本文選題：角膜 + 生物力學(xué)�。� 參考：《河北醫(yī)科大學(xué)》2012年碩士論文

【摘要】：角膜是位于眼球最前部的透明組織，，占整個眼球總屈光力的70%。同時，角膜因與外界直接接觸，最易受到外界損傷。研究表明，眼球外傷中角膜破裂傷占33.28%。，如果處理不當(dāng)可造成嚴(yán)重后果，甚至失明。因此，研究角膜穿通傷對于治療此類致盲性疾病具有重要的指導(dǎo)意義。 研究表明，角膜內(nèi)部膠原纖維的本質(zhì)是各向異性的，因此角膜是一種具有各向異性的生物材料，其內(nèi)部膠原纖維的排列肯定會影響角膜傷口愈合的承載能力以及角膜形狀。角膜傷口愈合過程中，角膜基質(zhì)細(xì)胞轉(zhuǎn)化為成纖維細(xì)胞或肌成纖維細(xì)胞，以促進(jìn)角膜傷口的愈合，同時因細(xì)胞特性會發(fā)生轉(zhuǎn)變，細(xì)胞會喪失其透明特性，進(jìn)而導(dǎo)致角膜瘢痕，影響視功能。 目前,角膜穿通傷多采用10-0尼龍縫線間斷縫合的方法。一般在術(shù)后45天至90天拆除角膜縫線。眾所周知，如果過早的拆除角膜縫線，可能造成角膜傷口未完全愈合，從而降低角膜韌性；如果拆除角膜縫線過晚，不但會因角膜縫線的長時間存在給患者帶來長時間的痛苦，還會因?yàn)榻悄たp線的刺激，使角膜瘢痕愈合，降低角膜的透明度，加劇角膜散光，影響術(shù)后視力。因此。探討角膜縫線最佳拆除時間是很必要的。然而，目前尚缺乏相關(guān)報道。 因此，本研究旨在探索角膜最佳拆線時間，以在增強(qiáng)角膜韌性的同時可以減少瘢痕形成及角膜散光。 第一部分兔角膜穿通傷愈合過程中生物力學(xué)的變化 目的：研究縫線縫合兔角膜角膜穿通傷術(shù)后不同時間點(diǎn)的角膜生物力學(xué)變化 方法：40只兔制作中央角膜穿通傷縫合術(shù)模型。隨機(jī)分為8組，每組5只，分別于術(shù)后1周、2周、3周、4周、5周、6周、7周、8周取5個傷口處角膜條試件在BOSE electroforce3220-AT生物力學(xué)試驗(yàn)機(jī)上進(jìn)行拉伸破壞實(shí)驗(yàn)。 結(jié)果：隨著術(shù)后時間的增長，兔角膜承受的拉力逐漸增大，到6周后逐漸穩(wěn)定。前6周任意兩個時間點(diǎn)相比較均有統(tǒng)計(jì)學(xué)意義(P＜0.01),6周與7周比較無明顯統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論：兔角膜穿通傷術(shù)后6周已達(dá)較大的抗拉力能力，是拆線的最佳時機(jī)。 第二部分兔角膜穿通傷愈合過程中角膜標(biāo)記物的變化 目的：檢測分析兔角膜穿通傷術(shù)后不同時間點(diǎn)角膜內(nèi)keratocan、vimentin、CD90和α-SMA標(biāo)記蛋白的變化。旨在觀察角膜達(dá)最大拉力時角膜瘢痕形成情況。為臨床角膜縫合術(shù)后最佳拆線時間提供參考。 方法：32只兔制作中央角膜穿通傷模型。隨機(jī)分為8組，每組4只，分別于術(shù)后1周、2周、3周、4周、5周、6周、7周、8周取完整角膜，采用RT-PCR方法檢測各時間點(diǎn)中角膜內(nèi)keratocan、vimentin、CD90、α-SMA基因的表達(dá)情況。然后，通過免疫組織化學(xué)實(shí)驗(yàn)測定角膜中vimentin、α-SMA蛋白的含量。 結(jié)果：RT-PCR結(jié)果表明：兔角膜穿通傷縫合術(shù)后傷口愈合過程中角膜基質(zhì)細(xì)胞標(biāo)記物keratocan隨著時間的增長其表達(dá)逐漸下降，前6周下降比較顯著，6周后表達(dá)逐漸平穩(wěn)。同時出現(xiàn)成纖維細(xì)胞和肌成纖維細(xì)胞標(biāo)記物CD90和α-SMA的表達(dá)，且表達(dá)隨著時間的延長逐漸增強(qiáng)，前6周增高比較明顯，6周后表達(dá)逐漸平穩(wěn)。上述蛋白各自從術(shù)后1周到6周之間兩兩比較均有統(tǒng)計(jì)學(xué)意義(P＜0.01)，6周到8周之間兩兩比較無明顯統(tǒng)計(jì)學(xué)意義(P0.05)。隨著術(shù)后時間的延長，vimentin的表達(dá)逐漸增多，術(shù)后1周到6周之間兩兩比較均有差異(P＜0.01)。6周與7周比較無明顯統(tǒng)計(jì)學(xué)意義(P0.05)。免疫組化數(shù)據(jù)顯示：隨著術(shù)后觀察時間的延長，肌成纖維細(xì)胞標(biāo)記物α-SMA的表達(dá)量逐漸增多，且術(shù)后6周時變化最顯著。與前五個時間點(diǎn)任意兩點(diǎn)比較均有統(tǒng)計(jì)學(xué)意義(P＜0.01)。間質(zhì)細(xì)胞標(biāo)記物vimentin的表達(dá)呈先增多然后逐漸降低趨勢，且術(shù)后3周時表達(dá)量最高。 結(jié)論：兔角膜穿通傷愈合過程中角膜基質(zhì)細(xì)胞逐漸減少且轉(zhuǎn)化為成纖維或肌成纖維細(xì)胞。術(shù)后6周時角膜基質(zhì)細(xì)胞轉(zhuǎn)化趨于穩(wěn)定。
[Abstract]:The cornea is the most transparent tissue of the eyeball in front of the eye, accounted for the total refractive power of the cornea 70%. at the same time, due to direct contact with the outside world, the most vulnerable to external damage. The study shows that the traumatic corneal rupture accounted for 33.28%., if improper handling can cause serious consequences, and even blindness. Therefore, the research of corneal penetrating injury an important guiding significance for the treatment of such disease causing blindness.
Research shows that the essence of internal corneal collagen fiber is anisotropic, so the cornea is an anisotropic biological material, its internal collagenous fibers will affect the bearing capability of corneal wound healing and corneal shape. The process of corneal wound healing, corneal stromal cells into fibroblasts or myofibroblasts, in order to promote corneal wound healing, at the same time because cells changed and the cells lose their transparent properties, leading to corneal scar, affect the visual function.
At present, corneal penetrating wound method using 10-0 nylon suture suture. In general, after 45 to 90 days removed corneal suture. As everyone knows, if the early removal of corneal suture may cause corneal wound not healed completely, so as to reduce the toughness of the cornea; if removed corneal suture late, will not only for a long time corneal suture the existence of patients brought long time pain, but also because the stimulation of corneal sutures to reduce corneal scar healing, corneal transparency, aggravation of corneal astigmatism, affect visual acuity. Therefore. To investigate the corneal suture the best removal time is necessary. However, there is still a lack of relevant reports.
Therefore, the aim of this study is to explore the best time for the disassembly of the cornea, which can reduce the formation of scar and corneal astigmatism while enhancing the toughness of the cornea.
Part one biomechanical changes during the healing process of corneal penetrating injury in rabbits
Objective: To study the changes of corneal biomechanics at different time points after corneal penetrating injury in rabbits with suture suture suture
Methods: a total of 40 rabbits were made to model the central corneal perforating injury suture. They were randomly divided into 8 groups, 5 in each group. They were respectively taken at 1 weeks, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks after operation, and 8 corneas were taken from 8 wounds. Tensile test was carried out on BOSE electroforce3220-AT biomechanical test machine.
Results: with the increase of postoperative time, the pulling force of rabbit corneas increased gradually, and then gradually stabilized after 6 weeks. There was statistically significant difference between any two time points at the first 6 weeks (P < 0.01), but there was no significant difference between 6 weeks and 7 weeks (P0.05).
Conclusion: 6 weeks after corneal penetrating injury in rabbits, it has reached a great ability to resist tension, and it is the best time to dismantle the line.
Changes of corneal markers during the healing process of corneal penetrating injury in the second part of rabbits
Objective: to detect and analyze the changes of keratocan, vimentin, CD90 and alpha -SMA marker proteins in cornea at different time points after penetrating injury of cornea in rabbits, so as to observe corneal scar formation at maximum pulling force, and provide reference for optimal suture time after clinical corneal suture.
Methods: 32 rabbits produced central corneal penetrating injury model were randomly divided into 8 groups, 4 rats in each group, respectively after 1 weeks, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks after complete cornea, using the method of RT-PCR keratocan in the cornea, to detect the time in vimentin, CD90, expression of -SMA alpha gene. Then, through the determination of vimentin in experimental corneal immunohistochemistry, content of alpha -SMA protein.
Results: RT-PCR results showed that the wound healing of corneal stromal cell markers in the process of keratocan with the growth of its expression gradually decreased in rabbit corneal penetrating wound suture after 6 weeks before significant decline, after 6 weeks the expression gradually stable. At the same time expression of fibroblasts and muscle fibroblast markers CD90 and alpha -SMA, and the expression gradually increased with the extension of time, the first 6 Zhou Zenggao obviously after 6 weeks. The protein expression gradually steadily from their respective after 1 to 6 weeks between the 22 was statistically significant (P < 0.01), 6 to 8 weeks between the 22 was not statistically significant (P0.05). With the extension the time after operation, the expression of vimentin increased gradually after 1 to 6 weeks between the 22 comparison difference (P < 0.01).6 weeks compared with 7 weeks of no statistical significance (P0.05). Immunohistochemical data show: with the observation after operation The extension of muscle fiber cell marker expression of alpha -SMA increased gradually, and at 6 weeks after operation. The most significant change compared with the former two arbitrary five time points were statistically significant (P < 0.01). The expression of mesenchymal cell marker vimentin was first increased and then decreased gradually and the trend. After 3 weeks the highest expression level.
Conclusion: corneal stromal cells gradually decrease and become fibroblasts or myofibroblasts in healing process of corneal penetrating injury. Corneal stromal cell transformation tends to be stable at 6 weeks after operation.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R772.2

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本文編號：1745108