Initial cell pre-cultivation can maximize ECM mineralization by human mesenchymal stem cells on silk fibroin scaffolds

Thimm, Benjamin; Wüst, Silke; Hofmann, Sandra; Hagenmüller, Henri; Müller, Ralph

doi:10.1016/j.actbio.2011.02.004

Cited by 30 publications

(22 citation statements)

References 48 publications

(71 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Paraffin wax sections of ASC microtissue spheroids were incubated with polyclonal rabbit anti-HTRA1 (1:50) or rabbit anti-osteocalcin (1:1000) using the conditions previously described [27]. Samples were then washed and incubated with goat anti-rabbit-Cy3 (1:400) for 1 h and mounted in DAPI containing Mowiol solution and images captured using the Leica DMI6000B system.…”

Section: Histologymentioning

confidence: 99%

See 1 more Smart Citation

Therapeutic potential of adipose-derived stromal cells in age-related osteoporosis

et al. 2014

Self Cite

View full text Add to dashboard Cite

a b s t r a c tAdipose-derived stromal cells (ASCs) are increasingly being used for orthopedic-based tissue engineering approaches due to their ability to readily undergo osteogenic differentiation. In the present study, we used in vitro and in vivo approaches to evaluate the use of ASCs as a treatment strategy for age-related osteoporosis. Molecular, histological and micro-computed tomography (micro-CT) based approaches confirmed that ASCs isolated from 18-week-old osteoporotic senescence-accelerated mice (SAMP6) were capable of undergoing osteogenesis when cultured in either silk fibroin (SF) scaffolds or scaffold-free microtissues (ASC-MT). A single intratibial injection of CM-Dil-labeled isogeneic ASCs or ASC-MT into SAMP6 recipients significantly improved trabecular bone quality after 6 weeks in comparison to untreated contralateral bones, as determined by micro-CT. Injected ASCs could be observed in paraffin wax bone sections at 24 h and 6 weeks post treatment and induced a significant increase in several molecular markers of bone turnover. Furthermore, a significant improvement in the osteogenic potential of osteoporotic patient-derived human bone marrow stromal cells (BMSCs) was observed when differentiated in conditioned culture media harvested from osteoporotic patient-derived human ASCs. These findings therefore support the use of ASCs as an autologous cell-based approach for the treatment of osteoporosis.

show abstract

Section: Histologymentioning

confidence: 99%

“…ASC-seeded SF scaffolds were prepared as previously described [26,27]. Briefly, silk cocoons from Bombyx Mori L. were cooked twice in sodium carbonate for 1 h to extract sericins.…”

Section: Osteogenesis Of Samp6-derived Ascs In 3-d Culture Systemsmentioning

confidence: 99%

Therapeutic potential of adipose-derived stromal cells in age-related osteoporosis

et al. 2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…This prompts the need to improve the mineralized matrix deposition throughout the scaffolds, for example, by perfusing BM through the scaffold in a controlled manner via a 3D perfusion bioreactor or by optimizing the scaffold properties through the study of the effect of the porous Ti-scaffold intrinsic properties (such as pore geometry, pore size, porosity, surface roughness, and chemistries) on mineralization of hPDCs. 63 …”

Section: Discussionmentioning

confidence: 99%

In VivoEctopic Bone Formation by Devitalized Mineralized Stem Cell Carriers Produced Under Mineralizing Culture Condition

Chai

Geris

Bolander

et al. 2014

BioResearch Open Access

View full text Add to dashboard Cite

Functionalization of tissue engineering scaffolds with in vitro–generated bone-like extracellular matrix (ECM) represents an effective biomimetic approach to promote osteogenic differentiation of stem cells in vitro. However, the bone-forming capacity of these constructs (seeded with or without cells) is so far not apparent. In this study, we aimed at developing a mineralizing culture condition to biofunctionalize three-dimensional (3D) porous scaffolds with highly mineralized ECM in order to produce devitalized, osteoinductive mineralized carriers for human periosteal-derived progenitors (hPDCs). For this, three medium formulations [i.e., growth medium only (BM1), with ascorbic acid (BM2), and with ascorbic acid and dexamethasone (BM3)] supplemented with calcium (Ca2+) and phosphate (PO43−) ions simultaneously as mineralizing source were investigated. The results showed that, besides the significant impacts on enhancing cell proliferation (the highest in BM3 condition), the formulated mineralizing media differentially regulated the osteochondro-related gene markers in a medium-dependent manner (e.g., significant upregulation of BMP2, bone sialoprotein, osteocalcin, and Wnt5a in BM2 condition). This has resulted in distinguished cell populations that were identifiable by specific gene signatures as demonstrated by the principle component analysis. Through devitalization, mineralized carriers with apatite crystal structures unique to each medium condition (by X-ray diffraction and SEM analysis) were obtained. Quantitatively, BM3 condition produced carriers with the highest mineral and collagen contents as well as human-specific VEGF proteins, followed by BM2 and BM1 conditions. Encouragingly, all mineralized carriers (after reseeded with hPDCs) induced bone formation after 8 weeks of subcutaneous implantation in nude mice models, with BM2-carriers inducing the highest bone volume, and the lowest in the BM3 condition (as quantitated by nano-computed tomography [nano-CT]). Histological analysis revealed different bone formation patterns, either bone ossicles containing bone marrow surrounding the scaffold struts (in BM2) or bone apposition directly on the struts' surface (in BM1 and BM3). In conclusion, we have presented experimental data on the feasibility to produce devitalized osteoinductive mineralized carriers by functionalizing 3D porous scaffolds with an in vitro cell-made mineralized matrix under the mineralizing culture conditions.

show abstract

“…12,28 In a recent study, pre-cultivation of hMSCs in growth media followed by the OI maximized the ECM mineralization in silk fibroin scaffolds, although the exact role of the ECM microenvironment remained unknown. 29 While these studies highlighted the influence of the ECM microenvironment on hMSC osteogenic differentiation, the…”

Section: ) (B)mentioning

confidence: 99%

Bioreactor Strategy in Bone Tissue Engineering: Pre-Culture and Osteogenic Differentiation Under Two Flow Configurations

Kim

2012

Tissue Engineering Part A

View full text Add to dashboard Cite

Since robust osteogenic differentiation and mineralization are integral to the engineering of bone constructs, understanding the impact of the cellular microenvironments on human mesenchymal stem cell (hMSCs) osteogenic differentiation is crucial to optimize bioreactor strategy. Two perfusion flow conditions were utilized in order to understand the impact of the flow configuration on hMSC construct development during both preculture (PC) in growth media and its subsequent osteogenic induction (OI). The media in the in-house perfusion bioreactor was controlled to perfuse either around (termed parallel flow [PF]) the construct surfaces or penetrate through the construct (termed transverse flow [TF]) for 7 days of the PC followed by 7 days of the OI. The flow configuration during the PC not only changed growth kinetics but also influenced cell distribution and potency of osteogenic differentiation and mineralization during the subsequent OI. While shear stress resulted from the TF stimulated cell proliferation during PC, the convective removal of de novo extracellular matrix (ECM) proteins and growth factors (GFs) reduced cell proliferation on OI. In contrast, the effective retention of de novo ECM proteins and GFs in the PC constructs under the PF maintained cell proliferation under the OI but resulted in localized cell aggregations, which influenced their osteogenic differentiation. The results revealed the contrasting roles of the convective flow as a mechanical stimulus, the redistribution of the cells and macromolecules in 3D constructs, and their divergent impacts on cellular events, leading to bone construct formation. The results suggest that the modulation of the flow configuration in the perfusion bioreactor is an effective strategy that regulates the construct properties and maximizes the functional outcome.

show abstract

Initial cell pre-cultivation can maximize ECM mineralization by human mesenchymal stem cells on silk fibroin scaffolds

Cited by 30 publications

References 48 publications

Therapeutic potential of adipose-derived stromal cells in age-related osteoporosis

Therapeutic potential of adipose-derived stromal cells in age-related osteoporosis

In VivoEctopic Bone Formation by Devitalized Mineralized Stem Cell Carriers Produced Under Mineralizing Culture Condition

Bioreactor Strategy in Bone Tissue Engineering: Pre-Culture and Osteogenic Differentiation Under Two Flow Configurations

Contact Info

Product

Resources

About