Abstract
Microcapsules with entrapped cells hold great promise for repairing bone defects. Unfortunately, the osteoinductivity of microcapsules has been restricted by many factors, among which the deficiency of functional proteins is a significant priority. We potentiated the osteoinductivity of microencapsulated cells via cotransfection with BMP-2 and VEGF genes. Various tissue-derived mesenchymal stem cells and cell lines were compared for BMP-2 and VEGF cotransfection. Ethidium bromide (EB)/Calcein AM staining revealed that all of the cell categories could survive for 4 weeks after microencapsulation. An ELISA assay indicated that all microencapsulated BMP-2 or VEGF transfected cells could secrete gene products constitutively for 1 month. Particularly, the recombinant microencapsulated C2C12 cells released the most desirable level of BMP-2 and VEGF. Further experiments demonstrated that microencapsulated BMP-2 and VEGF cotransfected C2C12 cells generated both BMP-2 and VEGF for 4 weeks. Additionally, the cotransfection of BMP-2 and VEGF in microencapsulated C2C12 cells showed a stronger osteogenic induction against BMSCs than individual BMP-2-transfected microencapsulated C2C12 cells. These results demonstrated that the cotransfection of BMP-2 and VEGF into microencapsulated C2C12 cells is of potent utility for the potentiation of bone regeneration, which would provide a promising clinical strategy for cellular therapy in bone defects.
Highlights
One major obstacle encountered by clinical orthopedic practice is the repair of bone defects caused by trauma, malignant disease, and prosthetic replacement [1]
Regarding the adipose-derived stem cells (ADSCs), after the first inoculation of 5–7 hr, some cells began to adhere and exhibited elongated pseudopodia, thereby contributing to a spindle-like appearance. 24 hr later, the cells completely adhered to the dishes, exhibiting a fibroblast-like cell growth with a proliferative peak at 3–5 days (Figure 1(a)-(B))
We explored the cultivation of microencapsulated bone marrow mesenchymal stem cells (BMSCs), ADSCs, synovium-derived mesenchymal stem cells (SMSCs), C3H10T1/2, C2C12, and NIH/3T3 cells, revealing that the microcapsules were able to provide sufficient living conditions for the inner cells
Summary
One major obstacle encountered by clinical orthopedic practice is the repair of bone defects caused by trauma, malignant disease, and prosthetic replacement [1]. The most common approaches to repair bone defects, such as the transplantation of autologous and allogeneic bone grafts or substitution of artificial bone, exhibit many disadvantages These disadvantages include the scarcity of supply resources, the risk of disease dissemination, and deficient osteogenesis, which lead to the delayed union or the nonunion of the bone [2,3,4]. The alginate-poly-L-lysinealginate (APA) microcapsules first reported by Lim and Sun [12] appear to exert an immune-protective effect on entrapped cells and form a spherical shape with a smooth surface and consistent uniformity. These were considered to be suitable standards for the use of microencapsulation in cell treatment research
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