See Original Article in : https://ijcc.chemoprev.org/index.php/ijcc/article/download/346/225
ABSTRACT
BACKGROUND Full-thickness wound are areas damage of skin associated with loss of epidermis and
dermis. The wound healing mechanism consists proliferation, migration and remodeling.
Hypoxic conditional medium of mesenchymal stem cells (HMSCs-CM) contains lots of soluble
molecules, such as protein growth factor and cytokine anti-inflammation. The soluble molecule of HMSCs-CM plays a critical role in wound healing by upregulation of VEGF and collagen synthesis. The objective of this study was to evaluate the effect of HMSCs-CM on VEGF and collagen concentrations in rats with incised wounds. The methods of this study were an experimental animal study with post-test only control group design was performed involving 24 Wistar rats. The rats were randomized into four groups consisting of sham, control and two treatment groups (gel of HMSCs-CM at doses of 200 μL and 400 μL). The VEGF levels and collagen density were analyses using ELISA assay and Masson-trichome
specific staining, respectively. One-way ANOVA and Post Hoc LSD were used to analyses the data. The results of this study showed that a VEGF levels was significant increased on day 6 with doses-dependent manner. Interestingly, the VEGF levels gradual decrease on day 9. In addition, the decreased of VEGF levels on day 9 in this study in line with our findings in which we found there was a trend in the decreased of collagen density, it indicated the completion of remodeling phase and there has been an acceleration in wound healing. This study demonstrated that HMSCs-CM were able to regulate VEGF levels and collagen synthesis in accelerate wound healing. The role of HMSCs-CM stimulate cutaneous wound
healing should be clarified further.
METHODS The Induction of Full-thickness Wound Model
Twenty-four wistar rat (four to six-month old, male, body weight 250-300 grams) were ob-
tained from local farmer at the Java Rat Labs, Tembalang, Semarang, Indonesia. The full-thick-ness wound model was generated as described previously (Kuntardjo, et al., 2019). In brief, after the hair removal from the dorsal surface and anaesthesiae 6- mm full- thickness skin wounds were created on one side of the midline.
RESULTS MSCs Characterization and HMSCs-CM Profile, The MSCs were successfully isolated
from the umbilical cord, based on their adherence to plastic. After reaching 80% confluence, the cells were harvested for fourth passages. The isolated cells showed peculiar fibroblast-like (spindle shape) morphology assessed the marker expression of MSC using flowcytometry (Figure 1). Flowcy-tometry analysis showed that the MSCs cells express an MSC-specific marker including positive expression of CD105 (95.9%), CD73 (99.2%), and CD90 (99.9%) and lack of Lin (2.0%).
Keywords: hypoxic conditional medium of mesenchymal stem cells (HMSCs-CM), vascular
endothelial growth factor, collagen synthesis, paracrine factors
