Culture on Tissue‐Specific Coatings Derived from α‐Amylase‐Digested Decellularized Adipose Tissue Enhances the Proliferation and Adipogenic Differentiation of Human Adipose‐Derived Stromal Cells

Shridhar, Arthi; Lam, Alan Y.L.; Sun, Yu; Simmons, Craig A.; Gillies, Elizabeth R.; Flynn, Lauren E.

doi:10.1002/biot.201900118

Cited by 12 publications

(19 citation statements)

References 58 publications

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“…There was no significant difference in the particle size distributions between the groups based on a Kolmogorov–Smirnov nonparametric test ( p < 0.05) ( Figure 3B ). To generate the coatings, applying methods previously developed by our group ( Shridhar et al, 2020 ), the particles were subsequently digested with α-amylase and homogenized in acetic acid.…”

Section: Resultsmentioning

confidence: 99%

“…SEM analysis was used to visualize the ultrastructure of the sieved and lyophilized DNP and COL particles, as well as the DNP and COL coatings, using established protocols ( Shridhar et al, 2020 ). To visualize their interior structure, the DNP and COL coatings were scratched using a scalpel blade and then lyophilized.…”

Section: Methodsmentioning

confidence: 99%

“…As such, there is great interest in the development of more customizable ECM-derived scaffold formats that incorporate the tissue-specific ECM composition within platforms that enable greater tuning of cell-cell and cell-ECM interactions. Using decellularized adipose tissue (DAT) as an ECM source, our team has previously established methods for fabricating novel ECM-derived coatings, microcarriers, foams, and hydrogel composites ( Cheung et al, 2014 ; Kornmuller et al, 2017 ; Yu et al, 2017 ; Shridhar et al, 2019 ; Shridhar et al, 2020 ). Importantly, we demonstrated that the pro-adipogenic properties of the DAT in directing the differentiation of human adipose-derived stromal cells (ASCs) were conserved in these various platforms ( Turner et al, 2012 ; Yu et al, 2013 ; Cheung et al, 2014 ; Shridhar et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

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Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus

Quijano

Sharma²,

Martin³

et al. 2022

Front. Bioeng. Biotechnol.

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Bioscaffolds derived from the extracellular matrix (ECM) have shown the capacity to promote regeneration by providing tissue-specific biological instructive cues that can enhance cell survival and direct lineage-specific differentiation. This study focused on the development and characterization of two-dimensional (2-D) and three-dimensional (3-D) cell culture platforms incorporating decellularized nucleus pulposus (DNP). First, a detergent-free protocol was developed for decellularizing bovine nucleus pulposus (NP) tissues that was effective at removing cellular content while preserving key ECM constituents including collagens, glycosaminoglycans, and the cell-adhesive glycoproteins laminin and fibronectin. Next, novel 2-D coatings were generated using the DNP or commercially-sourced bovine collagen type I (COL) as a non-tissue-specific control. In addition, cryo-milled DNP or COL particles were incorporated within methacrylated chondroitin sulphate (MCS) hydrogels as a 3-D cell culture platform for exploring the effects of ECM particle composition. Culture studies showed that the 2-D coatings derived from the DNP could support cell attachment and growth, but did not maintain or rescue the phenotype of primary bovine NP cells, which de-differentiated when serially passaged in monolayer culture. Similarly, while bovine NP cells remained highly viable following encapsulation and 14 days of culture within the hydrogel composites, the incorporation of DNP particles within the MCS hydrogels was insufficient to maintain or rescue changes in NP phenotype associated with extended in vitro culture based on gene expression patterns. Overall, DNP produced with our new decellularization protocol was successfully applied to generate both 2-D and 3-D bioscaffolds; however, further studies are required to assess if these platforms can be combined with additional components of the endogenous NP microenvironment to stimulate regeneration or lineage-specific cell differentiation.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus

Quijano

Sharma²,

Martin³

et al. 2022

Front. Bioeng. Biotechnol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…New strategies that can promote the regeneration of healthy host-derived adipose tissue and enable stable and predictable volume augmentation would be of great value for a broad range of applications in plastic and reconstructive surgery. With the long-term goal of advancing towards clinical translation, our lab has been developing a broad range of off-the-shelf bioscaffolds derived from the ECM of human adipose tissue 7,8,[35][36][37] . Our previous in vivo study exploring DAT scaffolds in a subcutaneous implant model in immunocompetent rats indicated that seeding the DAT with allogeneic rat ASCs significantly enhanced blood vessel formation and remodeling of the DAT into host-derived adipose tissue 13 .…”

Section: Discussionmentioning

confidence: 99%

Perfusion bioreactor culture of adipose-derived stromal cells on decellularized adipose tissue scaffolds enhancesin vivoregeneration

Han

Flynn

2020

Preprint

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Adipose tissue engineering holds promise to address the unmet need in plastic and reconstructive surgery for strategies that promote the stable and predictable regeneration of adipose tissue for volume augmentation applications. Previous studies have demonstrated that decellularized adipose tissue (DAT) scaffolds can provide a pro-adipogenic microenvironment, and that seeding with adipose-derived stromal cells (ASCs) can enhance in vivo angiogenesis and adipogenesis within DAT implants. Recognizing that bioreactor systems can promote cell expansion and infiltration on tissue-engineered scaffolds, this study evaluated the effects of culturing human ASCs on DAT scaffolds within a perfusion bioreactor. Using this system, the impact of both shear stress stimulation and hypoxic preconditioning were explored in vitro and in vivo. Initial studies compared the effects of 14 days of culture within the perfusion bioreactor under 2% O2 or ~20% O2 on human ASC expansion and hypoxia inducible factor 1 alpha (HIF-1α) expression in vitro relative to static cultured controls. The findings indicated that culturing within the bioreactor under 2% O2 significantly increased ASC proliferation on the DAT, with a higher cell density observed in the scaffold periphery. HIF-1α expression was significantly higher when the scaffolds were cultured under 2% O2. Subsequent characterization in a subcutaneous implant model in athymic nude mice revealed that in vivo angiogenesis and adipogenesis were markedly enhanced when the ASCs were cultured on the DAT within the perfusion bioreactor under 2% O2 for 14 days prior to implantation relative to the other culture conditions, as well as additional freshly-seeded and unseeded DAT control groups. Overall, dynamic culture within the perfusion bioreactor system under hypoxia represents a promising approach for preconditioning ASCs on DAT scaffolds to enhance their capacity to stimulate blood vessel formation and infiltration, as well as host-derived adipose tissue regeneration.

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“…The extracellular matrix plays an important role in modulating various cell behaviors including adhesion, proliferation and differentiation 10 . Decellularized adipose tissues have been used extensively as a promising scaffold material which provide a 3D complex ECM architecture for adipogenesis 11‐14 . Multiple strategies involving both synthetic and natural materials have been developed for soft tissue reconstruction 15,16 .…”

Section: Introductionmentioning

confidence: 99%

Hybrid adipose graft materials synthesized from chemically modified adipose extracellular matrix

Liu

McCann

et al. 2021

J Biomedical Materials Res

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Decellularized extracellular matrix (ECM) from tissues is a promising biomaterial that can provide a complex 3D microenvironment capable of modulating cell response and tissue regeneration. In this study, we have integrated the decellularized thiolated adipose‐derived ECM, at different concentrations, with polyethylene glycol (PEG) using Michael addition between thiol and acrylate moieties. The potential for this material to support adipogenic differentiation of human adipose‐derived stem cells was evaluated by encapsulating cells in hydrogels with increasing concentrations of chemically modified ECM (mECM). Our results demonstrated a positive correlation between the ECM content in the hydrogels and cell proliferation, adipogenic marker expression, and lipid formation and accumulation. Furthermore, we have shown host cell infiltration and enhanced adipogenesis in vivo after implantation. These findings support the graft as a potential alternative for adipose tissue regeneration.

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Culture on Tissue‐Specific Coatings Derived from α‐Amylase‐Digested Decellularized Adipose Tissue Enhances the Proliferation and Adipogenic Differentiation of Human Adipose‐Derived Stromal Cells

Cited by 12 publications

References 58 publications

Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus

Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus

Perfusion bioreactor culture of adipose-derived stromal cells on decellularized adipose tissue scaffolds enhancesin vivoregeneration

Hybrid adipose graft materials synthesized from chemically modified adipose extracellular matrix

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