Objective assessment of endogenous collagen in vivo during tissue repair by laser induced fluorescence.

Objective assessment of endogenous collagen in vivo during tissue repair by laser induced fluorescence.
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通过激光诱导的荧光在组织修复过程中内源性胶原蛋白的客观评估。

DOI:
10.1371/journal.pone.0098609
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发表时间:
2014
期刊:
影响因子:
3.7
通讯作者:
Mahato KK
Mahato KK
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Prabhu V;Rao SB;Fernandes EM;Rao AC;Prasad K;Mahato KK

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胶原蛋白是一种三螺旋蛋白,主要起机械作用,为皮肤提供拉伸强度,在组织修复中起关键作用。在组织再生过程中,胶原蛋白水平逐渐增加,因此,通过激光诱导荧光监测体内这种变化是本研究的主要目的。为了实现这一点,在6至8周龄的瑞士白化病小鼠上产生15 mm直径的切除伤口。用沿着激光诱导自体荧光技术,观察了在伤后即刻用2 J/cm ~ 2 He-Ne激光照射加速胶原沉积的情况。分别于伤后0、5、10、30、45和60天,用325 nm He-Cd激光激发肉芽组织/皮肤,记录各组动物的自体荧光光谱。根据年龄和性别检查来自小鼠肉芽组织/皮肤的平均胶原蛋白强度的变化。此外,伤口肉芽组织/未受伤皮肤组织中胶原蛋白合成的光谱发现通过Picro-Sirius红色偏振光显微镜以盲法通过相应胶原蛋白双折射的图像分析进行验证。体内自体荧光研究表明,与未照射的对照组相比,激光处理的动物中胶原蛋白合成显著增加。胶原双折射的图像分析进一步验证了自体荧光在体内胶原客观监测中的能力。我们的研究结果清楚地表明了激光诱导自体荧光在监测胶原合成组织再生过程中的潜力,这可能具有临床意义。
Collagen, a triple helical protein with the primary role of mechanical function, provides tensile strength to the skin, and plays a pivotal task in tissue repair. During tissue regeneration, collagen level increases gradually and therefore, monitoring of such changes in vivo by laser induced fluorescence was the main objective behind the present study. In order to accomplish this, 15 mm diameter excisional wounds were created on six to eight week old Swiss albino mice. The collagen deposition accelerated upon irradiation of single exposure of 2 J/cm2 He-Ne laser dose immediately after wounding was recorded by laser induced autofluorescence in vivo along with un-illuminated and un-wounded controls. Autofluorescence spectra were recorded for each animal of the experimental groups on 0, 5, 10, 30, 45 and 60 days post-wounding, by exciting the granulation tissue/skin with 325 nm He-Cd laser. The variations in the average collagen intensities from the granulation tissue/skin of mice were inspected as a function of age and gender. Further, the spectral findings of the collagen synthesis in wound granulation tissue/un-wounded skin tissues were validated by Picro-Sirius red- polarized light microscopy in a blinded manner through image analysis of the respective collagen birefringence. The in vivo autofluorescence studies have shown a significant increase in collagen synthesis in laser treated animals as compared to the un-illuminated controls. Image analysis of the collagen birefringence further authenticated the ability of autofluorescence in the objective monitoring of collagen in vivo. Our results clearly demonstrate the potential of laser induced autofluorescence in the monitoring of collegen synthesis during tissue regeneration, which may have clinical implications.
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