The Preparation PHBHOx/collagen Integrated Composite Scaffold Used In Osteochondral Repair And The Cellular Compatibility

Tissue engineering techniques was established in the1980s, which means thatmodern medicine step out of the scope of organ transplantation and enter into a newera of organ reconstruction. Cartilage tissue engineering as an important branch oftissue engineering, has made great progress in recent years. It has changed simplecartilage repair into osteochondral integrated repair. So it is necessary to developosteochondral integration scaffolds.This article, we initially selected Poly(3hydroxybutyrate co3hydroxyoctano)(PHBHOx) composited with type I collagen for preparation of osteochondralintegrated tissue engineering scaffold using solvent casting particulate leachingmethod. In order to meet the osteochondral tissue engineering requirements of thescaffold, we tested a variety of scaffoldsˊperformance and then assessed. To lay thefoundation for the use of tissue engineering technology to repair osteochondraldamage.We firstly made PHBHOx soluble in chloroform, and then added collagen typeⅠat a mass ratio of2%(collagen/PHBHOx), fully stirred well, added a certainamount of NaCl particles(100m), stirred well and loaded to the mold, extruded as thecartilage layer. Next we made PHBHOx soluble in chloroform, and then addedcollagen type Ⅰat a mass ratio of2%(collagen/PHBHOx), fully stirred well, addeda certain amount of NaCl particles(200m), stirred well and loaded to the mold,extruded as the bone layer. When it will be dried, connected cartilage layer and bonelayer. This allows the upper and lower layers firmly bonded. After the chloroformcompletely volatile，soaked in distilled water48h to remove porogen NaCl Particle,dried in the oven, called osteochondral integrated composite scaffold as theexperimental group. The control group scaffolds Simply made with PHBHOx，usingthe method as previously described.After that, we tested pore size and porosity of scaffolds. Isolated and culturedrabbit bone marrow mesenchymal stem cells (BMSCs), selected the secondarygeneration, seeded into scaffolds, cultured in vitro. Five days later, observe theadhesion of BMSCs through scanning electron microscopy. Respectively after2,4,6,8days, evaluate cell proliferation using MTT method and analysis results.The results shows that pore size of cartilage layer was80100m, porosity was90%, pore size of bone layer was200220m，porosity was70%. Porosity of thewhole scaffold was80%. The conditions above all meets the requirements of the osteochondral tissue engineering scaffold. Using the density gradient centrifugation toisolated BMSCs, is a reliable method with high cell survival rate and easy operation.The adhesion of BMSCs on scaffolds was very good, MTT method confirmed thatcells owns rapid proliferation.The results of the study show scaffolds made of PHBHOx composited withcollagen Ⅰ, meets the requirements of the osteochondral tissue engineering scaffoldsand is expected to be used in tissue engineering for osteochondral repair.