Assessment of Collagen-Based Nanostructured Biomimetic Systems with a Co-Culture of Human Bone-Derived Cells

Borciani, Giorgia; Montalbano, Giorgia; Melo, Priscila; Baldini, Nicola; Ciapetti, G.; Brovarone, Chiara Vitale

doi:10.3390/cells11010026

Cited by 3 publications

(3 citation statements)

References 48 publications

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Section: Methodsmentioning

confidence: 99%

“…Some of the assays used for the indirect co-culture with two collagen-based materials added with mesoporous bioactive glass particles (Coll_MBG) and nano-hydroxyapatite (Coll_HA) are reported underneath. These assays have been used for the 2D collagen-based hydrogels non-translucent due to the presence of genipin (blue-violet dye) as a crosslinker [21]. (b) ARS staining on OBs is performed as described in Section 3.4 "Cell B Cytochemical staining protocols" to be followed by stain elution: sample with 500 µ L of 10% cetylpyridinium solution at room tempera (b) ARS staining on OBs is performed as described in Section 3.4 "Cell Biochemistry-Cytochemical staining protocols" to be followed by stain elution: incubate each sample with 500 µL of 10% cetylpyridinium solution at room temperature for 5 min under orbital shaking.…”

Section: Materials Testing With the Indirect Co-culturementioning

confidence: 99%

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Protocol of Co-Culture of Human Osteoblasts and Osteoclasts to Test Biomaterials for Bone Tissue Engineering

Borciani

Montalbano

Baldini

et al. 2022

MPs

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New biomaterials and scaffolds for bone tissue engineering (BTE) applications require to be tested in a bone microenvironment reliable model. On this assumption, the in vitro laboratory protocols with bone cells represent worthy experimental systems improving our knowledge about bone homeostasis, reducing the costs of experimentation. To this day, several models of the bone microenvironment are reported in the literature, but few delineate a protocol for testing new biomaterials using bone cells. Herein we propose a clear protocol to set up an indirect co-culture system of human-derived osteoblasts and osteoclast precursors, providing well-defined criteria such as the cell seeding density, cell:cell ratio, the culture medium, and the proofs of differentiation. The material to be tested may be easily introduced in the system and the cell response analyzed. The physical separation of osteoblasts and osteoclasts allows distinguishing the effects of the material onto the two cell types and to evaluate the correlation between material and cell behavior, cell morphology, and adhesion. The whole protocol requires about 4 to 6 weeks with an intermediate level of expertise. The system is an in vitro model of the bone remodeling system useful in testing innovative materials for bone regeneration, and potentially exploitable in different application fields. The use of human primary cells represents a close replica of the bone cell cooperation in vivo and may be employed as a feasible system to test materials and scaffolds for bone substitution and regeneration.

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Section: Methodsmentioning

confidence: 99%

Section: Materials Testing With the Indirect Co-culturementioning

confidence: 99%

Protocol of Co-Culture of Human Osteoblasts and Osteoclasts to Test Biomaterials for Bone Tissue Engineering

Borciani

Montalbano

Baldini

et al. 2022

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“…The bulk samples were tested by employing an indirect co-culture of human osteoblasts and osteoclast precursors. Sr-enriched mesoporous BGs have structural and physicochemical properties that support the viability and proliferation of co-cultured human bone-derived cells, with multiple signals differently affecting the osteoblast and the osteoclast precursors [ 169 ].…”

Section: Biomaterials For Bone Tissue Engineering Approach and Their ...mentioning

confidence: 99%

Strontium Functionalization of Biomaterials for Bone Tissue Engineering Purposes: A Biological Point of View

et al. 2022

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Strontium (Sr) is a trace element taken with nutrition and found in bone in close connection to native hydroxyapatite. Sr is involved in a dual mechanism of coupling the stimulation of bone formation with the inhibition of bone resorption, as reported in the literature. Interest in studying Sr has increased in the last decades due to the development of strontium ranelate (SrRan), an orally active agent acting as an anti-osteoporosis drug. However, the use of SrRan was subjected to some limitations starting from 2014 due to its negative side effects on the cardiac safety of patients. In this scenario, an interesting perspective for the administration of Sr is the introduction of Sr ions in biomaterials for bone tissue engineering (BTE) applications. This strategy has attracted attention thanks to its positive effects on bone formation, alongside the reduction of osteoclast activity, proven by in vitro and in vivo studies. The purpose of this review is to go through the classes of biomaterials most commonly used in BTE and functionalized with Sr, i.e., calcium phosphate ceramics, bioactive glasses, metal-based materials, and polymers. The works discussed in this review were selected as representative for each type of the above-mentioned categories, and the biological evaluation in vitro and/or in vivo was the main criterion for selection. The encouraging results collected from the in vitro and in vivo biological evaluations are outlined to highlight the potential applications of materials’ functionalization with Sr as an osteopromoting dopant in BTE.

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Bioinspired Collagen/Hyaluronic Acid/Fibrin-Based Hydrogels for Soft Tissue Engineering: Design, Synthesis, and In Vitro Characterization

Bindi,

Perioli,

Melo

et al. 2023

JFB

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A major challenge for future drug development comprises finding alternative models for drug screening. The use of animal models in research is highly controversial, with an ongoing debate on their ethical acceptability. Also, animal models are often poorly predictive of therapeutic outcomes due to the differences between animal and human physiological environments. In this study, we aimed to develop a biomimetic hydrogel that replicates the composition of skin for potential use in in vitro modeling within tissue engineering. The hydrogel was fabricated through the crosslinking of collagen type I, hyaluronic acid, four-arm PEG succinimidyl glutarate (4S-StarPEG), and fibrinogen. Various ratios of these components were systematically optimized to achieve a well-interconnected porosity and desirable rheological properties. To evaluate the hydrogel’s cytocompatibility, fibroblasts were embedded within the matrix. The resulting hydrogel exhibited promising properties as a scaffold, also facilitating the growth of and proliferation of the cells. This biomimetic hydrogel holds great potential for tissue engineering applications, particularly in skin regeneration and cancer research. The study used melanoma spheroids fabricated using the 96-round bottom well plate method as a potential application. The results demonstrate that the developed hydrogels allowed the maintenance of spheroid integrity and viability, meaning it has a promising use as a three-dimensional in vitro model of melanoma for both tissue engineering and drug screening applications.

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Assessment of Collagen-Based Nanostructured Biomimetic Systems with a Co-Culture of Human Bone-Derived Cells

Cited by 3 publications

References 48 publications

Protocol of Co-Culture of Human Osteoblasts and Osteoclasts to Test Biomaterials for Bone Tissue Engineering

Protocol of Co-Culture of Human Osteoblasts and Osteoclasts to Test Biomaterials for Bone Tissue Engineering

Strontium Functionalization of Biomaterials for Bone Tissue Engineering Purposes: A Biological Point of View

Bioinspired Collagen/Hyaluronic Acid/Fibrin-Based Hydrogels for Soft Tissue Engineering: Design, Synthesis, and In Vitro Characterization

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