Osteoblastic Differentiation of hMSCs exposed to Alternating Current Electric Fields

Mesenchymal stem cells (MSCs) have become a major research area for the past decade. Their tremendous ability to regenerate and to differentiate into different cell types has given hope in tissue regeneration and wound healing. The ability to control their differentiation in vitro has made possible to develop various types of tissue engineered products. Osteoblastic differentiation of MSCs which is considered as the process involved in bone healing, can be achieved fast when exposed to alternating current (A.C) electric fields

Development of an experimental model of a decellularized kidney scaffold by perfusion mode and analyzing the three-dimensional extracellular matrix architecture by edge detection method

Renal transplant is treatment of choice for the patients with end stage renal disease. The kidney transplants are expensive and there are risks of immunological and infectious complications. We planned to develop an in vitro decellularized kidney scaffold model using sheep kidney. Kidney decellularization was carried out by perfusing chemical detergents such as sodium dodecyl sulfate (SDS), SDS and trypsin, and SDS and ethylenediaminetetraacetic acid solvent solution. Complete kidney was decellularized in 5 days by perfusing various chemical detergents in time-dependent intervals. Histological finding revealed the complete removal of cellular material in various regions of renal corpuscle, distal convoluted tubules, other cortex and medulla region. Details of interlobular veins and arteries were seen through naked eyes after trypan blue dye injection. We used edge detection technique for developing a three-dimensional (3-D) image (Image J software) for nephrological vasculature constructed of decellularized kidney scaffold specimen. This technique opens a gateway for the whole organ decellularization by perfusion technology and further imaging of its 3-D extracellular matrix texture by edge detection technique software

Regional Differentiation of Adipose-Derived Stem Cells Proves the Role of Constant Electric Potential in Enhancing Bone Healing

Mesenchymal stem cells (MSCs) have a great potential in the field of tissue engineering and regenerative medicine on account of their ability to self-renew and differentiate into various lineages. MSCs could be differentiated by a number of ways. Electric field is known to bring about differentiation, migration, proliferation, and reorientation of MSCs. Hence, we aim to create a bioreactor to attain osteodifferentiation of human-derived MSCs in the presence of osteoinduction medium (OIM) in combination with or without alternating current (AC) fields. A stimulation bioreactor was specially designed for the exposure of adipose-derived stem cells (ASCs) to an electric field of 20 mV/cm, 60 kHz. The electric field potential (E) within the chamber was simulated using COMSOL. The morphology, proliferation, and osteogenic differentiation of ASCs under the influence of electrical stimulation were studied. By week three, electrically stimulated ASCs exhibited their typical spindle-shaped morphology. Stimulated ASCs were more intensely stained with alkaline phosphatase and alizarin red, the markers of osteogenic differentiation, as compared to the unstimulated control groups. Darker stained regions correlated with the COMSOL simulation which showed constant electric potential at the same place. The results depicted a clear difference between the effect of constant and varying electric potential on osteodifferentiation of ASCs. Picogreen assay revealed lower DNA contents of electrically stimulated ASCs compared to the control group. In this study, we have additively enhanced the osteodifferentiation potential of ASCs by electrical stimulation and have proved that it is constant electric field potential which specifically augments osteogenic differentiation. We have successfully developed a bioreactor to improve the osteodifferentiation of ASCs by an electrical field, which could be applied in regenerative therapy strategies of bone fracture treatment