was evaluated by real time PCR.

    Results: PRP treated corneas exhibited reduced stromal cell apoptosis at day 3 and day 5 (p = 0.038, and <0.001, respectively) relative to controls. Interleukin-1β mRNA expression, however, was unchanged in PRP treated corneas relative to controls. Topical PRP treatment resulted in a higher proportion of αSMA-positive myofibroblasts recruited to the wound site relative to control corneas. PRP did not affect activation of SMAD2 but activation of SMAD3 was significantly reduced at day 1 (p=0.001) and dramatically increased at day 5 (p=0.032).

    Conclusions: PRP treatment resulted in suppressed stromal cell apoptosis followed by SMAD3 activation and a greater proportion of myofibroblasts present at the wound site. Suppression of stromal cell apoptosis after corneal wounding by use of a growth factor rich formulation may lead to myofibroblast accumulation by modulation of the TGF-β pathway.

  • 2.
    Lagali, Neil
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för neuro- och inflammationsvetenskap. Linköpings universitet, Medicinska fakulteten. Region Östergötland, Sinnescentrum, Ögonkliniken US/LiM. Sorlandet Hosp Arendal, Norway.
    Corneal Stromal Regeneration: Current Status and Future Therapeutic Potential2019Ingår i: Current Eye Research, ISSN 0271-3683, E-ISSN 1460-2202Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The corneal stroma comprises 90% of the corneal thickness and is critical for the corneas transparency and refractive function necessary for vision. When the corneal stroma is altered by disease, injury, or scarring, however, an irreversible loss of transparency can occur. Corneal stromal pathology is the cause of millions of cases of blindness globally, and although corneal transplantation is the standard therapy, a severe global deficit of donor corneal tissue and eye banking infrastructure exists, and is unable to meet the overwhelming need. An alternative approach is to harness the endogenous regenerative ability of the corneal stroma, which exhibits self-renewal of the collagenous extracellular matrix under appropriate conditions. To mimic endogenous stromal regeneration, however, is a challenge. Unlike the corneal epithelium and endothelium, the corneal stroma is an exquisitely organized extracellular matrix containing stromal cells, proteoglycans and corneal nerves that is difficult to recapitulate in vitro. Nevertheless, much progress has recently been made in developing stromal equivalents, and in this review the most recent approaches to stromal regeneration therapy are described and discussed. Novel approaches for stromal regeneration include human or animal corneal and/or non-corneal tissue that is acellular or is decellularized and/or re-cellularized, acellular bioengineered stromal scaffolds, tissue adhesives, 3D bioprinting and stromal stem cell therapy. This review highlights the techniques and advances that have achieved first clinical use or are close to translation for eventual therapeutic application in repairing and regenerating the corneal stroma, while the potential of these novel therapies for achieving effective stromal regeneration is discussed.

  • 3.
    Spinozzi, Daniele
    et al.
    Netherlands Inst Innovat Ocular Surg, Netherlands.
    Miron, Alina
    Netherlands Inst Innovat Ocular Surg, Netherlands.
    Bruinsma, Marieke
    Netherlands Inst Innovat Ocular Surg, Netherlands.
    Dapena, Isabel
    Netherlands Inst Innovat Ocular Surg, Netherlands; Melles Cornea Clin Rotterdam, Netherlands.
    Lavy, Itay
    Netherlands Inst Innovat Ocular Surg, Netherlands; Melles Cornea Clin Rotterdam, Netherlands.
    Binder, Perry S.
    Univ Calif Irvine, CA USA.
    Rafat, Mehrdad
    Linköpings universitet, Institutionen för medicinsk teknik, Avdelningen för medicinsk teknik. Linköpings universitet, Tekniska fakulteten. LinkoCare Life Sci AB, Linkoping, Sweden.
    Oellerich, Silke
    Netherlands Inst Innovat Ocular Surg, Netherlands.
    Melles, Gerrit R. J.
    Netherlands Inst Innovat Ocular Surg, Netherlands; Melles Cornea Clin Rotterdam, Netherlands; Amnitrans EyeBank Rotterdam, Netherlands.
    Evaluation of the Suitability of Biocompatible Carriers as Artificial Transplants Using Cultured Porcine Corneal Endothelial Cells2019Ingår i: Current Eye Research, ISSN 0271-3683, E-ISSN 1460-2202, Vol. 44, nr 3, s. 243-249Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose/Aim: Evaluating the suitability of bioengineered collagen sheets and human anterior lens capsules (HALCs) as carriers for cultivated porcine corneal endothelial cells (pCECs) and in vitro assessment of the cell-carrier sheets as tissue-engineered grafts for Descemet membrane endothelial keratoplasty (DMEK). Materials and Methods: pCECs were isolated, cultured up to P2 and seeded onto LinkCell (TM) bioengineered matrices of 20 mu m (LK20) or 100 mu m (LK100) thickness, and on HALC. During expansion, pCEC viability and morphology were assessed by light microscopy. ZO-1 and Na+/K+-ATPase expression was investigated by immunohistochemistry. Biomechanical properties of pCEC-carrier constructs were evaluated by simulating DMEK surgery in vitro using an artificial anterior chamber (AC) and a human donor cornea without Descemet membrane (DM). Results: During in vitro expansion, cultured pCECs retained their proliferative capacity, as shown by the positive staining for proliferative marker Ki67, and a high cell viability rate (96 +/- 5%). pCECs seeded on all carriers formed a monolayer of hexagonal, tightly packed cells that expressed ZO-1 and Na+/K+-ATPase. During in vitro surgery, pCEC-LK20 and pCEC-LK100 constructs were handled like Descemet stripping endothelial keratoplasty (DSEK) grafts, i.e. folded like a "taco" for insertion because of challenges related to rolling and sticking of the grafts in the injector. pCEC-HALC constructs behaved similar to the DMEK reference model during implantation and unfolding in the artificial AC, showing good adhesion to the bare stroma. Conclusions: In vitro DMEK surgery showed HALC as the most suitable carrier for cultivated pCECs with good intraoperative graft handling. LK20 carrier showed good biocompatibility, but required a DSEK-adapted surgical protocol. Both carriers might be notional candidates for potential future clinical applications.

  • 4.
    Sundelin, Staffan
    et al.
    Linköpings universitet, Institutionen för nervsystem och rörelseorgan, Oftalmologi. Linköpings universitet, Hälsouniversitetet.
    Wihlmark, Ulf
    Linköpings universitet, Institutionen för nervsystem och rörelseorgan, Oftalmologi. Linköpings universitet, Hälsouniversitetet.
    Nilsson, Sven Erik G.
    Linköpings universitet, Institutionen för nervsystem och rörelseorgan, Oftalmologi. Linköpings universitet, Hälsouniversitetet.
    Brunk, Ulf T.
    Linköpings universitet, Institutionen för nervsystem och rörelseorgan, Patologi. Linköpings universitet, Hälsouniversitetet.
    Lipofuscin accumulation in cultured retinal pigment epithelial cells reduces their phagocytic capacity1998Ingår i: Current Eye Research, ISSN 0271-3683, E-ISSN 1460-2202, Vol. 17, nr 8, s. 851-857Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose. Retinal pigment epithelial (RPE) cells slowly accumulate lipofuscin pigment within their acidic vacuolar apparatus as a result of extra- and/or intralysosomal oxidative alterations of phagocytosed photoreceptor outer segments (POS) with consequent imperfect degradation of these structures. In old age, lipofuscin accumulation may become quite substantial. It has been suggested that pronounced accumulation of lipofuscin is related to decreased RPE function and, possibly, to age-related macular degeneration. The aim of the present investigation was to study whether heavy loading with lipofuscin of RPE acidic lysosomes would affect the further phagocytic ability of the cells.

    Methods. In the first section of the investigation, cultures of rabbit RPE cells were exposed daily to bovine UV-irradiated POS (artificial lipofuscin) for 4 weeks, resulting in a pronounced lipofuscin accumulation of the cells. Fluorescent latex beads (labelled with a red fluorophore) were added to unloaded control cultures at 0 and 4 weeks after their establishment, and to lipofuscin loaded cells after 4 weeks of feeding with artificial lipofuscin. Cellular amounts of lipofuscin, and their phagocytotic activity, were quantified by static fluorometry measuring lipofuscin-specific and red bead-specific fluorescence, respectively. The intracellular location of the beads was verified by confocal laser scanning microscopy.

    Results. Unloaded, and thus almost lipofuscin-free, control cells exposed to latex beads showed numerous cytoplasmic particles emitting reddish fluorescence, while few particles were taken up by cells initially loaded with artificial, POS-derived, lipofuscin. Measurement of the latex bead-specific fluorescence showed significantly (p < 0.001) higher levels in unloaded control cells than in lipofuscin-loaded ones.

    In the second part of the investigation, unloaded control cultures and lipofuscin-loaded cultures were exposed to native bovine Texas Red-X-labelled POS 4 weeks after the establishment of the cultures. Unloaded control cells showed a large number of cytopiasmic POS emitting reddish fluorescence, while fewer POS were phagocytosed by cells loaded with artificial lipofuscin. Measurement of the Texas Red-X-specific fluorescence, thus quantifying the phagocytic ability of the cells, showed significantly (p < 0.001) higher levels in control cells than in lipofuscin-loaded ones.

    Conclusions. Severe lipofuscin accumulation of RPE cells appears to result in a greatly decreased phagocytic capacity. The resulting reduction in ability to cope with the needs of the overlying photoreceptor cells, in order to eliminate the obsolete tips of their POS, may well be of significance in the development of age-related macular degeneration.

  • 5.
    Utheim, Tor Paaske
    et al.
    Oslo Univ Hosp, Norway; Sorlandet Hosp Arendal, Norway.
    Chen, Xiangjun
    Sorlandet Hosp Arendal, Norway.
    Fricke, Otto
    Linköpings universitet, Institutionen för biomedicinska och kliniska vetenskaper. Linköpings universitet, Medicinska fakulteten.
    Bergersen, Linda Hildegard
    Univ Oslo, Norway.
    Lagali, Neil
    Linköpings universitet, Institutionen för biomedicinska och kliniska vetenskaper, Avdelningen för logopedi, otorhinolaryngologi och audiologi. Linköpings universitet, Medicinska fakulteten. Region Östergötland, Sinnescentrum, Ögonkliniken US/LiM. Sorlandet Hosp Arendal, Norway.
    Microdot Accumulation in the Anterior Cornea with Aging - Quantitative Analysis with in Vivo Confocal Microscopy2020Ingår i: Current Eye Research, ISSN 0271-3683, E-ISSN 1460-2202Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose: Degenerative microdot deposits in healthy and hypoxic corneas are believed to represent lipofuscin-like material aggregation in the stroma. To accurately assess microdot deposits in a clinical setting, we sought to quantify these deposits for the first time using the non-invasive clinical imaging technique of in vivo confocal microscopy (IVCM). Methods: The corneas of 102 healthy subjects aged 15-88 years were examined by IVCM and microdot density was quantified using a 6-point grading scale by two masked, trained examiners. Microdot density was analyzed with respect to age, sex and stromal depth, and inter-eye and inter-observer differences were evaluated. Results: In healthy subjects, microdot density decreased from the anterior to posterior stroma, with the greatest accumulation observed in the most anterior stroma (subepithelial region). In this region, microdot density correlated strongly with age (P amp;lt; .0001), with increased microdot deposition in older subjects (amp;gt;60 years) relative to younger ones (amp;lt;45 years) (P amp;lt; .001). Microdot density between eyes of the same subject was highly correlated (r = 0.92, P amp;lt; .0001), while no association with sex was noted (P amp;gt;= 0.05). The mean inter-observer difference in microdot assessment was 0.62 +/- 0.09 grades, with a high correlation of grading between observers (r = 0.77, P amp;lt; .0001). Conclusions: IVCM can be used to non-invasively quantify microdot deposits in the subepithelial corneal stroma with good inter-observer reproducibility. Microdot assessment may provide a novel means of quantifying age-related or pathologic degeneration of the corneal stroma in a clinical setting.

  • 6.
    Yazdani, Mazyar
    et al.
    Oslo Univ Hosp, Norway; Norwegian Dry Eye Clin, Norway.
    Chen, Xiangjun
    Norwegian Dry Eye Clin, Norway; Arendal Hosp, Norway; Univ Oslo, Norway; Univ Coll Southeast Norway, Norway.
    Tashbayev, Behzod
    Norwegian Dry Eye Clin, Norway; Univ Oslo, Norway.
    Utheim, Oygunn A.
    Oslo Univ Hosp, Norway; Norwegian Dry Eye Clin, Norway.
    Raeder, Sten
    Norwegian Dry Eye Clin, Norway.
    Lagali, Neil
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för neuro- och inflammationsvetenskap. Linköpings universitet, Medicinska fakulteten. Region Östergötland, Sinnescentrum, Ögonkliniken US/LiM.
    Stojanovic, Aleksandar
    Univ Hosp North Norway, Norway.
    Dartt, Darlene A.
    Harvard Med Sch, MA USA.
    Utheim, Tor P.
    Oslo Univ Hosp, Norway; Norwegian Dry Eye Clin, Norway; Arendal Hosp, Norway; Univ Oslo, Norway; Univ Coll Southeast Norway, Norway; Oslo Univ Hosp, Norway; Oslo Univ Hosp, Norway; Stavanger Univ Hosp, Norway; Univ Bergen, Norway.
    Tear Production Levels and Dry Eye Disease Severity in a Large Norwegian Cohort2018Ingår i: Current Eye Research, ISSN 0271-3683, E-ISSN 1460-2202, Vol. 43, nr 12, s. 1465-1470Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose: To determine if the Schirmer I test (without anesthesia) cut-off value is a predictor of dry eye severity in a large Norwegian cohort of dry eye disease (DED) patients, which are grouped into six levels of tear production. Methods: Patients (n = 1090) with DED of different etiologies received an extensive dry eye work-up: osmolarity (Osm), tear meniscus height (TMH), tear film break-up time (TFBUT), ocular protection index (OPI), ocular surface staining (OSS), Schirmer I test (ST), meibum expressibility (ME), and meibum quality (MQ). Classification of dry eye severity level (DESL) and diagnosis of meibomian gland dysfunction (MGD) were also included. The cohort was divided into six groups: below and above cut-off values of 5 (groups 1 and 2), 10 (groups 3 and 4), and 15 mm (groups 5 and 6) of ST. Mann-Whitney test and Chi-Square test were used for group comparison of parameters (p amp;lt;= 0.05). Results: The groups 1, 3, and 5 had values indicating more severe DED than the groups 2, 4, 6 with significant difference in DESL, Osm, TFBUT, OPI, OSS, and TMH. Regardless of the choice of cut-off values, there was no statistically significant difference in ME, MQ, and MGD between groups below and above selected cut-off value. When gender difference was considered in each group, significant difference was only observed for DESL (groups 2, 4, and 5), TFBUT (groups 2, 4, and 5), OPI (groups 2 and 6), and ME (group1). Conclusions: Schirmer I is a robust discriminator for DESL, Osm, TFBUT, OPI, OSS, and TMH, but not for ME, MQ, and MGD. Patients with lower tear production levels presented with more severe DED at all three defined cut-off values. Interestingly, the differences in the mean values of DESL were minimal although statistically significant. Thus, the clinical value of different Schirmer levels appears to be limited.

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