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The expression pattern of the subunit of chaperonin containing T-complex polypeptide 1 and its substrate, α-smooth muscle actin, during corneal wound healing
Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Ophthalmology. Östergötlands Läns Landsting, Reconstruction Centre, Department of Ophthalmology UHL.
Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Ophthalmology. Östergötlands Läns Landsting, Reconstruction Centre, Department of Ophthalmology UHL.
Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Ophthalmology. Östergötlands Läns Landsting, Reconstruction Centre, Department of Ophthalmology UHL.
2005 (English)In: Acta Ophthalmologica Scandinavica, ISSN 1395-3907, E-ISSN 1600-0420, Vol. 83, no 5, 543-548 p.Article in journal (Refereed) Published
Abstract [en]

Purpose: This study was designed to demonstrate the expression of the chaperonin containing T-complex polypeptide 1 (CCT) and α-smooth muscle actin (α-SMA), in normal corneas and corneas treated with ultraviolet radiation (UVR). The wound model chosen is previously characterized, the injury is mild and the cornea heals to transparency. Methods: Rabbit corneas were exposed to UVR at the dose producing keratitis. The corneas were allowed to heal for up to 5 days and the paraffin-embedded tissue specimens were double stained and examined morphologically and immunohistochemically. Expression of CCT and α-SMA genes was investigated by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Results: There was a front of repopulating keratocytes that showed positive staining for α-SMA after 3 days. The α-SMA mRNA was already strongly expressed after 1 day, whereas the expression level of CCT was increased after 2 days. After 5 days the levels were decreased. By this time the stroma was partly repopulated by keratocytes. Conclusion: In a mild wound, the expression of α-SMA mRNA is followed by expression of mRNA of at least one subunit of the complex folding α-SMA. At protein level, α-SMA is detected in the front line of repopulating keratocytes. Expression levels for both mRNAs decline as the stroma repopulation process progresses. Copyright © Acta Ophthalmol Scand 2005.

Place, publisher, year, edition, pages
2005. Vol. 83, no 5, 543-548 p.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-29987DOI: 10.1111/j.1600-0420.2005.00482.xLocal ID: 15425OAI: oai:DiVA.org:liu-29987DiVA: diva2:250806
Available from: 2009-10-09 Created: 2009-10-09 Last updated: 2017-12-13
In thesis
1. Corneal stromal cell responses to traumatic wounds and topical treatments
Open this publication in new window or tab >>Corneal stromal cell responses to traumatic wounds and topical treatments
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Background. The cornea has unique anatomic, cellular, molecular, and functional features that lead to important mechanistic differences in the process of repair in comparison with what occurs in skin and other organs. The first observable stromal response in corneal wound healing is keratocyte apoptosis. Shortly thereafter, remaining keratocytes in adjacent areas obtain a fibroblastic phenotype and begin to proliferate and to migrate, transforming into myofibroblasts, a phenotype associated with remodeling of stromal collagen. Return to normalcy following wound healing includes elimination of myofibroblasts and restoration of the quiescent state of the keratocytes. Often, however, a wound healing response results in the persistence of myofibroblasts and their subsequent production of fibrous scar tissue.

Aims. The overall aim is to understand the role of keratocytes, and their phenotypic variations in a cornea subjected to various types of trauma or treatments. More specific aims are to define expression pattern of alpha-smooth muscle actin (α-SMA) and chaperonin containing T-complex polypeptide 1 (CCT) in ultraviolet radiation wound model, to evaluate the effect of biglycan and platelet rich plasma (PRP) treatment during wound healing after corneal incision, and to characterize the structure of the bioengineered porcine construct and its interaction with stromal cells after implantation.

Methods. CCT and α-SMA expression level was evaluated by reverse transcription polymerase chain reaction (RT-PCR) in rabbit corneas subjected to ultraviolet radiation (UVR). Effect of biglycan and PRP on keratocyte phenotype and survival was evaluated by immunohistochemistry, and real time PCR using rat corneas after  incisional wounding. Bioengineered porcine construct (BPC) was implanted into rabbit corneas using femtosecond laser-enabled intrastromal keratoplasty (FLISK) and characterized by means of immunohistochemistry, electron microscopy, and in vivo confocal microscopy (IVCM).

Results and conclusions. In a mild wound, the expression of α-SMA mRNA is followed by expression of mRNA of at least one subunit of the complex folding α-SMA. At protein level, α-SMA is detected in the front line of repopulating keratocytes. Expression levels for both mRNAs decline as the stroma repopulation process progresses.

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

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.

A cost-effective BPC extracellular matrix equivalent can incorporate cells passively to initiate normal regenerative healing of the corneal stroma.

Taken together, results present an interesting possibility to combine BPC implantation and topical biglycan treatment to improve surgical outcome in future studies.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press, 2015. 88 p.
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 1451
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:liu:diva-114700 (URN)10.3384/diss.diva-114700 (DOI)978-91-7519-111-9 (ISBN)
Public defence
2015-03-27, Nils Holgersalen, Campus US, Linköping, 13:00 (English)
Opponent
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Available from: 2015-03-03 Created: 2015-03-03 Last updated: 2016-02-10Bibliographically approved

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Koulikovska, MarinaPodskochy, AlexanderFagerholm, Per

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