Research On The Effect Of Bio-oss On Osteogenesis Of Human Placenta-derived Mesenchymal Stem Cells In Vitro

Purpose and background:By trauma, tumor, alveolar bone resorption after tooth extraction and other causes of alveolar bone defect will directly affect the physical and mental health of patients, it has become a urgent problems for all dentists.Bone tissue engineering technology emerged in recent years has the potential to solve this problem, then how can developed vascularization,high efficiency osteogenesis, small toxic side effects on the body tissue engineered bone naturally became the focus of current research. In this study, we use human placenta-derived mesenchymal stem cells(HPMSCs)as seed cells, Bio-oss bone powder as a scaffold for tissue engineering bone structure. Recent studies on bone tissue engineering seed cells concentrated in bone marrow mesenchymal stem cells, few studies of human placental mesenchymal stem cells at home and abroad. It is through separation of the placenta in vitro extract a class of mesenchymal cells which have the characteristics of stem cells, besides, it has adequate supply,easily obtained, and is not bound by ethics. Bio-oss calf inorganic bone particles is a biological material that have been used in clinical treatment, it has been proved good porosity and biocompatibility.Therefore, this experiment combine human placenta-derived mesenchymal stem cells with Bio-oss bone powder,observing the proliferation and differentiation of HPMSCs after binding. To explore the possibilities of bio-derived bone that human placenta-derived stem cells(HPMSCs) as seed cells, Bio-oss bone powder as scaffolds for tissue engineering.Method:Human placenta-derived mesenchymal stem cells were dissociated under sterile conditions, conventional cell culture in CO2 incubator at room temperature. Tested respectively using the following methods:(1)the third generation of cells determined is used for eight consecutive days cell proliferation test by MTT method, observing morphology and growth characteristics of the cells.(2) Osteogenic in osteogenic medium, Alizarin Red staining after 21 days and observing.Dividing the cells into induced osteogenesis and basic training group, in the induction of 3 days, 7 days,14 days three time points detecting the alkaline phosphatase(ALP) of two groups of cells.(3)Inducing in adipogenic liquid,observing the results of red oil O staining after 21 days.(4) The third generation cells were divided into experimental group and control group, the experimental group cells and Bio-oss bone composite culture, control cells cultured alone, In 2, 4, 6, 8 days determined by MTT method to detect cell proliferation, the two groups with BCA method to detect the contents of protein, alkaline phosphatase(ALP).Results:1. Consecutive 8 days MTT results showed that: with the increase of cell culture time, OD value becomes larger, indicating that the cells increased.2. Red nodules of calcium are visible after osteogenic induction and ALP test results showed that the alkaline phosphatase of induced group more than non-induction group.Lipid droplets are visible clearly after adipogenic.3. The results determined by MTT、ALP and BCA show that there are no statistical difference between the experimental group and control group after cells combined with bone powder.Conclusions:There are no significant differences on proliferation, differentiation and other biological characteristics between the composite culture group and control group. Therefore, human placenta-derived mesenchymal stem cells as seed cells combined with Bio-oss bone powder for tissue engineered bone is feasible.