Method for staining hard tissue slices using picrosirius red and application of method

Abstract

The invention provides a method for staining hard tissue slices using picrosirius red and application of method. The method includes the following steps: sampling from a silk tissue engineering ligament, fixing and coating the sample using resin, performing hard tissue slicing, detecting the secretion condition of types I and III collagen through picrosirius red staining. According to the invention, the picrosirius red is used to stain the hard tissue slices, so that the sample processing time is shortened, the operation steps are simplified, and the types I and III collagen can be directly observed on the hard tissue slices.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for detecting type I and type III collagens of silk tissue engineering ligaments. Background technique [0002] The anterior cruciate ligament plays a very important role in the normal movement and stability of the knee joint. The ability to heal after an ACL injury is minimal and often requires graft reconstruction surgery. The sources of ligament grafts mainly include autologous or allogeneic ligament grafts and tissue engineered ligaments. Commonly used sources of autologous ligament grafts are insufficient, and allogeneic ligament grafts are prone to immune rejection. Silk tissue engineered ligaments have attracted more and more attention due to their good mechanical properties, degradability and low immune rejection. [0003] After the silk ligament graft is implanted in the body, it is often necessary to detect the production of type I and III collagen t...

AI Technical Summary

Problems solved by technology

So far, there are many methods for detecting collagen, such as immunofluorescence, radionuclide labeling, RT-PCR, Western blot, and immunohistochemical staining. Most of the above methods require paraffin sections or frozen sections after decalcification, so there are samples The processing cycle is long, the steps are cumbersome, and the cost is high, and it is difficult to complete the detection and observation of multiple types of collagen on the same slice

[0004] Immunohistochemical staining commonly used in silk tissue engineering ligaments requires decalcification of the tissue (about 1 month), paraffin embedding or frozen sectioning, staining requires steps such as repairing antigens and incubating secondary antibodies, the operation is complicated and time-consuming, and because silk is in After slicing, it becomes a small piece of silk, and the structure is very easy to fall off

So far, there has been no report on the detection of collagen types I and III by picric acid Sirius red staining in hard tissue sections

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