Biocompatibility Study On Tissue Engineering Human Acellular Amniotic Membrane Bioscaffold

Objective: To evaluate the biocompatibility and cytocompatibility of acellular amniotic membrane(HAAM) invitro. And to explore the potential possibility of HAAM to be an novel candidate of 3D adhension matrix for orthopedic tissue engineering. Methods: MC3T3-E1 cells were seeded on HAAM and cultured in a-MEM routinely. To observe the cells configuration, survival time, and the cells distribution, the invert phase contrast microscope and electron microscope was engaged. Expression and location of FAK, Integrin and Paxillin was detected by immunofluorescence staining and laser confocal scanning microscope for evaluate the cell-matrix adhesion manner. Results: Under invert phase contrast microscope MC3T3-E1 adhered completely in 12hours and became confluence on HAAM in three days. The cells configuration were miscellaneous, with bipolar, multipolar or rounded shape. Afer 7 days, they could secreted matrix. Cells migrated into clusters, there were cell beams among the clusters and knock the clusters together. Cells are gradually apoptosis until 21days routine culture. During the survival, integrin, FAK, paxllin were strong positive in immunofluorescence staining. Localization of integrin(green), FAK (red) and FAK (red) ,Paxillin (green) were double labled .The yellow represents the merged labeled of integrin(green) NFAK(red) and FAK(red) ^Paxillin(green). integrin(green)> FAK (red) and FAK (red) > Paxillin (green) strongly confocal expression.-Conclusion: 1.MC3T3-E1 cells can survival on the HAAM under routine culture for 3 weeks, and migrated into clusters with communication. 2. MC3T3-E1 cells can excrete biomolecules, and survival in different phase of cell circle. 3. MC3T3-E1 cells adhesion with HAMM in a manner of 3D adhension. HAMM is potential novel candidate of 3D matrix for orthopaedic tissue engineering.