Osteoblastic exosomes. A non-destructive quantitative approach of alkaline phosphatase to assess osteoconductive nanomaterials

Sánchez, María Dolores Martín; Felice, Betiana; Sappia, Luciano D.; Moura, Silio Lima; Pividori, M.I.

doi:10.1016/j.msec.2020.110931

Cited by 10 publications

(4 citation statements)

References 50 publications

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“…This could be due to the presence of the enzyme in the EVs. 51,52 On day 7, it increased further by 2.3-fold (p <0.001, n = 3) and 2.2-fold (p <0.01, n = 3) in MC3T3 cells treated with total EVs from complete medium (CM, naıve EVs) and osteogenic differentiation medium (DM, primed EVs), respectively. However, on day 14, the ALP activity decreased significantly by 4.7-fold (p <0.05, n = 3) in MC3T3 cells treated with primed EVs, compared to NC, but there was no significant difference (p >0.05) in ALP production in cells treated with naıve or primed EVs (Figure 7B).…”

Section: Resultsmentioning

confidence: 97%

“…This might be because of EVs carrying ALP enzyme via their cargo, which was also supported by internalization of these EVs by MC3T3 cells within 24 h. 51 A recent parallel study carried out on human fetal osteoblast cell line (hFOB 1.19) showed that these cells secrete exosomes containing ALP enzymes. 52 Decreased ALP secretion by MC3T3 cells treated with EVs harvested in the third week might be the effect of a simultaneous increase in the mineralization process observed on day 14 of culture.…”

Section: Discussionmentioning

confidence: 99%

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Hydrogel-Assisted 3D Model to Investigate the Osteoinductive Potential of MC3T3-Derived Extracellular Vesicles

Holkar

Kale

Ingavle

2021

ACS Biomater. Sci. Eng.

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Effective and rapid regeneration of bone defects often pose substantial challenges in severe accidental injuries and disabilities occurring due to diseases and/or advanced age, especially in patients having reduced tissue regeneration competence. The success of mesenchymal stromal cell (MSC)-based research strategies in improving bone regeneration was hampered not only due to the limited knowledge of therapeutic actions of MSCs but also due to difficulties as well as expenses related to cell manufacturing and time taken for ethical approvals for clinical use of living cells and engineered tissues. The recent trend indicated that there is a shift from the direct usage of MSCs toward the application of paracrine factors and extracellular vesicles (EVs) isolated from their MSC secretome in bone tissue regeneration. This shift has directed research into the development of "cell-free" therapeutics, which could be a better alternative due to its several advantages over the use of their parental MSCs. Furthermore, accumulating evidence suggested that the 3D microenvironment influences the paracrine effects of MSCs. Although the osteogenic role of EVs has been explored recently, the current study showed, for the first time, that encapsulation of EVs along with MC3T3 cells in a 3D hydrogel-assisted culture with a distinct porous microenvironment having meso and macro (0.05−200 μm) pore size distribution resulted in an improved osteogenic response in vitro. The present work was primarily focused on investigating the influence of EVs isolated under distinct priming conditions to enhance the osteogenic potential. In addition, in the current work, the osteogenic ability of different types of EVs (microvesicles and exosomes) and total EVs isolated at different time points was also examined when encapsulated with MC3T3 cells in an alginate gel. Using various biochemical assays, such as alkaline phosphatase (ALP) production and calcium secretion, it was observed that both microvesicles and exosomes collected from MC3T3 cells independently had osteogenic potential; however, their collective activity was found to be superior. We further showed that EVs induce an early osteogenic response in MC3T3 cells as indicated by ALP and calcium secretion at a much earlier time point, compared to the controls. Our data suggested that this 3D hydrogel-assisted system provides close proximity of cells and EVs, and thus, mimics the in vivo scenario, making it clinically useful for bone tissue engineering.

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Section: Resultsmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

Hydrogel-Assisted 3D Model to Investigate the Osteoinductive Potential of MC3T3-Derived Extracellular Vesicles

Holkar

Kale

Ingavle

2021

ACS Biomater. Sci. Eng.

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“…6d ). ALP activity was known as an early marker for osteoblastic differentiation [ 42 , 43 ]. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

Bioinspired laminated bioceramics with high toughness for bone tissue engineering

Huang

Zhai

Xue

et al. 2022

Regenerative Biomaterials

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For the research of biomaterials in bone tissue engineering, it is still a challenge to fabricate bioceramics that overcome brittleness while maintaining the great biological performance. Here, inspired by the toughness of natural materials with hierarchical laminated structure, we presented a directional assembly-sintering approach to fabricate laminated MXene/calcium silicate-based (L-M/CS) bioceramics. Benefiting from the orderly laminated structure, the L-M/CS bioceramics exhibited significantly enhanced toughness (2.23 MPa·m1/2) and high flexural strength (145 MPa), which were close to the mechanical properties of cortical bone. Furthermore, the L-M/CS bioceramics possessed more suitable degradability than traditional CaSiO3 bioceramics due to the newly formed CaTiSiO5 after sintering. Moreover, the L-M/CS bioceramics showed good biocompatibility and could stimulate the expression of osteogenesis-related genes. The mechanism of promoting osteogenic differentiation had been shown to be related to the Wnt signaling pathway. This work not only fabricated calcium silicate-based bioceramics with excellent mechanical and biological properties for bone tissue engineering, but also provided a strategy for the combination of bionics and bioceramics.

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“…Several studies Qi et al (2016) ; Zhang et al (2016) , and Ying et al (2020) reported that the combination of iPSC- or MSC-derived exosomes and a β-TCP scaffold promoted angiogenesis and osteogenesis. Three studies Sanchez et al (2020) ; Wang et al (2020d) , and Zha et al (2021) reported that the combination of exosomes and a PCL scaffold significantly enhanced osteogenic differentiation of MSCs. Gandolfi et al (2020) reported that the mineral-doped PLA porous scaffolds enriched with MSC-derived exosomes increased the osteogenic commitment of MSCs.…”

Section: Stem Cells (Scs)mentioning

confidence: 99%

Technological Advances of 3D Scaffold-Based Stem Cell/Exosome Therapy in Tissues and Organs

Feng

Waqas

et al. 2021

Front. Cell Dev. Biol.

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Osteoblastic exosomes. A non-destructive quantitative approach of alkaline phosphatase to assess osteoconductive nanomaterials

Cited by 10 publications

References 50 publications

Hydrogel-Assisted 3D Model to Investigate the Osteoinductive Potential of MC3T3-Derived Extracellular Vesicles

Hydrogel-Assisted 3D Model to Investigate the Osteoinductive Potential of MC3T3-Derived Extracellular Vesicles

Bioinspired laminated bioceramics with high toughness for bone tissue engineering

Technological Advances of 3D Scaffold-Based Stem Cell/Exosome Therapy in Tissues and Organs

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