Fabrication of Micro-tissues using Modules of Collagen Gel Containing Cells

Chamberlain, M. Dean; Butler, Mark J.; Ciucurel, Ema C.; Fitzpatrick, Lindsay E.; Khan, Omar F.; Leung, Brendan M.; Lo, Chuen; Patel, Rikinkumar S; Velchinskaya, Alexandra; Voice, Derek N.; Sefton, Michael V.

doi:10.3791/2177

Cited by 8 publications

(7 citation statements)

References 11 publications

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“…3,19,20 BmMSCs were suspended in the collagen at a concentration of 1.0 · 10 6 cells/mL. RAECs (2.5 · 10 6 ) were seeded dynamically onto the surface of 1 mL of modules (produced using 2.5 m of tubing) for 45 min on a low-speed shaker and incubated for 7 days prior to implantation.…”

Section: Module Fabricationmentioning

confidence: 99%

Bone Marrow-Derived Mesenchymal Stromal Cells Enhance Chimeric Vessel Development Driven by Endothelial Cell-Coated Microtissues

Chamberlain

Gupta

Sefton

2012

Tissue Engineering Part A

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Adding bone marrow-derived mesenchymal stromal cells (bmMSCs) to endothelialized collagen gel modules resulted in mature vessel formation, presumably caused in part by the observed display of pericyte-like behavior for the transplanted GFP(+) bmMSCs. A previous study determined that rat aortic endothelial cells (RAECs) delivered on the surface of small (∼0.8 mm long×0.5 mm diameter) collagen gel cylinders (microtissues, modular tissue engineering) formed vessels after transplantation into immunosuppressed Sprague-Dawley (SD) rats. Although the RAECs formed vessels in this allogeneic transplant model, there was a robust inflammatory response and the vessels that formed were leaky as shown by microcomputed tomography (microCT) perfusion studies. In vitro assays showed that SD rat bmMSCs embedded into the collagen gel modules increased the extent of EC proliferation and enhanced EC sprouting. In vivo, although vessel number was not affected, the new vessels formed by the bmMSCs and RAECs were more stable and leaked less in the microCT perfusion analysis than vessels formed by implanted RAECs alone. Addition of the bmMSCs also decreased the total number of CD68(+) macrophages that infiltrated the implant and changed the distribution of CD163(+) (M2) macrophages so that they were found within the newly developed vascularized tissue. Most interestingly, the bmMSCs became smooth muscle actin positive and migrated to surround the EC layer of the vessel, which is the location typical of pericytes. The combination of these two effects was presumed to be the cause of improved vascularity when bmMSCs were embedded in the EC-coated modules. Further exploration of these observations is warranted to exploit modular tissue engineering as a means of forming large vascularized functional tissues using microtissue components.

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Section: Module Fabricationmentioning

confidence: 99%

Bone Marrow-Derived Mesenchymal Stromal Cells Enhance Chimeric Vessel Development Driven by Endothelial Cell-Coated Microtissues

Chamberlain

Gupta

Sefton

2012

Tissue Engineering Part A

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“…Hence, our interests in using modular tissue engineering with endothelial cell-coated collagen gel modules to effect vascularized implant sites. [1][2][3][4][5] Beyond nutrient supply, the endothelial cells also influence the fate of cotransplanted cells and the resulting tissue, here, for example, fat.…”

Section: Introductionmentioning

confidence: 99%

Cotransplantation of Adipose-Derived Mesenchymal Stromal Cells and Endothelial Cells in a Modular Construct Drives Vascularization in SCID/bg Mice

Butler

Sefton

2012

Tissue Engineering Part A

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A modular approach to adipose tissue engineering was explored by embedding adipose-derived mesenchymal stromal cells (adMSC) in sub-mm-sized collagen rods or ''modules'' and coating with human microvascular endothelial cells (HMEC). After subcutaneous injection into a SCID/Bg mouse, HMEC on modules containing embedded adMSC appeared to detach from the modules to form vessels as early as day 3, as confirmed by the human EC-specific UEA-1 lectin stain, and these vessels persisted for up to 90 days. Vessel numbers decreased over 14 days, but vessel size increased suggesting a maturing of the vasculature. Vessel perfusion with the host was confirmed at 21 days by microCT. HMEC on modules without embedded adMSC remained attached to the module surface at day 3 and UEA-1 staining disappeared over 14 days suggesting cell death. It appeared that cotransplantation with adMSC had an anti-apoptotic and proangiogenic effect on HMEC. The early revascularization strategy may be successful in supporting adMSC viability and differentiation, as a preliminary study suggests progressive fat accumulation in the HMEC + adMSC implants: *60% of the implant area stained positive for Oil Red O by day 90. adMSC-embedded modules without HMEC surface coating did not show similar levels of Oil Red O staining. All implant volumes decreased over the time course of the experiment, yet HMEC + adMSC module implants were larger than adMSC-only implants at day 90. Collagen gel is mechanically weak and contracts in vivo making it unsuitable as a biomaterial for adipose tissue engineering where volume maintenance is critical. When combined with an appropriate biomaterial, the modular approach to adipose tissue engineering may represent a successful strategy to engineer soft tissue substitutes of clinical relevance.

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“…A range of different module shapes, that have been fabricated, is shown in (b). In (c) we present a diagram showing the different module structures and organization that could be used to represent the various periodontium tissues structurally and compositionally in a very basic representation to mimic the structural environments necessary for gingival epithelium development, subepithelial connective tissue formation and the stacked layers of cementum, ligament and the alveolar bone lining (McGuigan and Sefton, 2006 ; McGuigan et al, 2006 , 2008 ; Nichol and Khademhosseini, 2009 ; Chamberlain et al, 2010 ; Reproduced with kind permission from PLoS, MacMillan Publishing, MYJoVE Corporation, American Association for the Advancement of Science, Royal Society of Chemistry). (B) (a) Showing the arrangement of printed cells in a distribution reminiscent of skin tissue; (b) Red staining to highlight the distinct keratinocyte cell region; (c) Fluorescently labeled red fibroblasts and green keratinocytes printed using the laser technique into tight and clear layers; (d) Phenotype related fluorescence tagging of fibroblasts (red) and keratinocytes for cytokeratin 14 expression.…”

Section: Framework To Support Periodontal Tissue Regenerationmentioning

confidence: 99%

Small-Scale Fabrication of Biomimetic Structures for Periodontal Regeneration

2016

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The periodontium is the supporting tissues for the tooth organ and is vulnerable to destruction, arising from overpopulating pathogenic bacteria and spirochaetes. The presence of microbes together with host responses can destroy large parts of the periodontium sometimes leading tooth loss. Permanent tissue replacements are made possible with tissue engineering techniques. However, existing periodontal biomaterials cannot promote proper tissue architectures, necessary tissue volumes within the periodontal pocket and a “water-tight” barrier, to become clinically acceptable. New kinds of small-scale engineered biomaterials, with increasing biological complexity are needed to guide proper biomimetic regeneration of periodontal tissues. So the ability to make compound structures with small modules, filled with tissue components, is a promising design strategy for simulating the anatomical complexity of the periodotium attachment complexes along the tooth root and the abutment with the tooth collar. Anatomical structures such as, intima, adventitia, and special compartments such as the epithelial cell rests of Malassez or a stellate reticulum niche need to be engineered from the start of regeneration to produce proper periodontium replacement. It is our contention that the positioning of tissue components at the origin is also necessary to promote self-organizing cell–cell connections, cell–matrix connections. This leads to accelerated, synchronized and well-formed tissue architectures and anatomies. This strategy is a highly effective preparation for tackling periodontitis, periodontium tissue resorption, and to ultimately prevent tooth loss. Furthermore, such biomimetic tissue replacements will tackle problems associated with dental implant support and perimimplantitis.

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Fabrication of Micro-tissues using Modules of Collagen Gel Containing Cells

Cited by 8 publications

References 11 publications

Bone Marrow-Derived Mesenchymal Stromal Cells Enhance Chimeric Vessel Development Driven by Endothelial Cell-Coated Microtissues

Bone Marrow-Derived Mesenchymal Stromal Cells Enhance Chimeric Vessel Development Driven by Endothelial Cell-Coated Microtissues

Cotransplantation of Adipose-Derived Mesenchymal Stromal Cells and Endothelial Cells in a Modular Construct Drives Vascularization in SCID/bg Mice

Small-Scale Fabrication of Biomimetic Structures for Periodontal Regeneration

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