Evaluation of BMP-2 Enhances the Osteoblast Differentiation of Human Amnion Mesenchymal Stem Cells Seeded on Nano-Hydroxyapatite/Collagen/Poly(l-Lactide)

Abstract

Background: The aim of this study is to evaluate the effects of recombinant human bone morphogenetic protein 2 (rhBMP-2), human amnion mesenchymal stem cells (hAMSCs), and nanohydroxyapatite/collagen/poly(l-lactide) (nHAC/PLA) in tissue engineering to provide potential approaches for periodontal bone regeneration. Methods: hAMSCs were isolated from discarded amniotic membrane samples and cultured in vitro. Alkaline phosphatase (ALP) staining and alizarin red staining were performed to evaluate the osteoblast (OB) differentiation ability of hAMSCs. Three groups were divided: the experimental group (cells transfected with pcDNA3.1-rhBMP-2), the blank group (cells without gene transfection), and the control group (cells transfected with empty plasmid). RT-PCR and western blot were used to examine whether rhBMP-2 has been successfully expressed. 3-(4,5)-dimethylthiahiazol(-z-y1)-3,5-di-phenytetrazo-liumromide assay (MTT) was done to detect the effect of rhBMP-2 on hAMSCs seeded on nHAC/PLA. ALP activity, mineral formation assay, calcium, phosphate and osteocalcin (OCN) content, and OCN and RUNX2 expression of hAMSCs were detected to evaluate osteogenic differentiation capability of rhBMP-2 on hAMSCs seeded on nHAC/PLA. Results: hAMSCs exhibited intense ALP staining, obvious calcium deposition, and mineralization nodules, and rhBMP-2 were highly expressed in the experimental group. The proliferation of the hAMSCs with rhBMP-2 on nHAC/PLA was significantly higher than the cells without rhBMP-2, and the cells all increased in a time-dependent manner. rhBMP-2 significantly increased the OCN and phosphate content, mineral formation, ALP activity, osteogenic biomarkers OCN, and Runx2, and decreased calcium content in hAMSCs seeded on the nHAC/PLA scaffold. Conclusions: This finding demonstrated that hAMSCs has an ideal OB differentiation ability. rhBMP-2 facilitates the proliferation and osteogenesis of hAMSCs. The nHAC/PLA could act as a good scaffold for hAMSCs seeding, proliferation, and osteogenic differentiation. The application of rhBMP-2, nHAC/PLA, and hAMSCs in tissue engineering may offer promising possibilities for periodontal bone regeneration.

Hydroxylapatite-collagen hybrid scaffold induces human adipose-derived mesenchymal stem cells to osteogenic differentiation in vitro and bone regrowth in patients

“…Epigenetic analyses carried out in 84 genes of the osteogenetic pathway, in hASCs grown on biomaterial, indicate that among DEGs BMP2 is induced and upregulated. Accumulating studies proved that BMP‐2 is involved in bone formation, bone remodeling, bone development, and osteoblast differentiation . BMP‐2, belonging to the BMPs family, is a potent osteoinductive cytokine from the transforming growth factor beta (TGF‐β) family, and it is currently the most commonly used protein‐based bone graft substitute .…”

“…BMP‐2, belonging to the BMPs family, is a potent osteoinductive cytokine from the transforming growth factor beta (TGF‐β) family, and it is currently the most commonly used protein‐based bone graft substitute . Recombinant human bone morphogenetic protein 2 (rhBMP‐2), in human amnion MSCs, with nano‐HA/Coll/poly ( l ‐lactide), provides a potential approaches in tissue engineering for periodontal bone regeneration . Furthermore, in vitro and in vivo results using a rat cranial bone defect model demonstrated that the 3D nanofibrous scaffolds incorporated with nano‐hydroxylapatite (nHA) and BMP‐2 peptides exhibited favorable biocompatibility and osteoinductivity .…”

“…Accumulating studies proved that BMP-2 is involved in bone formation, bone remodeling, bone development, and osteoblast differentiation. 31,32 BMP-2, belonging to the BMPs family, is a potent osteoinductive cytokine from the transforming growth factor beta (TGF-β) family, and it is currently the most commonly used protein-based bone graft substitute. 33 Recombinant human bone morphogenetic protein 2 (rhBMP-2), in human amnion MSCs, with nano-HA/Coll/poly (L-lactide), provides a potential approaches in tissue engineering for periodontal bone regeneration.…”

“…33 Recombinant human bone morphogenetic protein 2 (rhBMP-2), in human amnion MSCs, with nano-HA/Coll/poly (L-lactide), provides a potential approaches in tissue engineering for periodontal bone regeneration. 31 Furthermore, in vitro and in vivo results using a rat cranial bone defect model demonstrated that the 3D nanofibrous scaffolds incorporated with nano-hydroxylapatite (nHA) and BMP-2 peptides exhibited favorable biocompatibility and osteoinductivity. 34 In addition, the DEG analysis performed by RT 2 Profiler PCR array allowed us to detect early epigenetic modifications induced in hASCs by scaffold.…”

Wnt and BMP signaling pathways co‐operatively induce the differentiation of multiple myeloma mesenchymal stem cells into osteoblasts by upregulating EMX2

“…22 2.14 | Statistical analysis Statistical analysis was performed using SPSS 21.0 Software (IBM Corp. Armonk, NY). After 24 hours of culture, cells were treated with trypsin and centrifuged at 157g for 5 minutes, and resuspended with PBS containing 3% penicillin.…”