肌成纤维细胞和转化生长因子- β 1在CO2激光促进大鼠牙槽愈合过程中的动态变化

H. Fukuoka, Y. Daigo, M. Ishikawa, H. Amano, Hiroshi Koga, N. Enoki, K. Taniguchi, Hironobu Sato
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However, the mechanism of healing promotion by CO 2 laser irradiation remains unclear, so we investigated the emergence of myofibroblasts ( α -smooth muscle actin ( α -SMA)) involved in granulation tissue scar contracture in extraction wounds and changes in the expression of transforming growth factor-beta 1 (TGF- β 1) associated with myofibroblast differentiation and apoptosis. Subjects and Methods: Seventy-two 5-week-old male Wistar rats were used. The first molar was extracted to prepare an extraction socket model. The rats were divided into CO 2 laser-irradiated and non-irradiated (control) groups and pathological-ly compared. In the CO 2 laser-irradiated group, HLLT was performed immediately after tooth extraction, followed by LLLT after one day, corresponding to clinical cases. 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摘要

目的:最近报道了激光照射在眼眶保存中的临床应用,并探讨了激光照射对拔牙创面愈合的有效条件。据我们实验室报道,使用co2激光进行拔牙窝的HLLT和LLLT治疗导致大鼠特有的新骨形成和维持高牙槽嵴。然而,co2激光照射促进愈合的机制尚不清楚,因此我们研究了参与拔伤创面肉芽组织瘢痕挛缩的肌成纤维细胞(α -平滑肌肌动蛋白(α - sma))的出现以及与肌成纤维细胞分化和凋亡相关的转化生长因子- β 1 (TGF- β 1)表达的变化。对象与方法:选用5周龄雄性Wistar大鼠72只。拔除第一磨牙,制作拔牙槽模型。将大鼠分为co2激光照射组和未照射组(对照组)进行病理比较。co2激光照射组,拔牙后立即行HLLT, 1天后按临床病例进行LLLT。治疗后6小时,3、5、7、10、21天切除包括拔牙窝在内的辐照组织,4%多聚甲醛固定,10% EDTA脱钙,标准方法石蜡包埋,制作连续矢状切片。采用抗α - sma抗体和抗tgf - β 1抗体对拔牙窝浅层肉芽组织进行免疫组织学观察。测定α - sma阳性细胞数和TGF- β 1阳性面积,并检验差异的显著性。结果:抗α - sma抗体免疫染色,第3、7天未照射组α - sma阳性细胞较多,而co2激光照射组α - sma阳性细胞较少(p < 0.05)。在抗TGF- β 1抗体免疫染色上,co2激光照射组与未照射组比较,TGF- β 1阳性不明显,治疗5 d后差异有统计学意义。结论:co2 HLLT与LLLT联合治疗拔牙槽能抑制拔牙槽内瘢痕形成,提示与TGF- β 1有关。
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Dynamics of Myofibroblasts and Transforming Growth Factor-beta 1 in CO2 Laser-enhanced Healing Process of Rat Tooth Sockets
: Objective: The usefulness of laser irradiation for socket preservation in clinical cases has recently been reported, and the effective laser irradiation conditions for healing of the extraction wound have also been investigated. It was reported by our laboratory that HLLT and LLLT therapies using a CO 2 laser for extraction sockets resulted in characteristic new bone formation and the maintenance of a high alveolar crest in rats. However, the mechanism of healing promotion by CO 2 laser irradiation remains unclear, so we investigated the emergence of myofibroblasts ( α -smooth muscle actin ( α -SMA)) involved in granulation tissue scar contracture in extraction wounds and changes in the expression of transforming growth factor-beta 1 (TGF- β 1) associated with myofibroblast differentiation and apoptosis. Subjects and Methods: Seventy-two 5-week-old male Wistar rats were used. The first molar was extracted to prepare an extraction socket model. The rats were divided into CO 2 laser-irradiated and non-irradiated (control) groups and pathological-ly compared. In the CO 2 laser-irradiated group, HLLT was performed immediately after tooth extraction, followed by LLLT after one day, corresponding to clinical cases. The irradiated tissue including the extraction socket was excised 6 hours and 3, 5, 7, 10, and 21 days after treatment, fixed in 4% paraformaldehyde, decalcified in 10% EDTA solution, and paraffin-embedded employing the standard method, and serial sagittal sections were prepared. Granulation tissue of the superficial layer of the extraction socket was immunohistologically investigated using anti- α -SMA and anti-TGF- β 1 antibodies. The number of α -SMA-positive cells and TGF- β 1-positive area were measured, and the significance of differences was tested. Results: On immunostaining with anti- α -SMA antibody, there were many α -SMA-positive cells in the non-irradiated group at 3 and 7 days, whereas significantly fewer positive cells were noted in the CO 2 laser-irradiated group (p < 0.05). On immunostaining with anti-TGF- β 1 antibody, TGF- β 1-positivity was not marked the CO 2 laser-irradiated group, compared to that in the non-irradiated group, significant difference was noted 5 days treatment Conclusion: The combination of CO 2 HLLT and LLLT for the extraction sockets inhibited scar formation in the sockets, and the involvement of TGF- β 1 suggested.
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