激光照射软骨对软骨细胞基因表达及胶原基质的影响。
The effects of laser irradiation of cartilage on chondrocyte gene expression and the collagen matrix.
作者信息
Holden Paul K, Li Chao, Da Costa Victor, Sun Chung-Ho, Bryant Susan V, Gardiner David M, Wong Brian J F
机构信息
Department of Otolaryngology-Head and Neck Surgery, University of California, Irvine, California 92612, USA.
出版信息
Lasers Surg Med. 2009 Sep;41(7):487-91. doi: 10.1002/lsm.20795.
OBJECTIVES
Laser reshaping of cartilage is an emerging technology aimed at replacing conventional techniques for aesthetic and reconstructive surgery. Little is known about the mechanisms of wound healing following the photothermal heating during laser reshaping and, ultimately, how collagen remodels in the irradiated tissue. Healthy hyaline and elastic cartilage as found in the ear, nose, larynx, and trachea does not express collagen type I which is characteristic of fibro-cartilage and scar tissue. The aim of the study was to determine if collagen I and II gene expression occurs within laser irradiated rabbit septal cartilage.
METHODS
Nasal septum harvested from freshly euthanized New Zealand White rabbits were irradiated with an Nd:YAG laser. After 2 weeks in culture, the laser spot and surrounding non-irradiated regions were imaged using immunofluorescence staining and evaluated using reverse transcription polymerase chain reaction (RT-PCR) to determine the presence of collagen I and II, and ascertain collagen I and II gene expression, respectively.
RESULTS
All laser irradiated specimens showed a cessation in collagen II gene expression within the center of the laser spot. Collagen II was expressed in the surrounding region encircling the laser spot and within the non-irradiated periphery in all specimens. Immunohistochemistry identified only type II collagen. Neither collagen I gene expression nor immunoreactivity were identified in any specimens regardless or irradiation parameters.
CONCLUSIONS
Laser irradiation of rabbit septal cartilage using dosimetry parameters similar to those used in laser reshaping does not result in the detection of either collagen I gene expression or immunoreactivity. Only collagen type II was noted after laser exposure in vitro following cell culture, which suggests that the cellular response to laser irradiation is distinct from that observed in conventional wound healing. Laser irradiation of cartilage can leave an intact collagen matrix which likely allows chondrocyte recovery on an intact scaffold.
目的
软骨激光重塑是一项新兴技术，旨在替代美学和重建手术中的传统技术。关于激光重塑过程中光热加热后的伤口愈合机制，以及最终照射组织中胶原蛋白如何重塑，目前知之甚少。在耳朵、鼻子、喉和气管中发现的健康透明软骨和弹性软骨不表达I型胶原蛋白，而I型胶原蛋白是纤维软骨和瘢痕组织的特征。本研究的目的是确定I型和II型胶原蛋白基因表达是否会在激光照射的兔鼻中隔软骨中出现。
方法
从刚安乐死的新西兰白兔身上采集鼻中隔，用Nd:YAG激光进行照射。培养2周后，使用免疫荧光染色对激光光斑及其周围未照射区域进行成像，并使用逆转录聚合酶链反应（RT-PCR）进行评估，分别确定I型和II型胶原蛋白的存在，并确定I型和II型胶原蛋白基因的表达情况。
结果
所有激光照射的标本在激光光斑中心均显示II型胶原蛋白基因表达停止。在所有标本中，II型胶原蛋白在环绕激光光斑的周围区域以及未照射的周边区域均有表达。免疫组织化学仅鉴定出II型胶原蛋白。无论照射参数如何，在任何标本中均未鉴定出I型胶原蛋白基因表达或免疫反应性。
结论
使用与激光重塑中相似的剂量参数对兔鼻中隔软骨进行激光照射，未检测到I型胶原蛋白基因表达或免疫反应性。细胞培养后体外激光照射后仅发现II型胶原蛋白，这表明细胞对激光照射的反应与传统伤口愈合中观察到的不同。软骨的激光照射可留下完整的胶原蛋白基质，这可能使软骨细胞在完整的支架上得以恢复。
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