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Cultured Autologous Oral Mucosal Epithelial Cell- Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS, Boston, april 2010 C.Burillon, O.Damour Tissus et Cells HCL Bank The authors have no financial interest in the subject matter of this poster Ophthalmology department
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Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Jan 19, 2016

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Page 1: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS)

One-year follow-up for 17 patients/25 included

ASCRS, Boston, april 2010

C.Burillon, O.Damour

Tissus et Cells HCL Bank

The authors have no financial interest in the subject matter of this poster

Ophthalmology department

Page 2: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Limbal epithelial deficiency (LSCD)

Population Consequences

Thermal or chemical Burns

Stevens Johnson and Lyell Syndrom

Ocular pemphigoid

Aniridia

Severe dry eye

Limbal stem-cell deficiency after cornea transplantations

Other bilateral disorders of the ocular surface

Ulcers

pain

Dryness

Neovascularisation

Conjunctivalisation

Symblepharon

Total bilateral LSCD gives rise to blindness

Page 3: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

What do we have to do in LSCD?

To Bring an Epithelium

Autologous limbus graftCultured Autologous

limbal EpitheliumOral Mucosa Graft

For unilateral deficiencyAutograft from controlateral

safe eyeBrings stem cells to the

corneal epithelium

1-3 mm² Biopsy

For unilateral deficiency

For bilateral deficiencyOral mucosa very closed to cornea epithelium +++

Page 4: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

PurposeHypothesis of CAOMECS use:

1 – To restore the ocular surface

2 – To obtain sufficient Cornea transparency when stroma is healthy.

3 – To allow secondary donor cornea graft when stroma is opaque.

Page 5: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Materials and methodsWe use a specific support for the culture: an UpCell ®

Insert which is a thermosensitive polymer

- The temperature responsive polymer (PIPAAm) is immobilized by covalent binding on the surface.

- The surface change between hydrophobic and hydrophilic is temperature-dependent and reversible.

- At the temperature 20~25 , cells can't keep attachment because the surface turned to ℃hydrophilic to liberate hydrophobic interaction.

- The surface antigen and extra cellular matrix (ECM) of harvested cells are intact, because trypsin or dispase is not necessary to harvest the cells.

37°C 20°C

UpCell Basal membrane

hydrophobic hydrophilic

Page 6: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Biopsy and Culture using UpCell Insert ®

UpCell®-Insert

Biopsy

Culture on Upcell Insert at 37°C: 3 weeks

Isolation

NIH/3T3 in storage

Temperature reduced to 20ºC

PVDF doughnut ring as a white ring

Page 7: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

After three weeks, we obtain in 100% cases: • Robust, viable, multilayered epithelial cell sheets, • Transparent to read a letter Arial size 10.• Holoclones is garant of their functionality. • Keratin 3, p63 and b1 integrin and Laminin 5 expression on the sheet

Final Product Validation

Cytokeratin 3

Laminin 5 Integrin beta-1

p63

EfficiencyCFE > 2.1 ± 0.9%

Qualitykeratin 3/76, p63, β1 integrin

Microbiological SafetyOn biopsy transport medium On the culture medium4 days before harvest the sheet Graft day: on the medium just before harvest the sheetTransport medium

Clinical Trial Hospices Civils de Lyon, France prospective, open, non comparative and monocentric study

Stage I/II study by Gehan study design

Page 8: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Results: safety and efficacy at 12 monthsGrade changes improvement in blue worsening in red

# Primary endpoint Secondary endpoint

1

Corneal burn / 4 keratoplasties cataract surgery, Trabeculectomy + Mitomycine

No Ulcer anymore No more vessels

Comfort (dryness or watering, pain and photophobia)

2Neuroparalytic keratitis 1 lamellar keratoplasty

PEK (2-level improvement)No vessels anymore(3 to 0)

visual acuity, Comfort (pain, photophobia, dryness watering)

3Rosacea keratitis / no surgical treatment

No Ulcer anymore,vascularisationRecurrence of few localized vessels

Visual acuity Comfort (dryness or watering and pain)

4 Corneal burn /1 amniotic membrane

Blood vessels’ activitiesRecurrence Localized conjunctivalisation since at 6 months, due to surgery or contact lens

Comfort (dryness or watering) Visual acuity

5 Lyell syndrome /1 lamellar keratoplasty

SAECorneal Perforation at 6 month, but not related with CAOMECS

6Corneal burn / 1 lamellar keratoplasty, 1 amniotic membrane

No more PEKCorneal vascularisation (uncountable to 1), blood vessels activity

Comfort (dryness and pain) 1 level decrease of visualAcuity

7Corneal burn /1 keratoplasty1 amniotic membrance

No more UlcerNo more PEKCorneal vascularisation (5 to 1)

Comfort (Photophobia, dryness, pain)

Page 9: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Follow up of safety and efficacy at 12 monthsGrade changes improvement in blue worsening in red

# Primary endpoint Secondary endpoint

8Trachoma / Several corneal grafts

No Ulcer anymore neither PEKNo vascularisation anymore(20 to 0)

Visual acuityComfort (no dryness Photophobia,)

9 Lyell syndrome / 1 lamellar keratoplasty

No more ulcerPEKCorneal vascularisation (20 to 2)

Visual acuityComfort (no dryness, no photophobia)

11Corneal burn /1 keratoplasty

No Ulcer anymore PEKNo vascularisation anymore(12 to 0)

Visual acuityBut Corneal opacification Comfort (no photophobia, no dryness)

12

Corneal burn / 1 amniotic membrane, 1 lamellar keratoplasty

No Ulcer anymoreRecurrence Vessel (firstly decrease 20 to 0, but shown recurrence to 5 at 2-month and 7 at 12 month post-graftRecurrence PEK (firstly decreased grade to 0, but shown recurrence to + since 3 –month post-graft)

Comfort (no photophobia and no pain)but Comfort (dryness was appeared after 1 month.)

Patient No 13 could not come for the follow-up assessments since 2-month time.

Page 10: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Follow up of safety and efficacy at 12 months Grade changes improvement in blue worsening in red

# Primary endpoint Secondary endpoint

10

Rosacea keratitis/3 keratoplasties, Cataract surgery

No Ulcer anymore PEK Corneal vascularisation (15 to 3), vessels activity

visual acuity, Comfort (no photophobia, no dryness)

14

Corneal burn/1 Palpebral reconstruction1 amniotic membrance

KPS de ++ to 0Recurrence of

neovasculatisation at 8months

visual acuity, Comfort (no pain)

15Aphakia post-congenital cataract + corneal anoxia induce by contact lens/no surgical treatment

PEK Corneal vascularisation(40 to 2), Blood vessels activity

visual acuity, Comfort (no pain, photophobia, no dryness)

16

Corneal burn/1 keratoplasty, 1 amniotic membrane

No more Ulcer PEK Corneal vascularisation (30 to 3

visual acuity, Comfort (no pain, no dryness, photophobia)

17Lyell syndrome/1 amniotic membrane

PEK Corneal vascularisation (30 to 8) , vessels activity

Comfort (dryness +++ to +)

Page 11: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Without fluorescein with fluorescein

Inclusion One year follow up Inclusion One year follow up

Some examples:Primary criteria : Ocular Surface observed by Slit Lamp Examination

After corneal graft

Page 12: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Conclusions• Significant regenerated epithelium on the ocular surface:

– new good functional epithelium– less vascularisation and vessels activities

• When stroma and endothelium are healthy, visual acuity can be improved

• When stroma and endothelium are damaged, visual acuity is difficult to be improved with only CAOMECS treatment but it allows a conventional donor graft without rejections and visual acuity is improved (5 cases up to day)

Histology of corneaAfter CAOMECS + corneal graft

Page 13: Cultured Autologous Oral Mucosal Epithelial Cell-Sheet for Corneal Epithelial Reconstruction (CAOMECS) One-year follow-up for 17 patients/25 included ASCRS,

Aknowlegments

GCS/CTC tissus and cells bankOdile DamourChantal HéloireEric VenetPascale PascalCéline Auxenfans

Laboratoire ImmunocytologySerge Nataf

CellSeedYukio HasegawaNao NakamuraAyumi MurayamaVirginie Justin

OphthalmologyCarole Burillon

Preliminary Human StudyNishida et al , N. Engl.J.Med, 2004, 351,1187-1196