Title : Tissue Engineering Approach to Decrease Complications after Surgical Treatment of Pressure Injuries in Spinal Cord Injured Patients
Pressure injuries (PI) are chronic wounds caused by increased pressure over a prolonged period of time. Interruption of blood circulation leads to a shortage of oxygen, nutrient supplies, accumulation of toxic by-products and tissue necrosis. Patients with spinal cord injury (SCI) are at high risk of PI because of impaired mobility, and in many cases lack of protective sensory perception. Currently, treatment of PI is associated with long hospitalization and recurrence of lesions remains a major problem. We hypothesized that collagen scaffolds, designed to precisely fit the wound and populated with autologous ADSC, re-suspended in autologous platelet-rich plasma (PRP), will accelerate and improve wound healing. Here, we aim to test and optimize in vitro various combinations of adipose-derived mesenchymal stem cells (ADSC), PRP and collagen scaffolds to promote faster PI recovery after surgical intervention. As reference, we also included in our experimental protocols cells isolated from the PI tissues, such as endothelial cells and fibroblasts. ADSC monolayers and ADSC-constructs were differentiated for 14 days towards adipogenic lineage using adipogenic medium supplemented with or without PRP, as well as different scaffolds. Outcomes were measures by cell proliferation assay, lipid quantification, DNA quantification and the expression of adipogenic genes, evaluated via qRT-PCR. ADSC could be successfully differentiated towards adipogenic lineage on the 3D cell-constructs, especially on porous scaffolds. Regarding the effects of PRP on the proliferation of ADSC, our results showed enhancement of cell proliferation, however it had an inhibitory effect on adipogenesis. ADSC and fibroblasts supported the formation of new blood vessel in 3D cultures by endothelial cells, detected by immunohistological staining with CD31 antibody. Based on the collection of these in vitro results, we plan to optimize a protocol which can be in future implemented into surgical treatment of PI, to reduce their recurrence in SCI people.