PMID- 31559928
OWN - NLM
STAT- MEDLINE
DCOM- 20210527
LR  - 20210527
IS  - 1937-335X (Electronic)
IS  - 1937-3341 (Linking)
VI  - 26
IP  - 3-4
DP  - 2020 Feb
TI  - Exogenous Keratinocyte Growth Factor Is Not Required for Pigmentation of Skin 
      Substitutes with Three Isogeneic Cell Types.
PG  - 214-224
LID - 10.1089/ten.TEA.2019.0203 [doi]
AB  - Engineered skin substitutes (ESS) containing human fibroblasts (hF) and human 
      keratinocytes (hK) provide significant medical benefits for treatment of acute 
      and chronic skin wounds, including, but not limited to, burns, burn scars, 
      congenital skin lesions, and cutaneous ulcers. However, anatomic deficiencies, 
      such as lack of pigment, can contribute to long-term morbidity, including 
      hypopigmentation and reduced solar protection. To address the deficiency of 
      hypopigmentation, ESS were populated sequentially with cultured hF, human 
      melanocytes (hM), and hK to generate ESS with pigment (ESS-P). Constructs were 
      incubated in media containing 0.0, 1.5, or 5.0 ng/mL keratinocyte growth factor 
      (KGF), which promotes survival and differentiation of hM in ESS-P, and had media 
      changed at 24 or 48 h intervals. ESS-P were evaluated in vitro for surface 
      hydration, surface color, and distribution of hM. Proliferation was assessed by 
      measuring incorporation of 5-bromo-2'-deoxyuridine into replicating DNA in basal 
      epidermal cells. ESS-P from test conditions were grafted to immunodeficient mice, 
      and were assessed over 12 weeks for pigmented area, pigment density, and 
      distribution of hM in healed human grafts. The in vitro data showed differences 
      among test groups, including increase in hydration of the epidermal surface with 
      higher KGF, increase of surface pigmentation with 24 h media changes, increase of 
      hM density with higher KGF and 24 h media changes, and time-dependent decrease of 
      proliferation. At 12 weeks after grafting, differences among groups were found 
      for pigment density, but not for distribution of hM or percentage of pigmented 
      area. These differences demonstrate that a higher concentration of KGF (5 ng/mL) 
      in the maturation medium of ESS-P and more frequent media changes (24 h interval) 
      promote higher viability and hM differentiation of ESS-P before grafting, but are 
      not required for full pigmentation (pigmented area, pigment density, hM 
      distribution) of grafted wounds. Based on these results, reductions of the 
      concentration of KGF (i.e., 1.5 ng/mL) in the maturation medium, and of the 
      frequency of medium changes (48 h intervals) would be expected to support 
      survival, continued replication, and restoration of skin color by hM in 
      therapeutic transplantation of ESS-P. Impact Statement Restoration of skin color 
      after traumatic injury affects personal identity and provides protection from 
      exposure to solar radiation. Keratinocyte growth factor (KGF) and nutrient supply 
      are known to regulate survival of melanocytes before transplantation in 
      engineered skin substitutes with pigment (ESS-P). This report demonstrates that 
      exogenous KGF is not required to restore skin color and that replacement of the 
      nutrient medium at lower frequency (48 versus 24 h) does not inhibit development 
      of skin color after melanocyte transplantation. These results offer new 
      alternatives to conserve resources in fabrication of ESS-P and to maintain 
      efficacy for restoration of skin color.
FAU - Boyce, Steven T
AU  - Boyce ST
AD  - Research Department, Shriners Hospitals for Children, Cincinnati, Ohio.
AD  - Department of Surgery, University of Cincinnati, Cincinnati, Ohio.
FAU - Supp, Dorothy M
AU  - Supp DM
AD  - Research Department, Shriners Hospitals for Children, Cincinnati, Ohio.
AD  - Department of Surgery, University of Cincinnati, Cincinnati, Ohio.
FAU - Lloyd, Christopher M
AU  - Lloyd CM
AD  - Department of Surgery, University of Cincinnati, Cincinnati, Ohio.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20191101
PL  - United States
TA  - Tissue Eng Part A
JT  - Tissue engineering. Part A
JID - 101466659
SB  - IM
MH  - Cells, Cultured
MH  - Female
MH  - Humans
MH  - Keratinocytes/*cytology/*metabolism
MH  - Melanocytes/*cytology/*metabolism
MH  - Regenerative Medicine/*methods
MH  - *Skin, Artificial
MH  - Tissue Engineering/*methods
OTO - NOTNLM
OT  - engineered skin
OT  - epidermis
OT  - regenerative medicine
OT  - tissue engineering
OT  - wounds
EDAT- 2019/09/29 06:00
MHDA- 2021/05/28 06:00
CRDT- 2019/09/28 06:00
PHST- 2019/09/29 06:00 [pubmed]
PHST- 2021/05/28 06:00 [medline]
PHST- 2019/09/28 06:00 [entrez]
AID - 10.1089/ten.TEA.2019.0203 [doi]
PST - ppublish
SO  - Tissue Eng Part A. 2020 Feb;26(3-4):214-224. doi: 10.1089/ten.TEA.2019.0203. Epub 
      2019 Nov 1.