Efficacy of the therapy of 5-aminolevulinic acid photodynamic therapy combined with human umbilical cord mesenchymal stem cells on methicillin-resistant Staphylococcus aureus-infected wound in a diabetic mouse model

Abstract

BackgroundA distressing issue of diabetic ulcer (DU) is its poor healing feature with limited clinical solutions. We have previously shown that 5-aminolevulinic acid photodynamic therapy (ALA-PDT) is a promising alternative to the currently limited measures for DU. Mesenchymal stem cells (MSCs) transplantation has been believed to impose certain therapeutic effect on restoration of injury. Thus, this study aims to explore whether the combination of MSCs and ALA-PDT will exert a more advanced curative effect on DU. MethodsDiabetic mice were induced by intraperitoneal injection of streptozotocin (STZ, 60 mg/kg/d) for consecutive 5 days. A full-thickness skin injury (diameter 6 mm) was created in the center of the back of each mouse, and then 10 μl of methicillin-resistant Staphylococcus aureus (MRSA) suspension was added to establish an infected DU model. All DU models were randomly divided into four groups: Untreated group, MSCs group, ALA-PDT group, and ALA-PDT combined with human umbilical cord mesenchymal stem cells (hUC-MSCs) (ALA-PDT + MSCs) group. The wound sizes were recorded by a digital camera, and the healing rates were calculated using Image J software. Bacterial loads on wounds were measured using CFU (Colony forming units) analysis. The epithelialization, inflammatory cells infiltration and granulation tissue formation were monitored by Haematoxylin and eosin (H&E) staining, and the corresponding semi-quantitative score was matched. Growth and pro-inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA). ResultsEither ALA-PDT or injection of hUC-MSCs resulted in a rapid wound closure compared with the untreated, while their combination brought about the most prominent healing. On day 12, healing rates of the untreated, MSCs, ALA-PDT and ALA-PDT + MSCs were 40.56% ± 7.06%, 74.23 ± 4.83%, 84.03 ± 3.53%, 99.67 ± 0.49%, respectively. The bacterial burden reductions were approximately 1.58 logs (97.36%, P < 0.05), 2.34 logs (99.54%, P < 0.01), 4.50 logs (nearly 100%, P < 0.001) for MSCs, ALA-PDT and ALA-PDT + MSCs, respectively. Histology revealed reduced inflammatory cells and improved collagen precipitation and angiogenesis after hUC-MSCs and ALA-PDT treatment compared to the untreated. The combined therapy leaded to a more intact epithelium, similar to the healthy. Finally, ELISA revealed that the property of ALA-PDT to stimulate transforming growth factor-β1 (TGF-β1) and vascular endothelial growth factor (VEGF) and inhibit IL (interleukin) -1β and IL-6 outweighed that of hUC-MSCs, and this function of the combination overwhelmed that of any single therapy. ConclusionsOur findings indicated that the strategy of combining ALA-PDT with hUC-MSCs possessed a significantly enhanced therapeutic effect over either single therapy, providing a promising innovative therapeutic candidate for refractory wounds.