基于超快光致破裂效應的飛秒激光鞏膜造瘺微創(chuàng)手術基礎研究
發(fā)布時間：2018-08-17 15:25
【摘要】： 目的探索利用飛秒激光的光致破裂效應實施微創(chuàng)性鞏膜造瘺術的可行性,并尋求適宜的激光參數和切割方式。 方法飛秒激光(800nm/50fs／1kHz)經0.1數值孔徑(numerical aperture, NA)的物鏡聚焦后掃描離體兔眼水合鞏膜。通過計算機控制的精密三維平臺的移動,嘗試應用不同脈沖能量和曝光時間的飛秒激光對鞏膜片的多種切割方式。應用掃描電鏡對飛秒激光作用后的鞏膜組織進行形態(tài)學觀察和測量。 結果飛秒激光能夠在離體兔眼鞏膜上完成包括線性切割,柱形孔隙和矩形空腔在內的三種切口。當脈沖能量在37.5-150μJ的范圍內變化時,飛秒激光以0.025-0.1mm/s的速度線性掃描鞏膜后均能在鞏膜表面產生光致破裂效應。而當脈沖能量低于31.25μJ,功率密度低于4.06×1014 W／cm2的閾值水平時,受飛秒激光輻射的鞏膜組織在掃描電鏡下不會產生任何形態(tài)學改變。飛秒激光在鞏膜表面產生的線性切口深度與激光的脈沖能量以及曝光時間呈正相關。掃描電鏡顯示飛秒激光在鞏膜表面產生的切口具有精確的幾何形狀并且切口邊緣光滑整齊,切口周圍組織未見明顯的附帶損傷。 結論飛秒激光對離體兔眼鞏膜高精度、微創(chuàng)傷的光致破裂效應,預示了它在抗青光眼的鞏膜造瘺術中有著潛在的應用價值。 目的探索飛秒激光經外路鞏膜造瘺的可行性,評價手術的有效性和安全性。 方法對20只青紫藍兔右眼后房注射α-糜蛋白酶,制備兔慢性高眼壓模型。將兔隨機分為實驗組和對照組,每組10只兔。實驗組右眼行飛秒激光鞏膜造瘺術,對照組右眼不做手術治療。飛秒激光脈寬50fs,波長800nm,重復頻率1kHz,脈沖能量0.4mJ。觀察術后1個月內濾過泡形態(tài)、眼壓及并發(fā)癥情況,組間眼壓比較采用重復測量方差分析。分別于術后第3、7、14、30天對濾過道組織進行光鏡觀察,術后14天對濾過道組織進行掃描電鏡觀察。 結果10只兔眼鞏膜均被飛秒激光一次性穿透性切除,激光作用時間為15s-16s,鞏膜切口約2mm×1mm2。術后第1、3、7、14、21、30天,實驗組和對照組眼壓比較,差異具有統計學意義(F=117.46,39.96,15.17,11.62,15.31,11.10；P0.01)。除2眼因激光在切穿鞏膜時損傷虹膜根部導致前房出血外,未見其他明顯的手術并發(fā)癥。術后第1天所有實驗眼均有濾過泡形成,到第3天濾過泡隆起度最高。隨后濾過泡逐漸縮小,泡壁增厚,第3周時濾過泡逐漸扁平消失。病理學檢查顯示飛秒激光鞏膜造瘺的切口光滑銳利,對周圍組織的附帶損傷小,術后濾過道的修復反應主要表現為成纖維細胞的輕度增生和薄層疏松膠原纖維的形成。 結論飛秒激光經外路鞏膜造瘺術簡便快捷,安全有效,有可能成為治療青光眼的一種新的手術方法。 目的比較經外路飛秒激光鞏膜造瘺術和機械刀鞏膜造瘺術的手術效果和濾過道創(chuàng)傷修復反應,探討飛秒激光鞏膜造瘺術的有效性和優(yōu)越性。 方法實驗研究。20只青紫藍兔隨機分為兩組,每組各10只兔。一組右眼行飛秒激光鞏膜造瘺術,另一組右眼行機械刀鞏膜造瘺術,左眼作為非手術對照眼。觀察術后1個月內濾過泡表現、眼壓變化及手術并發(fā)癥情況。另有6只兔,隨機選擇右眼行飛秒激光鞏膜造瘺術,左眼行機械刀鞏膜造瘺術,術后第14天處死,采用HE染色、Masson三色染色、免疫組化和掃描電鏡技術,觀察濾過道的創(chuàng)傷修復反應。飛秒激光脈寬50fs,波長800nm,重復頻率1kHz,脈沖能量0.4mJ。本研究中的等級資料和分類計數資料采用兩獨立樣本非參數檢驗,手術成功率的比較采用Kaplan-Meier法。 結果16只兔眼鞏膜均被飛秒激光一次性穿透性切除,激光作用時間為15s-16s。飛秒激光組19%(3／16)的術眼因激光誤傷虹膜導致前房少量出血,50%(8／16)的術眼手術區(qū)周邊角膜出現水腫混濁。飛秒激光鞏膜造瘺術后功能性濾過泡的生存時間以及眼壓下降的持續(xù)時間均較機械刀組明顯延長,差異有統計學意義(Mann-WhitneyU值=24.000,20.000；P=0.025,0.016)。飛秒激光組的手術成功率明顯高于機械刀組,差異有統計學意義(χ2=7.785,P=0.005)。病理學檢查顯示術后第14天,機械刀組的鞏膜切口及結膜下間隙已被致密的纖維結締組織完全阻塞,而飛秒激光組的濾過道則基本保持通暢,結膜下仍見大范圍濾過間隙存在。術后第14天,飛秒激光組濾過道內新生膠原纖維量、成纖維細胞量和新生血管量均少于機械刀組,差異均有統計學意義(Mann-WhitneyU值=7.000,5.500,2.500；P=0.036,0.045,0.013)。 結論飛秒激光鞏膜造瘺術有可能成為一種安全有效,方便快捷的新型微創(chuàng)性抗青光眼手術方式。
[Abstract]:Objective To explore the feasibility of minimally invasive scleral fistula with femtosecond laser photorupture effect, and to find suitable laser parameters and cutting methods.
Methods Femtosecond laser (800 nm/50 fs/1 kHz) was focused by 0.1 numerical aperture (NA) objective lens and then scanned the hydration sclera of rabbit eyes in vitro. The sclera tissue after second laser treatment was observed and measured.
Results Femtosecond laser can perform three kinds of incisions including linear incision, cylindrical cavity and rectangular cavity on the sclera of rabbits in vitro. When the pulse energy changes in the range of 37.5-150 uJ, the sclera can be linearly scanned by femtosecond laser at the speed of 0.025-0.1 mm/s and the sclera surface can produce photorupture effect when the pulse energy is low. The scleral tissue irradiated by femtosecond laser has no morphological changes under scanning electron microscope when the power density is below the threshold level of 4.06 *1014 W/cm 2 at 31.25 uJ. The linear incision depth produced by femtosecond laser on the scleral surface is positively correlated with the laser pulse energy and exposure time. The scleral incision has precise geometry and smooth edge, and there is no obvious incidental damage to the surrounding tissue.
Conclusion Femtosecond laser has high precision and micro-traumatic photorupture effect on isolated rabbit sclera, which indicates that it has potential application value in anti-glaucoma scleral fistula.
Objective to explore the feasibility of femtosecond laser through external scleral fistulas and evaluate the effectiveness and safety of the operation.
Methods Twenty rabbits were injected with alpha-chymotrypsin into the posterior chamber of the right eye and divided into experimental group and control group randomly, 10 rabbits in each group. The morphology of filtering blebs, intraocular pressure and complications were observed one month after operation. The intraocular pressure between groups was compared by repeated analysis of variance.
Results The sclera of all 10 rabbits were resected by femtosecond laser. The duration of laser treatment was 15s-16s, and the scleral incision was about 2mm *1mm 2. The intraocular pressure of the experimental group and the control group was significantly different (F = 117.46, 39.96, 15.17, 11.62, 15.31, 11.10; P 0.01). There were no significant surgical complications except for anterior chamber hemorrhage at the iris root. On the first day after operation, all the experimental eyes had bleb formation, and the bleb swelling was the highest on the third day. The repairing reaction of the filter passage was mainly manifested by mild proliferation of fibroblasts and formation of thin-layer loose collagen fibers.
Conclusion Femtosecond laser sclerostomy is a simple, rapid, safe and effective method for the treatment of glaucoma.
Objective To compare the effect of external femtosecond laser sclerostomy with mechanical scalpel sclerostomy and repair reaction of filtering tract wound, and to explore the effectiveness and superiority of femtosecond laser sclerostomy.
Methods Twenty blue and blue rabbits were randomly divided into two groups, 10 rabbits in each group. One group underwent femtosecond laser sclerostomy on the right eye, the other underwent mechanical scalpel sclerostomy on the right eye, and the other underwent non-operative control on the left eye. Femtosecond laser sclerostomy was performed on the left eye and mechanical scalpel sclerostomy was performed on the 14th day after operation. HE staining, Masson trichrome staining, immunohistochemistry and scanning electron microscopy were used to observe the wound healing response of the filter passage. Two independent sample nonparametric tests were used to count data. Kaplan-Meier method was used to compare the success rate of operation.
Results The sclera of all the 16 rabbits were resected by femtosecond laser for 15s-16s. In the femtosecond laser group, 19% (3/16) of the eyes suffered from a small amount of bleeding in the anterior chamber and edema and turbidity in the cornea around the operation area of 50% (8/16) of the eyes. The survival time of the functional filtering blebs after femtosecond laser scleral fistulation. The duration of IOP decrease was significantly longer in the femtosecond laser group than in the mechanical knife group (Mann-Whitney U = 24.000,20.000; P = 0.025,0.016). The success rate in the femtosecond laser group was significantly higher than that in the mechanical knife group (2 = 7.785, P = 0.005). Pathological examination showed sclerectomy in the mechanical knife group on the 14th day after surgery. The oral cavity and subconjunctival space were completely blocked by dense fibrous connective tissue, while the filtering passage of femtosecond laser group remained unobstructed, and large-scale subconjunctival filtering space was still found. Academic significance (Mann-WhitneyU value =7.000,5.500,2.500; P=0.036,0.045,0.013).
Conclusion Femtosecond laser sclerostomy may be a safe, effective, convenient and new minimally invasive surgery for glaucoma.
【學位授予單位】：華中科技大學
【學位級別】：博士
【學位授予年份】：2010
【分類號】：R779.6
本文編號：2188067
[Abstract]:Objective To explore the feasibility of minimally invasive scleral fistula with femtosecond laser photorupture effect, and to find suitable laser parameters and cutting methods.
Methods Femtosecond laser (800 nm/50 fs/1 kHz) was focused by 0.1 numerical aperture (NA) objective lens and then scanned the hydration sclera of rabbit eyes in vitro. The sclera tissue after second laser treatment was observed and measured.
Results Femtosecond laser can perform three kinds of incisions including linear incision, cylindrical cavity and rectangular cavity on the sclera of rabbits in vitro. When the pulse energy changes in the range of 37.5-150 uJ, the sclera can be linearly scanned by femtosecond laser at the speed of 0.025-0.1 mm/s and the sclera surface can produce photorupture effect when the pulse energy is low. The scleral tissue irradiated by femtosecond laser has no morphological changes under scanning electron microscope when the power density is below the threshold level of 4.06 *1014 W/cm 2 at 31.25 uJ. The linear incision depth produced by femtosecond laser on the scleral surface is positively correlated with the laser pulse energy and exposure time. The scleral incision has precise geometry and smooth edge, and there is no obvious incidental damage to the surrounding tissue.
Conclusion Femtosecond laser has high precision and micro-traumatic photorupture effect on isolated rabbit sclera, which indicates that it has potential application value in anti-glaucoma scleral fistula.
Objective to explore the feasibility of femtosecond laser through external scleral fistulas and evaluate the effectiveness and safety of the operation.
Methods Twenty rabbits were injected with alpha-chymotrypsin into the posterior chamber of the right eye and divided into experimental group and control group randomly, 10 rabbits in each group. The morphology of filtering blebs, intraocular pressure and complications were observed one month after operation. The intraocular pressure between groups was compared by repeated analysis of variance.
Results The sclera of all 10 rabbits were resected by femtosecond laser. The duration of laser treatment was 15s-16s, and the scleral incision was about 2mm *1mm 2. The intraocular pressure of the experimental group and the control group was significantly different (F = 117.46, 39.96, 15.17, 11.62, 15.31, 11.10; P 0.01). There were no significant surgical complications except for anterior chamber hemorrhage at the iris root. On the first day after operation, all the experimental eyes had bleb formation, and the bleb swelling was the highest on the third day. The repairing reaction of the filter passage was mainly manifested by mild proliferation of fibroblasts and formation of thin-layer loose collagen fibers.
Conclusion Femtosecond laser sclerostomy is a simple, rapid, safe and effective method for the treatment of glaucoma.
Objective To compare the effect of external femtosecond laser sclerostomy with mechanical scalpel sclerostomy and repair reaction of filtering tract wound, and to explore the effectiveness and superiority of femtosecond laser sclerostomy.
Methods Twenty blue and blue rabbits were randomly divided into two groups, 10 rabbits in each group. One group underwent femtosecond laser sclerostomy on the right eye, the other underwent mechanical scalpel sclerostomy on the right eye, and the other underwent non-operative control on the left eye. Femtosecond laser sclerostomy was performed on the left eye and mechanical scalpel sclerostomy was performed on the 14th day after operation. HE staining, Masson trichrome staining, immunohistochemistry and scanning electron microscopy were used to observe the wound healing response of the filter passage. Two independent sample nonparametric tests were used to count data. Kaplan-Meier method was used to compare the success rate of operation.
Results The sclera of all the 16 rabbits were resected by femtosecond laser for 15s-16s. In the femtosecond laser group, 19% (3/16) of the eyes suffered from a small amount of bleeding in the anterior chamber and edema and turbidity in the cornea around the operation area of 50% (8/16) of the eyes. The survival time of the functional filtering blebs after femtosecond laser scleral fistulation. The duration of IOP decrease was significantly longer in the femtosecond laser group than in the mechanical knife group (Mann-Whitney U = 24.000,20.000; P = 0.025,0.016). The success rate in the femtosecond laser group was significantly higher than that in the mechanical knife group (2 = 7.785, P = 0.005). Pathological examination showed sclerectomy in the mechanical knife group on the 14th day after surgery. The oral cavity and subconjunctival space were completely blocked by dense fibrous connective tissue, while the filtering passage of femtosecond laser group remained unobstructed, and large-scale subconjunctival filtering space was still found. Academic significance (Mann-WhitneyU value =7.000,5.500,2.500; P=0.036,0.045,0.013).
Conclusion Femtosecond laser sclerostomy may be a safe, effective, convenient and new minimally invasive surgery for glaucoma.
【學位授予單位】：華中科技大學
【學位級別】：博士
【學位授予年份】：2010
【分類號】：R779.6
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