The Potential Of ERβ For Bone Marrow Mesenchyme Stem Cells Differentiating Chondrogenesis

Objective To explore the different of normal and ERβ^-\-mouse’ BMMSCs chondrogenicdifferentiation by culture both of the BMMSCs in vitro. And observe the process of impairedcartilage and subchondral bone healing by histopathology and Mirco CT scan: a drill-holefull-thickness cartilage and subchondral bone defect of the unilateral trochlear of femoral.Methods The BMMSCs were obtained by centrifugated with percoll solution and the differentspeed of adherence to plastic bottle. Each group of3rdpassage cells were subjected to FACSanalysis to determine cell phenotype as CD29、CD44、CD105、CD34、CD45、CD11b. And drewgrowth curve by CCK-8method. We used3rdpassage cells for chondrogenic differentiation withserum free and HG-DMEM containing10ng/mL TGF-β1,100nmol/L decaesadril and50ug/mLantiscorbic acid. After21days induced, the cells were detected for type-Ⅱcollagen byimmunohistochemical analysis and RT-PCR. Observe the process of impaired subchondral boneand cartilage healing by Mirco CT scan and histopathology after the surgery of a drill-holefull-thickness cartilage defect of the unilateral trochlear of femoral at1、2、3、4weeks.Results The C57and ERβ^-\-mouse’ BMMSCs were stained positive for MSCs marker CD29、CD44、CD105, but were negative for hematopoietic stem cell marker CD34、CD45、CD11b.After21days induced, Immunocytochemistry showed that type-Ⅱ collagen in both C57andERβ^-\-mouse’ induced cells. And the differentiation rates were （56.30±5.25）%for C57,（42.61±6.72）%for ERβ^-\-（P＜0.05）. The RT-PCR showed that the C57had higher mRNA oftype-Ⅱ collagen than ERβ^-\-with （43.08±8.73）%，（P＜0.01）. Mirco CT scan each group showedthat there been new bone in the hole at7days and healing at28days. And the histopathologyshowed that there had been new hyaline cartilage which in the drill-hole.Conclusions1The high purity BMMSCs were obtained by centrifugated with percoll solution and thedifferent speed of adherence to plastic bottle.2The BMMSCs could differentiate into chondyocyte with free serum and HG-DMEMcontaining10ng/mL TGF-β1,100nmol/L decaesadril and50ug/mL antiscorbic acid.3The BMMSCs lack of ERβ gene had diminished capacity to differentiate into chondyocyte, and the ability of secretion typeⅡcollagen was reduced.40.6mm in diameter of the full-thickness cartilage defect which in the unilateral trochlear offemoral could been heal.And there had been new hyaline cartilage.