Monitoring Wound Healing Of Rat Based On Multiphoton Microscopy And Image Texture Analysis

The skin is the largest organ of the body and the largest protective barrier.It is necessary to resist the damage of the external environment all the time.If the structural integrity of this barrier is damaged,it will form a wound.The causes of trauma are many and very complex,such as fire,radiation,high temperature burns and so on.Once the wound is formed,it will often affect the patient’s life to varying degrees if it is not healing in time.In mammals,collagen is the most abundant protein which is essential to repair the skin after injury.However,there is still no satisfactory method to imaging and estimating the wound healing process base on rat skin collagen in vivo.The clinical assessment of healing progress generally relies on the doctor’s many years of diagnostic experience for visual observation.This non-quantitative assessment may produce large errors,so we need find a new technology to diagnose the wound healing process.The purpose of this paper is to use multiphoton microscope(MPM)technology based on nonlinear optical effects to image collagen in vivo during wound healing,to enrich the understanding of the healing mechanism by capturing a series of dynamic changes in collagen during healing,and establish a new evaluation method.First,we using the MPM imaging technology to microscopically visualize the skin tissue structure of normal rats,and then to compare the imaging quality of collagen in vitro and in vivo skin,we conclude that MPM requires a high resolution setting of 16 scan averages for in vivo imaging of rat parameter.Secondly,2D experimental imaging and evaluation of total wound healing were performed at different time points after wound healing.Combined with the Gray Level Run Length Matrix to quantify the morphological changes of collagen in different stages.The experimental results allow us to understand more clearly that the distribution of collagen in skin repair is gradually transformed from the original disorder to an ordered arrangement,and the collagen shrinks from the periphery to the center during the healing process.Finally,we use the inherent 3D resolution of MPM imaging technology to perform threedimensional quantitative analysis of full-thickness wounds,and through the 3D texture analysis program to study the spatial orientation and spatial distribution of collagen during the healing process,the results prove that 3D monitoring can The spatial distribution of collagen is more reduced,and the 3D characteristics of collagen can be quantified from a more stereoscopic perspective,indicating that the combination of 3D quantitative analyses and MPM technology can more accurately assess the wound healing process.The above results prove that the MPM is an ideal tool for monitoring the wound healing process in vivo.It can obtain high-resolution and high-quality collagen micrographs in vivo,and a series of dynamic changes in collagen during the healing process were quantitatively characterized by image texture extraction.This will be a powerful tool for monitoring wound healing with minimally invasive method.