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OPTIMIZATION OF COLLAGEN-ELASTIN-LIKE POLYPEPTIDE-BIOGLASS SCAFFOLD COMPOSITION FOR OSTEOGENIC DIFFERENTIATION OF ADIPOSE-DERIVED STEM CELLS.

ABSTRACT: We have developed a multicomponent hydrogel scaffold that can mimic the bone extracellular matrix by incorporating collagen, elastin-like polypeptide (ELP), and Bioglass. We examined the effects of Bioglass addition to collagen-ELP scaffolds on mechanical properties, physical characteristics, and in vitro osteogenic differentiation, by varying the Bioglass amount and particle size. Response surface methodology with a central composite design predicted 5 mg (6.6 mg/mL) Bioglass with a particle size of 142 ± 5 ?m as the optimal amount and particle size to be mixed with 6 mg/mL collagen and 18 mg/mL ELP to obtain a combination of maximized compressive properties. Swelling ratio and FTIR spectroscopy indicated lower hydrophilicity and the presence of hydrophobic and secondary interactions between collagen, ELP, and Bioglass. Scanning electron microscopy showed a nanofibrous morphology of intermingled collagen-ELP-Bioglass network. In vitro osteogenic characterization using human adipose-derived stem cells revealed increased cell attachment and proliferation with increased ALP activity, osteocalcin content, and mineralized deposit formation during a three-week culture. Numerous mineralized deposits composed of calcium and phosphorous were shown by energy dispersive spectroscopy. Overall, our results show that the collagen-ELP-Bioglass multicomponent composites have enhanced mechanical properties with adequate physical features and cell culture properties for bone tissue engineering.

SUBMITTER: Gurumurthy B

PROVIDER: S-EPMC7055731 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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OPTIMIZATION OF COLLAGEN-ELASTIN-LIKE POLYPEPTIDE-BIOGLASS SCAFFOLD COMPOSITION FOR OSTEOGENIC DIFFERENTIATION OF ADIPOSE-DERIVED STEM CELLS.

Gurumurthy Bhuvaneswari B Pal Pallabi P Griggs Jason A JA Janorkar Amol V AV

Materialia 20200124

We have developed a multicomponent hydrogel scaffold that can mimic the bone extracellular matrix by incorporating collagen, elastin-like polypeptide (ELP), and Bioglass. We examined the effects of Bioglass addition to collagen-ELP scaffolds on mechanical properties, physical characteristics, and <i>in vitro</i> osteogenic differentiation, by varying the Bioglass amount and particle size. Response surface methodology with a central composite design predicted 5 mg (6.6 mg/mL) Bioglass with a part ...[more]

PMID: 32133439

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Project description:Craniomaxillofacial bone defects can occur as a result of congenital, post-oncologic, and high-energy impact conditions. The scale and irregularity of such defects motivate new biomaterials to promote regeneration of the damaged bone. We have recently described a mineralized collagen scaffold capable of instructing stem cell osteogenic differentiation and new bone infill in the absence of traditional osteogenic supplements. Herein, we report the integration of a millimeter-scale reinforcing poly (lactic acid) frame fabricated via 3D-printing into the mineralized collagen scaffold with micron-scale porosity to form a multi-scale mineralized collagen-PLA composite. We describe modifications to the PLA frame design to increase the compressive strength (Young's Modulus, ultimate stress and strain) of the composite. A critical challenge beyond increasing the compressive strength of the collagen scaffold is addressing challenges inherent with the irregularity of clinical defects. As a result, we examined the potential for modifying the frame architecture to render the composite with increased compressive strength in one axis or radial compressibility and shape-fitting capacity in an orthogonal axis. A library of mineralized collagen-PLA composites was mechanically characterized via compression testing and push-out test to describe mechanical performance and shape-fitting capacity. We also report in vitro comparison of the bioactivity of porcine adipose derived stem cells in the mineralized collagen-PLA composite versus the mineralized collagen scaffold via metabolic activity, gene expression, and functional matrix synthesis. The results suggest that incorporation of the PLA reinforcing frame does not negatively influence the osteoinductive nature of the mineralized collagen scaffold. Together, these findings suggest a strategy to address often competing bioactivity, mechanical strength, and shape-fitting design requirements for biomaterials for craniomaxillofacial bone regeneration.

Project description:Biomineralized collagen with intrafibrillar calcium phosphate mineral provides an excellent mimic of the composition and structure of the extracellular matrix of bone, from nano- to micro-scale. Scaffolds prepared from this material have the potential to become the next-generation of synthetic bone graft substitutes, as their unique properties make them closer to the native tissue than synthetic alternatives currently available to clinicians. To understand the interaction between biomineralized collagen and cells that are relevant in the context of bone regeneration, we studied the growth and osteogenic differentiation of bone marrow derived human mesenchymal stromal cells (hMSCs) cultured on biomineralized collagen membranes, and compared it to the cell behavior on collagen membranes without mineral. Cells proliferated normally on both biomimetic membranes, and were more triggered to differentiate toward the osteogenic lineage by the biomineralized collagen. This was shown by the elevated mRNA levels of RUNX2, SPP1, ENPP1, and OCN after 3 days of culture, and COL1A1 after 14 days of culture on mineralized collagen. The mRNA levels of the tested markers of osteogenesis were lower on collagen membranes without mineral, with the exception of OCN, which was more highly expressed on collagen than on biomineralized collagen membranes. Expression by hMSCs of OPG, a gene involved in inhibition of osteoclastogenesis, was higher on biomineralized collagen at day 3, while M-CSF, involved in osteoblast-osteoclast communication, was upregulated on both membranes at day 3 and 14 of culture. Alkaline phosphatase activity of hMSCs was high on both biomimetic membranes when compared with cells cultured on tissue culture plastic. Cell-induced mineralization was observed on collagen membranes, while the high mineral content of the biomineralized membranes prohibited a reliable analysis of cell-induced mineralization on these membranes. In conclusion, we have identified that both collagen and biomineralized collagen support proliferation, osteogenic differentiation and mineralization of hMSCs, with biomineralized membranes having a more pronounced positive effect. These findings support the existing evidence that biomineralized collagen is a promising material in the field of bone regeneration.

Dataset Information

OPTIMIZATION OF COLLAGEN-ELASTIN-LIKE POLYPEPTIDE-BIOGLASS SCAFFOLD COMPOSITION FOR OSTEOGENIC DIFFERENTIATION OF ADIPOSE-DERIVED STEM CELLS.

Publications

OPTIMIZATION OF COLLAGEN-ELASTIN-LIKE POLYPEPTIDE-BIOGLASS SCAFFOLD COMPOSITION FOR OSTEOGENIC DIFFERENTIATION OF ADIPOSE-DERIVED STEM CELLS.

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