The Effect of Mummy Substance on Matrix Protein Synthesis by Human Adipose-Derived Stem Cells and Dermal Fibroblast and Their Behavior on Plated PCL Scaffold

Abstract

Objectives: Wounds repair has always been regarded as a challenge in health world due to its high prevalence characteristics. Therefore, there are many proposed therapeutic strategies for repairing wounds in which the use of herbal medicines is highlighted because of the low costs and complications. In traditional medicine, 'Mummy' is regarded as almost a first choice for some treatments such as bone fractures, bleeding control, poisoning treatment, headache relief and wound repair. This study seeks to provide a scientific venture for the effect of mummy on wound healing. Materials and Methods: Human fetal foreskin fibroblast cells (HFFF-2) were purchased from Pasteur Institute (Tehran, Iran). The fibroblast cell lines and stem cells derived from adipose-derived stem cells (ASCs) were isolated by means of explant culture. Synthesis of components of extracellular matrix (ECM) such as collagen type I, type III and fibronectin 1 (FN1) were determined by real-time polymerase chain reaction (PCR) and cells attachments were examined by scaffold and cell proliferation determined by MTT assay. ASCs and HFFF-2, were investigated under mono-culture experimental conditions, then the other two-cell co-culture cells were also investigated under laboratory condition with 50-50 and 30-70 ratio respectively. They were all in an experimental group including culture medium and mummy; on the other side, in the control group there were just culture medium. Results: The results suggested that in fibroblasts, the level of mRNA expression of fibronectin was up-regulated in the treatment group (P<0.0001), it was up-regulated Col type I (P < 0.0001) and Col type III (P<0.01) in ASCs. In co-culture, mRNA expression of Col type I, III, and fibronectin increased (P<0.0001). The cells were successfully penetrated and adhered, and spread on PCL (polycaprolactone) scaffolds in all groups. Higher proliferation rate of fibroblasts was observed in Mummy-treated cells compared to the control group after 24 hours (P<0.0001). Increased proliferation rate in ASCs was seeded on scaffolds and treated with mummy compared to control groups for 24 hours (P< 0.01) and 96 hours (P<0.0001). Proliferation rate in co-cultured ASCs and fibroblasts (proportion of 50-50 and treated with mummy) was higher after 24 hours compared to the control cells (P<0.0001); but in co-cultured ASCs and fibroblasts, proportion of 70-30 was significantly lower in Mummy-treated group compared to the control group (P <0.0001) at both lengths of time. Conclusions: mummy may improve wound healing through synthesizing ECM attachment and proliferating cells on PCL scaffold.