Fukui Tsuyoshi, Kawaguchi Akira T, Takekoshi Susumu, Miyasaka Muneo, Sumiyoshi Hideaki, Tanaka Rica
Department of Plastic Surgery, Tokai University School of Medicine, Kanagawa.
Department of Cell Transplantation and Regenerative Medicine, Tokai University School of Medicine, Kanagawa.
Artif Organs. 2017 Apr;41(4):319-326. doi: 10.1111/aor.12864. Epub 2017 Mar 21.
Since liposome-encapsulated hemoglobin with high O affinity (h-LEH, P O = 10 mm Hg) has been reported to accelerate skin wound healing in normal mice, it was tested in dB/dB mice with retarded wound healing, as seen in human diabetics. Two full-thickness dorsal wounds 6 mm in diameter encompassed by silicone stents were created in dB/dB mice. Two days later (day 2), the animals were randomly assigned to receive intravenous h-LEH (2 mL/kg, n = 7) or saline (2 mL/kg, n = 7). The same treatment was repeated 4 days after wounding (day 4), and the size of the skin lesions was analyzed by photography, surface perfusion was detected by Laser-Doppler imager, and plasma cytokines and chemokines were determined on days 0, 2, 4, and 7, when all animals were euthanized for morphological studies. The size of the ulcer compared to the skin defect or silicone stent became significantly reduced on days 4 and 7 in mice treated with h-LEH (47 ± 8% of original size), similar to the level in wild-type mice, compared to saline-treated dB/dB mice (68 ± 18%, P < 0.01). Mice treated with h-LEH had significantly attenuated inflammatory cytokines, increased surface perfusion, and increased Ki67 expression on day 7 in accordance with the ulcer size reduction, while there was no significant difference in chemokines, histological granulation, epithelial thickness, and granulocyte infiltration detected by immunohistochemical staining in the ulcer between the treatment groups. The results suggest that h-LEH (2 mL/kg) early after wounding may accelerate skin wound healing in dB/dB mice to levels equivalent to wild-type mice probably via mechanism(s) involving reduced hypoxia, increased surface perfusion, suppressed inflammation, accelerated in situ cell proliferation and protein synthesis.
由于据报道，具有高氧亲和力的脂质体包裹血红蛋白（h-LEH，P50 = 10 mmHg）可加速正常小鼠的皮肤伤口愈合，因此在伤口愈合延迟的db/db小鼠中进行了测试，这种情况在人类糖尿病患者中也可见。在db/db小鼠的背部制造两个直径6毫米、由硅胶支架包围的全层伤口。两天后（第2天），将动物随机分为两组，分别接受静脉注射h-LEH（2 mL/kg，n = 7）或生理盐水（2 mL/kg，n = 7）。在受伤后4天（第4天）重复相同的治疗，通过拍照分析皮肤损伤的大小，用激光多普勒成像仪检测表面灌注情况，并在第0、2、4和7天测定血浆细胞因子和趋化因子，此时所有动物均处死后进行形态学研究。与生理盐水处理的db/db小鼠相比（68 ± 18%，P < 0.01），用h-LEH处理的小鼠在第4天和第7天，溃疡相对于皮肤缺损或硅胶支架的大小显著减小（为原始大小的47 ± 8%），与野生型小鼠的水平相似。与溃疡大小减小一致，用h-LEH处理的小鼠在第7天炎症细胞因子显著减弱，表面灌注增加，Ki67表达增加，而治疗组之间在趋化因子、组织学肉芽组织、上皮厚度以及通过免疫组织化学染色检测的溃疡中的粒细胞浸润方面没有显著差异。结果表明，受伤后早期给予h-LEH（2 mL/kg）可能通过减少缺氧、增加表面灌注、抑制炎症、加速原位细胞增殖和蛋白质合成等机制，将db/db小鼠的皮肤伤口愈合加速至与野生型小鼠相当的水平。
