Osteogenic medium is superior to growth factors in differentiation of human adipose stem cells towards bone-forming cells in 3D culture

Tirkkonen, Laura; Haimi, Suvi; Huttunen, Sanna; Wolff, Jan; Pirhonen, Eija; Sándor, GK; Miettinen, Susanna

doi:10.22203/ecm.v025a10

Cited by 56 publications

(65 citation statements)

References 69 publications

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“…On the other hand, TGF-β1 protects pre-osteocytes from apoptosis [63]. Similar results regarding the osteogenic medium were obtained by Tirkkonen et al [64] in an adipose stem cell osteogenic differentiation study that compared the efficiency of BMP-2 and BMP-7, VEGF and osteogenic medium. Stem cells were grown on commercially available bioactive glass scaffolds and biphasic granules.…”

Section: Discussionsupporting

confidence: 61%

“…They did not find a benefit after the addition ofgrowth factors in comparison with plain osteogenic medium. Moreover, BMP-7 inhibited the proliferation and osteogenic differentiation of adipose stem cells [64]. The lack of improvement of the osteogenic differentiation of stem cells through the addition of BMP-2 or BMP-6 was also reported by other authors [65-67].…”

Section: Discussionmentioning

confidence: 80%

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Comparative in vitro study regarding the biocompatibility of titanium-base composites infiltrated with hydroxyapatite or silicatitanate

et al. 2014

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BackgroundThe development of novel biomaterials able to control cell activities and direct their fate is warranted for engineering functional bone tissues. Adding bioactive materials can improve new bone formation and better osseointegration. Three types of titanium (Ti) implants were tested for in vitro biocompatibility in this comparative study: Ti6Al7Nb implants with 25% total porosity used as controls, implants infiltrated using a sol–gel method with hydroxyapatite (Ti HA) and silicatitanate (Ti SiO2). The behavior of human osteoblasts was observed in terms of adhesion, cell growth and differentiation.ResultsThe two coating methods have provided different morphological and chemical properties (SEM and EDX analysis). Cell attachment in the first hour was slower on the Ti HA scaffolds when compared to Ti SiO2 and porous uncoated Ti implants. The Alamar blue test and the assessment of total protein content uncovered a peak of metabolic activity at day 8–9 with an advantage for Ti SiO2 implants. Osteoblast differentiation and de novo mineralization, evaluated by osteopontin (OP) expression (ELISA and immnocytochemistry), alkaline phosphatase (ALP) activity, calcium deposition (alizarin red), collagen synthesis (SIRCOL test and immnocytochemical staining) and osteocalcin (OC) expression, highlighted the higher osteoconductive ability of Ti HA implants. Higher soluble collagen levels were found for cells cultured in simple osteogenic differentiation medium on control Ti and Ti SiO2 implants. Osteocalcin (OC), a marker of terminal osteoblastic differentiation, was most strongly expressed in osteoblasts cultivated on Ti SiO2 implants.ConclusionsThe behavior of osteoblasts depends on the type of implant and culture conditions. Ti SiO2 scaffolds sustain osteoblast adhesion and promote differentiation with increased collagen and non-collagenic proteins (OP and OC) production. Ti HA implants have a lower ability to induce cell adhesion and proliferation but an increased capacity to induce early mineralization. Addition of growth factors BMP-2 and TGFβ1 in differentiation medium did not improve the mineralization process. Both types of infiltrates have their advantages and limitations, which can be exploited depending on local conditions of bone lesions that have to be repaired. These limitations can also be offset through methods of functionalization with biomolecules involved in osteogenesis.

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Section: Discussionsupporting

confidence: 61%

Section: Discussionmentioning

confidence: 80%

Comparative in vitro study regarding the biocompatibility of titanium-base composites infiltrated with hydroxyapatite or silicatitanate

et al. 2014

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“…Contrary to the results of our study, D'Alimonte et al 42) showed that the addition of VEGF to the DSCs (10-40 ng/ml for 8-day dose-and timedependently) in osteogenic condition could enhance both osteogenesis and angiogenesis, while results of the current study disagreed with these fi ndings. The work of Tirkkonen et al 43) strongly agreed with our study as he confi rmed that using high dose of VEGF protein in free released did not enhance osteogenesis process due to diff erentiation of HASC by meant BMP-2 inhibition. BMP-2 is a growth factor that has a high ability to induce osteogenesis by inducing the signaling control osteogenesis 44) .…”

Section: Discussionsupporting

confidence: 90%

Osteoinductive Activity of Bone Scaffold Bioceramic Companied with Control Release of VEGF Protein Treated Dental stem cells as A New Concept for Bone Regeneration: Part II

Enezei

Afsari

Takeuchi

et al. 2018

J. Hard Tissue Biology.

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In bone tissue engineering, angiogenesis is closely associated with osteogenesis where reciprocal interactions between endothelial and osteoblast cells play an important role in bone regeneration. Over-expression of the angiogenesisrelated gene due to a higher dose of vascular endothelial growth factor (VEGF) protein can inhibit osteogenesis process at mRNA level. To study the eff ect of controlled released of the VEGF protein incorporating fi brin glue (FG) treated with fabricated porous biphasic calcium phosphate (BCP) on osteogenesis gene (BMP-2) and angiogenesis gene (VEGF) on dental stem cells (DSCs) at mRNA level. DSCs were treated with two diff erent modalities; VEGF protein incorporated FG, and VEGF protein incorporated FG added-BCP treated media. The cells were harvested at four diff erent time intervals (day 3, day 7, day 10 and day 14) and were subjected to RNA isolation using the RNA extraction kit. This was followed by performing one step-reverse transcriptase-PCR (RT-PCR) to amplify the osteogenesis BMP-2 gene, angiogenesis VEGF gene and the osteoblast-specifi c transcription factor expression Osterix (Osx) with and without the controlled release of VEGF protein. The RT-PCR products were then electrophoresed. The gel image was captured using Image Analyser. Controlled release of VEGF protein using FG as a natural delivery system, using a single growth factor, show a signifi cantly enhanced osteogenesis BMP-2 gene and angiogenesis VEGF gene with a high expression of Osx compare with non-delivered free VEGF protein treated groups. FG is a biocompatible material that could be employed as a delivery vehicle for controlled release of VEGF protein single or dual release in bone tissue engineering strategy and design of the study. Application of this method for using FG is mixing with a porous ceramic scaff old loading with the growth factors is a convenient and promising strategy for improving osteogenesis and angiogenesis processes of reconstruction critical-sized bone defects and might change the scope of modern surgery.

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“…The current finding showed that only angiogenic enhances with VEGF-BCP combined group but not osteogenic. The work of Tirkkonen et al 31) strongly agreed with our study fi ndings as he confi rmed that VEGF did not enhance osteogenesis diff erentiation of human adipose stem cells (HASC) by meant BMP-2 inhibition. From this information, it is, therefore, clear that VEGF-BCP culture conditions had signifi cant eff ects in these experiments, but a defi nitive conclusion on angiogenesis and osteogenesis will need to be confi rmed by further studies.…”

Section: Discussionsupporting

confidence: 88%

Enhanced Osteogenic and Angiogenic-Related Gene Expression of Human Dental Stem Cells on Biphasic Calcium Phosphate Scaffold Treated with Vascular Endothelial Growth Factor: Part I

Enezei

Ahmad

Khamis

et al. 2017

J. Hard Tissue Biology.

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To study the eff ectiveness of vascular endothelial growth factor (VEGF) added with biphasic calcium phosphate (BCP) on the expression of osteogenesis and angiogenesis-related gene in dental stem cells (DSCs). The cells were treated with three diff erent modalities; BCP group, VEGF group, and VEGF-added-BCP. The optimal BCP and VEGF concentrations were determined. The cells were harvested at four diff erent time intervals (day 3, day 7, day 10 and day 14) and were subjected to RNA isolation. Osteogenesis and angiogenesis-regulated genes was amplifi ed using reverse transcriptase-PCR (RT-PCR). The RT-PCR products were then electrophoresed. The gel images were captured using Image Analyser AlphaEaseFC™. Angiogenesis and osteogenesis genes were clearly expressed in DSCs in response to treatments with 75mg/ml BCP and 5ng/ml VEGF. Angiogenesis gene VEGF was highly expressed by VEGF treatment group but showed some changes when added with BCP. Osteogenesis genes (BMP-2 and OPN) were positively aff ected by both BCP and VEGF. Some genes were expressed at an earlier time interval compared to the other genes depending on the type of treatments. BCP treatment induced high expression of initial-regulated osteogenesis genes (BMP-2 and OPN). Combination of BCP and VEGF modality on DSCs was suggested to initiate osteogenesis and angiogenesis-related gene expressions earlier than the other modalities. In our design, uncontrolled release of VEGF protein showed inhibition of BMP-2 at mRNA level in VEGF-BCP combined group.

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Osteogenic medium is superior to growth factors in differentiation of human adipose stem cells towards bone-forming cells in 3D culture

Cited by 56 publications

References 69 publications

Comparative in vitro study regarding the biocompatibility of titanium-base composites infiltrated with hydroxyapatite or silicatitanate

Comparative in vitro study regarding the biocompatibility of titanium-base composites infiltrated with hydroxyapatite or silicatitanate

Osteoinductive Activity of Bone Scaffold Bioceramic Companied with Control Release of VEGF Protein Treated Dental stem cells as A New Concept for Bone Regeneration: Part II

Enhanced Osteogenic and Angiogenic-Related Gene Expression of Human Dental Stem Cells on Biphasic Calcium Phosphate Scaffold Treated with Vascular Endothelial Growth Factor: Part I

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