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The Tracer Study of the Adipose-derived Stem Cells (ADSCs) Grafted by Intravenous in Femoral Head Steroid-induced Avascular Necrosis Models on Rabbits

Author: LiZheng
Tutor: WangWanMing
School: Fujian College of Traditional Chinese Medicine
Course: TCM orthopedics and traumatology
Keywords: Steroid-induced Avascular Necrosis of the Femoral Head Animal mode Adipose-derived stem cells 5-bromo-2’-deoxyuridine bromodeoxyuridine Transfection Tissue engineering
CLC: R681.8
Type: Master's thesis
Year: 2009
Downloads: 69
Quote: 0
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Abstract


Objectives:To observe the biological characteristics of the adipose-derived stem cells from rabbits cultured in vitro.To explore the feasibility study of the establishment of femoral head steroid-induced avascular necrosis models on rabbits by small doses of hormone combined with horse serum.To observe that if the ADSCs grafted into the animal by intravenous could survive and proliferate in the femoral head steroid-induced avascular necrosis models and explore the tracer of the adipose-derived stem cells in steroid-induced femoral head necrosis models.Methods:①.Isolate and purify the rabbit ADSCs with density gradient solution and attachment-independent culture system.Analyse the surface antigens with CD44 immunohistochemistry.Detect that whether the ADSCs were differentiated inducedly into osteoblasts or adipogenic cells by Alkaline phosphatase staining、Von kossa staining and Oil red "O"staining after the ADSCs were cultured in inductive basic medium containing osteogenic induction agent and adipogenic inducers for four weeks.②.Establish the femoral head steroid-induced avascular necrosis models on rabbits by small doses of hormone combined with horse serum and observe the situation of avascular necrosis of femoral head by X-ray、hematoxylin eosin and electron microscopy at 2、4 and 8 weeks.③.Grafted the ADSCs marked with BrdU into the femoral head steroid-induced avascular necrosis models by intravenous and observe that if the ADSCs grafted into the animal by intravenous could survive and proliferate in the femoral head steroid-induced avascular necrosis models by BrdU immunohistochemistry after 4 and 8 weeks.Results:①.The ADSCs have the capacity of rapid proliferation that can amplify to tissue-engineered seed cell quantity in a short period of time.The CD44 of the ADSCs express positive by immunohistochemistry and the cells maintain the undifferentiated state;In special induced conditions,the ADSCs were differentiated into osteoblasts or adipogenic cells.②It proves that we suceedly establish the femoral head steroid-induced avascular necrosis models on rabbits by X-ray、hematoxylin and electron microscopy.③The BrdU marker rate in vitro has reached to 90%,and the activities and growth trends between marked cells and non-tag ones have no significant difference;The marrow,liver and other tissues in the model group can be observed cells by BrdU immunohistochemistry at different time after transplantation,. Conclusions:①The ADSCs may serve as a valuable resource in cell transplantation therapy and tissue engineering;②It proves that we suceedly establish the femoral head steroid-induced avascular necrosis animal models on rabbits by long-term small doses of hormone combined with horse serum.③BrdU marker is a good cell tracer method in vitro and in vivo,the ADSCs grafted into the animal by intravenous could survive and proliferate in the femoral head steroid-induced avascular necrosis models.

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CLC: > Medicine, health > Surgery > Orthopaedic Surgery ( movement system diseases,orthopedic surgery ) > Bone diseases > Lower limb
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