Modified perfusion bioreactor enhances proliferation of MC3T3-E1 seeded on PLGA/Β-TCP/PCL scaffold

IntroductionBone tissue engineering, an important field in contemporary clinical practices, aims to repair or replace bone defects using cells, scaffolds, and bioreactors. This study investigated whether modified perfusion bioreactor at 5 ml/min flow rate enhances proliferation, distribution and extracellular matrix formation of MC3T3-E1 cells cultured on a porous PLGA/β-TCP which PCL porous hollow fibers was embedded in.MethodsMC3T3-E1 pre-osteoblastic cells were seeded onto the porous PLGA/β-TCP/PCL scaffolds (2 cm in diameter and 1 cm in thickness) by adding 200 μl of cell suspension media with 1×106 cells to each scaffold in a dropwise manner and incubated for 2 hr (200 μl of culture medium were added to each scaffold every 30 min). After 2 hr, 10 ml of culture medium was added to each scaffold and incubated in a humidified atmosphere at 37 °C and 5% CO2. After an overnight static culture, scaffolds were transferred to the modified perfusion bioreactors (one scaffold for each bioreactor) with flow rates 5 ml/min.ResultsOur data showed that this new designed perfusion bioreactor with flow rate of 5 ml/min increased proliferation 1.8-fold after 5 days of dynamic culture. The homogeneous cell distribution and extracellular matrix formation within the PLGA/β-TCP scaffolds were demonstrated through DAPI staining and Alizarin red staining, respectively.ConclusionIn conclusion, modified perfusion bioreactor promotes cell proliferation, cell distribution and extracellular matrix formation in bone tissue-engineered constructs which is a promising approach to treat patients with large bone defects.

Keywords:

Bone , Modified perfusion bioreactor , proliferation , Cell distribution , Extracellular matrix