An axial distribution of seeding, proliferation, and osteogenic differentiation of XXXX cells and @entity35 bone marrow-derived mesenchymal stem cells across a 3D Thai silk fibroin/gelatin/ @entity692 scaffold in a perfusion bioreactor.

@entity19 @entity26 @entity692 @entity2146 @entity460 @entity35
In cell culture, a perfusion bioreactor provides effective transportation of nutrients, @entity26 , and waste removal to and from the core of the scaffold. In addition, it provides mechanical stimuli for enhancing osteogenic differentiation. In this study, we used an axial distribution of cell numbers, alkaline phosphatase (ALP) enzyme activity, and @entity460 content across 4 cross-sections of 10mm thick scaffold, made of Thai silk fibroin (SF)/gelatin (G)/ @entity692 (HA), as a tool to evaluate the suitable perfusion flow rate. These evaluations cover all cellular developmental phases starting from seeding, to proliferation, and later osteogenic differentiation. @entity19 pre-osteoblastic @entity19 cell lines were used as a cell model during seeding and proliferation. The bioreactor seeded scaffold provided more uniform cell distribution across the scaffold compared to centrifugal and @entity2146 seeding, while the overall number of adhered cells from bioreactor seeding was slightly lower than @entity2146 seeding. The dynamic culture using 1ml/min perfusion flow rate (initial shear stress of 0.1dyn/cm(2)) enabled statistically higher MC3T3-E1 proliferation, ALP activity, and @entity460 deposition than those observed in the static-culturing condition. However, the perfusion flow rate of 1ml/min seemed not to be enough for enhancing ALP expression across all sections of the scaffold. @entity35 bone marrow derived stromal cells (rMSC) were used in the detachment test and osteogenic differentiation. It was found that perfusion flow rate of 5ml/min caused statistically higher cell detachment than that of 1 and 3ml/min. The perfusion flow rate of 3ml/min gave the highest rMSC osteogenic differentiation on a SF/G/HA scaffold than other flow rates, as observed from the significantly highest number of ALP enzyme activity and the @entity460 content without any significant cell growth. In addition, all of these parameters were evenly distributed across all scaffold sections.
