Angiogenesis in newly regenerated bone by secretomes of human mesenchymal stem cells

Abstract

BackgroundFor an effective bone graft for reconstruction of the maxillofacial region, an adequate vascular network will be required to supply blood, osteoprogenitor cells, and growth factors. We previously reported that the secretomes of bone marrow-derived mesenchymal stem cells (MSC-CM) contain numerous growth factors such as insulin-like growth factor (IGF)-1, transforming growth factor (TGF)-β1, and vascular endothelial growth factor (VEGF), which can affect the cellular characteristics and behavior of regenerating bone cells. We hypothesized that angiogenesis is an important step for bone regeneration, and VEGF is one of the crucial factors in MSC-CM that would enhance its osteogenic potential. In the present study, we focused on VEGF in MSC-CM and evaluated the angiogenic and osteogenic potentials of MSC-CM for bone regeneration.MethodsCytokines in MSC-CM were measured by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were cultured with MSC-CM or MSC-CM with anti-VEGF antibody (MSC-CM + anti-VEGF) for neutralization, and tube formation was evaluated. For the evaluation of bone and blood vessel formation with micro-computed tomography (micro-CT) and for the histological and immunohistochemical analyses, a rat calvarial bone defect model was used.ResultsThe concentrations of IGF-1, VEGF, and TGF-β1 in MSC-CM were 1515.6 ± 211.8 pg/mL, 465.8 ± 108.8 pg/mL, and 339.8 ± 14.4 pg/mL, respectively. Tube formation of HUVECs, bone formation, and blood vessel formation were increased in the MSC-CM group but decreased in the MSC-CM + anti-VEGF group. Histological findings suggested that new bone formation in the entire defect was observed in the MSC-CM group although it was decreased in the MSC-CM + anti-VEGF group. Immunohistochemistry indicated that angiogenesis and migration of endogenous stem cells were much more abundant in the MSC-CM group than in the MSC-CM + anti-VEGF group.ConclusionsVEGF is considered a crucial factor in MSC-CM, and MSC-CM is proposed to be an adequate therapeutic agent for bone regeneration with angiogenesis.

Modifiers of mesenchymal stem cell quantity and quality

“…In contrast, BM‐MSCs secreted only two (IL‐8 and RANTES) cytokines at higher amounts at P3 compared to P1, and secretion of six cytokines and growth factors (fractalkine, PGDF‐BB, IL‐2, IL‐6, IL‐8, and RANTES) was significantly increased at P5 compared to P1. Multiple studies indicate that the biologic functions mediated by MSCs are dependent on cytokine release by the MSC . Since the cytokine secretion profiles of both AD‐MSC and BM‐MSC seem to be passage dependent, it stands to reason that MSC passage number would also influence their biological potency.…”

“…Multiple studies indicate that the biologic functions mediated by MSCs are dependent on cytokine release by the MSC. [14][15][16] Since the cytokine secretion profiles of both AD-MSC and BM-MSC seem to be passage dependent, it stands to reason that MSC passage number would also influence their biological potency. When we compared overall cytokine and growth factor secretion by AD-MSCs with BM-MSCs at P3, we found that AD-MSCs significantly secreted a higher number of cytokines and growth factors at greater concentrations.…”

Angioplasty Using 4-Hexylresorcinol-Incorporated Silk Vascular Patch in Rat Carotid Defect Model

“…Moreover, the platelet-derived growth factor gene, which plays a significant role in angiogenesis, is also overexpressed in the protein solution from the inner silk layer [57]. These factors are believed to have contributed to the increased expression level of VEGF-A, which plays a critical role in angiogenesis, that was observed in the 4-HR silk patch group (Figure 1) [59].…”

Role of Mesenchymal Stem Cell-Conditioned Medium (MSC-CM) in the Bone Regeneration: A Systematic Review from 2007-2018

“…There were two studies which evaluated the importance of angiogenesis capability hence the vascular endothelial growth factor (VEGF) in CM-MSC during acceleration of bone regeneration. In 2017, Katagiri et al [8]. examined the value VEGF in the CM-MSC to accelerate the bone healing by comparing four groups of treatment consisting of MSC-CM, MSC-CM + anti-VEGF antibody, phosphate buffer saline (PBS), and control group with an unfilled defect.…”