Chung Eui-Sang, Saban Daniel R, Chauhan Sunil K, Dana Reza
Schepens Eye Research Institute, Boston, Massachusetts 02114, USA.
Invest Ophthalmol Vis Sci. 2009 Apr;50(4):1613-8. doi: 10.1167/iovs.08-2212. Epub 2008 Nov 21.
In the present study, the authors developed novel models to stimulate blood vessel formation (hemangiogenesis) versus lymphatic vessel formation (lymphangiogenesis) in the cornea.
Micropellets loaded with high-dose (80 ng) or low-dose (12.5 ng) basic fibroblast growth factor (bFGF) were placed in BALB/c corneas. Angiogenic responses were analyzed by immunohistochemistry to quantify blood neovessels (BVs) and lymphatic neovessels (LVs) to 3 weeks after implantation. Areas covered by BV and LV were calculated and expressed as a percentage of the total corneal area (percentage BV and percentage LV). Hemangiogenesis (HA) and lymphangiogenesis (LA) were also assessed after antibody blockade of VEGFR-2 or VEGFR-3
Although high-dose bFGF stimulation induced a more potent angiogenic response, the relative LV (RLV=percentage LV/percentage BV x 100) was nearly identical with high- and low-doses of bFGF. Delayed LA responses induced 3 weeks after implantation of high-dose bFGF resulted in a lymphatic vessel-dominant phenotype. Interestingly, the blockade of VEGFR-2 significantly suppressed BV and LV. However, the blockade of VEGFR-3 inhibited only LV (P=0.0002) without concurrent inhibition of BV (P=0.79), thereby resulting in a blood vessel-dominant phenotype
An HA-dominant corneal phenotype can be obtained in BALB/c mice 2 weeks after implantation of an 80-ng bFGF micropellet with VEGFR-3 blockade. Alternatively, an LA-dominant corneal phenotype can be obtained 3 weeks after implantation of an 80-ng bFGF micropellet without supplementary modulating agents. These models will be useful in evaluating the differential contribution of BV and LV to a variety of corneal abnormalities, including transplant rejection, wound healing and microbial keratitis.
在本研究中，作者开发了新模型以刺激角膜中的血管生成（血管新生）与淋巴管生成（淋巴管新生）。
将装载高剂量（80 ng）或低剂量（12.5 ng）碱性成纤维细胞生长因子（bFGF）的微颗粒置于BALB/c小鼠角膜中。通过免疫组织化学分析血管生成反应，以量化植入后3周内的新生血管（BV）和新生淋巴管（LV）。计算BV和LV覆盖的面积，并表示为角膜总面积的百分比（BV百分比和LV百分比）。在对VEGFR-2或VEGFR-3进行抗体阻断后，也评估了血管新生（HA）和淋巴管新生（LA）。
尽管高剂量bFGF刺激诱导了更强的血管生成反应，但高剂量和低剂量bFGF的相对LV（RLV = LV百分比/BV百分比×100）几乎相同。高剂量bFGF植入3周后诱导的延迟LA反应导致淋巴管占主导的表型。有趣的是，VEGFR-2的阻断显著抑制了BV和LV。然而，VEGFR-3的阻断仅抑制LV（P = 0.0002），而未同时抑制BV（P = 0.79），从而导致血管占主导的表型。
在植入80 ng bFGF微颗粒并阻断VEGFR-3的情况下，BALB/c小鼠在植入后2周可获得HA占主导的角膜表型。或者，在不使用补充调节剂的情况下，植入80 ng bFGF微颗粒3周后可获得LA占主导的角膜表型。这些模型将有助于评估BV和LV对各种角膜异常（包括移植排斥、伤口愈合和微生物性角膜炎）的不同作用。
