Wound-healing potential of Cultured Epidermal Sheets is unaltered after lyophilization: a preclinical study in comparison to cryopreserved CES.
Lyophilized Cultured Epidermal Sheets (L-CES) have been reported to be as effective as the cryopreserved CES (F-CES) in treating skin ulcers. However, unlike F-CES, no preclinical study assessing wound-healing effects has been conducted for L-CES. The present study was set out to investigate the microstructure, cytokine profile, and wound-healing effects of L-CES in comparison to those of F-CES. Keratinocytes were cultured to prepare CES, followed by cryopreservation at -70 °C and lyophilization. Under microscopic observation, intact cells with apparent intracellular junctions were observed in L-CES. The L-CES, like fresh CES, consisted of three to four well-maintained epidermal layers, as shown by the expression of keratins, involucrin, and p63. There were no differences in the epidermal layer or protein expression between L-CES and F-CES, and both CES were comparable to fresh CES. TGF-α, EGF, VEGF, IL-1α, and MMPs were detected in L-CES at levels similar to those in F-CES. In a mouse study, wounds treated with L-CES or F-CES completely healed at least 4 days faster than untreated wounds. CES-treated wounds completely healed by day 10, while the untreated wounds did not heal by day 14. Masson's trichrome staining showed that collagen deposition in the CES-treated wounds was highly increased in the dermis of the wound center compared to that in the control wounds. Thus, this study demonstrates that L-CES is as clinically effective as F-CES for wound treatment.
Cutigen Research Institute, Tego Science Inc., Daerung Technotown III, 448 Gasan-Dong, Gumcheon-Gu, Seoul 153-772, Republic of Korea.
H
H
Jang
Cutigen Research Institute, Tego Science Inc., Daerung Technotown III, 448 Gasan-Dong, Gumcheon-Gu, Seoul 153-772, Republic of Korea.
Y H
YH
Kim
Department of Biosystems & Biomaterials Science and Engineering, Seoul National University, Seoul 151-921, Republic of Korea.
M K
MK
Kim
Department of Biosystems & Biomaterials Science and Engineering, Seoul National University, Seoul 151-921, Republic of Korea.
K H
KH
Lee
Cutigen Research Institute, Tego Science Inc., Daerung Technotown III, 448 Gasan-Dong, Gumcheon-Gu, Seoul 153-772, Republic of Korea.
S
S
Jeon
D000818:Animals; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D015925:Cryopreservation; D000078404:Epidermal Cells; D005612:Freeze Drying; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000637:transplantation*; D051379:Mice; D012883:Skin Ulcer / Q000473:pathology / Q000628:therapy*; D059467:Transcriptome; D014945:Wound Healing