In Vitro Evaluation of a Composite Gelatin–Hyaluronic Acid–Alginate Porous Scaffold with Different Pore Distributions for Cartilage Regeneration

Although considerable achievements have been made in the field of regenerative medicine, since self-repair is not an advanced ability of articular cartilage, the regeneration of osteochondral defects is still a challenging problem in musculoskeletal diseases. Cartilage regeneration aims to design a scaffold with appropriate pore structure and biological and mechanical properties for the growth of chondrocytes. In this study, porous scaffolds made of gelatin, hyaluronic acid, alginate, and sucrose in different proportions of 2 g (SL2) and 4 g (SL4) were used as porogens in a leaching process. Sucrose with particle size ranges of 88–177 μm (Hμ) and 44–74 μm (SHμ) was added to the colloid, and the individually cross-linked hydrogel scaffolds with controllable pore size for chondrocyte culture were named Hμ-SL2, Hμ-SL4, SHμ-SL2 and SHμ-SL4. The perforation, porosity, mechanical strength, biocompatibility, and proliferation characteristics of the hydrogel scaffold and its influence on chondrocyte differentiation are discussed. Results show that the addition of porogen increases the porosity of the hydrogel scaffold. Conversely, when porogens with the same particle size are added, the pore size decreases as the amount of porogen increases. The perforation effect of the hydrogel scaffolds formed by the porogen is better at 88–177 μm compared with that at 44–74 μm. Cytotoxicity analysis showed that all the prepared hydrogel scaffolds were non-cytotoxic, indicating that no cross-linking agent residues that could cause cytotoxicity were found. In the proliferation and differentiation of the chondrocytes, the SHμ-SL4 hydrogel scaffold with the highest porosity and strength did not achieve the best performance. However, due to the compromise between perforation pores, pore sizes, and strength, as well as considering cell proliferation and differentiation, Hμ–SL4 scaffold provided a more suitable environment for the chondrocytes than other groups; therefore, it can provide the best chondrocyte growth environment for this study. The development of hydrogels with customized pore properties for defective cartilage is expected to meet the requirements of the ultimate clinical application.

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Preparation and Characterization of Ha/Gel/β-TCP Microspheres Composite Porous Scaffold

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.782.103 ◽

2018 ◽

Vol 782 ◽

pp. 103-115

Author(s):

Yang Zi Zhao ◽

You Fa Wang

Keyword(s):

Tissue Engineering ◽

Compressive Strength ◽

Hyaluronic Acid ◽

Pore Size ◽

Porous Scaffold ◽

Three Dimensional ◽

Swelling Ratio ◽

Cell Compatibility ◽

Hydrogel Scaffolds

Being one of the three elements of tissue engineering, three-dimensional porous structure scaffold plays an important role in tissue engineering. As it not only prvovide cells for the life, but also serves as a template to guide tissue regeneration and control of organizational structure and other functions. In this study, hyaluronic acid and gelatin are successfully cross-linked by 1-ethyl- (3-dimethylaminopropyl) -carbodiimide hydrochloride (EDC) , and compound β-TCP microspheres to prepare porous hydrogel scaffolds. The microspheres were analyzed by X-ray diffraction (XRD). The scaffolds were characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). At the same time, the compressive strength, swelling ratio, degradation of the scaffold were tested. To assess the in vitro cell compatibility of the scaffolds, mouse L929 fibroblasts were seeded onto scaffolds for cell morphology and cell viability studies. The results showed that the pore size of the porous scaffold can be adjusted by changing the ratio of gelatin to hyaluronic acid (HA), increasing the proportion of hyaluronic acid in a certain range, pore size will be significantly increased. With the increase of the proportion of hyaluronic acid in the scaffold, the swelling ratio and the degradation rate also increased. The compressive strength of the scaffold increased with the increase of the proportion of gelatin. The appropriate ratio of β-TCP can promote cell growth and proliferation.

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A Novel Cross-Linked Hyaluronic Acid Porous Scaffold for Cartilage Repair

Cartilage ◽

10.1177/1947603515611949 ◽

2015 ◽

Vol 7 (3) ◽

pp. 265-273 ◽

Cited By ~ 10

Author(s):

Christoph Bauer ◽

Manuela Berger ◽

Renate R. Baumgartner ◽

Sonja Höller ◽

Hannes Zwickl ◽

...

Keyword(s):

Cell Proliferation ◽

Hyaluronic Acid ◽

Articular Chondrocytes ◽

Porous Scaffold ◽

Cartilage Regeneration ◽

Cell Number ◽

Cell Content ◽

3 Dimensional ◽

Chondrogenic Phenotype ◽

Differentiation Status

Purpose An important feature of biomaterials used in cartilage regeneration is their influence on the establishment and stabilization of a chondrocytic phenotype of embedded cells. The purpose of this study was to examine the effects of a porous 3-dimensional scaffold made of cross-linked hyaluronic acid on the expression and synthesis performance of human articular chondrocytes. Materials and Methods Osteoarthritic chondrocytes from 5 patients with a mean age of 74 years were passaged twice and cultured within the cross-linked hyaluronic acid scaffolds for 2 weeks. Analyses were performed at 3 different time points. For estimation of cell content within the scaffold, DNA-content (CyQuant cell proliferation assay) was determined. The expression of chondrocyte-specific genes by embedded cells as well as the total amount of sulfated glycosaminoglycans produced during the culture period was analyzed in order to characterize the synthesis performance and differentiation status of the cells. Results Cells showed a homogenous distribution within the scaffold. DNA quantification revealed a reduction of the cell number. This might be attributed to loss of cells from the scaffold during media exchange connected with a stop in cell proliferation. Indeed, the expression of cartilage-specific genes and the production of sulfated glycosaminoglycans were increased and the differentiation index was clearly improved. Conclusions These results suggest that the attachment of osteoarthritic P2 chondrocytes to the investigated material enhanced the chondrogenic phenotype as well as promoted the retention.

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Novel Method to Fabricate Porous n-HA/PVA Hydrogel Scaffolds

Materials Science Forum ◽

10.4028/www.scientific.net/msf.510-511.878 ◽

2006 ◽

Vol 510-511 ◽

pp. 878-881 ◽

Cited By ~ 6

Author(s):

Yuan Hua Mu ◽

Yu Bao Li ◽

Ming Bo Wang ◽

Feng Lan Xu ◽

Xiang Zhang ◽

...

Keyword(s):

Pore Size ◽

Freeze Drying ◽

Normal Pressure ◽

Drying Method ◽

Hydrogel Scaffold ◽

Hydrogel Scaffolds ◽

Novel Method ◽

Porous Hydrogel ◽

Pva Hydrogel

Porous n-HA/PVA hydrogel composite was prepared through in-situ hydrothermal treatment under normal pressure and emulsion foam freeze-drying method, which was used to fabricate porous hydrogel. The pores exhibited interconnection-pore structure owing to the injection of air bubbles and the removal of emulsifier (OP). The porous hydrogels were investigated by using IR, XRD, TEM and SEM. The results indicated that n-HA in the composite could disperse uniformly, and there were chemical bonding with PVA. In addition, nano-hydroxyapatite existed in the composite in the shape of short-rod. The pores were interconnection with narrowly pore size and highly porosity. And the pore size and size distribution were influenced by the weight of OP. The emulsion foam freeze-drying method can be used to prepare porous hydrogel scaffold for tissue engineering, or to contain proteins scaffold, because of operating at a low temperature. The method displayed a vast potential of applied foreground.

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Bioactive Hydrogel Scaffolds - Advances in Cartilage Regeneration Through Controlled Drug Delivery

Current Pharmaceutical Design ◽

10.2174/1381612821666150115150712 ◽

2015 ◽

Vol 21 (12) ◽

pp. 1545-1555 ◽

Cited By ~ 14

Author(s):

Roberta Censi ◽

Alessandra Dubbini ◽

Pietro Matricardi

Keyword(s):

Drug Delivery ◽

Controlled Drug Delivery ◽

Cartilage Regeneration ◽

Hydrogel Scaffolds

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Polylactic acid-based electrospun fiber and hyaluronic acid-valsartan hydrogel scaffold for chronic wound healing

Beni-Suef University Journal of Basic and Applied Sciences ◽

10.1186/s43088-020-00057-9 ◽

2020 ◽

Vol 9 (1) ◽

Author(s):

Margaret O. Ilomuanya ◽

Prosper S. Okafor ◽

Joyce N. Amajuoyi ◽

John C. Onyejekwe ◽

Omotunde O. Okubanjo ◽

...

Keyword(s):

Wound Healing ◽

Hyaluronic Acid ◽

Polylactic Acid ◽

Chronic Wound ◽

Electrospun Fiber ◽

Hydrogel Scaffold

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

Gels ◽

10.3390/gels6040044 ◽

2020 ◽

Vol 6 (4) ◽

pp. 44

Author(s):

Marfa N. Egorikhina ◽

Yulia P. Rubtsova ◽

Diana Ya. Aleynik

Keyword(s):

Tissue Engineering ◽

Proliferative Activity ◽

Experimental Protocol ◽

Hydrogel Scaffold ◽

Hybrid Hydrogel ◽

Hydrogel Scaffolds ◽

High Viability ◽

Scaffold Structure ◽

Difficult Issue

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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Digital Light Processing Bioprinted Human Chondrocyte-Laden Poly (γ-Glutamic Acid)/Hyaluronic Acid Bio-Ink towards Cartilage Tissue Engineering

Biomedicines ◽

10.3390/biomedicines9070714 ◽

2021 ◽

Vol 9 (7) ◽

pp. 714

Author(s):

Alvin Kai-Xing Lee ◽

Yen-Hong Lin ◽

Chun-Hao Tsai ◽

Wan-Ting Chang ◽

Tsung-Li Lin ◽

...

Keyword(s):

United States ◽

Tissue Engineering ◽

Hyaluronic Acid ◽

Glutamic Acid ◽

Cellular Proliferation ◽

Cartilage Tissue Engineering ◽

Cartilage Regeneration ◽

The United States ◽

Cartilage Injury ◽

Cartilage Tissue

Cartilage injury is the main cause of disability in the United States, and it has been projected that cartilage injury caused by osteoarthritis will affect 30% of the entire United States population by the year 2030. In this study, we modified hyaluronic acid (HA) with γ-poly(glutamic) acid (γ-PGA), both of which are common biomaterials used in cartilage engineering, in an attempt to evaluate them for their potential in promoting cartilage regeneration. As seen from the results, γ-PGA-GMA and HA, with glycidyl methacrylate (GMA) as the photo-crosslinker, could be successfully fabricated while retaining the structural characteristics of γ-PGA and HA. In addition, the storage moduli and loss moduli of the hydrogels were consistent throughout the curing durations. However, it was noted that the modification enhanced the mechanical properties, the swelling equilibrium rate, and cellular proliferation, and significantly improved secretion of cartilage regeneration-related proteins such as glycosaminoglycan (GAG) and type II collagen (Col II). The cartilage tissue proof with Alcian blue further demonstrated that the modification of γ-PGA with HA exhibited suitability for cartilage tissue regeneration and displayed potential for future cartilage tissue engineering applications. This study built on the previous works involving HA and further showed that there are unlimited ways to modify various biomaterials in order to further bring cartilage tissue engineering to the next level.

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