Predominant control of PDGF/PDGF receptor signaling in the migration and proliferation of human adipose­derived stem cells under culture conditions with a combination of growth factors.

Human adipose-derived stem cells (hASCs) play important roles in regenerative medicine and tissue engineering. However, their clinical applications are limited because of their instability during cell culture. Platelet lysates (PLTs) contain large amounts of growth factors that are useful for manufacturing cellular products. Platelet-derived growth factor (PDGF) is a major growth factor in PLTs and a potent mitogen in hASCs. To optimize growth conditions, the effects of a combination of growth factors on the promotion of hASC proliferation were investigated. Moreover, PDGF-BB combined with vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) markedly enhanced the viability of hASCs compared with the effects of PDGF-BB alone. Neither VEGF nor HGF had any effect alone. All growth factor receptor inhibitors inhibited cell proliferation. Wound healing assays revealed that VEGF and HGF stimulated PDGF-dependent cell migration. The effects of these growth factors on the activation of their cognate receptors and signaling enzymes were assessed using immunoblotting. Phosphorylation of PDGF receptor (PDGFR)ß, VEGF receptor (VEGFR)2 and MET proto-oncogene and receptor tyrosine kinase was induced by PDGF-BB treatment, and was further increased by treatment with PDGF-BB/VEGF and PDGF-BB/HGF. The levels of phospho-ERK1/2 and phospho-p38MAPK were increased by these treatments in parallel. Furthermore, the expression levels of SRY-box transcription factor 2 and peroxisome proliferator-activated receptor g were increased in PDGF-BB-treated cells, and PDGF-BB played a dominant role in spheroid formation. The findings of the present study highlighted that PDGF/PDGFR signaling played a predominant role in the proliferation and migration of hASCs, and suggested that PDGF was responsible for the efficacy of other growth factors when hASCs were cultured with PLTs.

Beneficial Impact of "Supercharged" Pectoralis Major Musculocutaneous Flap With Indocyanine Green Angiography on Reconstruction in a Patient at High Risk for Necrosis.

Background: The pectoralis major musculocutaneous (PMMC) flap is a versatile and commonly used technique for reconstruction of the head and neck in patients at high risk in free tissue transfer procedures. In this report, a "supercharged" PMMC flap, in which the lateral thoracic artery was cut, preserved, and anastomosed to the cervical vessels, was developed to stabilize blood flow. Methods: The supercharged PMMC flap was introduced in a patient who was at high risk for partial necrosis of the flap and underwent surgery for reconstruction of the tongue. Intraoperative perfusion monitoring using laser-assisted indocyanine green angiography (ICGA) was also performed to verify the status of blood flow in the flap. Results: ICGA monitoring of blood flow in the flap revealed instability. Subsequently, a supercharged PMMC flap was applied, which prevented partial necrosis of the flap. Conclusions: Combined with ICGA, a supercharged PMMC flap contributed to successful reconstruction in a patient at high risk for necrosis of the flap region.

A case report of the first application of culture epithelial autograft (JACE®) for giant congenital melanocytic nevus after its approval in Japan.

In Japan, the JACE® cultured epidermal autograft (CEA) was approved and covered by public healthcare insurance for use in the treatment of giant congenital melanocytic nevus (GCMN) in 2016. We herein report the results of the application of JACE® after curettage and Q-switched ruby laser therapy. The current patient was the first patient with GCMN to be treated with JACE® since its approval. A 3-month-old girl had a hairy GCMN of 9.5 cm in diameter from her cheek to her temple on the left side of her face. We first performed curettage of the nevus on the temple and applied irradiation using a Q-switched ruby laser; however, erosion relapsed at 2 months after first surgery. After preparing JACE®, we performed curettage a second time at 7 months with irradiation of a Q-switched ruby laser and the application of the CEA. The CEA took successfully and the wound was completely epithelized at 1 week after grafting. Re-pigmentation is an important issue that remains to be solved; however, overcoming this would allow for a deeper abrasion or more intense laser irradiation to be performed in cases in which CEA will be subsequently applied.