Effective Wound Healing in Streptozotocin-Induced Diabetic Rats by Adipose-Derived Stromal Cell Transplantation in Plasma-Gel Containing Fragmin/Protamine Microparticles

Abstract

We investigated the effectiveness of the application of inbred adipose-derived stromal cells (IR-ASCs) in high inbred rat plasma (IRP) (6%)-Dulbecco modified Eagle medium (DMEM) gel with fragmin/protamine microparticles (F/P MPs) (IR-ASCs + IRP-DMEM gel + F/P MPs) on wound healing in streptozotocin-induced diabetic rats. F/P MPs have previously been used as a cell carrier for IR-ASCs in inbred Fisher 344 rats and for preservation and controlled release of various cytokines in IRP-DMEM gel. We applied IR-ASCs + IRP-DMEM gel + F/P MPs to full-thickness skin excisions on the backs of the diabetic rats. The statistical significance of wound closure was evaluated on postwounding days 3, 7, 10, and 14, and the skin area surrounding the wound was removed for histological examination on days 7 and 14. The wound closure rate and histological examination of wounds treated with IR-ASCs + IRP-DMEM gel + F/P MPs demonstrated significantly advanced epithelialization, capillary formation, and granulation tissue formation. When DiI-labeled IR-ASCs + IRP-DMEM gel + F/P MPs were applied to full-thickness skin wounds on the backs of the diabetic rats, histological observation at 2 weeks showed appearances of both DiI-labeled granulation tissue and CD31-immunostained microvessels in the transplant areas. A portion of the transplanted IR-ASCs + IRP-DMEM gel + F/P MPs had been taken up into the granulation tissues to promote wound healing. Thus, IR-ASCs + IRP-DMEM gel + F/P MPs were effective for repairing healing-impaired wounds such as those arising in the diabetic rats.

Enhanced healing of mitomycin C-treated healing-impaired wounds in rats with PRP-containing fragmin/protamine microparticles (PRP&F/P MPs)

“…Although the mechanism of well-known healing-impaired wounds such as pressure or diabetic ulcer in humans may be different from those in mitomycin C-treated wounds, the PRP&F/P MPs-treatment enhanced the healing of mitomycin C-treated wounds in this study. In addition, recently we demonstrated effective wound healing in streptozotocin-induced diabetic rats by PRP&F/P MPs-treatment [26]. Thus, we believe that mytomycin Ctreated wounds in rats are relevant to general healing-impaired wounds.…”

Biomedical Application of Low Molecular Weight Heparin/Protamine Nano/Micro Particles as Cell- and Growth Factor-Carriers and Coating Matrix

“…The 3D culture methods presented required no animal serum, because the low concentration (3%) of autologous plasma was sufficient. Furthermore, we have successfully applied 3D-cultured IR-ADSCs using injectable low IRP (3%)-DMEM gel with LMWH/P N/MPs to in vivo cell transplantation treatment for repairing healing-impaired wounds such as those occurring in diabetes [58,59]. …”

“…Figure 8B showed the application of IR-ADSCs using IRP (6%)-DMEM gel with LMWH/P N/MPs/FGF-2 to full thickness skin excisions as healing-impaired wounds on the backs of STZ-induced diabetic rats [59]. The wound closures treated with IR-ADSCs using IRP-DMEM gel with LMWH/P N/MPs/FGF-2 were significantly enhanced on post-wounding [59]. The histological examination of wounds treated with IR-ADSCs using IRP-DMEM gel with LMWH/P N/MPs/FGF-2 demonstrated significantly advanced epithelialization, capillary formation and granulation tissue formation.…”

“…The histological examination of wounds treated with IR-ADSCs using IRP-DMEM gel with LMWH/P N/MPs/FGF-2 demonstrated significantly advanced epithelialization, capillary formation and granulation tissue formation. Those results indicate that a portion of the transplanted IR-ADSCs using IRP-DMEM gel with LMWH/P N/MPs/FGF-2 had been taken up by the granulation tissues to promote wound healing [59]. Thus, those studies suggest that IR-ADSCs using IRP-DMEM gel with LMWH/P N/MPs is effective for repairing healing-impaired wounds such as those occurring in diabetes.…”

Heparinoid Complex-Based Heparin-Binding Cytokines and Cell Delivery Carriers

“…We previously applied 3D-cultured IR-ADSCs that were derived from inbred male Fisher 344 rats while using injectable low IRP (3%)-DMEM gel with LMWH/P N/MPs for cell transplantation [175]. Furthermore, IR-ADSCs using IRP (6%)-DMEM gel with LMWH/P N/MPs/FGF-2 were applied to full thickness skin excisions as healing-impaired wounds on the backs of STZ-induced diabetic rats [176]. The wound closures that were treated with IR-ADSCs while using IRP-DMEM gel with LMWH/P N/MPs/FGF-2 were significantly enhanced in post-wounding [176] (Table 4).…”