Amniotic fluid-derived multipotent stromal cells drive diabetic wound healing through modulation of macrophages

Abstract

Background Cutaneous wounds in patients with diabetes exhibit impaired healing due to physiological impediments and conventional care options are severely limited. Multipotent stromal cells (MSCs) have been touted as a powerful new therapy for diabetic tissue repair owing to their trophic activity and low immunogenicity. However, variations in sources and access are limiting factors for broader adaptation and study of MSC-based therapies. Amniotic fluid presents a relatively unexplored source of MSCs and one with wide availability. Here, we investigate the potential of amniotic fluid-derived multipotent stromal cells (AFMSCs) to restore molecular integrity to diabetic wounds, amend pathology and promote wound healing. Method We obtained third trimester amniotic fluid from term cesarean delivery and isolated and expanded MSCs in vitro. We then generated 10 mm wounds in Leprdb/db diabetic mouse skin, and splinted them open to allow for humanized wound modeling. Immediately after wounding, we applied AFMSCs topically to the sites of injuries on diabetic mice, while media application only, defined as vehicle, served as controls. Post-treatment, we compared healing time and molecular and cellular events of AFMSC-treated, vehicle-treated, untreated diabetic, and non-diabetic wounds. A priori statistical analyses measures determined significance of the data. Result Average time to wound closure was approximately 19 days in AFMSC-treated diabetic wounds. This was significantly lower than the vehicle-treated diabetic wounds, which required on average 27.5 days to heal (p < 0.01), and most similar to time of closure in wild type untreated wounds (an average of around 18 days). In addition, AFMSC treatment induced changes in the profiles of macrophage polarizing cytokines, resulting in a change in macrophage composition in the diabetic wound bed. We found no evidence of AFMSC engraftment or biotherapy induced immune response. Conclusion Treatment of diabetic wounds using amniotic fluid-derived MSCs encourages cutaneous tissue repair through affecting inflammatory cell behavior in the wound site. Since vehicle-treated diabetic wounds did not demonstrate accelerated healing, we determined that AFMSCs were therapeutic through their paracrine activities. Future studies should be aimed towards validating our observations through further examination of the paracrine potential of AFMSCs. In addition, investigations concerning safety and efficacy of this therapy in clinical trials should be pursued.

Collagen scaffolds derived from bovine skin loaded withMSCoptimizedM1macrophages remodeling and chronic diabetic wounds healing

“…From Days 0 to 7, no difference was recorded in the residual wounds among the groups; however, the exudate in the control (gauze) group had increased significantly. Previous studies also showed that the time of chronic wound healing in db/db mice was significantly longer than that of normal wounds 46 . Until Day 10, new granulation tissues (GTs) and epithelialization degrees were significantly increased in CBS‐MSCs when compared with those in the other groups.…”

“…Previous studies also showed that the time of chronic wound healing in db/db mice was significantly longer than that of normal wounds. 46 Until Day 10, new granulation tissues (GTs) and epithelialization degrees were significantly increased in CBS‐MSCs when compared with those in the other groups. The residual wound area (43.26 ± 4.60%) on Day 10 in CBS‐MSCs was significantly lower than that in the control (100.10 ± 0.05%, p < 0.05) (Figure 6c ).…”

Perinatal Stem Cell Therapy to Treat Type 1 Diabetes Mellitus: A Never-Say-Die Story of Differentiation and Immunomodulation

“…AFSCs can differentiate along adipogenic, osteogenic, myogenic, endothelial, neurogenic, and hepatic pathways [ 126 ]. Moreover, Mu and co-workers [ 129 ] fully differentiated AFSCs into IPCs though a multi-step protocol. The differentiated cells expressed insulin, Glut2, Nkx6.1, and glucokinase.…”

“…Administration of human AFSCs in a diabetic mouse model brought a reduction in the time of wound healing and a reduction of local inflammation, because of the increased secretion of IL-6, which is responsible for polarizing of macrophages in a pro-repair status [ 129 ]. Likewise, Sato, Y. et al [ 130 ] reported the ability of AFSCs to switch macrophages M1 to M2 in an animal model of perinatal sepsis, reducing inflammation.…”

Panthenol Citrate Biomaterials Accelerate Wound Healing and Restore Tissue Integrity

“…Various approaches have been developed to treat impaired diabetic wounds, such as the administration of cytokines and growth factors, [ 12–14 ] cell‐based therapies, [ 15,16 ] or negative pressure wound therapy (NPWT). [ 17 ] In recent years, many studies have focused on the development of biomimetic scaffold systems that are associated with peptide‐, cytokine‐, or growth factor‐based therapeutics.…”