The Experimental Study Of Combination Of Cholorogenic Acid And Mesenchymal Stem Cells Orthotopic Transplantation As Treatment For The Early Stage Osteonecrosis Of The Femoral Heads

Objective: To study the usefulness to treat the early stage osteonecrosis of thefemoral head(ONFH) with Chlorogenic acid and bone marrow mesenchymal stemcell orthotopic transplantation, and provide the better theoretical basis for moreextensive clinical application．Methods: The first part: Establishment of a minimally invasive model of theearly stage osteonecrosis of the femoral head in rabbits induced by focal alternativecooling and heating insults. nine adult New Zealand white rabbits minimally invasivemodels of the early stage ONFH on stochastic side were established by focalalternative cooling and heating insults. the animals were killed respectively at1,2and4weeks after operation three rabbits once. X-ray, gross observation and histologyexamination were made periodically to evaluate necrosis and the subsequent repair.The second part: the animal experimental study of combination of chlorogenicacid and mesenchymal stem cells orthotopic transplantation as treatment for the earlystage osteonecrosis of the femoral head,twenty-four adult New Zealand white rabbitsminimally invasive models of the early stage ONFH on stochastic side wereestablished by focal alternative cooling and heating Insults. which were randomlyclassified into three groups after1week, group A were conducted Chlorogenic acidand bone marrow mesenchymal stem cells orthotopic transplantation, group B wereconducted simple Chlorogenic acid orthotopic injection， and group C wereconducted nothing,the health side of three groups were treated as self-contro1.X-ray,gross observation and histology examination were made at12weeks after operation toevaluate necrosis and the subsequent repair.Result: The first part: one week after operation：X-ray and gross observedshowed no obvious abnormalities, histopathologic results shows that necrosis ofchondrocytes and osteocytes occurred, and empty osteocyte lacunae appearedapparently; two weeks after operation: X-ray showed no obvious abnormal-lities,edema and hemorrhage articular cartilage could be observed, histopathologicresults shows that necrosis of chondrocytes and osteocytes occurred, and empty osteocyte lacunae appeared apparently, the histological observations showed that thetrabecular bone state: thinning, irregular arrangement and portion fracture, four weeksafter opration: X-ray showed uneven density of the femoral head, low-density cysticshadow, no mild femoral head collapse, partial defected articular cartilage could beobserved, histopathologic results showed that the trabecular bone became thinning,sparse and fracture and percentage of empty lacunae counts increased.The second part:12weeks after operation: X-ray showed different states: noabnormal femoral head shape, normal femoral head shape, the drilled holes weredisappear for group A, almost normal femoral head shape, low-density subchondralbone the drilled holes were smaller for group B, femoral head collapse, defectedsubchondral bone, significant low-density cystic shadow and hip joint space narrow-wing for group C.Gross observation: no obvious abnormalities for group A, visible pinhole defects,formal cartilage darken yellow and some defects for group B, femoral head areacollapse, pinhole defects increases, cartilage defects and peeling for group C.Group A: trabecular bone of the defect area became wide, closely arranged,abundant osteoid, less empty bone lacunae, visible mature bone trabecular andmarrow cavity formation. group B: immature trebecular bone in the defect area, lessosteoid, more empty bone lacunae, no mature trabecular bone and marrow cavityformation. Group C: trabecular bone thin and arranged in a sparse, osteoid rare and alarge number of empty lacunae. the percentage of trabecular and empty lacunaecounts area, the percentage of empty lacunae counts in group A is lower than groupB,C(p<0.05)while the difference between in operation side and normal side is notobvious (p<0.05), the percentage of trabecular area in group A is higher than groupB,C (p<0.05)and the difference between in operation side and normal side is notobvious (p<0.05).Conclusions: The first part: minimally invasive approach and focal alternativecooling and heating insults was be used to the establishment of the earlyosteonecrosis of the femoral head in rabbits, its pathological change process and thehuman early and mid-stage avascular necrosis of the femoral head pathologyevloution rule is very similar, the model is an ideal model to study the early and mid-femoral head necrosis treatment.The second part: Chlorogenic acid combined with bone marrow mesenchymalstem cells orthotopic transplantation for treatment of the early stage osteonecrosis ofthe femoral head has very significant effect. The repairing effect of chlorogenic acidcombined with bone marrow mesenchymal stem cells on rabbits early stageosteonecrosis of the femoral head is better than that of applying bone marrow mesen-chymal stem cells alone.