Therapeutic Effect Of Adipose-derived Stem Cells To Treat Chemotherapy-induced Oral Mucositis Of Golden Syrian Hamsters

Objective:Chemotherapy-induced OM is an acute or chronic inflammation disease of the oral mucosa or the surrounding soft tissue,which due to the toxic effects of chemotherapy drugs.It is often characterized by dry mouth,pain,ulcers and influences the quality of patients'life,even leads to tumor treatment interruption.Therefore,clinical doctors hope can effectively prevent the disease or quick treatment.This study by building an animal model of Golden Syrian hamsters'chemotherapy-induced OM,cultivate and identify ADSCs,and mark the ADSCs,then inject it into the the cheek pouch around the ulcer.Analysis the survivability of ADSCs in animal and study the result of ADSCs to treat OM.To provide the experimental basis for treat oral mucositis this caused by chemotherapy,and to lay a certain foundation for repairing oral mucosa tissue by ADSCs injecting in the future.Methods:1.The reference of literature and the preliminary experiments determine the drug dose and frequency when modeling,set up the animal model of chemotherapy-induced OM and compare with the control group,then analysis the results.2.To extract the hamsters'ADSCs and subculture,and test its ability to differentiation.3.To culture the hamsters'ADSCs to the third-generation,mark the cells with the CM-Dil probe and injecte into the cheek pouch around the ulcer,injection amount is 2×10~6.Observe the survival situation of ADSCs in animals and compare the healing with the control group.Results:1.There is no obvious change of intaking in control group,and the weight increase gradually over time.From the 5~thh day,the food intaking and body weight decreased obviously,after the 14~thh days,eating recover and weight increase gradually.Compared with control group,the cheek pouch mucosa congests,local scratch form a typical ulcer in the 5~thh day.Ulcer heals at the 14~thh day.Generally until the 21~stt day,the ulcer heals basically.HE staining showed,in experimental group,a large number of inflammatory cells infiltrate below epithelial beneath,epithelial has been destroyed,lamina collagen fiber edema,to 21~stt day,most of the lesions were closed,the cortex thickness is differ,and individual submucous muscle layer scar repair,HE staining found the oral mucosa of the control group has a integrity epithelial,clear,no inflammatory cell infiltration.2.To observe under optical microscope,the cells wall and in sample fibroblasts form,double cell proliferation after 3~5d,and cell fusion could reach 80%after 6~7d,after sub culturing to the third-generation,cell morphology uniform.Represented more than 8 times,each generation of cells while retain the original form;cell proliferation rate has no obvious slow down.When inducing the third generation of ADSCs after 3 days,a lipid droplets in the cytoplasm and gradually gathering,induced 2 weeks later found that increased cell volume,rounded lipid droplets are scattered throughout the cytoplasm,grape bunches of oil red O staining positive,round cell intrinsic optical microscope can see clear bright red particles,not induced cells is still normal ADSCs forms,after oil red O staining has no obvious red granules in the cell.3.CM-Dil tags still dark spindle cells after in vitro culture and in a state of proliferation,grow well,good pervious to light,form a complete,less cellular debris.Marked cells by fluorescence microscopy exert a red fluorescence,consistenting with membrane dye annular red and nucleus is not fluorescent tags.After cell transplantation,all hamsters are alive,and no adverse reactions,in 3~rdd days after transplantation ulcer surface of the experimental group began to shrink,ulcer is almost entirely disappeared after the 7~thh days.The average healing time is 20 days in the control group.Observation the specimens of frozen section in the 3~rdd and 7~thh day found that ADSCs had a good activity after transplantation in 3~rdd and 7~thh day,and gathered around the ulcer.Conclusion:1.A stable and reliable animal model can be established by intraperitoneal injection of 5-fluorouracil and scraping cheek pouch,and have obvious ulcer.2.ADSCs extraction is simple and fast,and the ability of proliferation and differentiation is strong in vitro,which can be used as a treatment for OM,a seed cell for repairing the mucosa damage.3.ADSCs transplantation can significantly shorten the ulcer healing time,and has a certain therapeutic effect for chemotherapy-induced OM.