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Effects of Hair Follicle Dermal Sheath Cells in Skin Wound Healing

Author: LiYouChen
Tutor: WangDeWen;LiuJie
School: PLA Military Academy of Medical Sciences
Course: Pathology and Pathophysiology
Keywords: hair follicle dermal sheath cells wound healing scaring
CLC: R363
Type: PhD thesis
Year: 2007
Downloads: 133
Quote: 1
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Abstract


Objective How to improve wound healing and reduce scaring is the aim of studies on wound therapeutics. A relation exists between wound healing and hair follicle numbers in local skin. It has been proved presently that the hair follicle dermal sheath cells have extensive adult stem cells activities. Therefore, it is necessary to reveal the role that the dermal sheath cells play in wound healing. The purpose of the study is to investigate the effects of dermal sheath cells in skin wound healing and hope to provide new ideas in wound therapeutics.Methods Rat vibrissa follicle dermal sheath cells were cultured by the method of tissue adherence and identified by immunocytochemistry and immunofluorescence. The growth characteristics and ultrastructural features of these cells were observed by MTT assay, light microscopy and electron microscopy. The experimental model was established by choosing the optimizing methods of cell labeling with BrdU and the style of cell transplantation. Scar volume quantity by three-dimensional reconstruction and \ultrastructure of scar observed by using scanning electron microscope (SEM) act as scar assessment system. Allogeneic dermal sheath cells labeled with BrdU were implanted into skin wounds of rat back. The process of wound healing and scar formation was estimated by pathomorphologicl observation qualitatively and image analysis quantitatively. The expression of some important protein related with healing in dermal sheath cells was detected by RT-PCR, immunocytochemistry, immunofluorescence and immunoblotting.Results (1) The dermal sheath cells grow slowly in primary culture and proliferate quickly after subculture. The cells are grown as clone-like cluster. The cells adhering on the dermal sheath look like spindles with big cell body. The cultured cells have a big nucleus with prominent nucleolus. Bundles of fine filament were noticed beneath the cytoplasmic membrane. (2) The experimental model of labeled cells transplantation in skin wound had been established. The cells labeled with 10μmol/L BrdU were implanted before cutting skin. The cells could be traced in healing tissues 14 days later. (3) Scar assessment system established in this study included qualitative assessment on collagenous fibers and fibrils of scar and quantitative assessment on scar volume. (4) The dermal sheath cells were implanted in wound. After operation, the rate of wound contraction of implanted group was higher than that of control group significantly (P<0.01). 7 days after operation, implanted cells were found in granulation tissues hyperdispersionally. After 14 days, labeled cells were reduced in healing tissues and showed apoptosis. The results of pathology studies showed that the wound healing process of implanted group was faster than that of control group, including granulation tissue formation, epidermis regeneration, collagen deposition, scar formation and tissue reconstruction. The scar volume had decreased tendency in implanted group. (5) The mRNA of procollagen- I was high expressed in dermal sheath cells. From protein level, dermal sheath cells expressed more bFGF, EGFR and TIMP1 than fibroblast from dermal.Conclusions The hair follicle dermal sheath cells are immature mesenchymal cells with potential proliferation activity. In skin wound healing, dermal sheath cells can form granulation tissue, synthesize collagen, accelerate wound contraction, promote healing process and reduce scar formation. These capabilities are related with some proteins participating healing high expressed in dermal sheath cells. The results of this study have revealed the biological characteristics of dermal sheath cells and provided the experimental evidence to their clinical application in wound healing.

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