The Investigation of Viability and Proliferation Potential of Mesenchymal Stem Cells Entrapped in Chitosan Hydrogel

Introduction: Diabetic ulcer is classified as chronic ulcer due to hyperglycemic conditions, growth of bacteria and lack of blood supply, and unfortunately, today's treatment methods have not achieved the desired results. In this research, we used Adipose-Derived Mesenchymal Stromal/Stem Cells (AD-MSCs) to investigate the growth, proliferation and survival of these cells encapsulated chitosan hydrogel, due to their unique characteristics such as high accessibility with minimal invasion and no ethical restrictions and their immunomodulatory potential. One of the remarkable features of chitosan hydrogel is its intrinsic antimicrobial property, which causes microbial defense in the wound area. The innovation of this research in the synthesis of chitosan hydrogel is due to the presence of abundant water and the simulation of extracellular matrix space for these cells. By combining these cells with hydrogel, we examine the adhesion and functional properties of these cells.Methods and Results:To isolate AD-MSCs, we washed the sample obtained from liposuction three times with PBS solution containing antibiotics, and then we used a fresh solution of collagenase supplemented with bovine serum albumin, and calcium chloride. In order to digest adipose tissues, they were added to the above solution. After incubation and centrifugation, the cell pellet was suspended in the DMEM culture medium (%۱۰ fetal bovine serum) and transferred to the flask to be maintained in the incubator at ۳۷. For the synthesis of chitosan hydrogel, we combined ۳.۳۳% chitosan solution with ۰.۵ M β-GP solution. In order to evaluate the adhesion and viability of the cells after ۱, ۳ and ۵ days, we used ۲۴-well plates to culture AD-MSCs in combination with chitosan hydrogel. At each time point, the survival and proliferation of cells were examined by the following tests: FDA/PI staining and evaluation by a fluorescent microscope were performed and the result showed ۹۲% of survival. The electron microscope. (FE-SEM) images also indicated the distribution of highly proliferated adhering cells. Conclusion:This research shows that chitosan hydrogel provides optimal conditions for the growth, proliferation and survival of AD-MSCs in vitro and can be introduced as a suitable option as a biological dressing in wound healing.

کلیدواژه ها:

Wound healing ، Chitosan hydrogel ، AD-MSCs