| In 2010,Popescu’s team named a new interstitial cell----Telocytes and brought it into public vision.At present,people have a systematic understanding of the ultrastructure of TCs:its cell body is oval which is about 1/4 size of the cell with a small perinuclear amount of cytoplasm.The TCs cytoplasm is rich in mitochondria,Golgi complex,endoplasmic reticulum and vacuolae.TCs showed long cytoplasmic processes known as telopodes,typically 1 to 5 in numbers which extends in multi-directions and ranging from tens to hundreds of microns in length.Still so far,transmission electron microscopy is the best method used to identify and demonstrate the TCs.At the same time,immunohistochemistry and immunofluorescence labeling can be used to explain the TCs in various organs of different animals.Since the TCs were discovered,it has been found in a variety of organs of many animals,but its distribution and function in the skin system of canine and other companion animals are still needed to study.The skin is one of the largest organ in canine,which is in direct contact with the external environment and is the most important first barrier structure of the body.In addition,the skin is rich in skin appendages,connective tissues,blood vessels and nerve tissue.Importantly,the external stimulation can be transmitted to the internal organs of the body to cause the corresponding physiological reactions.Whereas,the pathological changes and abnormalities of the internal tissues and organs of the body can also be reflected through the skin.Therefore,this study takes the dog skin as the research object to explore the distribution and ultrastructural characteristics of telocytes in normal state and wound healing by immunolabeling and transmission electron microscopy.Present study comprehensively provides the TCs distribution in the mechanism of skin wound healing and offer a morphological basis of TCs as a regulatory signal during skin wound healing.ExperimentI Identification and distribution of Telocytes in normal canine skinTelocytes is a new type of interstitial cells which discovered and named in past few years.Telocytes have been confirmed in different tissues and organs of many animals.With the development of the times,canine as companion pets are more and more appearing in the public’s vision and become a member of human families.However,in clinical practices,the research about TCs in canine and other companion animals’ skin system is still in its infancy.In current study,hematoxylin and eosin staining,immunohistochemistry,immunofluorescence,and transmission electron microscopy were used to observe and identify the morphology and distribution of TCs in canine skin.The results showed that TCs were distributed in the dermis and subcutaneous tissues of canine skin,but there are some differences in morphology and distribution of cells.In dermis and subcutaneous tissues,the elongated protuberances of TCs are usually parallel to the epidermis.Also,TCs are usually distributed between the hair follicles,around the sweat gland,and also in the middle of the connective tissue.Under transmission electron microscopy,TCs surrounded hair follicles,sweat glands and blood vessels,forming a three-dimensional network structure with adjacent tissues in the skin.In this study,the distribution and ultrastructure of TCs in canine skin were studied,which provided a cellular basis of TC in skin for clinical research.ExperimentⅡ Observation and identificationon on the Telocytes in the process of skin wound healing in canineSkin injury is one of the most common clinical symptoms in canine.It can be seen in many cases,such as bruise,knife hurt,bite and so on.Mild skin trauma can usually be self-healing,but severe wound skin defects are serious,and even affect the health and appearance of canine.In pet clinic,there are many ways to treat skin trauma.But the mechanism of skin wound healing,and how to promote wound healing faster and better,has been one of the most hot spots in clinical research.In this study,the morphological structure and distribution of TCs in the skin tissue around the wound on the third day,the seventh day and the eleventh day after wound healing were observed by various staining methods.The results showed that in the healing process of wounds,the wounds almost recovered on the eleventh day,and the number of TCs in the epidermis increased gradually,and formed a complete TCs layer on the eleventh day.With the increase of healing time,TCs around hair follicles and sweat glands gradually formed a complete ring around the outer layer,and TCs network like structure gradually formed in subcutaneous tissue.According to the results,we speculate that TCs play an important role in skin metabolism and homeostasis.At the same time,TCs may play a positive role in the process of wound healing as a regulatory signal of skin wound healing,and also provide new ideas for the treatment of clinical skin trauma.ExperimentⅢ Optimization on the procedure of paraffin section of canine skinParaffin section is one of the most commonly used methods in histological research,and also one of the most commonly used methods to study TCs in canine skin.The main production process of canine skin tissue section usually includes sampling,fixation,embedding,sectioning,etc.However,due to the loose tissue structure and thick subcutaneous tissue layer of canine skin,the paraffin section is often broken and incomplete in the production process,and the paraffin section is easy to be detached,which seriously affects the test.In this experiment,the procedure of making paraffin section of canine skin was improved to optimize the result of TCs staining.The process of paraffin section was optimized by changing the thickness of paraffin section and the treatment time of different reagents.The results showed that when the transparent time of xylene was 5min and the thickness of the section was set to 7μm,the staining results of canine skin paraffin section were.better,and the structure of TCs observed was more complete and clear. |
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