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Koulikovska, Marina
Publications (10 of 15) Show all publications
Rafat, M., Xeroudaki, M., Koulikovska, M., Sherrell, P., Groth, F., Fagerholm, P. & Lagali, N. (2016). Composite core-and-skirt collagen hydrogels with differential degradation for corneal therapeutic applications. Biomaterials, 83, 142-155
Open this publication in new window or tab >>Composite core-and-skirt collagen hydrogels with differential degradation for corneal therapeutic applications
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2016 (English)In: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 83, p. 142-155Article in journal (Refereed) Published
Abstract [en]

Scarcity of donor tissue to treat corneal blindness and the need to deliver stem cells or pharmacologic agents to ensure corneal graft survival are major challenges. Here, new composite collagen-based hydrogels are developed as implants to restore corneal transparency while serving as a possible reservoir for cells and drugs. The composite hydrogels have a centrally transparent core and embedded peripheral skirt of adjustable transparency and degradability, with the skirt exhibiting faster degradation in vitro. Both core and skirt supported human epithelial cell populations in vitro and the skirt merged homogeneously with the core material to smoothly distribute a mechanical load in vitro. After in vivo transplantation in rabbit corneas over three months, composites maintained overall corneal shape and integrity, while skirt degradation could be tracked in vivo and non-invasively due to partial opacity. Skirt degradation was associated with partial collagen breakdown, thinning, and migration of host stromal cells and macrophages, while the central core maintained integrity and transparency as host cells migrated and nerves regenerated.

IMPACT:

This study indicates the feasibility of a collagen-based composite hydrogel to maintain corneal stability and transparency while providing a degradable peripheral reservoir for cell or substance release.

Keywords
Composite; Cornea; Degradation; Femtosecond laser; Keratoplasty; Porcine collagen
National Category
Cell and Molecular Biology
Identifiers
urn:nbn:se:liu:diva-125229 (URN)10.1016/j.biomaterials.2016.01.004 (DOI)000371651700012 ()26773670 (PubMedID)
Note

Funding agencies:  Abbott Medical Optics Inc, Solna, Sweden

Available from: 2016-02-16 Created: 2016-02-16 Last updated: 2018-01-22
Koulikovska, M., Rafat, M., Petrovski, G., Veréb, Z., Akhtar, S., Fagerholm, P. & Lagali, N. (2015). Enhanced Regeneration of Corneal Tissue Via a Bioengineered Collagen Construct Implanted by a Nondisruptive Surgical Technique. Tissue Engineering. Part A, 21(5-6), 1116-1130
Open this publication in new window or tab >>Enhanced Regeneration of Corneal Tissue Via a Bioengineered Collagen Construct Implanted by a Nondisruptive Surgical Technique
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2015 (English)In: Tissue Engineering. Part A, ISSN 1937-3341, E-ISSN 1937-335X, Vol. 21, no 5-6, p. 1116-1130Article in journal (Refereed) Published
Abstract [en]

Severe shortage of donor corneas for transplantation, particularly in developing countries, has prompted the advancement of bioengineered tissue alternatives. Bioengineered corneas that can withstand transplantation while maintaining transparency and compatibility with host cells, and that are additionally amenable to standardized low-cost mass production are sought. In this study, a bioengineered porcine construct (BPC) was developed to function as a biodegradable scaffold to promote corneal stromal regeneration by host cells. Using high-purity medical-grade type I collagen, high 18% collagen content and optimized EDC-NHS cross-linker ratio, BPCs were fabricated into hydrogel corneal implants with over 90% transparency and four-fold increase in strength and stiffness compared with previous versions. Remarkably, optical transparency was achieved despite the absence of collagen fibril organization at the nanoscale. In vitro testing indicated that BPC supported confluent human epithelial and stromal-derived mesenchymal stem cell populations. With a novel femtosecond laser-assisted corneal surgical model in rabbits, cell-free BPCs were implanted in vivo in the corneal stroma of 10 rabbits over an 8-week period. In vivo, transparency of implanted corneas was maintained throughout the postoperative period, while healing occurred rapidly without inflammation and without the use of postoperative steroids. BPC implants had a 100% retention rate at 8 weeks, when host stromal cells began to migrate into implants. Direct histochemical evidence of stromal tissue regeneration was observed by means of migrated host cells producing new collagen from within the implants. This study indicates that a cost-effective BPC extracellular matrix equivalent can incorporate cells passively to initiate regenerative healing of the corneal stroma, and is compatible with human stem or organ-specific cells for future therapeutic applications as a stromal replacement for treating blinding disorders of the cornea.

Place, publisher, year, edition, pages
Mary Ann Liebert, 2015
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:liu:diva-114699 (URN)10.1089/ten.tea.2014.0562 (DOI)000350549500025 ()25412075 (PubMedID)
Available from: 2015-03-03 Created: 2015-03-03 Last updated: 2018-01-22Bibliographically approved
Koulikovska, M., Szymanowski, O., Lagali, N. & Fagerholm, P. (2015). Platelet Rich Plasma Prolongs Myofibroblast Accumulation in Corneal Stroma with Incisional Wound. Current Eye Research, 40(11), 1102-1110
Open this publication in new window or tab >>Platelet Rich Plasma Prolongs Myofibroblast Accumulation in Corneal Stroma with Incisional Wound
2015 (English)In: Current Eye Research, ISSN 0271-3683, E-ISSN 1460-2202, Vol. 40, no 11, p. 1102-1110Article in journal (Refereed) Published
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.

Place, publisher, year, edition, pages
Taylor & Francis, 2015
Keywords
Platelet rich plasma, corneal wound healing, α-smooth muscle actin, apoptosis, keratocytes
National Category
Cell and Molecular Biology Medical Biotechnology
Identifiers
urn:nbn:se:liu:diva-114698 (URN)10.3109/02713683.2014.978478 (DOI)000369891500004 ()
Note

Funding agencies:Swedish Research Council, Konung Gustaf V:s och Drottning Victorias Frimurarestiftelse, County Council of Ostergotland 

Vid tiden för disputation förelåg publikationen endast som manuskript

Available from: 2015-03-03 Created: 2015-03-03 Last updated: 2018-01-22Bibliographically approved
Koulikovska, M., Szymanowski, O., Lagali, N. & Fagerholm, P. (2015). Topical Biglycan Modulates Stromal Cell Apoptosis in Corneal Incisional Wound Model.
Open this publication in new window or tab >>Topical Biglycan Modulates Stromal Cell Apoptosis in Corneal Incisional Wound Model
2015 (English)Manuscript (preprint) (Other academic)
Abstract [en]

Purpose: The purpose of this study was to determine whether exogenous topicallyapplied biglycan has an effect on corneal stromal cells during wound healing.

Methods: Enzyme-linked immunosorbent assay (ELISA) was used to determine the effect of biglycan on cell survival in vitro following IL-1β induced cell death. In a corneal incisional wound model, 40 rats were wounded and treated with either topically administered biglycan or sodium chloride (sham control). At 4 hours and 1, 2, and 5 days after incision, α-smooth muscle actin (SMA) expression and apoptosis in stromal cells were evaluated by immunohistochemistry.

Results: In vitro, biglycan significantly enhanced IL-1β-induced apoptosis of myofibroblasts (p = 0.038), but not corneal fibroblasts. Biglycan treated corneas exhibited reduced stromal cell apoptosis at 4 hours, day 1 and day 5 (p = 0.012, 0.040, and 0.048, respectively) and increased apoptosis at day 3 (p = 0.003) relative to controls. In wounded corneas, biglycan appeared to promote early accumulation of myofibroblasts and initiate an earlier subsequent apoptosis of these cells, relative to controls.

Conclusion: Biglycan appears to accelerate corneal wound healing in vivo by modulating myofibroblast apoptosis, resulting in removal of myofibroblasts that may otherwise compromise corneal transparency.

Keywords
Corneal wound healing; biglycan; keratocytes; IL-1β; α-SMA
National Category
Cell and Molecular Biology Medical Biotechnology
Identifiers
urn:nbn:se:liu:diva-114697 (URN)
Available from: 2015-03-03 Created: 2015-03-03 Last updated: 2018-01-22
Mirabelli, P., Bourghardt Peebo, B., Xeroudaki, M., Koulikovska, M. & Lagali, N. (2014). Early effects of dexamethasone and anti-VEGF therapy in an inflammatory corneal neovascularization model. Experimental Eye Research, 125, 118-127
Open this publication in new window or tab >>Early effects of dexamethasone and anti-VEGF therapy in an inflammatory corneal neovascularization model
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2014 (English)In: Experimental Eye Research, ISSN 0014-4835, E-ISSN 1096-0007, Vol. 125, p. 118-127Article in journal (Refereed) Published
Abstract [en]

Inflammatory angiogenesis is the pathogenic mechanism of various sight-threatening eye diseases, among them corneal neovascularization. Current treatment options include steroids which have undesirable side effects, or anti-VEGF which has only limited efficacy. In an inflammatory environment, however, angiogenesis can be stimulated by numerous factors not directly targeted by anti-VEGF therapy. The aim of this study was to induce corneal inflammation leading to angiogenesis, and investigate the early, differential effects of steroid and anti-VEGF therapy at the cellular, tissue, and gene expression levels. Fifty-two Wistar rats received a single intrastromal corneal suture to induce a controlled inflammatory angiogenic response. Rats were subsequently treated with dexamethasone, rat specific anti-VEGF, or goat IgG (control), topically 4 times daily for 7 days. In vivo confocal microscopy of the cornea was performed longitudinally from 5 h up to 7 d to investigate morphology at the cellular and tissue-level. In vivo photographic vessel analysis and immunohistochemistry were also performed. RT-PCR for VEGF-A, FGF-2, IL-6, TNF-alpha, CXCL2, CCL2, CCL3 and DLL4 was performed at 24 h, and for VEGF-A, IL-6, TNF-alpha, FGF-2, CXCL2, CCL2, and CCL3 at 7 days. Early infiltration of CD11b + myeloid cells into the cornea at 5 h post-suture was delayed by both treatments relative to controls; however neither treatment was able to suppress accumulation of myeloid cells at day 2 or 7. Limbal vessel dilation was inhibited at 5 h by both treatments, but only dexamethasone showed sustained effect until day 2. Early macrophage recruitment was also suppressed by dexamethasone (but not by anti-VEGF) until day 2. Dexamethasone furthermore suppressed corneal neovascularization at day 7 by over 90%, whereas suppression by anti-VEGF was 14%. Despite differential suppression of vessel dilation, macrophage recruitment, and vascular invasion, anti-VEGF and dexamethasone both down-regulated VEGF-A and IL-6 expression at 24 h with sustained effect to 7 d. They also both down regulated FGF-2 and TNF-alpha at 24 h and CCL2 at 7 d. In conclusion, anti-angiogenic treatments influence early, pre-angiogenic tissue activity such as limbal vessel dilation, inflammatory cell infiltration of the stroma, and macrophage recruitment. Importantly, the differential effects of steroids and anti-VEGF treatment in suppressing neovascular growth could not be attributed to differential inhibition of several major angiogenic and inflammatory factors in the early pre-sprouting phase, including IL-6, VEGF-A, FGF-2, TNF-alpha, CCL2, CCL3, CXCL2, or DLL4.

Place, publisher, year, edition, pages
Elsevier, 2014
Keywords
angiogenesis; neovascularization; cornea; inflammation; dexamethasone; anti-VEGF; confocal microscopy; rat
National Category
Clinical Medicine
Identifiers
urn:nbn:se:liu:diva-110279 (URN)10.1016/j.exer.2014.06.006 (DOI)000340079300014 ()24933712 (PubMedID)
Note

Funding Agencies|Crown Princess Margaretas Foundation; County Council of Ostergotland; Swedish Research Council [2012-2472]

Available from: 2014-09-05 Created: 2014-09-05 Last updated: 2019-04-18Bibliographically approved
Rafat, M., Lagali, N., Koulikovska, M. & Fagerholm, P. (2013). Development of a Highly Elastic Bioengineered Cornea: From Research to Commercialization. In: : . Paper presented at 2nd Biotechnology World Conference, February 18-21 2013, Dubai, UAE.
Open this publication in new window or tab >>Development of a Highly Elastic Bioengineered Cornea: From Research to Commercialization
2013 (English)Conference paper, Oral presentation with published abstract (Other academic)
Abstract [en]

Background: Despite the promising clinical results that we previously reported on biosynthetic corneas, more elastic materials are required for surgical manipulation and withstanding the adverse host conditions faced by high risk corneal transplants.

Purpose: The overall objective was to develop novel bioengineered materials that can replace the damaged corneal tissue. Another objective was to evaluate the in vivo integration of the materials in rabbit models using a femtosecond laser intrastromal surgical technique.

Methods: Bioengineered corneas were prepared using porcine collagen cross-linked by carbodiimides at various compositions and pH. Promising formulations were tested for their mechanical, optical, and enzymatic and thermal degradation properties as well as for interactions with corneal cells, and in vivo implantation in rabbit’s eyes. A femtosecond laser was used to cut 100 mircon thick discs of mid-stromal tissue from corneas of 15 rabbits and replaced with the bioengineered materials.

Results: The new material demonstrated improved mechanical properties while maintaining its clarity and biocompatibility. The bioengineered implant retained its shape, thickness, and clarity 8 weeks post-surgery in rabbits.  

Conclusions: The bioengineered cornea developed in this work has the potential to be used and commercialized as corneal implants to replace the damaged tissue or for corrective surgery applications.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-101524 (URN)
Conference
2nd Biotechnology World Conference, February 18-21 2013, Dubai, UAE
Available from: 2013-11-22 Created: 2013-11-22 Last updated: 2018-01-22Bibliographically approved
Rafat, M., Lagali, N., Koulikovska, M., Griffith, M. & Fagerholm, P. (2013). In vivo integrity of intra‐corneal bioengineered discs in rabbit models. In: Acta Ophthalmologica; Special Issue: Abstracts from the 2013 European Association for Vision and Eye Research Conference, August 2013 Volume 91, Issue Supplement s252: . Paper presented at European Association for Vision and Eye Research (EVER-2013), 18-21 September, Nice, France. John Wiley & Sons
Open this publication in new window or tab >>In vivo integrity of intra‐corneal bioengineered discs in rabbit models
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2013 (English)In: Acta Ophthalmologica; Special Issue: Abstracts from the 2013 European Association for Vision and Eye Research Conference, August 2013 Volume 91, Issue Supplement s252, John Wiley & Sons, 2013Conference paper, Oral presentation with published abstract (Other academic)
Abstract [en]

Background: We have previously reported the successful integration and safety of bioengineered materials as corneal substitutes in human models. Despite the promising results as corneal implants, more elastic and robust materials are required for use as thin intra-corneal lenses to withstand surgical manipulation for corrective surgery and improved vision. Most of the existing corneal inlays are made of synthetic materials. Here we describe the potential of bioengineerd materials for vision correction. Objectives: to develop bioengineered materials as inlays within the corneal tissue as well as evaluating the in vivo integrity and integration of the materials in rabbit models. Methods: Bioengineered inlays were prepared from collagen and tested for their physical and biological propertis. A femtosecond laser was used to cut 100 mircon thick discs of mid-stromal tissue from corneas of 20 rabbits and replaced with bioengineered inlays. Results: The new materials demonstrated improved mechanical properties while maintaining their clarity and biocompatibility. The bioengineered inlays retained their shapes, thickness, and clarity 8 weeks post-surgery in rabbits.

Place, publisher, year, edition, pages
John Wiley & Sons, 2013
National Category
Medical and Health Sciences Health Sciences
Identifiers
urn:nbn:se:liu:diva-101516 (URN)10.1111/j.1755-3768.2013.4733.x (DOI)
Conference
European Association for Vision and Eye Research (EVER-2013), 18-21 September, Nice, France
Available from: 2013-11-22 Created: 2013-11-22 Last updated: 2018-01-22Bibliographically approved
Germundsson, J., Fagerholm, P., Koulikovska, M. & Lagali, N. (2012). An Accurate Method to Determine Bowmans Layer Thickness In Vivo in the Human Cornea. Investigative Ophthalmology and Visual Science, 53(4), 2354-2359
Open this publication in new window or tab >>An Accurate Method to Determine Bowmans Layer Thickness In Vivo in the Human Cornea
2012 (English)In: Investigative Ophthalmology and Visual Science, ISSN 0146-0404, E-ISSN 1552-5783, Vol. 53, no 4, p. 2354-2359Article in journal (Refereed) Published
Abstract [en]

PURPOSE. To determine an accurate value for Bowmans layer (BL) thickness in vivo in humans. less thanbrgreater than less thanbrgreater thanMETHODS. Seventeen corneal transplant patients were examined preoperatively by laser-scanning in vivo confocal microscopy (IVCM), and corneal buttons were removed post-operatively and sectioned for light microscopy (LM). Nine corneas with uniformly thick BL by LM were used for thickness measurement. In the uniformly thick samples, probable overestimation of BL thickness in vivo by a first in vivo method (Method 1) led to the development of a revised in vivo method (Method 2). Method 2 was used to measure BL thickness in 20 healthy volunteers. less thanbrgreater than less thanbrgreater thanRESULTS. In nine patients, mean BL thickness prior to transplantation was 13.7 +/- 1.6 mu m by IVCM (Method 1) while BL thickness of the removed corneal button was 9.7 +/- 1.7 mu m by LM (P andlt; 0.001). The correlation of BL thickness between IVCM (Method 1) and LM was poor (P = 0.226). In 20 right eyes of 20 normal corneas, both in vivo methods were used to determine BL thickness. Mean BL thickness by Method 1 was 13.2 +/- 1.6 mu m and by Method 2 was 9.1 +/- 1.4 mu m (P andlt; 0.001). BL thickness measurements by both in vivo methods were highly correlated (P andlt; 0.001). less thanbrgreater than less thanbrgreater thanCONCLUSION. BL thickness by a revised in vivo method was close to LM values in this study and to values reported in fixed tissue in other studies. The authors believe this revised method provides the most accurate estimates of BL thickness in vivo to date.

Place, publisher, year, edition, pages
Association for Research in Vision and Ophthalmology (ARVO), 2012
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-78278 (URN)10.1167/iovs.11-8565 (DOI)000303669400076 ()
Note

Funding Agencies|European Union||

Available from: 2012-06-08 Created: 2012-06-08 Last updated: 2018-01-22
Koulikovska, M., Sharipo, A. & Fagerholm, P. (2007). Interleukin1-Induced apoptosis of keratocytes: effect of biglycan. In: European Association for Vision and Eye Research,2007.
Open this publication in new window or tab >>Interleukin1-Induced apoptosis of keratocytes: effect of biglycan
2007 (English)In: European Association for Vision and Eye Research,2007, 2007Conference paper, Published paper (Other academic)
Abstract [en]

  

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-39231 (URN)47546 (Local ID)47546 (Archive number)47546 (OAI)
Available from: 2009-10-10 Created: 2009-10-10
Bourghardt Peebo, B., Koulikovska, M. & Fagerholm, P. (2007). The suppression of early angiogenic markers by the antiangiogenic aptamer Macugen R is dose dependent. In: European Association for Vision and Eye Research,2007.
Open this publication in new window or tab >>The suppression of early angiogenic markers by the antiangiogenic aptamer Macugen R is dose dependent
2007 (English)In: European Association for Vision and Eye Research,2007, 2007Conference paper, Published paper (Other academic)
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-39234 (URN)47588 (Local ID)47588 (Archive number)47588 (OAI)
Available from: 2009-10-10 Created: 2009-10-10
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