人工晶状体的自动压缩测试
Summary
我们提出了一种使用压缩测试表征人工晶状体有效弹性模量的自动化方法。
Abstract
人工晶状体的生物力学特性对于其作为可变功率光学元件的功能至关重要。这些特性随着人类晶状体年龄的增长而发生巨大变化，导致近视力丧失，称为老花眼。然而，这些变化的机制仍然未知。晶状体压缩提供了一种相对简单的方法，用于定性地评估晶状体的生物力学刚度，并且当与适当的分析技术相结合时，可以帮助量化生物力学特性。迄今为止，已经进行了各种镜片压缩测试，包括手动和自动，但这些方法应用生物力学测试的关键方面不一致，例如预处理、加载速率和测量间隔时间。本文描述了一种全自动镜头压缩测试，其中电动载物台与相机同步，以在整个预编程的加载协议中捕获镜头的力、位移和形状。然后可以从这些数据中计算出特征弹性模量。虽然这里使用猪镜片进行演示，但该方法适用于任何物种的镜片压缩。
Introduction
晶状体是眼睛中发现的透明且灵活的器官，它可以通过改变屈光度来聚焦在不同的距离上。这种能力被称为适应。由于睫状肌的收缩和松弛，屈光力会改变。当睫状肌收缩时，晶状体增厚并向前移动，增加其屈光力1,2。屈光率的增加使镜头能够聚焦在附近的物体上。随着人类年龄的增长，晶状体会变得更硬，这种适应能力会逐渐丧失;这种情况被称为老花眼。变硬的机制仍然未知，至少部分原因是与晶状体的生物力学表征相关的困难。
已经采用了多种方法来估计晶状体刚度和生物力学性能。这些方法包括透镜旋转3,4,5，声学方法6,7,8，光学方法，如布里渊显微镜9，压痕10,11和压缩12,13。压缩是最容易获得的实验技术，因为它可以使用简单的仪器（例如，玻璃盖玻片14,15）或单个电动载物台进行。我们之前已经展示了如何从压缩测试中严格估计晶状体的生物力学特性16.这个过程在技术上具有挑战性，需要专门的软件，而对相对刚度测量感兴趣的镜头研究人员不容易使用。因此，在本研究中，我们专注于在考虑透镜尺寸的同时估计透镜弹性模量的可用方法。弹性模量是与其变形性相关的固有材料特性：高弹性模量对应于较硬的材料。
该测试本身是平行板压缩测试，因此可以在合适的商业机械测试系统上进行。在这里，构建了一个由电机、线性平台、运动控制器、称重传感器和放大器组成的定制仪器。这些都是使用定制软件控制的，该软件还定期记录时间、位置和负载。猪镜片不适应，但易于获得且价格低廉17.开发了以下方法来逐步压缩眼睛晶状体并量化其弹性模量。这种方法可以很容易地复制，并且在晶状体刚度的研究中很有用。
Protocol
Representative Results
Discussion
镜片压缩是一种用于估计镜片刚度的通用方法。上述程序允许比较不同物种和不同尺寸的镜片。所有变形都根据透镜尺寸进行归一化，弹性模量的计算大致考虑透镜尺寸。有效模量远高于之前报道的猪透镜4、7、11、19 的模量，至少部分原因是使用了厚度而不是曲率半径：猪透镜的极曲半径明显大于厚…
Declarações
The authors have nothing to disclose.
Acknowledgements
由美国国立卫生研究院资助 R01 EY035278 （MR） 支持。
Materials
| Curved Medium Point General Purpose Forceps | Fisherbrand | 16-100-110 | |
| Galil COM Libraries | Galil Motion Control | ||
| High Precision Scalpel Handle | Fisherbrand | 12-000-164 | |
| Linear Stage | McMaster-Carr | 6734K4 0.125" | |
| Load Cell | FUTEK | LSB200-FSH03869 | |
| Load Cell Amplifier | FUTEK | IAA300-FSH03931 | |
| MATLAB | The Mathworks, Inc. | ||
| Microprobe | Surgical Design | 22-079-740 | |
| Miniature Self Opening Precision Scissors | Excelta | 63042-004 | |
| Motion Controller | Galil Motion Control | DMC-31012 | |
| Motor | Galil Motion Control | BLM-N23-50-1000-B | |
| Straight Hemastats | Fine Science | NC9247203 | stainless steel, 14cm |
Referências
- Gullstrand, A. Helmholtz’s treatise on physiological optics. translated edn. The Optical Society of America. , (1924).
- Helmholtz, H. Uber die akkommodation des auges. Arch Ophthalmol. 1, 1-74 (1855).
- Burd, H. J., Wilde, G. S., Judge, S. J. An improved spinning lens test to determine the stiffness of the human lens. Exp Eye Res. 92 (1), 28-39 (2011).
- Reilly, M. A., Martius, P., Kumar, S., Burd, H. J., Stachs, O. The mechanical response of the porcine lens to a spinning test. Z Med Phys. 26 (2), 127-135 (2016).
- Fisher, R. F. The elastic constants of the human lens. J Physiol. 212 (1), 147-180 (1971).
- Erpelding, T. N., Hollman, K. W., O’Donnell, M. Spatially mapping the elastic properties of the lens using bubble-based acoustic radiation force. IEEE Ultrasonics Symp. 1, 613-616 (2005).
- Erpelding, T. N., Hollman, K. W., O’Donnell, M. Mapping age-related elasticity changes in porcine lenses using bubble-based acoustic radiation force. Exp Eye Res. 84 (2), 332-341 (2007).
- Yoon, S., Aglyamov, S., Karpiouk, A., Emelianov, S. A high pulse repetition frequency ultrasound system for the ex vivo measurement of mechanical properties of crystalline lenses with laser-induced microbubbles interrogated by acoustic radiation force. Phys Med Biol. 57 (15), 4871-4884 (2012).
- Scarcelli, G., Kim, P., Yun, S. H. In vivo measurement of age-related stiffening in the crystalline lens by Brillouin optical microscopy. Biophys J. 101 (6), 1539-1545 (2011).
- Weeber, H. A., Gabriele, E., Wolfgang, P. Stiffness gradient in the crystalline lens. Graefes Arch Clin Exp Ophthalmol. 245 (9), 1357-1366 (2007).
- Reilly, M. A., Ravi, N. Microindentation of the young porcine ocular lens. J Biomech Eng. 131 (4), 044502 (2009).
- Gu, S., et al. Connexin 50 and AQP0 are essential in maintaining organization and integrity of lens fibers. Invest Ophthalmol Vis Sci. 60 (12), 4021-4032 (2019).
- Sharma, P. K., Busscher, H. J., Terwee, T., Koopmans, S. A., van Kooten, T. G. A comparative study on the viscoelastic properties of human and animal lenses. Exp Eye Res. 93 (5), 681-688 (2011).
- Cheng, C., Gokhin, D. S., Nowak, R. B., Fowler, V. M. Sequential application of glass coverslips to assess the compressive stiffness of the mouse lens: strain and morphometric analyses. J Vis Exp. (111), e53986 (2016).
- Baradia, H., Negin, N., Adrian, G. Mouse lens stiffness measurements. Exp Eye Res. 91 (2), 300-307 (2010).
- Reilly, M. A., Cleaver, A. Inverse elastographic method for analyzing the ocular lens compression test. J Innov Opt Health Sci. 10 (06), 1742009 (2017).
- Hahn, J., et al. Measurement of ex vivo porcine lens shape during simulated accommodation, before and after fs-laser treatment. Invest Ophthalmol Vis Sci. 56 (9), 5332-5343 (2015).
- Parreno, J., Cheng, C., Nowak, R. B., Fowler, V. M. The effects of mechanical strain on mouse eye lens capsule and cellular microstructure. Mol Biol Cell. 29 (16), 1963-1974 (2018).
- Yoon, S., Aglyamov, S., Karpiouk, A., Emelianov, S. The mechanical properties of ex vivo bovine and porcine crystalline lenses: age-related changes and location-dependent variations. Ultrasound Med Biol. 39 (6), 1120-1127 (2013).
- Reilly, M. A., Hamilton, P., Gavin, P., Nathan, R. Comparison of the behavior of natural and refilled porcine lenses in a robotic lens stretcher. Exp Eye Res. 88 (3), 483-494 (2009).
- Mekonnen, T., et al. The lens capsule significantly affects the viscoelastic properties of the lens as quantified by optical coherence elastography. Front Bioeng Biotechnol. 11, 1134086 (2023).
- Wilde, G. S., Burd, H. J., Judge, S. J. Shear modulus data for the human lens determined from a spinning lens test. Exp Eye Res. 97 (1), 36-48 (2012).
