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19 pages, 12306 KiB

Open AccessArticle

Towards Complex Tissues Replication: Multilayer Scaffold Integrating Biomimetic Nanohydroxyapatite/Chitosan Composites

by Barbara Palazzo, Stefania Scialla, Amilcare Barca, Laura Sercia, Daniela Izzo, Francesca Gervaso and Francesca Scalera

Bioengineering 2024, 11(5), 471; https://doi.org/10.3390/bioengineering11050471 - 9 May 2024

Viewed by 1483

Abstract

This study explores an approach to design and prepare a multilayer scaffold mimicking interstratified natural tissue. This multilayer construct, composed of chitosan matrices with graded nanohydroxyapatite concentrations, was achieved through an in situ biomineralization process applied to individual layers. Three distinct precursor concentrations [...] Read more.

This study explores an approach to design and prepare a multilayer scaffold mimicking interstratified natural tissue. This multilayer construct, composed of chitosan matrices with graded nanohydroxyapatite concentrations, was achieved through an in situ biomineralization process applied to individual layers. Three distinct precursor concentrations were considered, resulting in 10, 20, and 30 wt% nanohydroxyapatite content in each layer. The resulting chitosan/nanohydroxyapatite (Cs/n-HAp) scaffolds, created via freeze-drying, exhibited nanohydroxyapatite nucleation, homogeneous distribution, improved mechanical properties, and good cytocompatibility. The cytocompatibility analysis revealed that the Cs/n-HAp layers presented cell proliferation similar to the control in pure Cs for the samples with 10% n-HAp, indicating good cytocompatibility at this concentration, while no induction of apoptotic death pathways was demonstrated up to a 20 wt% n-Hap concentration. Successful multilayer assembly of Cs and Cs/n-HAp layers highlighted that the proposed approach represents a promising strategy for mimicking multifaceted tissues, such as osteochondral ones. Full article

(This article belongs to the Special Issue Biomaterials for Cartilage and Bone Tissue Engineering)

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21 pages, 7235 KiB

Open AccessArticle

Photobiomodulation Therapy Improves Repair of Bone Defects Filled by Inorganic Bone Matrix and Fibrin Heterologous Biopolymer

by Maria Fernanda Rossi Vigliar, Lais Furlaneto Marega, Marco Antonio Hungaro Duarte, Murilo Priori Alcalde, Marcelie Priscila de Oliveira Rosso, Rui Seabra Ferreira Junior, Benedito Barraviera, Carlos Henrique Bertoni Reis, Daniela Vieira Buchaim and Rogerio Leone Buchaim

Bioengineering 2024, 11(1), 78; https://doi.org/10.3390/bioengineering11010078 - 13 Jan 2024

Cited by 1 | Viewed by 1802

Abstract

Biomaterials are used extensively in graft procedures to correct bone defects, interacting with the body without causing adverse reactions. The aim of this pre-clinical study was to analyze the effects of photobiomodulation therapy (PBM) with the use of a low-level laser in the [...] Read more.

Biomaterials are used extensively in graft procedures to correct bone defects, interacting with the body without causing adverse reactions. The aim of this pre-clinical study was to analyze the effects of photobiomodulation therapy (PBM) with the use of a low-level laser in the repair process of bone defects filled with inorganic matrix (IM) associated with heterologous fibrin biopolymer (FB). A circular osteotomy of 4 mm in the left tibia was performed in 30 Wistar male adult rats who were randomly divided into three groups: G1 = IM + PBM, G2 = IM + FB and G3 = IM + FB + PBM. PBM was applied at the time of the experimental surgery and three times a week, on alternate days, until euthanasia, with 830 nm wavelength, in two points of the operated site. Five animals from each group were euthanized 14 and 42 days after surgery. In the histomorphometric analysis, the percentage of neoformed bone tissue in G3 (28.4% ± 2.3%) was higher in relation to G1 (24.1% ± 2.91%) and G2 (22.2% ± 3.11%) at 14 days and at 42 days, the percentage in G3 (35.1% ± 2.55%) was also higher in relation to G1 (30.1% ± 2.9%) and G2 (31.8% ± 3.12%). In the analysis of the birefringence of collagen fibers, G3 showed a predominance of birefringence between greenish-yellow in the neoformed bone tissue after 42 days, differing from the other groups with a greater presence of red-orange fibers. Immunohistochemically, in all experimental groups, it was possible to observe immunostaining for osteocalcin (OCN) near the bone surface of the margins of the surgical defect and tartrate-resistant acid phosphatase (TRAP) bordering the newly formed bone tissue. Therefore, laser photobiomodulation therapy contributed to improving the bone repair process in tibial defects filled with bovine biomaterial associated with fibrin biopolymer derived from snake venom. Full article

(This article belongs to the Special Issue Biomaterials for Cartilage and Bone Tissue Engineering)

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14 pages, 6185 KiB

Open AccessArticle

Retention of Human iPSC-Derived or Primary Cells Following Xenotransplantation into Rat Immune-Privileged Sites

by Thomas Später, Giselle Kaneda, Melissa Chavez, Julia Sheyn, Jacob Wechsler, Victoria Yu, Patricia Del Rio, Dave Huang, Melodie Metzger, Wafa Tawackoli and Dmitriy Sheyn

Bioengineering 2023, 10(9), 1049; https://doi.org/10.3390/bioengineering10091049 - 6 Sep 2023

Cited by 1 | Viewed by 1785

Abstract

In regenerative medicine, experimental animal models are commonly used to study potential effects of human cells as therapeutic candidates. Although some studies describe certain cells, such as mesenchymal stromal cells (MSC) or human primary cells, as hypoimmunogenic and therefore unable to trigger strong [...] Read more.

In regenerative medicine, experimental animal models are commonly used to study potential effects of human cells as therapeutic candidates. Although some studies describe certain cells, such as mesenchymal stromal cells (MSC) or human primary cells, as hypoimmunogenic and therefore unable to trigger strong inflammatory host responses, other studies report antibody formation and immune rejection following xenotransplantation. Accordingly, the goal of our study was to test the cellular retention and survival of human-induced pluripotent stem cell (iPSCs)-derived MSCs (iMSCs) and primary nucleus pulposus cells (NPCs) following their xenotransplantation into immune-privileged knee joints (14 days) and intervertebral discs (IVD; 7 days) of immunocompromised Nude and immunocompetent Sprague Dawley (SD) rats. At the end of both experiments, we could demonstrate that both rat types revealed comparably low levels of systemic IL-6 and IgM inflammation markers, as assessed via ELISA. Furthermore, the number of recovered cells was with no significant difference between both rat types. Conclusively, our results show that xenogeneic injection of human iMSC and NPC into immunoprivileged knee and IVD sites did not lead to an elevated inflammatory response in immunocompetent rats when compared to immunocompromised rats. Hence, immunocompetent rats represent suitable animals for xenotransplantation studies targeting immunoprivileged sites. Full article

(This article belongs to the Special Issue Biomaterials for Cartilage and Bone Tissue Engineering)

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17 pages, 3240 KiB

Open AccessEditor’s ChoiceArticle

Ex Vivo Model to Evaluate the Antibacterial and Anti-Inflammatory Effects of Gelatin–Tricalcium Phosphate Composite Incorporated with Emodin and Lumbrokinase for Bone Regeneration

by Wen-Ling Wang, Yuan-Man Hsu, Meng-Liang Lin, Shih-Shun Chen, Yi-Hui Lai, Chiung-Hua Huang and Chun-Hsu Yao

Bioengineering 2023, 10(8), 906; https://doi.org/10.3390/bioengineering10080906 - 31 Jul 2023

Cited by 2 | Viewed by 1941

Abstract

Tricalcium phosphate (TCP) has gained attention due to its interconnected porous structures which promote fibrovascular invasion and bony replacement. Moreover, when gelatin is added and crosslinked with genipin (GGT), TCP exhibits robust biocompatibility and stability, making it an excellent bone substitute. In this [...] Read more.

Tricalcium phosphate (TCP) has gained attention due to its interconnected porous structures which promote fibrovascular invasion and bony replacement. Moreover, when gelatin is added and crosslinked with genipin (GGT), TCP exhibits robust biocompatibility and stability, making it an excellent bone substitute. In this study, we incorporated emodin and lumbrokinase (LK) into GGT to develop an antibacterial biomaterial. Emodin, derived from various plants, possesses antibacterial and anti-inflammatory properties. LK comprises proteolytic enzymes extracted from the earthworm Lumbricus rubellus and exhibits fibrinolytic activity, enabling it to dissolve biofilms. Additionally, LK stimulates osteoblast activity while inhibiting osteoclast differentiation. GGT was combined with emodin and lumbrokinase to produce the GGTELK composite. The biomedical effects of GGTELK were assessed through in vitro assays and an ex vivo bone defect model. The GGTELK composite demonstrated antibacterial properties, inhibiting the growth of S. aureus and reducing biofilm formation. Moreover, it exhibited anti-inflammatory effects by reducing the secretion of IL-6 in both in vivo cell experiments and the ex vivo model. Therefore, the GGTELK composite, with its stability, efficient degradation, biocompatibility, and anti-inflammatory function, is expected to serve as an ideal bone substitute. Full article

(This article belongs to the Special Issue Biomaterials for Cartilage and Bone Tissue Engineering)

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19 pages, 4697 KiB

Open AccessArticle

Osteochondral Regeneration Ability of Uncultured Bone Marrow Mononuclear Cells and Platelet-Rich Fibrin Scaffold

by Tung Nguyen-Thanh, Bao-Song Nguyen-Tran, Sara Cruciani, Thuy-Duong Nguyen-Thi, Thuan Dang-Cong and Margherita Maioli

Bioengineering 2023, 10(6), 661; https://doi.org/10.3390/bioengineering10060661 - 31 May 2023

Cited by 3 | Viewed by 2363

Abstract

Objectives: Platelet-rich fibrin (PRF) and bone marrow mononuclear cells are potential scaffolds and cell sources for osteochondral regeneration. The main aim of this paper is to examine the effects of PRF scaffolds and autologous uncultured bone marrow mononuclear cells on osteochondral regeneration in [...] Read more.

Objectives: Platelet-rich fibrin (PRF) and bone marrow mononuclear cells are potential scaffolds and cell sources for osteochondral regeneration. The main aim of this paper is to examine the effects of PRF scaffolds and autologous uncultured bone marrow mononuclear cells on osteochondral regeneration in rabbit knees. Materials and Methods: Three different types of PRF scaffolds were generated from peripheral blood (Ch-PRF and L-PRF) and bone marrow combined with uncultured bone marrow mononuclear cells (BMM-PRF). The histological characteristics of these scaffolds were assessed via hematoxylin–eosin staining, PicroSirius red staining, and immunohistochemical staining. Osteochondral defects with a diameter of 3 mm and depth of 3 mm were created on the trochlear groove of the rabbit’s femur. Different PRF scaffolds were then applied to treat the defects. A group of rabbits with induced osteochondral defects that were not treated with any scaffold was used as a control. Osteochondral tissue regeneration was assessed after 2, 4, and 6 weeks by macroscopy (using the Internal Cartilage Repair Society score, X-ray) and microscopy (hematoxylin—eosin stain, safranin O stain, toluidine stain, and Wakitani histological scale, immunohistochemistry), in addition to gene expression analysis of osteochondral markers. Results: Ch-PRF had a heterogeneous fibrin network structure and cellular population; L-PRF and BMM-PRF had a homogeneous structure with a uniform distribution of the fibrin network. Ch-PRF and L-PRF contained a population of CD45-positive leukocytes embedded in the fibrin network, while mononuclear cells in the BMM-PRF scaffold were positive for the pluripotent stem cell-specific antibody Oct-4. In comparison to the untreated group, the rabbits that were given the autologous graft displayed significantly improved healing of the articular cartilage tissue and of the subchondral bone. Regeneration was gradually observed after 2, 4, and 6 weeks of PRF scaffold treatment, which was particularly evident in the BMM-PRF group. Conclusions: The combination of biomaterials with autologous platelet-rich fibrin and uncultured bone marrow mononuclear cells promoted osteochondral regeneration in a rabbit model more than platelet-rich fibrin material alone. Our results indicate that autologous platelet-rich fibrin scaffolds combined with uncultured bone marrow mononuclear cells applied in healing osteochondral lesions may represent a suitable treatment in addition to stem cell and biomaterial therapy. Full article

(This article belongs to the Special Issue Biomaterials for Cartilage and Bone Tissue Engineering)

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19 pages, 10334 KiB

Open AccessArticle

PCL/Graphene Scaffolds for the Osteogenesis Process

by Silvia Anitasari, Ching-Zong Wu and Yung-Kang Shen

Bioengineering 2023, 10(3), 305; https://doi.org/10.3390/bioengineering10030305 - 28 Feb 2023

Cited by 7 | Viewed by 2471

Abstract

This study aims to characterize the osteoconductivity, optimal bioresorbable, biodegradability, biocompatibility, and mechanical properties of Poly-ε-caprolactone (PCL)/graphene (G) scaffolds at concentrations of 0.5, 1, 1.5, 2, 2.5, and 3 wt%, which are used to support bone regeneration through solvent casting and [...] Read more.

This study aims to characterize the osteoconductivity, optimal bioresorbable, biodegradability, biocompatibility, and mechanical properties of Poly-ε-caprolactone (PCL)/graphene (G) scaffolds at concentrations of 0.5, 1, 1.5, 2, 2.5, and 3 wt%, which are used to support bone regeneration through solvent casting and particulate leaching. The water contact angle measurement revealed a transition from a hydrophobic to a hydrophilic surface after incorporating various G concentrations. The scaffolds with 0.5 wt% G had smaller pores compared to those produced using 3 wt% G. Furthermore, numerous pores were connected, particularly those with larger diameters in the 2 and 3 wt% G samples. The proportion of water absorption varied between 50% and 350% for 4 months, with large percentages of scaffolds containing high G concentrations. Raman spectroscopy and X-ray diffraction, which were used to confirm the presence of nanofiller by increasing the ratios of I_D/I_G, I_2D/I_G, and band 2θ = 26.48°. The mechanical properties were improved by the addition of G, with a Young’s modulus of 3 wt% G, four times that of PCL. Measuring cell biocompatibility, adhesion, proliferation, and differentiation with osteoblast-like (MG-63) cells revealed that PCL/G scaffolds with higher concentrations were more biocompatible than PCL as well as those with lower concentrations. Full article

(This article belongs to the Special Issue Biomaterials for Cartilage and Bone Tissue Engineering)

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16 pages, 4338 KiB

Open AccessArticle

Bioactive Scaffold Fabricated by 3D Printing for Enhancing Osteoporotic Bone Regeneration

by Xiaoting Zhang, Xinluan Wang, Yuk-wai Lee, Lu Feng, Bin Wang, Qi Pan, Xiangbo Meng, Huijuan Cao, Linlong Li, Haixing Wang, Shanshan Bai, Lingchi Kong, Dick Ho Kiu Chow, Ling Qin, Liao Cui, Sien Lin and Gang Li

Bioengineering 2022, 9(10), 525; https://doi.org/10.3390/bioengineering9100525 - 5 Oct 2022

Cited by 6 | Viewed by 3232

Abstract

We develop a poly (lactic-co-glycolic acid)/β-calcium phosphate (PLGA/TCP)-based scaffold through a three-dimensional (3D) printing technique incorporating icaritin (ICT), a unique phytomolecule, and secretome derived from human fetal mesenchymal stem cells (HFS), to provide mechanical support and biological cues for stimulating bone defect healing. [...] Read more.

We develop a poly (lactic-co-glycolic acid)/β-calcium phosphate (PLGA/TCP)-based scaffold through a three-dimensional (3D) printing technique incorporating icaritin (ICT), a unique phytomolecule, and secretome derived from human fetal mesenchymal stem cells (HFS), to provide mechanical support and biological cues for stimulating bone defect healing. With the sustained release of ICT and HFS from the composite scaffold, the cell-free scaffold efficiently facilitates the migration of MSCs and promotes bone regeneration at the femoral defect site in the ovariectomy (OVX)-induced osteoporotic rat model. Furthermore, mechanism study results indicate that the combination of ICT and HFS additively activates the Integrin–FAK (focal adhesion kinase)–ERK1/2 (extracellular signal-regulated kinase 1/2)–Runx2 (Runt-related transcription factor 2) axis, which could be linked to the beneficial recruitment of MSCs to the implant and subsequent osteogenesis enhancement. Collectively, the PLGA/TCP/ICT/HFS (P/T/I/S) bioactive scaffold is a promising biomaterial for repairing osteoporotic bone defects, which may have immense implications for their translation to clinical practice. Full article

(This article belongs to the Special Issue Biomaterials for Cartilage and Bone Tissue Engineering)

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Review

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19 pages, 2796 KiB

Open AccessReview

Cartilage Defect Treatment Using High-Density Autologous Chondrocyte Implantation (HD-ACI)

by Pedro Guillén-García, Isabel Guillén-Vicente, Elena Rodríguez-Iñigo, Marta Guillén-Vicente, Tomás Fernando Fernández-Jaén, Ramón Navarro, Lucía Aboli, Raúl Torres, Steve Abelow and Juan Manuel López-Alcorocho

Bioengineering 2023, 10(9), 1083; https://doi.org/10.3390/bioengineering10091083 - 13 Sep 2023

Cited by 8 | Viewed by 2197

Abstract

Hyaline cartilage’s inability to self-repair can lead to osteoarthritis and joint replacement. Various treatments, including cell therapy, have been developed for cartilage damage. Autologous chondrocyte implantation (ACI) is considered the best option for focal chondral lesions. In this article, we aimed to create [...] Read more.

Hyaline cartilage’s inability to self-repair can lead to osteoarthritis and joint replacement. Various treatments, including cell therapy, have been developed for cartilage damage. Autologous chondrocyte implantation (ACI) is considered the best option for focal chondral lesions. In this article, we aimed to create a narrative review that highlights the evolution and enhancement of our chondrocyte implantation technique: High-Density-ACI (HD-ACI) Membrane-assisted Autologous Chondrocyte Implantation (MACI) improved ACI using a collagen membrane as a carrier. However, low cell density in MACI resulted in softer regenerated tissue. HD-ACI was developed to improve MACI, implanting 5 million chondrocytes per cm², providing higher cell density. In animal models, HD-ACI formed hyaline-like cartilage, while other treatments led to fibrocartilage. HD-ACI was further evaluated in patients with knee or ankle defects and expanded to treat hip lesions and bilateral defects. HD-ACI offers a potential solution for cartilage defects, improving outcomes in regenerative medicine and cell therapy. HD-ACI, with its higher cell density, shows promise for treating chondral defects and advancing cartilage repair in regenerative medicine and cell therapy. Full article

(This article belongs to the Special Issue Biomaterials for Cartilage and Bone Tissue Engineering)

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