Long-Term Cryostorage of Mesenchymal Stem Cell-Containing Hybrid Hydrogel Scaffolds Based on Fibrin and Collagen

The most difficult issue when using tissue engineering products is enabling the ability to store them without losing their restorative capacity. The numbers and viability of mesenchymal stem cells encapsulated in a hydrogel scaffold after cryostorage at −80 °C (by using, individually, two kinds of cryoprotectors—Bambanker and 10% DMSO (Dimethyl sulfoxide) solution) for 3, 6, 9, and 12 months were determined, with subsequent assessment of cell proliferation after 96 h. The analysis of the cellular component was performed using fluorescence microscopy and the two fluorochromes—Hoechst 3334 and NucGreenTM Dead 488. The experimental protocol ensured the preservation of cells in the scaffold structure, retaining both high viability and proliferative activity during storage for 3 months. Longer storage of scaffolds led to their significant changes. Therefore, after 6 months, the proliferative activity of cells decreased. Cryostorage of scaffolds for 9 months led to a decrease in cells’ viability and proliferative activity. As a result of cryostorage of scaffolds for 12 months, a decrease in viability and proliferative activity of cells was observed, as well as pronounced changes in the structure of the hydrogel. The described scaffold cryostorage protocol could become the basis for the development of storage protocols for such tissue engineering products, and for helping to extend the possibilities of their clinical use while accelerating their commercialization.

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Mineralization of 3D Osteogenic Model Based on Gelatin-Dextran Hybrid Hydrogel Scaffold Bioengineered with Mesenchymal Stromal Cells: A Multiparametric Evaluation

Materials ◽

10.3390/ma14143852 ◽

2021 ◽

Vol 14 (14) ◽

pp. 3852

Author(s):

Federica Re ◽

Luciana Sartore ◽

Elisa Borsani ◽

Matteo Ferroni ◽

Camilla Baratto ◽

...

Keyword(s):

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Bone Repair ◽

Active Role ◽

Ionization Mass ◽

Hydrogel Scaffold ◽

Hybrid Hydrogel ◽

Hydrogel Scaffolds ◽

Human Platelet Lysate ◽

Identical Composition

Gelatin–dextran hydrogel scaffolds (G-PEG-Dx) were evaluated for their ability to activate the bone marrow human mesenchymal stromal cells (BM-hMSCs) towards mineralization. G-PEG-Dx1 and G-PEG-Dx2, with identical composition but different architecture, were seeded with BM-hMSCs in presence of fetal bovine serum or human platelet lysate (hPL) with or without osteogenic medium. G-PEG-Dx1, characterized by a lower degree of crosslinking and larger pores, was able to induce a better cell colonization than G-PEG-Dx2. At day 28, G-PEG-Dx2, with hPL and osteogenic factors, was more efficient than G-PEG-Dx1 in inducing mineralization. Scanning electron microscopy (SEM) and Raman spectroscopy showed that extracellular matrix produced by BM-hMSCs and calcium-positive mineralization were present along the backbone of the G-PEG-Dx2, even though it was colonized to a lesser degree by hMSCs than G-PEG-Dx1. These findings were confirmed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), detecting distinct lipidomic signatures that were associated with the different degree of scaffold mineralization. Our data show that the architecture and morphology of G-PEG-Dx2 is determinant and better than that of G-PEG-Dx1 in promoting a faster mineralization, suggesting a more favorable and active role for improving bone repair.

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Hydrogel Scaffolds - Emerging Biomaterial for Regenerative Medicine

International Journal of Pharmaceutical Sciences and Nanotechnology ◽

10.37285/ijpsn.2016.9.2.2 ◽

1970 ◽

Vol 9 (2) ◽

pp. 3156-3168

Author(s):

Prajeesh Kumar ◽

Shivansh Swamy ◽

Raj K. Narang

Keyword(s):

Tissue Engineering ◽

Growth Factors ◽

Regenerative Medicine ◽

Tissue Regeneration ◽

Cellular Proliferation ◽

Hydrogel Scaffold ◽

Hydrogel Scaffolds ◽

Cellular Environment ◽

Direct Incorporation ◽

Unique Ability

The regenerative medicine field has led to the development of various biomaterials. One such development is in the form of Hydrogel Scaffold for Tissue regeneration. This review describes the biomedical advances in Hydrogel Scaffolds as emerging biomaterial for regenerative medicine. Their unique ability to mimic the extra cellular environment, biocompatibility, flexible method of synthesis, desirable framework for cellular proliferation and survival has made them the material of choice. Hydrogels have demonstrated features which exemplify many of the broad-based manifestations of tissue engineering, providing realized as well as potential commercial value. Direct incorporation of cells and growth factors has led to efficient and more promising results in regenerative medicine. This review gives an overview of the various kinds of hydrogels, fabrication methods with specific features and few of the recent applications of hydrogels in the field of regenerative medicine.

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Biocompatibility and Biological Efficiency of Inorganic Calcium Filled Bacterial Cellulose Based Hydrogel Scaffolds for Bone Bioengineering

International Journal of Molecular Sciences ◽

10.3390/ijms19123980 ◽

2018 ◽

Vol 19 (12) ◽

pp. 3980 ◽

Cited By ~ 10

Author(s):

Probal Basu ◽

Nabanita Saha ◽

Radostina Alexandrova ◽

Boyka Andonova-Lilova ◽

Milena Georgieva ◽

...

Keyword(s):

Tissue Engineering ◽

Calcium Phosphate ◽

Bone Tissue Engineering ◽

Bacterial Cellulose ◽

Bone Tissue ◽

Human Fibroblasts ◽

Biological Efficiency ◽

Hydrogel Scaffold ◽

Hydrogel Scaffolds ◽

Depth Analysis

The principal focus of this work is the in-depth analysis of the biological efficiency of inorganic calcium-filled bacterial cellulose (BC) based hydrogel scaffolds for their future use in bone tissue engineering/bioengineering. Inorganic calcium was filled in the form of calcium phosphate (β-tri calcium phosphate (β-TCP) and hydroxyapatite (HA)) and calcium carbonate (CaCO3). The additional calcium, CaCO3 was incorporated following in vitro bio-mineralization. Cell viability study was performed with the extracts of BC based hydrogel scaffolds: BC-PVP, BC-CMC; BC-PVP-β-TCP/HA, BC-CMC-β-TCP/HA and BC-PVP-β-TCP/HA-CaCO3, BC-CMC-β-TCP/HA-CaCO3; respectively. The biocompatibility study was performed with two different cell lines, i.e., human fibroblasts, Lep-3 and mouse bone explant cells. Each hydrogel scaffold has facilitated notable growth and proliferation in presence of these two cell types. Nevertheless, the percentage of DNA strand breaks was higher when cells were treated with BC-CMC based scaffolds i.e., BC-CMC-β-TCP/HA and BC-CMC-β-TCP/HA-CaCO3. On the other hand, the apoptosis of human fibroblasts, Lep-3 was insignificant in BC-PVP-β-TCP/HA. The scanning electron microscopy confirmed the efficient adhesion and growth of Lep-3 cells throughout the surface of BC-PVP and BC-PVP-β-TCP/HA. Hence, among all inorganic calcium filled hydrogel scaffolds, ‘BC-PVP-β-TCP/HA’ was recommended as an efficient tissue engineering scaffold which could facilitate the musculoskeletal (i.e., bone tissue) engineering/bioengineering.

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A pH-Triggered, Self-Assembled, and Bioprintable Hybrid Hydrogel Scaffold for Mesenchymal Stem Cell Based Bone Tissue Engineering

ACS Applied Materials & Interfaces ◽

10.1021/acsami.8b19094 ◽

2019 ◽

Vol 11 (9) ◽

pp. 8749-8762 ◽

Cited By ~ 22

Author(s):

Chen Zhao ◽

Nader Taheri Qazvini ◽

Monirosadat Sadati ◽

Zongyue Zeng ◽

Shifeng Huang ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Hydrogel Scaffold ◽

Hybrid Hydrogel ◽

Self Assembled

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Hybrid Methacrylated Gelatin and Hyaluronic Acid Hydrogel Scaffolds. Preparation and Systematic Characterization for Prospective Tissue Engineering Applications

International Journal of Molecular Sciences ◽

10.3390/ijms22136758 ◽

2021 ◽

Vol 22 (13) ◽

pp. 6758

Author(s):

B. Velasco-Rodriguez ◽

T. Diaz-Vidal ◽

L. C. Rosales-Rivera ◽

C. A. García-González ◽

C. Alvarez-Lorenzo ◽

...

Keyword(s):

Tissue Engineering ◽

Hyaluronic Acid ◽

Engineering Application ◽

Chemical Properties ◽

Tissue Engineering Application ◽

Hydrogel Scaffolds ◽

Hybrid Hydrogels ◽

Physico Chemical ◽

Scaffold Structure ◽

The Relationship

Hyaluronic acid (HA) and gelatin (Gel) are major components of the extracellular matrix of different tissues, and thus are largely appealing for the construction of hybrid hydrogels to combine the favorable characteristics of each biopolymer, such as the gel adhesiveness of Gel and the better mechanical strength of HA, respectively. However, despite previous studies conducted so far, the relationship between composition and scaffold structure and physico-chemical properties has not been completely and systematically established. In this work, pure and hybrid hydrogels of methacroyl-modified HA (HAMA) and Gel (GelMA) were prepared by UV photopolymerization and an extensive characterization was done to elucidate such correlations. Methacrylation degrees of ca. 40% and 11% for GelMA and HAMA, respectively, were obtained, which allows to improve the hydrogels’ mechanical properties. Hybrid GelMA/HAMA hydrogels were stiffer, with elastic modulus up to ca. 30 kPa, and porous (up to 91%) compared with pure GelMA ones at similar GelMA concentrations thanks to the interaction between HAMA and GelMA chains in the polymeric matrix. The progressive presence of HAMA gave rise to scaffolds with more disorganized, stiffer, and less porous structures owing to the net increase of mass in the hydrogel compositions. HAMA also made hybrid hydrogels more swellable and resistant to collagenase biodegradation. Hence, the suitable choice of polymeric composition allows to regulate the hydrogels´ physical properties to look for the most optimal characteristics required for the intended tissue engineering application.

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A microfluidic-based cell encapsulation platform to achieve high long-term cell viability in photopolymerized PEGNB hydrogel microspheres

Journal of Materials Chemistry B ◽

10.1039/c6tb02551j ◽

2017 ◽

Vol 5 (1) ◽

pp. 173-180 ◽

Cited By ~ 26

Author(s):

Zhongliang Jiang ◽

Bingzhao Xia ◽

Ralph McBride ◽

John Oakey

Keyword(s):

Tissue Engineering ◽

Polyethylene Glycol ◽

Regenerative Medicine ◽

Cell Viability ◽

Cell Encapsulation ◽

Cell Delivery ◽

Cellular Behavior ◽

Hydrogel Scaffolds

Cell encapsulation within photopolymerized polyethylene glycol (PEG)-based hydrogel scaffolds has been demonstrated as a robust strategy for cell delivery, tissue engineering, regenerative medicine, and developing in vitro platforms to study cellular behavior and fate.

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Synthesis and characterization of site selective photo-crosslinkable glycidyl methacrylate functionalized gelatin-based 3D hydrogel scaffold for liver tissue engineering

Materials Science and Engineering C ◽

10.1016/j.msec.2020.111694 ◽

2021 ◽

Vol 123 ◽

pp. 111694 ◽

Cited By ~ 1

Author(s):

Md Moniruzzaman Sk ◽

Prativa Das ◽

Amit Panwar ◽

Lay Poh Tan

Keyword(s):

Tissue Engineering ◽

Liver Tissue ◽

Glycidyl Methacrylate ◽

Synthesis And Characterization ◽

Hydrogel Scaffold ◽

Liver Tissue Engineering ◽

Site Selective

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Superabsorbent poly(acrylic acid) and antioxidant poly(ester amide) hybrid hydrogel for enhanced wound healing

Regenerative Biomaterials ◽

10.1093/rb/rbaa059 ◽

2021 ◽

Vol 8 (2) ◽

Author(s):

Jianhua Zhang ◽

Junfei Hu ◽

Baoshu Chen ◽

Tianbao Zhao ◽

Zhipeng Gu

Keyword(s):

Wound Healing ◽

Acrylic Acid ◽

Protein Denaturation ◽

Healing Process ◽

Antioxidant Properties ◽

Skin Damage ◽

Hybrid Hydrogel ◽

Hydrogel Scaffolds ◽

Hybrid Hydrogels ◽

Poly Acrylic Acid

Abstract Wound healing dressing is increasingly needed in clinical owing to the large quantity of skin damage annually. Excessive reactive oxygen species (ROS) produced through internal or external environmental influences can lead to lipid peroxidation, protein denaturation, and even DNA damage, and ultimately have harmful effects on cells. Aiming to sufficiently contact with the wound microenvironment and scavenge ROS, superabsorbent poly (acrylic acid) and antioxidant poly (ester amide) (PAA/PEA) hybrid hydrogel has been developed to enhance wound healing. The physical and chemical properties of hybrid hydrogels were studied by Fourier-transform infrared (FTIR) absorption spectrum, compression, swelling, degradation, etc. Besides, the antioxidant properties of hybrid hydrogels can be investigated through the free radical scavenging experiment, and corresponding antioxidant indicators have been tested at the cellular level. Hybrid hydrogel scaffolds supported the proliferation of human umbilical vein endothelial cells and fibroblasts, as well as accelerated angiogenesis and skin regeneration in wounds. The healing properties of wounds in vivo were further assessed on mouse skin wounds. Results showed that PAA/PEA hybrid hydrogel scaffolds significantly accelerated the wound healing process through enhancing granulation formation and re-epithelialization. In summary, these superabsorbent and antioxidative hybrid hydrogels could be served as an excellent wound dressing for full-thickness wound healing.

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14-3-3ε protein-loaded 3D hydrogels favor osteogenesis

Journal of Materials Science Materials in Medicine ◽

10.1007/s10856-020-06434-1 ◽

2020 ◽

Vol 31 (11) ◽

Author(s):

Ana A. Aldana ◽

Marina Uhart ◽

Gustavo A. Abraham ◽

Diego M. Bustos ◽

Aldo R. Boccaccini

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

3D Printing ◽

Osteogenic Differentiation ◽

Alginate Hydrogel ◽

Hydrogel Scaffolds ◽

Future Developments ◽

Positive Effect ◽

Cell Adhesion And Proliferation

Abstract3D printing has emerged as vanguard technique of biofabrication to assemble cells, biomaterials and biomolecules in a spatially controlled manner to reproduce native tissues. In this work, gelatin methacrylate (GelMA)/alginate hydrogel scaffolds were obtained by 3D printing and 14-3-3ε protein was encapsulated in the hydrogel to induce osteogenic differentiation of human adipose-derived mesenchymal stem cells (hASC). GelMA/alginate-based grid-like structures were printed and remained stable upon photo-crosslinking. The viscosity of alginate allowed to control the pore size and strand width. A higher viscosity of hydrogel ink enhanced the printing accuracy. Protein-loaded GelMA/alginate-based hydrogel showed a clear induction of the osteogenic differentiation of hASC cells. The results are relevant for future developments of GelMA/alginate for bone tissue engineering given the positive effect of 14-3-3ε protein on both cell adhesion and proliferation.

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