A quantitative method to determine osteogenic differentiation aptness of scaffold

Trivedi, Shilpa; Srivastava, Kirti; Gupta, Arobinda; Saluja, Tajindra Singh; Kumar, Sumit; Mehrotra, Divya; Singh, Satyendra Kumar

doi:10.1016/j.jobcr.2020.04.006

Cited by 32 publications

(18 citation statements)

References 10 publications

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“…As an early indicator of osteoblast differentiation, ALP regulates the extracellular matrix mineralization. , In this work, the ALP activity of MC3T3-E1 cells was evaluated after cultivation for 7 days. The results showed that the ALP activity on the Ti2448-PDA-ECM scaffolds was noticeably higher than that from the Ti2448 scaffolds (Figure c and Table S2).…”

Section: Resultsmentioning

confidence: 99%

Improving Biological Functions of Three-Dimensional Printed Ti2448 Scaffolds by Decoration with Polydopamine and Extracellular Matrices

Bai

Misra

et al. 2022

ACS Appl. Bio Mater.

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Extracellular matrices (ECMs) provide important cues for cell proliferation and differentiation in the complex environment, which show a significant influence on cell functions. Herein, cell-derived ECMs were deposited on the polydopamine (PDA)-decorated porous Ti-24Nb-4Zr-8Sn (Ti2448) scaffolds fabricated by the electron beam melting method in order to improve biological functions. The influence of PDA-ECM coatings on cell functions was further investigated. The results demonstrated that the PDA-ECM coating facilitated adhesion, proliferation, and migration of MC3T3-E1 cells on Ti2448 scaffolds. Moreover, Ti2448-PDA-ECM scaffolds promoted osteogenesis differentiation of cells indicated by greater alkaline phosphatase activity and further mineralization, compared to the plain Ti2448 group. Meanwhile, Ti2448-PDA-ECM scaffolds enhanced bone growth after implantation for one month in rabbit femoral bone defects. Our findings suggest that the bioinspired PDA-ECM coating can be implemented on the porous Ti2448 scaffolds, which significantly improve the biological functions of orthopedic implants.

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Section: Resultsmentioning

confidence: 99%

Improving Biological Functions of Three-Dimensional Printed Ti2448 Scaffolds by Decoration with Polydopamine and Extracellular Matrices

Bai

Misra

et al. 2022

ACS Appl. Bio Mater.

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“…To gain further insight into the tissue-specificity of ECM, relative mRNA expression of ALP and ACAN was investigated. ALP is recognized as an early marker of osteoblast differentiation as it is a by-product of osteoblast activity, thereby suggesting a correlation between increased expression and active bone formation [ 36 ]. ACAN is recognized as a hallmark of articular chondrocytes as its covalently attached sulfated glycosaminoglycan [ 10 ] chains exert water-attracting characteristics and generate a fixed negative charge of cartilage [ 8 , 31 ].…”

Section: Discussionmentioning

confidence: 99%

Degenerative tendon matrix induces tenogenic differentiation of mesenchymal stem cells

Hwang

Lee

2023

J. exp. orthop.

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Purpose Mesenchymal stem cells (MSCs) react dynamically with the surrounding microenvironment to promote tissue-specific differentiation and hence increase targeted regenerative capacity. Extracellular matrix (ECM) would be the first microenvironment to interact with MSCs injected into the tissue lesion. However, degenerative tissues would have different characteristics of ECM in comparison with healthy tissues. Therefore, the influence of degenerative ECM on tissue-specific differentiation of MSCs and the formation of matrix composition need to be considered for the sophisticated therapeutic application of stem cells for tissue regeneration. Methods Human degenerative tendon tissues were obtained from patients undergoing rotator cuff repair and finely minced into 2 ~ 3 mm fragments. Different amounts of tendon matrix (0.005 g, 0.01 g, 0.025 g, 0.05 g, 0.1 g, 0.25 g, 0.5 g, 1 g, and 2 g) were co-cultured with bone marrow MSCs (BM MSCs) for 7 days. Six tendon-related markers, scleraxis, tenomodulin, collagen type I and III, decorin, and tenascin-C, osteogenic marker, alkaline phosphatase (ALP), and chondrogenic marker, aggrecan (ACAN), were analyzed by qRT-PCR. Cell viability and senescence-associated beta-galactosidase assays were performed. The connective tissue growth factor was used as a positive control. Results The expressions of six tendon-related markers were significantly upregulated until the amount of tendon matrix exceeded 0.5 g, the point where the mRNA expressions of all six genes analyzed started to decrease. The tendon matrix exerted an inhibitory effect on ACAN expression but had a negligible effect on ALP expression. Cell viability did not change significantly over the culture period. The amount of tendon matrix exceeding 0.01 g significantly increased the SA-βgal activity of BM MSCs. Conclusion This study successfully demonstrated tendon ECM-stimulated tenogenesis of BM MSCs through an indirect co-culture system without the use of exogenous growth factors and the alteration of cellular viability. In contrast to the initial hypothesis, the tenogenesis of BM MSCs induced with the degenerative tendon matrix accompanied cellular senescence.

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“…The primary marker of osteoblasts, alkaline phosphatase, is a membrane enzyme that hydrolyzes phosphate ions, allowing the formation of hydroxyapatite crystals and stimulating mineralization. ALP plays a crucial role in bone regeneration and differentiation [ 110 ]. Compared to pure gelatin scaffold, Li-doped bioactive glass, PRGF, and a combination group showed a substantial increase in ALP activity with increasing culture time.…”

Section: Discussionmentioning

confidence: 99%

Preparation and In Vitro Osteogenic Evaluation of Biomimetic Hybrid Nanocomposite Scaffolds Based on Gelatin/Plasma Rich in Growth Factors (PRGF) and Lithium-Doped 45s5 Bioactive Glass Nanoparticles

et al. 2022

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Bone tissue engineering is an emerging technique for repairing large bone lesions. Biomimetic techniques expand the use of organic–inorganic spongy-like nanocomposite scaffolds and platelet concentrates. In this study, a biomimetic nanocomposite scaffold was prepared using lithium-doped bioactive-glass nanoparticles and gelatin/PRGF. First, sol–gel method was used to prepare bioactive-glass nanoparticles that contain 0, 1, 3, and 5%wt lithium. The lithium content was then optimized based on antibacterial and MTT testing. By freeze-drying, hybrid scaffolds comprising 5, 10, and 20% bioglass were made. On the scaffolds, human endometrial stem cells (hEnSCs) were cultured for adhesion (SEM), survival, and osteogenic differentiation. Alkaline phosphatase activity and osteopontin, osteocalcin, and Runx2 gene expression were measured. The effect of bioactive-glass nanoparticles and PRGF on nanocomposites' mechanical characteristics and glass-transition temperature ( T g ) was also studied. An optimal lithium content in bioactive glass structure was found to be 3% wt. Nanoparticle SEM examination indicated grain deformation due to different sizes of lithium and sodium ions. Results showed up to 10% wt bioactive-glass and PRGF increased survival and cell adhesion. Also, Hybrid scaffolds revealed higher ALP-activity and OP, OC, and Runx2 gene expression. Furthermore, bioactive-glass has mainly increased ALP-activity and Runx2 expression, whereas PRGF increases the expression of OP and OC genes. Bioactive-glass increases scaffold modulus and T g continuously. Hence, the presence of both bioactive-glass and nanocomposite scaffold improves the expression of osteogenic differentiation biomarkers. Subsequently, it seems that hybrid scaffolds based on biopolymers, Li-doped bioactive-glass, and platelet extracts can be a good strategy for bone repair. Graphical Abstract

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A quantitative method to determine osteogenic differentiation aptness of scaffold

Cited by 32 publications

References 10 publications

Improving Biological Functions of Three-Dimensional Printed Ti2448 Scaffolds by Decoration with Polydopamine and Extracellular Matrices

Improving Biological Functions of Three-Dimensional Printed Ti2448 Scaffolds by Decoration with Polydopamine and Extracellular Matrices

Degenerative tendon matrix induces tenogenic differentiation of mesenchymal stem cells

Preparation and In Vitro Osteogenic Evaluation of Biomimetic Hybrid Nanocomposite Scaffolds Based on Gelatin/Plasma Rich in Growth Factors (PRGF) and Lithium-Doped 45s5 Bioactive Glass Nanoparticles

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