Use of Multipotent Mesenchymal Stromal Cells, Fibrin, and Scaffolds in the Production of Clinical Grade Bone Tissue Engineering Products

Vives, Joaquim; Rodríguez, Laura; Coca, María Isabel; Reales, Laura; Cabrera-Pérez, Raquel; Martorell, L.

doi:10.1007/7651_2020_280

Cited by 6 publications

(3 citation statements)

References 17 publications

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“…An experimental matrix was defined to test suitable combinations of hydrogels, which were chosen based on prior preclinical and clinical experience in the generation of (non‐3D printed) osteogenic MSC‐based TEP, as reported elsewhere (García de Frutos et al, 2020; Prat et al, 2018; Vivas et al, 2020; Vives et al, 2021). Particularly: clinical grade fibrin (Tissucol Duo; Baxter) as a result from mixing fibrinogen and 1:100 diluted thrombin at 5 UI/mL final concentration in saline solution (Viaflo Plasmalyte 148; Baxter) supplemented with 2% human serum albumin (HSA; Albutein®; Grifols); clinical grade HA (Bioiberica) dissolved either at 1% or 5% (w/v) in warm saline solution and sterilized by filtration with a 0.45 µm Millex‐HV filter (Merck Millipore) or by autoclaving, respectively; and research grade polymer precursors of gelatin and alginate methacryloyl (GelMA and AlgMA, respectively) (Sigma‐Aldrich) were altered to a 40% methacrylation degree.…”

Section: Methodsmentioning

confidence: 99%

Preservation of critical quality attributes of mesenchymal stromal cells in 3D bioprinted structures by using natural hydrogel scaffolds

Martorell

López-Fernández

García-Lizarribar

et al. 2023

Biotech & Bioengineering

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Three dimensional (3D) bioprinting is an emerging technology that enables complex spatial modeling of cell‐based tissue engineering products, whose therapeutic potential in regenerative medicine is enormous. However, its success largely depends on the definition of a bioprintable zone, which is specific for each combination of cell‐loaded hydrogels (or bioinks) and scaffolds, matching the mechanical and biological characteristics of the target tissue to be repaired. Therefore proper adjustment of the bioink formulation requires a compromise between: (i) the maintenance of cellular critical quality attributes (CQA) within a defined range of specifications to cell component, and (ii) the mechanical characteristics of the printed tissue to biofabricate. Herein, we investigated the advantages of using natural hydrogel‐based bioinks to preserve the most relevant CQA in bone tissue regeneration applications, particularly focusing on cell viability and osteogenic potential of multipotent mesenchymal stromal cells (MSCs) displaying tripotency in vitro, and a phenotypic profile of 99.9% CD105+/CD45,− 10.3% HLA‐DR,+ 100.0% CD90,+ and 99.2% CD73+/CD31− expression. Remarkably, hyaluronic acid, fibrin, and gelatin allowed for optimal recovery of viable cells, while preserving MSC's proliferation capacity and osteogenic potency in vitro. This was achieved by providing a 3D structure with a compression module below 8.8 ± 0.5 kPa, given that higher values resulted in cell loss by mechanical stress. Beyond the biocompatibility of naturally occurring polymers, our results highlight the enhanced protection on CQA exerted by bioinks of natural origin (preferably HA, gelatin, and fibrin) on MSC, bone marrow during the 3D bioprinting process, reducing shear stress and offering structural support for proliferation and osteogenic differentiation.

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Section: Methodsmentioning

confidence: 99%

Preservation of critical quality attributes of mesenchymal stromal cells in 3D bioprinted structures by using natural hydrogel scaffolds

Martorell

López-Fernández

García-Lizarribar

et al. 2023

Biotech & Bioengineering

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“…No previous sample size calculation was made because of the pilot nature of this study. Patients were randomised to either one of the two study treatments described next: 1) Treatment A (experimental): mechanical stabilisation (if required) associated with a Tissue Engineering Product (TEP composed of ex vivo expanded autologous Mesenchymal Stromal Cells (MSC) loaded onto allogeneic cancellous bone graft described previously ( Prat et al, 2018 ; García de Frutos et al, 2020 ) and prepared in accordance with current Good Manufacturing Practices as reported elsewhere ( Codinach et al, 2016 ; Vives et al, 2021 ; García-Muñoz and Vives, 2021 ); and 2) Treatment B (control): mechanical stabilisation, if required, associated with autologous iliac crest graft (gold standard) ( Fig. 1 , Fig.…”

Section: Methodsmentioning

confidence: 99%

A pilot study of circulating levels of TGF-β1 and TGF-β2 as biomarkers of bone healing in patients with non-hypertrophic pseudoarthrosis of long bones

et al. 2022

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Background Pseudoarthrosis or non-union is a complication with an incidence of 5–10% of bone fractures, most frequently located in the diaphysis of long bones. The management of this complication is addressed by means of complex surgical procedures and is a concern for orthopaedic and trauma surgeons nowadays. The use of biomarkers for diagnosing patients at risk of non-union would help us to establish special measures for early corrective treatment. Methods Prospective exploratory pilot study with a cohort of 20 patients diagnosed of non-hypertrophic pseudoarthrosis of long bones who were treated surgically with either autologous bone graft or a Tissue Engineering Product composed of bone marrow-derived Mesenchymal Stromal Cells. Patients were followed for 12 months and plasma blood samples were obtained to determine circulating levels of Transforming Growth Factor Beta 1 and Beta 2 (TGF-β1 and TGF-β2, respectively) at inclusion, and at 1 week, 2 weeks, and months 1, 2, 3, 6 and 12 after surgery. Radiological bone healing was evaluated by the Tomographic Union Score (TUS). Results Basal levels of TGF-β1 and TGF-β2 were determined in the twenty patients (26,702 ± 14,537 pg/mL and 307.8 ± 83.1 pg/mL, respectively). Three of them withdrew from the study, so complete follow-up was conducted on 17 patients (9 successfully healed vs. 8 that did not heal). Statistically significant differences between the bone healing group and the non-union group were found at month 12 for both TGF-β1 ( p = 0.005) and TGF-β2 ( p = 0.02). Conclusions TGF-β1 and TGF-β2 are biomarkers that correlate with clinical evidence of bone regeneration and may be used to monitor patients, although early predictive value after intervention needs to be further studied in combination with other molecules.

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“…monocytes), but the MSC fraction is efficiently enriched over subsequent passaging (Bieback et al 2008 ). This methodology was successfully used in our laboratory for the production of BM-MSC in a Phase I/IIa clinical trial of spinal fusion in degenerative disk disease (García de Frutos et al 2020 ; Vives et al 2020 ).…”

Section: Msc Isolation Techniquesmentioning

confidence: 99%

Towards the standardization of methods of tissue processing for the isolation of mesenchymal stromal cells for clinical use

García-Muñoz

Vives

2021

Cytotechnology

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Multipotent mesenchymal stromal cells (MSCs) are currently the most extensively studied type of adult stem cells in advanced stages of development in the field of regenerative medicine. The biological properties of MSCs have generated great hope for their therapeutic use in degenerative and autoimmune conditions that, at present, lack effective treatment options. Over the last decades, MSCs have been typically obtained from adult bone marrow, but the extraction process is highly invasive and the quality and numbers of isolated cells is drastically influenced by patient age, medication and associated comorbidities. Therefore, there is currently an open discussion on the convenience of allogeneic over autologous treatments, despite potential disadvantages such as rejection by the host. This shift to the allogeneic setting entails the need for high production of MSCs to ensure availability of sufficient cell numbers for transplantation, and therefore making the search for alternative tissue sources of highly proliferative MSC cultures with low levels of senescence occurrence, which is one of the greatest current challenges in the scale up of therapeutic cell bioprocessing. Herein we (i) present the main isolation protocols of MSCs from bone marrow, adipose tissue and Wharton’s jelly of the umbilical cord; and (ii) compare their qualities from a bioprocess standpoint, addressing both quality and regulatory aspects, in view of their anticipated clinical use.

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Use of Multipotent Mesenchymal Stromal Cells, Fibrin, and Scaffolds in the Production of Clinical Grade Bone Tissue Engineering Products

Cited by 6 publications

References 17 publications

Preservation of critical quality attributes of mesenchymal stromal cells in 3D bioprinted structures by using natural hydrogel scaffolds

Preservation of critical quality attributes of mesenchymal stromal cells in 3D bioprinted structures by using natural hydrogel scaffolds

A pilot study of circulating levels of TGF-β1 and TGF-β2 as biomarkers of bone healing in patients with non-hypertrophic pseudoarthrosis of long bones

Towards the standardization of methods of tissue processing for the isolation of mesenchymal stromal cells for clinical use

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