Download OwN ExPERIENcE fROm thE uSE Of A SubStItutE Of

POLSKI
PRZEGLĄD CHIRURGICZNY
2015, 87, 10, 513–521
10.1515/pjs-2015-0097
Own experience from the use of a substitute of an
allogeneic acellular dermal matrix revitalized with
in vitro cultured skin cells in clinical practice
Wojciech Łabuś1, Marek Kawecki1, 2, Justyna Glik1,3, Mariusz Maj1,
Diana Kitala1, Marcelina Misiuga, Agnieszka Klama-Baryła1,
Małgorzata Kraut1, Mariusz Nowak1
Dr Stanisław Sakiel Centre for Burn Treatment in Siemianowice Śląskie1
Dyrektor: dr n. med. M. Nowak
Faculty of Health Sciences, University of Bielsko-Biała2
Dziekan Wydziału: dr hab. M. Mikulska, prof. ATH
Medical University of Silesia in Katowice, Faculty of Health Sciences in Katowice, Department of
Chronic Wound Treatment Organisation3
Dziekan Wydziału: prof. dr hab. V. Skrzypulec-Plinta
As a result of the removal of cells from human allogeneic dermis, a collagen scaffold is obtained, which
can be populated de novo with autologous/allogeneic skin cells and transplanted onto the area of skin
loss. The optimal method for production of acellular dermal matrices (ADM) has been selected. Three
female patients (a mean age of 54 years) were subjected to the transplantation of either autologous or
allogeneic keratinocytes and fibroblasts into the holes of acellular dermal matrix (ADM) mesh graft.
The method for burn wound treatment based on the use of a viable dermal-epidermal skin substitute
(based on ADM and in vitro cultured fibroblasts and keratinocytes) may be the optimal method of burn
treatment.
Key words: acellular dermal matrix ADM, burn, fibroblasts, keratinocytes, tissue engineering
The optimal method for the treatment of
extensive full thickness burn wounds (3rd
degree) is the resection of necrotic tissue and
the use of autologous free split-thickness skin
grafts (STSG), harvested from an intact area
(1-7), for skin closure. The main limitation of
this method is the scarcity of healthy, intact
skin to be used as a graft. Moreover, donor
areas constitute an additional wound putting
a burden on the body (8). The sites from which
skin has been harvested for a graft are painful,
therefore require analgesic therapy, and leave
scars while healing (5, 8). Limitations associated with the use of autologous free splitthickness skin grafts necessitate searching for
new methods and agents for the treatment of
severely burned patients.
An alternative clinical standard in the
therapy of extensive full-thickness burns in
patients with the scarcity of donor areas, al-
ready available for over three decades, is the
use of autologous keratinocytes (CEA) cultured
under in vitro conditions (4, 5). This method
results in the acceleration of integument regeneration process, while in the long run, hypertrophied scars unacceptable for functional and
aesthetic reasons may occur (4, 5, 9, 10).
Another example of an alternative method
for burn treatment is the use of allogeneic skin
grafts. This method has been known for over
seventy years. The main clinical indications
for the use of allogeneic skin grafts include the
protection against drying of the wound bed,
protection against infection, and a mechanical
barrier protecting against the loss of heat,
electrolytesand proteins. The use of allogeneic
skin grafts is, however, a temporary solution.
A graft of this type is a temporary biological
dressing which is rejected by the recipient’s
body after a certain time period (6, 7, 11).
Unauthenticated
Download Date | 6/18/17 6:34 PM
514
W. Łabuś et al.
In order to improve graft properties and
extend their stability, tissue engineering
procedures, primarily including methods for
the removal of cells from tissues, are used.
The goal of using these methods is to obtain
a non-immunogenic graft which, following
transplantation, would be populated de novo
with the recipient’s cells to become an integral
part of the body (4, 5, 12-15). Since the immune response is mainly triggered against
proteins and lipids of the cell membrane, the
removal of cells from tissues is a promising
method aimed at the elimination of the immune response in the recipient’s body after
the transplantation has been performed (16,
17).
The methods of cell removal from tissues
result in obtaining and isolation of the structures of extracellular matrix (ECM). The extracellular matrix is composed of proteins,
glycosaminoglycans (GAGs), proteoglycans,
and growth factors. ECM is a natural bioprosthesis that structurally strengthens the damaged tissues. Numerous tissues have been
subjected to the processes of cell removal, e.g.
the tissues of the submucosal layer of the intestine (13), oesophagus (14), urinary bladder
(15), placenta (16), pericardium (17), heart
valve (18-21), and dermis (22, 23).
Skin substitutes containing no cells, known
as acellular dermal matrices (ADM) or acellular dermal grafts (ADG), serve as a scaffold
stimulating the body to initiate natural regenerative and repairing mechanisms. Such dressings play a significant role in minimizing both
the scarring process and the deformation of a
healing wound (24).
Cell culturing on bioprostheses devoid of
skin cells, obtained by using tissue engineering
methods, is a promising strategy in the production of “new tissues” i.e. a viable, stable skin
graft (25). The essence of this method is the
use of elements of extracellular matrix of allogeneic skin, serving as a scaffold for in vitro
cultured skin cells. The main benefits offered
by this technique include the introduction of
a scaffold for the newly forming tissue, ensuring mechanical protection that is able to withstand the forces occurring in vivo until the
developed skin substitute becomes an integral
part of the body (25-28).
The aim of the study was to evaluate methods for burn wound treatment using an independently produced viable skin substitute
based on ADM and either autologous or allogeneic skin cells.
Material and methods
The proposed research programme has been
approved by the Bioethics Committee of the
Silesian Chamber of Physicians in Katowice.
For the production of acellular dermal matrices (ADM), allogeneic and biostatic human
skin grafts deposited at the Tissue Bank of the
Dr Stanisław Sakiel Centre for Burn Treatment (Polish: Centrum Leczenia Oparzeń,
CLO) were used. Allogeneic skin was harvested from deceased donors during multiorgan harvests, and then developed in accordance with applicable regulations.
Based on an analysis of the results obtained
in the research work at the CLO, the optimal
method for the production of stable, viable skin
substitutes based on independently produced
acellular dermal matrices (ADM) and in vitro
cultured skin cells. The research protocol in
question proposed an enzymatic method for
cell removal (0.05% trypsin solution in EDTA
and 0.025% trypsin solution in EDTA), a
chemical method (0.1% solution of sodium
dodecyl sulphate (SDS), and a combined
method which assumes the use of an enzyme
at the first stage, and a chemical agent at the
second stage (0.05% trypsin solution in EDTA
and 0.1% SDS, and 0.025% trypsin solution in
EDTA and 0.1% SDS). The test material was
analysed using an atomic force microscope
(AFM), and in histological preparations (hematoxylin and eosin staining). Based on the
obtained results, the optimal method for the
removal of cells from allogeneic dermis was
selected. The procedure of cell removal from
allogeneic dermis was based on 24-hour, onestage incubation of human dermis in 0.05%
trypsin solution in 0.2% EDTA (PAA Laboratories GmBH) (PhD thesis, 2014).
Prior to the proper part of the ADM production, human allogeneic skin obtained from the
CLO’s Tissue Bank was defrosted and rinsed
twice in normal saline solution. The separation
of dermis and epidermis resulted from the 1
to 3-hour incubation in a solution of dispase
enzyme at a concentration of 2.4 U/ml (Gibco)
at a temperature of 37˚C.Thus obtained fragments of human dermis were incubated in
0.05% trypsin solution in 0.2% EDTA (PAA
Unauthenticated
Download Date | 6/18/17 6:34 PM
Substitute of an acellular allogeneic dermal matrix in clinical practice
Laboratories GmBH) for 24 hours at a temperature of 37˚C in order to remove undesirable cellular elements.Following processing,
the obtained ADMs were thoroughly rinsed
three times in a buffered normal saline solution PBS (Sigma). Prior to the performance of
transplantation, slits are cut in the obtained
ADMs using a mesh dermatome at various
degrees of meshing. Allogeneic and biostatic
human skin grafts subjected to no procedures
of cell removal were used as a control. Allogeneic skin grafts were meshed to an extent
analogous to that of the used acellular matrix
grafts.
A living donor was qualified for the establishment of an in vitro skin cell culture based
on the donor’s direct consent and the statutory eligibility criteria.
In order to establish an in vitro culture of
fibroblasts and keratinocytes, a fragment of
split-thickness skin measuring approx. 2 x 2
cm was harvested from a living donor. The skin
was harvested under operating theatre conditions. Following the harvest, the fragment of
skin was incubated in 20 mL of culture medium
KGM-CD with addition of antibiotics and antimycotics (to 500 mL of culture medium KGMCD, 5 mL of antibiotic and antimycotic solution
composed of penicillin 10,000 U/mL, streptomycin sulphate 10 mg/mL, and amphotericin B 25
µg/mL was added) (PAA Laboratories GmBH)
for 20 minutes at a room temperature. Culturing procedures were followed in accordance with
the Good Manufacturing Practice (GMP) requirements. The isolation of cells comprised the
stage of separating dermis from epidermis by
a 2-3 hour long incubation in 2.4 U/mL of dispase enzyme solution (Gibco) at a temperature
of 37˚C. At a further stage, cells were isolated
from the obtained fragments of epidermis and
dermis by a 2-minute incubation in 0.05%
trypsin solution in 0.2% EDTA (PAA Laboratories GmBH) at a temperature of 37˚C. The
process was repeated 3-4 times. The obtained
cell suspension was centrifuged, and then
suspended in culture medium: KGM-CD for
keratinocytes, and DMEM for fibroblasts. The
cells were cultured in an incubator (HeraCell)
at 37˚C, 5% of CO2 content, and 95% humidity.
The culture medium was replaced every 24
hours. After confluence has been reached, the
cells were isolated using 0.05% trypsin solution. The cells were suspended in PBS, and
transported to the operating theatre.
515
Cultured skin cells were transplanted in
Platelet Leukocyte Rich Gel (PLRG). PLRG
was obtained from the patient’s peripheral
blood using an Angel System device (CZM) in
accordance with the manufacturer’s instructions. The isolated cells were suspended in
PLRG, and then transplanted into the holes
of the previously meshed ADM graft.
The wound under study was divided into
two equal areas. Acellular dermal matrices
(ADM) and cultured skin cells were transplanted onto one area, while allogeneic and
biostatic human skin grafts and cultured skin
cells were transplanted onto the other as a
control of the treatment process.
On days 3, 5, 14 and 21 after the transplantation, a visual clinical evaluation of the wound
was performed. Photographic documentation
was produced. The obtained observations were
confirmed in histopathological examinations.
Fragments of tissue intended for microscopic
examinations were harvested from an area
located as close as possible to the edge of the
wound, and then placed in 10% formaldehyde
solution. Then, histological preparations were
made using the hematoxylin and eosin staining
method. In addition, in order to evaluate the
microbial purity of the wound environment,
samples were taken using the swab method.
For the proposed research programme,
three female patients (a mean age of 54) were
selected:
Case 1. Female patient, aged 63, admitted
with a thermal burn of 34% of the TBSA, including 24% of 3rd/4th degree burns. Patient
with history of myocardial infarction, who also
underwent angioplasty of the right coronary
artery with stent implantation. Transplantation of ADM and allogeneic skin (control)
meshed 1:2 was performed, and then in vitro
cultured autologous fibroblasts and keratinocytes suspended in PLRG were transplanted.
On days 0, 3 and 5, the condition of wounds
after the transplantation was assessed, histological preparations were made, and microbiological examinations were carried out. Photographic documentation was produced.
Case 2. Female patient, aged 36, admitted
with a thermal burn of 24% of the TBSA, including 24% of 3rd/4th degree burns. Transplantation of ADM and allogeneic skin (control)
meshed 1:2 was performed, and then in vitro
cultured autologous fibroblasts and keratinocytes suspended in PLRG were transplanted
Unauthenticated
Download Date | 6/18/17 6:34 PM
516
W. Łabuś et al.
onto poorly healing donor areas. On days 0, 3,
5, 14 and 21, histological preparations were
made, and swabs from wounds were taken.
Photographic documentation was produced.
Case 3. Female patient, aged 63, admitted
with a 2a/2b degree thermal burn of 20% of the
TBSA. Transplantation of ADM meshed 1:1.5
was performed, and then in vitro cultured allogeneic fibroblasts and keratinocytes suspended in normal saline were transplanted.
On day 0, a histological preparation was made.
The patient gave no consent to further participation in the study. On days 3, 5, 19 and
21, swabs from wounds were taken, and photographic documentation was produced.
Results
Case 1. The patient was transplanted with
a total of 2800 cm2 allogeneic skin meshed 1:2,
of which 1063 cm2 were ADMs. Autologous
keratinocytes and fibroblasts in PLRG were
inoculated into the graft holes. Figure 1 presents the procedure of cleansing burn wounds
using a hydrosurgery system (VersaJet), and
the transplantation of acellular dermal matrices (ADM) and allogeneic skin (control). Figure
2. presents the appearance of wounds on day
3. Figure 3 shows the clinical picture of wounds
on day 5. The patient was in a very poor, and
worsening, general condition. On day 6 following the transplantation, the patient died due
to multiple organ dysfunction syndrome. The
photograph was taken in the autopsy room (fig.
4). In all presented histological pictures, fragments of skin with epidermal necrosis, noticeable detachment of epidermis, and purulent
exudate were observed. Granulocytic exudate
in the subcutaneous tissue (fig. 1-3).
Case 2. The patient was transplanted with
a total of 657 cm2 allogeneic skin meshed 1:1.5,
of which 462 cm2 were ADMs. Autologous keratinocytes and fibroblasts in PLRG were inoculated into the graft holes (fig. 5). The histological picture obtained on the day of surgery
(day 0) shows a granulating wound with oedema of connective tissue (fig. 5). Figure 6
shows the clinical picture of wounds on day 3
following the surgery. Figure 6A presents the
Fig. 1. Case 1. Wound appearance at day 0.
Fig. 2. Case 1. Wound appearance at day 3
Fig. 3. Case 1. Wound appearance at day 5
Unauthenticated
Download Date | 6/18/17 6:34 PM
Substitute of an acellular allogeneic dermal matrix in clinical practice
Fig. 4. Wound appearance at day 6. Patient’s death
appearance of the wound onto which ADM and
autologous skin cells were transplanted. As
compared to the control (fig. 6B), the appearance of the wound treated with ADM and cells
reveals the initiated incorporation of acellular
matrix into the wound bed. Clinical observations are confirmed in microscopic tests. An
analysis of the results of a histopathological
examination carried out on day 3 following the
surgery for the area under study revealed the
presence of epidermis with signs of hyperkeratosis and parakeratosis. Under the epidermis, mature granulation tissue was revealed
(fig. 6A). As regards the control area, as early
as on day 3, self-renewing epidermis with signs
of parakeratosis was observed. Epidermis was
locally separated from the surface layer, with
granulocytic exudate (fig. 6B). Observation of
wounds on day 5 following the surgery reveals
further progress in regeneration processes for
wounds treated with ADM (fig. 7A). The appearance of wounds treated with allogeneic
skin (control) shows allogeneic grafts during
rejection (fig. 7B). The histological picture of
the area under study on day 5 shows a fragment of skin covered with mature epidermis.
B
A
Fig. 6. Case 2. Wound appearance at day 3
517
Fig. 5. Case 2. Wound appearance at day 0
In the dermis, numerous small blood vessels
were observed. A mild inflammatory reaction
was observed (fig. 7A). In the histological picture of the control area on day 5 small strips
of epidermis and a profuse inflammatory response in the dermis were observed (fig. 7B).
Figure 8A shows the appearance of wounds
treated with ADM on day 14 following the
transplantation. The photograph reveals the
treated wound with few residual areas (fig.
8A). The picture of the control area on day 14
reveals progressive regeneration processes, yet
at a slightly earlier stage of progress (fig. 8B).
On the other hand, histological pictures of both
the area under study and the control area,
obtained on day 14, revealed mature epidermis
with signs of pareketosis (fig. 8). The patient
was discharged from hospital on day 21 following the transplantation. Photographic documentation produced on the day of discharge
presents the complete closure of the wound,
which is confirmed in microscopic examinations (fig. 9).
Based on microbiological tests carried out
on particular days, the presence of two Staphylococcus aureus strains in the wound, name-
B
A
Fig. 7. Case 2. Wound appearance at day 5
Unauthenticated
Download Date | 6/18/17 6:34 PM
518
W. Łabuś et al.
B
A
Fig. 8. Case 2. Wound appearance at day 14
Fig. 9. Case 2. Wound appearance at day 21. Discharge
from hospital
ly methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA), was found.
Case 3. The patient was transplanted with
92 cm2 ADM. Autologous keratinocytes and
fibroblasts in 0.9% NaCl were inoculated into
the graft holes. Eventually, the patient was
transplanted with an autologous free splitthickness skin graft, which resulted in the
closure of the wound. The clinical picture of
the healing process on the successive days following the transplantation is presented in Fig.
10. Based on the microbiological tests carried
out on particular days, the presence of a methicillin-resistant Staphylococcus aureus
(MRSA) strain in the wound was found.
Discussion
Rapid development of tissue engineering
over the recent years has led to the emergence
of new therapeutic methods based on the use
of natural bioprostheses populated with autologous cells. This improvement is a promising strategy in the production of “new tissues”
(29-32).
The currently conducted research is focused
on the development of a treatment strategy
involving the combination of in vitro cultured
skin equivalents, e.g. fibroblasts, with a matrix
obtained through the removal of cells from the
dermis. This project may possibly result in
obtaining a viable, stable skin substitute
(32).
In the proposed protocols for the production
of viable skin grafts based on ADM and in
vitro cultured fibroblasts, most research teams
used the commercially available acellular allograft dermal matrix, namely AlloDerm (2940). On the other hand, in this study, due to
Fig. 10. Case 3. Wound appearance in sequence at day
0, 3. 5, 19 and 24
the wide availability of allogeneic human skin
grafts deposited at the Tissue Bank of the
Centre for Burn Treatment, this particular
material was used and subjected to an original
procedure of cell removal from tissues, followed
by the repopulation with the patient’s autologous or allogeneic cells. Acquiring biomedical
material from the Bank Tissue is particularly
beneficial for severely burned patients, since
it allows the performance of transplantation
and closure of the burn wound without the
need for an adverse interference with the patient’s body (harvesting an autologous skin
graft).
Based on the results obtained from the research carried out at the CLO, it was concluded that fibroblasts cultured on the surface
of ADM develop a single- or a few-layered cell
cover. However, they are not able to migrate
within the collagen matrices, as is the case in
natural skin. Similar observations were made
in studies carried out by other researchers,
inter alia Rodrigues et al. (38), Sorrell and
Caplan (39), and Ng et al. (40). A probable
cause of this phenomenon has been sought in
the compact, cross-linked structure of the collagen forming the extracellular dermal matrix
Unauthenticated
Download Date | 6/18/17 6:34 PM
Substitute of an acellular allogeneic dermal matrix in clinical practice
(40). In the clinical context, this unfavourable
phenomenon may significantly contribute to
the failure of therapy based on the transplantation of thus regenerated dermal matrices.
During the meshing of such a graft, the integrity of the layer of cells cultured on the surface
of ADM can deteriorate, or these cells can be
completely removed. Bearing this in mind, a
decision was taken on the production of a
meshed ADM graft in the first place, followed
by the transplantation of a culture cell suspension into the graft holes.
So far, three female patients have been
invited to participate in the research programme concerned. The research programme
is at the early stage of development, and there
is no doubt that in order to perform a complete
and valuable analysis of the obtained results,
significantly more research material need to
be collected. In addition, as regards the clinical
cases in question, only one (case 2) can be
considered highly valuable in terms of the
analysis of obtained results.
Case 2 – a female patient aged 34, who was
transplanted with ADM and autologous fibroblasts and keratinocytes suspended in PLRG
onto poorly healing wounds on the donor areas
on both upper legs. On the successive days
following the surgery, progressive and accelerated healing of wounds treated with regenerated ADM was observed, as compared to the
control areas. The therapeutic effect was confirmed in microscopic tests.
Case 1 – a 63 year old female patient with
extensive burns and numerous concomitant
diseases (history of myocardial infarction). The
patient’s condition continually deteriorated on
successive days. On day 6 the patient died due
to multiple organ dysfunction syndrome.
Case 3 – another 63 year old female patient
with a poorly healing burn wound on the right
forearm. Following the transplantation, the
woman gave no consent to further participation in the study, therefore the performance of
519
activities scheduled in the study were withdrawn from. However, photographic documentation was produced. The transplantation of
ADM and allogeneic skin cells failed to result
in a complete closure of the wound, and the
patient needed the transplantation of an autologous split-thickness skin graft, which resulted in the healing of the wound. It can be
assumed that the use of ADM and allogeneic
fibroblasts and keratinocytes led to the optimal
preparation of the wound to the transplantation of the patient’s own skin.
There is a serious difficulty in referring the
obtained results to the achievements of other
researchers, as there are no available literature resources directly dealing with the clinical
use of biotechnologically developed skin substitutes in the treatment of wounds in humans.
All the more, one should not doubt that the
introduction of stable and viable skin grafts to
the clinical practice could produce positive and
socially acceptable outcomes. A positive outcome of the use of such grafts can be a significant shortening of the time spent in hospital,
and a more rapid return to full fitness. It
should be assumed that the next step in the
research on obtaining a viable skin graft will
be the performance of the procedure of multicultural culture of all types of cells occurring
in natural skin on an acellular dermal matrix.
Such a research programme could possibly
result in obtaining a skin graft of full value.
Conclusions
The procedure of burn wound treatment
based on the use of a viable dermal-epidermal
skin substitute (based on ADM and in vitro
cultured fibroblasts and keratinocytes) may be
the optimal method of burn treatment. However, it is required that the sample size be
increased in order to perform a statistical
analysis of the obtained results.
references
1.Stanton RA, Billmire DA: Skin resurfacing for the
burned patent. Clin Plast Surg 2002; 29: 29-51.
2.Andreassi A, Bilenchi R, Biagioli M et al.: Clasisification and pathophysiology of skin grafts. Clin
Dermatol 2005; 23: 332-37.
3.Supp DM, Boyce ST: Engeenered skin substitutes: practices and potentials. Clin Dermatol 2005;
23: 403-12.
4.Klama-Baryła A, Glik J, Kawecki M et al.: Skin
substitutes – the application of tissue engineering
Unauthenticated
Download Date | 6/18/17 6:34 PM
520
W. Łabuś et al.
in burn treatment Part 1. J Orthop TraumaSurg
Rel Res 2008; 11: 96-103.
5.Łabuś W, Kawecki M, Nowak M: The role of tissue engineering in the treatment of burn wounds.
Pol Przegl Chir 2012; 3: 167-71.
6.Chen B, Hu DH, Jia CY et al.: Management of
a patient with massive and deep burns: early care
and reconstruction after convalescence. Zhonghua
Shao Shang Za Zhi 2007; 23: 112-16.
7.Kagan RJ, Robb EC, Plessinger RT: Human Skin
Banking. Clin Lab Med 2005; 25: 587-605
8.Kim JJ, Evans GR: Applications of biomaterials
in plastic surgery. Clin Plast Surg 2012; 4: 35976.
9.Gosk J: Skin substitutes – the present and the
future. Polim Med 2012; 42: 109-14.
10.Sood R, Roggy DE, Zieger MJ et al.: A comparative study of spray keratinocytes and autologous meshed split-thickness skin graft in the treatment of acute burn injuries. Wounds 2015; 27:
31-40.
11.Shevchenko RV, James SL, James SE: A review
of tissue-engineered skin bioconstructs available
for skin reconstruction. J R Soc Interface 2010; 7:
229-58.
12.Carson AE, Barker TH: Emerging concepts in
engineering extracellular matrix variants for directing cell phenotype. Regen Med 2009; 4: 593600.
13.Badylak SF, Record R, Lindberg K: Small intestinal submucosa: a substrate for in vitro cell
growth. J Biomater Sci Polym Ed 1998; 9: 86378.
14.Bhrany AD, Beckstead BL, Lang TC et al.: Development of an esophagus acellular matrix tissue
scaffold. Tissue Eng 2006; 12: 319-30.
15.Bolland F, Korossis S, Wilshaw SP et al.: Development and characterisation of a full-thickness
acellular porcine bladder matrix for tissue engineering. Biomaterials 2007; 28: 1061-70.
16.Schmidt Ch, Baier J.M: Acellular vascular tissues: natural biomaterials for tissue engineering.
Biomaterials 2000; 21: 2215-31.
17.Graus RW, Hazekamp MG, Vliet van S et al.:
Decellulrization of rat aortic valve allografts reduces leaflet destruction and extracellular matrix
remodeling. J Thorac Cardiovasc Surg 2003; 126:
2003-10.
18.Hopper RA, Woodhouse K, Semple JL: Acellularization of human placenta with preservation of
the basement membrane: a potential matrix for
tissue engineering. Ann Plast Surg 2003; 51: 598602.
19.Mirsadraee S,Wilcox HE, Korossis SA et al.:
Development and characterization of an acellular
human pericardial matrix for tissue engineering,
Tissue Eng 2006; 12: 763-73.
20.Curtil A, Pegg DE, Wilson A: Freeze drying of
cardiac valves in preparation for cellular repopulation. Cryobiology 1997; 34: 13-22.
21.Kasimir MT, Rieder E, Seebacher G et al.: Comparison of different decellularization procedures of
porcine heart valves. Int J Artif Organs 2003; 26:
421-27.
22.Chen RN, Ho HO, Tsai YT et al.: Process development of an acellular dermal matrix (ADM) for
biomedical applications. Biomaterials 2004; 25:
2679-86.
23.Buinewicz B, Rosen B: Acellular cadaveric dermis (AlloDerm): a new alternative for abdominal
hernia repair. Ann Plast Surg 2004; 52: 188-94.
24.Priya SG, Jungvid H, Kumar A: Skin tissue
engineering for tissue repair and regeneration.
Tissue Eng Part B Rev 2008; 1: 105-18.
25.Vacanti JP, Langer R, Upton J, Marler JJ:
Transplantation of cells in matrices for tissue regeneration. Adv Drug Deliv Rev 1998; 33: 165-82.
26.Kim BS, Mooney DJ: Development of biocompatible synthetic extracellular matrices for tissue
engineering. Trends Biotechnol 1998; 16: 224-30.
27.Seland H, Gustafson CJ, Johnson H et al.:
Transplantation of acellular dermis and keratinocytes cultured on porous biodegradable microcarriers into full-thickness skin injuries on athymic
rats. Burns 2011; 37: 99-108.
28.Bannasch H, Stark GB, Knam F et al.: Decellularized dermis in combination with cultivated
keratinocytes in a short- and long-term animal
experimental investigation. JEADV 2008; 22, 4149.
29.Butterfield JL: 440 Consecutive Immediate,
Implant-Based, Single-Surgeon Breast Reconstructions in 281 Patients: A Comparison of
Early Outcomes and Costs between SurgiMend
Fetal Bovine and AlloDerm Human Cadaveric
Acellular Dermal Matrices. Plast Reconstr Surg
2013; 5: 940-51.
30.Jansen LA, De Caigny P, Guay NA et al.: The
evidence base for the acellular dermal matrix AlloDerm: a systematic review. Ann Plast Surg 2013;
5: 587-94.
31.Callcut RA, Sshurr MJ, Sloan M et al.: Clinical
experience with Alloderm: a one-staged composite
dermal/epidermal replacement utilizing processed
cadevar dermis and thing autograft. Burns 2006;
32: 583-88.
32.Wainwright DJ: Use of an acellular allograft
dermal matrix (AlloDerm) in the management
of full-thickness burns. Burns 1995; 21: 24348.
33.Mirastschijski U, Kerzel C, Schnabel R et al.:
Complete horizontal skin cell resurfacing and delayed vertical cell infiltration into porcine reconstructive tissue matrix compared to bovine collagen
matrix and human dermis. Plast Reconstr Surg
2013; 4: 861-69.
34.Khodadadi L, Shafieyan S, Sotoudeh M et al.:
Intraepidermal injection of dissociated epidermal
cell suspension improves vitiligo. Arch Dermatol
Res 2010; 8: 593-99.
35.Brown AC, Rowe JA, Barker TH: Guiding epithelial cell phenotypes with engineered integrinspecific recombinant fibronectin fragments. Tissue
Eng Part A 2011,1-2: 139-50.
Unauthenticated
Download Date | 6/18/17 6:34 PM
Substitute of an acellular allogeneic dermal matrix in clinical practice
36.Marcelo D, Beatriz PM, Jussara R, Fabiana B:
Tissue therapy with autologous dermal and epidermal culture cells for diabetic foot ulcers. Cell Tissue
Bank 2012; 2: 241-49.
37.Kulig KM, Luo X, Finkelstein EB et al.: Biologic
properties of surgical scaffold materials derived from
dermal ECM. Biomaterials 2013; 23: 5776-84.
38.Rodrigues AZ, de Oliveira PT, Novaes Jr AB et
al.: Evaluation of in vitro human gingival fibroblast
521
seeding an Acellular Dermal Matrix. Braz Dent J
2010,21: 179-185.
39.Sorrel JM, Caplan AI: Fibroblast heterogeneity:
more than skin deep. J Cell Science 2004; 117: 667675.
40.Ng KW, Khor HL, Hutmacher DW: In vitro
characterisation of natural and synthetic dermal
matrices cultured with human dermal fibroblasts.
Biomaterials 2004; 25: 2807-2818.
Received: 20.10.2015 r.
Adress correspondence: 41-100 Siemianowice Śląskie, ul. Jana Pawła II 2
Unauthenticated
Download Date | 6/18/17 6:34 PM