A mouse perfusion model using fluorescently labeled dextran has been developed to investigate the functionality of blood vessels during cutaneous wound healing. By immunostaining cryostat sections of perfused wounds with antibodies that identify vessels, we were able to assess their functionality. There was an increase in the proportion of CD31(+)-perfused vessels in all wound regions with time, although the vessels of the wound margins and superficial granulation tissue (GT) took the longest to become perfused. More than 50% of the latter vessels were not perfused at 10 days postwounding. This is consistent with the growth of functional vessels from the wound base proceeding to the more superficial GT. The CD34 marker was expressed by a subpopulation of CD31(+) vessels. However, in contrast to CD31(+) vessels, the functionality of CD34(+) vessels did not change significantly with time and 50-75% of CD34(+) vessels in the GT and wound margins were nonfunctional. This might be explained either by apoptosis of the CD34(+) vessels or the loss of the marker with time. This study has important implications for assays of wound-healing angiogenesis based on histology and immunohistochemical markers for vessels, because vessel functionality differs both spatially and temporally during wound healing.

译文

:已经开发了使用荧光标记的葡聚糖的小鼠灌注模型,以研究皮肤伤口愈合过程中血管的功能。通过用识别血管的抗体免疫染色灌注伤口的低温恒温器切片,我们能够评估其功能。尽管伤口边缘的血管和浅表肉芽组织(GT)的灌注时间最长,但所有伤口区域中CD31()灌注血管的比例均随时间增加。受伤后10天,没有再灌注50%以上的血管。这与功能性血管从伤口基部到更浅层的GT的生长是一致的。 CD34标记由CD31()血管的亚群表达。然而,与CD31()血管相反,CD34()血管的功能并未随时间而显着变化,GT中50-75%的CD34()血管和伤口边缘均无功能。这可能是由于CD34()血管凋亡或标记随时间流逝而造成的。这项研究对基于组织学和血管免疫组织化学标记的伤口愈合血管生成的测定具有重要意义,因为在伤口愈合过程中血管功能在空间和时间上都不同。

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