Iontophoresis Transcorneal Delivery Technique for Transepithelial Corneal Collagen Crosslinking With Riboflavin in a Rabbit Model

We compared an iontophoresis riboflavin delivery technique for transepithelial corneal collagen crosslinking (I-CXL) with a conventional CXL (C-CXL). We designed three experimental sets using 152 New Zealand rabbits to study riboflavin application by iontophoresis using charged riboflavin solution (...

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Vydáno v:Investigative ophthalmology & visual science Ročník 57; číslo 2; s. 594 - 603
Hlavní autoři: Cassagne, Myriam, Laurent, Camille, Rodrigues, Magda, Galinier, Anne, Spoerl, Eberhard, Galiacy, Stéphane D., Soler, Vincent, Fournié, Pierre, Malecaze, François
Médium: Journal Article
Jazyk:angličtina
Vydáno: United States Association for Research in Vision and Ophthalmology 01.02.2016
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ISSN:1552-5783, 0146-0404, 1552-5783
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Abstract We compared an iontophoresis riboflavin delivery technique for transepithelial corneal collagen crosslinking (I-CXL) with a conventional CXL (C-CXL). We designed three experimental sets using 152 New Zealand rabbits to study riboflavin application by iontophoresis using charged riboflavin solution (Ricrolin+) with a 1-mA current for 5 minutes. The first set was to compare riboflavin concentration measured by HPLC in corneas after iontophoresis or conventional riboflavin application. The second set was to analyze autofluorescence and stromal collagen modification immediately and 14 days after I-CXL or C-CXL, by using nonlinear two-photon microscopy (TP) and second harmonic generation (SHG). In the third set, physical modifications after I-CXL and C-CXL were evaluated by stress-strain measurements and by studying corneal resistance against collagenase digestion. Based on HPLC analysis, we found that iontophoresis allowed riboflavin diffusion with 2-fold less riboflavin concentration than conventional application (936.2 ± 312.5 and 1708 ± 908.3 ng/mL, respectively, P < 0.05). Corneal TP and SHG imaging revealed that I-CXL and C-CXL resulted in a comparable increased anterior and median stromal autofluorescence and collagen packing. The stress at 10% strain showed a similar stiffness of corneas treated by I-CXL or C-CXL (631.9 ± 241.5 and 680.3 ± 216.4 kPa, respectively, P = 0.908). Moreover, we observed an increased resistance against corneal collagenase digestion after I-CXL and C-CXL (61.90% ± 5.28% and 72.21% ± 4.32% of remaining surface, respectively, P = 0.154). This experimental study suggests that I-CXL is a promising alternative methodology for riboflavin delivery in crosslinking treatments, preserving the epithelium.
AbstractList We compared an iontophoresis riboflavin delivery technique for transepithelial corneal collagen crosslinking (I-CXL) with a conventional CXL (C-CXL).PURPOSEWe compared an iontophoresis riboflavin delivery technique for transepithelial corneal collagen crosslinking (I-CXL) with a conventional CXL (C-CXL).We designed three experimental sets using 152 New Zealand rabbits to study riboflavin application by iontophoresis using charged riboflavin solution (Ricrolin+) with a 1-mA current for 5 minutes. The first set was to compare riboflavin concentration measured by HPLC in corneas after iontophoresis or conventional riboflavin application. The second set was to analyze autofluorescence and stromal collagen modification immediately and 14 days after I-CXL or C-CXL, by using nonlinear two-photon microscopy (TP) and second harmonic generation (SHG). In the third set, physical modifications after I-CXL and C-CXL were evaluated by stress-strain measurements and by studying corneal resistance against collagenase digestion.METHODSWe designed three experimental sets using 152 New Zealand rabbits to study riboflavin application by iontophoresis using charged riboflavin solution (Ricrolin+) with a 1-mA current for 5 minutes. The first set was to compare riboflavin concentration measured by HPLC in corneas after iontophoresis or conventional riboflavin application. The second set was to analyze autofluorescence and stromal collagen modification immediately and 14 days after I-CXL or C-CXL, by using nonlinear two-photon microscopy (TP) and second harmonic generation (SHG). In the third set, physical modifications after I-CXL and C-CXL were evaluated by stress-strain measurements and by studying corneal resistance against collagenase digestion.Based on HPLC analysis, we found that iontophoresis allowed riboflavin diffusion with 2-fold less riboflavin concentration than conventional application (936.2 ± 312.5 and 1708 ± 908.3 ng/mL, respectively, P < 0.05). Corneal TP and SHG imaging revealed that I-CXL and C-CXL resulted in a comparable increased anterior and median stromal autofluorescence and collagen packing. The stress at 10% strain showed a similar stiffness of corneas treated by I-CXL or C-CXL (631.9 ± 241.5 and 680.3 ± 216.4 kPa, respectively, P = 0.908). Moreover, we observed an increased resistance against corneal collagenase digestion after I-CXL and C-CXL (61.90% ± 5.28% and 72.21% ± 4.32% of remaining surface, respectively, P = 0.154).RESULTSBased on HPLC analysis, we found that iontophoresis allowed riboflavin diffusion with 2-fold less riboflavin concentration than conventional application (936.2 ± 312.5 and 1708 ± 908.3 ng/mL, respectively, P < 0.05). Corneal TP and SHG imaging revealed that I-CXL and C-CXL resulted in a comparable increased anterior and median stromal autofluorescence and collagen packing. The stress at 10% strain showed a similar stiffness of corneas treated by I-CXL or C-CXL (631.9 ± 241.5 and 680.3 ± 216.4 kPa, respectively, P = 0.908). Moreover, we observed an increased resistance against corneal collagenase digestion after I-CXL and C-CXL (61.90% ± 5.28% and 72.21% ± 4.32% of remaining surface, respectively, P = 0.154).This experimental study suggests that I-CXL is a promising alternative methodology for riboflavin delivery in crosslinking treatments, preserving the epithelium.CONCLUSIONSThis experimental study suggests that I-CXL is a promising alternative methodology for riboflavin delivery in crosslinking treatments, preserving the epithelium.
We compared an iontophoresis riboflavin delivery technique for transepithelial corneal collagen crosslinking (I-CXL) with a conventional CXL (C-CXL). We designed three experimental sets using 152 New Zealand rabbits to study riboflavin application by iontophoresis using charged riboflavin solution (Ricrolin+) with a 1-mA current for 5 minutes. The first set was to compare riboflavin concentration measured by HPLC in corneas after iontophoresis or conventional riboflavin application. The second set was to analyze autofluorescence and stromal collagen modification immediately and 14 days after I-CXL or C-CXL, by using nonlinear two-photon microscopy (TP) and second harmonic generation (SHG). In the third set, physical modifications after I-CXL and C-CXL were evaluated by stress-strain measurements and by studying corneal resistance against collagenase digestion. Based on HPLC analysis, we found that iontophoresis allowed riboflavin diffusion with 2-fold less riboflavin concentration than conventional application (936.2 ± 312.5 and 1708 ± 908.3 ng/mL, respectively, P < 0.05). Corneal TP and SHG imaging revealed that I-CXL and C-CXL resulted in a comparable increased anterior and median stromal autofluorescence and collagen packing. The stress at 10% strain showed a similar stiffness of corneas treated by I-CXL or C-CXL (631.9 ± 241.5 and 680.3 ± 216.4 kPa, respectively, P = 0.908). Moreover, we observed an increased resistance against corneal collagenase digestion after I-CXL and C-CXL (61.90% ± 5.28% and 72.21% ± 4.32% of remaining surface, respectively, P = 0.154). This experimental study suggests that I-CXL is a promising alternative methodology for riboflavin delivery in crosslinking treatments, preserving the epithelium.
Purpose: We compared an iontophoresis riboflavin delivery technique for transepithelial corneal collagen crosslinking (I-CXL) with a conventional CXL (C-CXL).Methods: We designed three experimental sets using 152 New Zealand rabbits to study riboflavin application by iontophoresis using charged riboflavin solution (Ricrolin+) with a 1-mA current for 5 minutes. The first set was to compare riboflavin concentration measured by HPLC in corneas after iontophoresis or conventional riboflavin application. The second set was to analyze autofluorescence and stromal collagen modification immediately and 14 days after I-CXL or C-CXL, by using nonlinear two-photon microscopy (TP) and second harmonic generation (SHG). In the third set, physical modifications after I-CXL and C-CXL were evaluated by stress-strain measurements and by studying corneal resistance against collagenase digestion. Results: Based on HPLC analysis, we found that iontophoresis allowed riboflavin diffusion with 2-fold less riboflavin concentration than conventional application (936.2 ± 312.5 and 1708 ± 908.3 ng/mL, respectively, P < 0.05). Corneal TP and SHG imaging revealed that I-CXL and C-CXL resulted in a comparable increased anterior and median stromal autofluorescence and collagen packing. The stress at 10% strain showed a similar stiffness of corneas treated by I-CXL or C-CXL (631.9 ± 241.5 and 680.3 ± 216.4 kPa, respectively, P = 0.908). Moreover, we observed an increased resistance against corneal collagenase digestion after I-CXL and C-CXL (61.90% ± 5.28% and 72.21% ± 4.32% of remaining surface, respectively, P = 0.154).Conclusions: This experimental study suggests that I-CXL is a promising alternative methodology for riboflavin delivery in crosslinking treatments, preserving the epithelium.
Author Cassagne, Myriam
Soler, Vincent
Galiacy, Stéphane D.
Laurent, Camille
Spoerl, Eberhard
Rodrigues, Magda
Galinier, Anne
Malecaze, François
Fournié, Pierre
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  surname: Cassagne
  fullname: Cassagne, Myriam
  organization: Department of Ophthalmology Purpan Hospital, Toulouse, France 2Laboratory of Pathology, Purpan Hospital, Toulouse, France 3Department of Biochemistry, Rangueil Hospital, Toulouse, France 4Department of Ophthalmology, Technische Universität Dresden, Dresde
– sequence: 2
  givenname: Camille
  surname: Laurent
  fullname: Laurent, Camille
  organization: Laboratory of Pathology, Purpan Hospital, Toulouse, France 3Department of Biochemistry, Rangueil Hospital, Toulouse, France 4Department of Ophthalmology, Technische Universität Dresden, Dresden, Germany 5INSERM U1043, Center of Physiopathology, Dynamique
– sequence: 3
  givenname: Magda
  surname: Rodrigues
  fullname: Rodrigues, Magda
  organization: Laboratory of Pathology, Purpan Hospital, Toulouse, France 3Department of Biochemistry, Rangueil Hospital, Toulouse, France 4Department of Ophthalmology, Technische Universität Dresden, Dresden, Germany 5INSERM U1043, Center of Physiopathology, Dynamique
– sequence: 4
  givenname: Anne
  surname: Galinier
  fullname: Galinier, Anne
  organization: Department of Biochemistry, Rangueil Hospital, Toulouse, France
– sequence: 5
  givenname: Eberhard
  surname: Spoerl
  fullname: Spoerl, Eberhard
  organization: Department of Ophthalmology, Technische Universität Dresden, Dresden, Germany
– sequence: 6
  givenname: Stéphane D.
  surname: Galiacy
  fullname: Galiacy, Stéphane D.
  organization: Department of Ophthalmology Purpan Hospital, Toulouse, France 2Laboratory of Pathology, Purpan Hospital, Toulouse, France 3Department of Biochemistry, Rangueil Hospital, Toulouse, France 4Department of Ophthalmology, Technische Universität Dresden, Dresde
– sequence: 7
  givenname: Vincent
  surname: Soler
  fullname: Soler, Vincent
  organization: Department of Ophthalmology Purpan Hospital, Toulouse, France 2Laboratory of Pathology, Purpan Hospital, Toulouse, France 3Department of Biochemistry, Rangueil Hospital, Toulouse, France 4Department of Ophthalmology, Technische Universität Dresden, Dresde
– sequence: 8
  givenname: Pierre
  surname: Fournié
  fullname: Fournié, Pierre
  organization: Department of Ophthalmology Purpan Hospital, Toulouse, France
– sequence: 9
  givenname: François
  surname: Malecaze
  fullname: Malecaze, François
  organization: Department of Ophthalmology Purpan Hospital, Toulouse, France 2Laboratory of Pathology, Purpan Hospital, Toulouse, France 3Department of Biochemistry, Rangueil Hospital, Toulouse, France 4Department of Ophthalmology, Technische Universität Dresden, Dresde
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Keywords collagen
UVA
crosslinking
riboflavin
iontophoresis
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Snippet We compared an iontophoresis riboflavin delivery technique for transepithelial corneal collagen crosslinking (I-CXL) with a conventional CXL (C-CXL). We...
We compared an iontophoresis riboflavin delivery technique for transepithelial corneal collagen crosslinking (I-CXL) with a conventional CXL (C-CXL).PURPOSEWe...
Purpose: We compared an iontophoresis riboflavin delivery technique for transepithelial corneal collagen crosslinking (I-CXL) with a conventional CXL...
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SubjectTerms Animals
Collagen - administration & dosage
Cross-Linking Reagents
Disease Models, Animal
Epithelium, Corneal
Female
Human health and pathology
Infectious diseases
Iontophoresis - methods
Keratoconus - drug therapy
Keratoconus - metabolism
Keratoconus - pathology
Life Sciences
Rabbits
Riboflavin - administration & dosage
Riboflavin - pharmacokinetics
Vitamin B Complex - administration & dosage
Vitamin B Complex - pharmacokinetics
Title Iontophoresis Transcorneal Delivery Technique for Transepithelial Corneal Collagen Crosslinking With Riboflavin in a Rabbit Model
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