Research On Tendon Stem Cells To Repair The Animal Model Of Hurdler's Tendon Injury

In track and field sports,short span class projects hurdler Achilles tendon(hereinafter referred to as the body's biggest tendon,tendon)prone to tissue damage and restore slower,the serious influence the athletes' athletic achievements and sports career,at present the clinical medicine and sports medicine mainly to conquer the difficulty of this kind of damage there are three main mechanisms: tear,tensile overload and contusion.The traditional methods to treat tendon injury include physical therapy,surgical therapy,cryotherapy,etc.,but these therapies have a long treatment cycle,slow repair and easy repair.After repair,the physiological function and mechanical properties of the tendon decline,which is easy to cause re-injury.In the outline of the national program for medium-and long-term scientific and technological development(2006-2020),the state council stressed that it is necessary to vigorously promote the experimental system of stem cells for sports injury tissue repair in an all-round way.Six ministries and commissions jointly issued the "13th five-year" health industry science and technology innovation special plan,which clearly requires to accelerate the clinical application of stem cell and regenerative sports medicine.The self-renewal,high-growth and multi-differentiation potential of stem cells provide important theoretical and experimental basis for the development of individual tissues and the rerepair of damaged tissues.The discovery of tendon stem cells and the study of their biological characteristics provide a theoretical basis for the exploration of the pathological mechanism of tendon tissue and an experimental basis for stem cells to repair tendon injuries.This study using the literature material method,mathematical statistics method,expert interview method and experimental research methods such as main stem cell cultivation in vitro in duck embryo tendons,optimize the tendon cells best cultivation system and establish the in vitro amplification technology system,explores the tendons of stem cell proliferation and differentiation,and a series of biological characteristics and multiple differentiation potential,build the animal tendons stem cell transplantation to repair damage model and tendons,explore after transplantation in vivo stem cell migration and repair function,got the following conclusion:1.The use of type I collagen enzyme,the neutral protease and trypsin by mechanical separation separation duck embryo sources of tendon cells,and by addingdifferent factor of whole cell culture medium purification,purification of cells under the optimal cultivation system to 31 generation by generation time in cells before recovery and cryopreserved morphology observation,cell still has the very strong appreciation ability;By flow cytometry and immunohistochemical identification,RT-PCR specificity for the identification of tendon stem cell surface markers,immunofluorescence identification showed that cells of CD44,CD105 and Collagen I,Collagen III,Oct4,Tenascin-C are positive;Flow cytometry showed that the positive rates of CD44,CD166,CollagenI,CollagenIII and tenascin-c were all above 90%.Rt-pcr showed that the cells were positively expressed in CollagenI,CollagenIII,Nucleostemin,CD44,tenascin-c specific genes and surface antigens,indicating that the isolated cells were tendon stem cells with stem cell characteristics.Tendon stem cells were identified by cloning ability formation and growth curve mapping.Tendon stem cells were induced to differentiate into adipoblast cells,osteoblasts,and chondroblasts through different induction systems.The induced adipoblast cells were stained with oil red O to present red lipid droplets.RT-PCR showed positive expressions of specific molecular markers LPL and ppar-gamma of adipocytes.After successful induction of osteoblasts,osteocyte clusters appeared and were positive for alizarin red staining.RT-PCR results showed that OPN and Runx2,specific molecular markers of osteoblasts,were positive for expression.Chondrocyte-specific genes Sox9 and ACAN were detected by RT-PCR.While 90 percent of hESCs differentiate into other cell types or die,only 10 percent retain the ability to form all cell types,maintaining their so-called "dry" nature.These results suggest that tendon stem cells derived from Achilles tendon cultured in vitro have "dry" characteristics,self-proliferation ability and multiple induced differentiation potential.2.By type I collagen enzyme tendon tissue injury in mice model was constructed,and the sample preparation of paraffin and frozen section,the results on paraffin section showed serious damage and disordered arrangement of fiber bundles,use CM-Dil tagged tendon injury tendon stem cells transplanted into mice,tendon stable stem cells can migrate to the damaged tissue around the proliferation and differentiation,different time gradient sampling analysis and serological related indicators of continuous observation showed that cell transplantation in treatment group damage tendons have good repair;Compared with the normal group,the CK value of the tendon stem cell transplantation group decreased to a certain extent,indicating that the tendon stem cellscan repair the damaged tendon tissue.In conclusion,this experiment studied tendon tissue injury of hurdler,and isolated and cultured tendon stem cells in vitro,tested and identified the biological characteristics of tendon stem cells,and explored the optimal culture system.Multiple experiments on the differentiation potential of tendon stem cells have proved that tendon stem cells have the characteristics of "dry",and an animal model of tendon stem cells repairing tendon injury in mice has been constructed,which provides an experimental basis for the clinical application of cell therapy based on stem cell tamping to treat sports injury and promote tissue regeneration.