2. Comparative Histological ProfilesComparative Histological Profiles
Apligraf® OrCel®
Fully Differentiated and
Stratified Keratinocytes
Partially Differentiated,
Non-Stratified “Wound Healing”
(Migrating & Replicating) Keratinocytes
Low Density Dermal Fibroblasts,
Many Undergoing Apoptosis
Medium Density, Vigorously
Growing Dermal Fibroblasts
(≥15% “Dividing” Cells)
3. In Vitro expression of KGF and GMCSF
by cultured LSEand ORCEL
1
1000
1000000
KGF GM-CSF
CytokineOutputPerunitArea,pg/cm2/day
LSE
ORCEL
Living Skin Equivalent (LSE) is a generic name for Apligraf
5. OrCel® in Donor Site Healing
(Independent Physician Evaluation:
Donor Site Treated with both OrCel® & Apligraf®)
OrCel®
Apligraf®
OrCel®: Faster healing, less inflammation
6. Days to
100% Healing
(Median)
OrCel
Standard of
Care
DONOR SITES
Pivotal: 2/01
Based on Photography
0
2
4
6
8
10
12
14
16
18
20
22
22 Days
15 Days
PMA Claims
•Accelerated
healing
•Less Scarring
•Reduced time to
recropping
7. OrCelOrCel ™™ HealsHeals
Wounds FasterWounds Faster
47%
23%
0
10
20
30
40
50
60
Diabetic Foot Ulcers Pilot Trial
% of Patients
Achieving 100%
Wound Closure
By 12 Weeks
OrCel
OrCel
Standard of Care
Standard
Of Care
8. Summary of OrCel® vs. Apligraf® Features
Feature OrCel® Apligraf®
Shape/Size Square 6.5 x 6.5 x 0.09 cm (42 cm2
) Round 75mm diam x
0.75mm (44cm2
)
Matrix Preformed porous cross-linked
bovine collagen sponge, very pliant
Contracted, dense, stiff
bovine collagen gel
Keratinocytes (K) Immature mono-/bilayer
(submerged culture condition)
Stratified multilayer (air
interface culture condition)
Fibroblasts (F) Healthy vigorously dividing Dormant or apoptotic
Cell type ratio Nominal ~90% F ~10% K Nominal ~30% F ~70% K
Cytokine profile F (& F/K co-culture) dominated;
activated “wound healing” profile
K (& F/K co-culture)
dominated; normal skin
steady state dormant profile
Ease of use Semi-auto thaw/rinse (~20 min) &
direct application
Remove from dish, cut to
size, fenestrate (~20 min)
Shelf life 7 mo (clin. trial); expect 12 – 18 mo 10 days in sealed pkg.
Current
reimbursement
$1,100./42 cm2
$1,200/44 cm2
9. Summary: OrCel® vs. Apligraf® Data
• Pilot ORCEL (fresh) VLU trial data achieved closure in 12
weeks (59% vs. 36%) comparable to adjusted (Cox) Apligraf
results at 6 months (56.8% vs. 39.8%); pivotal ORCEL
(cryopreserved) clinical data are similar (50% vs. 31%).
• Pilot ORCEL DFU trial data achieve results within 12 weeks
(47% vs. 23%) comparable to Apligraf (56% vs. 39%) at 12
weeks (SOC differed)
• ORCEL is easier to apply
• ORCEL, since cryopreserved, has far superior shelf life (7-
12 months+ (projected) vs. 10 days) and can also be stored
at the clinical site for “off-the-shelf” availability
Notas do Editor
Note that Apligraf, shown in Trichrome stain, is top-heavy, containing a multi-layered “mature” stratum corneum with a high concentration of keratinocytes, but a rather sparse fibroblast population by comparison. Apligraf is about ¾ keratinocytes and has almost no dividing cells. OrCel, by comparison (shown in Hematoxylin/Eosin, or H&E stain), is over ¾ fibroblasts and has about 15% dividing cells. Apligraf has cells in a dense gel matrix, while OrCel cells are more sparse, are still growing in a very porous sponge matrix. In short, Apligraf cells are relatively quiescent, while OrCel cells are dynamic. The collateral effects of these differences can be seen in the two products’ relative cytokine expression levels in vitro (next slide).
In vitro Cytokine Expression by OrCel vs. LSE: Cultures of equivalent sized pieces of OrCel (freshly thawed and rinsed in saline solution) and Living Skin Equivalent, or LSE (Apligraf, freshly removed from its packaging at room temperature) in OrCel growth medium (Ortec International, Inc.) were incubated for 48 hr, and media were sampled daily; assay of KGF-I and GMCSF were by commercial ELISA kits (Quantikine, R&D Systems) following manufacturer’s recommendations. Results are expressed as picograms (pg) of cytokine secreted per square centimeter area of tissue per day.
This graph illustrates the relative output of specific cytokines, growth factors and Matrix Metalloproteinases by pieces of OrCel and Apligraf of identical area cultured in vitro under identical “transwell” (the Apligraf culture method) conditions. Note that the left side of the graph shows growth and migration stimulatory factors, while the right side shows pro-inflammatory factors. In terms of potency, GM-CSF, a keratinocyte mitogen and migration factor, is believed to be highly potent at >1000 pg/cm2/day, as above in the ORCEL bar, while the angiogenic response of the wound to VEGF is believed to be proportional to concentration above 1000 pg/cm2/day. The practical potency levels of the other stimulatory cytokines are not known. The higher MMP-9 and PGE-2 production by APLIGRAF imply that it is significantly more pro-inflammatory -- as evidenced clinically in the example on the next slide.
As you may know, there are two commercially available tissue engineered products which contain both epidermal and dermal cells…OrCel and Apligraf
This photograph represents a direct head to head comparison of these two products on a single donor site wound.
This is a burn patient who was treated at Doctors Hospital in Augusta, GA who had OrCel placed on her donor sites. The OrCel was placed on the right medial thigh…. And Apligraf was placed on the same donor site on the lateral aspect creating a direct side by side comparison of the two products. Note that ORCEL is “square” while Apligraf is “round.” This photo was taken 13 days after the original treatment.
As you can see the OrCel treated area has re-epithelialized in a uniform fashion with an acceptable cosmetic outcome. Additionally, pigment is already returning to the OrCel treated area
As you can see the Apligraf treated areas are not yet fully epithelialized and appear red slightly inflamed and not fully healed. This is significant not just from a cosmetic visual point of view but also because the earlier one can heal any open surface area on a burn patient the better it is for that patient.
These are the results from the Diabetic Ulcer Pilot Study. As you can see from the graph, 47% of OrCel treated patients achieved 100% wound closure by Week 12 compared to 23% of Standard Care treated patients.