In Vivo Regeneration of Large Bone Defects by Cross-Linked Porous Hydrogel: A Pilot Study in Mice Combining Micro Tomography, Histological Analyses, Raman Spectroscopy and Synchrotron Infrared Imaging

Adachi, Tetsuya; Boschetto, Francesco; Miyamoto, Nao; Yamamoto, Toshiro; Marin, Elia; Zhu, Wenliang; Kanamura, Narisato; Tahara, Yoshiro; Akiyoshi, Kazunari; Mazda, Osam; Nishimura, Ichiro; Pezzotti, Giuseppe

doi:10.3390/ma13194275

Cited by 14 publications

(17 citation statements)

References 43 publications

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“…Using Raman spectroscopy, we developed a technology for the real-time monitoring of the differentiation state of regenerated cartilage tissue and of the “quality” of the cartilage matrix at the molecular level, including information on its steric construction. The data presented in this study build upon previous studies of cross-linked pullulan polysaccharide-based nanogels and the subsequent development of a nanogel-cross-linked porous freeze-dried (NanoCliP-FD) gel scaffold [ 7 , 8 , 9 , 10 , 11 ]. As a supplementary output, this study completes our previous Raman spectroscopic characterizations of human cartilage tissue explanted from both healthy and osteoarthritic patients and osteoarthritic cartilage regeneration by mRNA therapeutics [ 12 , 13 , 14 ].…”

Section: Introductionmentioning

confidence: 53%

“…To characterize the content and distribution of collagen and proteoglycan in the cartilage tissue, SR-FTIR spectromicroscopy was performed with the BL15 beam line at the Ritsumeikan University SR Center (Kusatsu-shi, Shiga-ken, Japan). Sample preparation and SR-FTIR spectral analysis were performed according to a previously described procedure [ 11 , 73 ]. Briefly, paraffin-embedded scaffolds were sagittally sectioned into 5 μm slices, which were placed onto BaF 2 substrates (Pier Optics Co., Ltd., Gunma, Japan) immediately after slicing.…”

Section: Methodsmentioning

confidence: 99%

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Three-Dimensional Culture of Cartilage Tissue on Nanogel-Cross-Linked Porous Freeze-Dried Gel Scaffold for Regenerative Cartilage Therapy: A Vibrational Spectroscopy Evaluation

Adachi

Miyamoto

Imamura

et al. 2022

IJMS

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This study presents a set of vibrational characterizations on a nanogel-cross-linked porous freeze-dried gel (NanoCliP-FD gel) scaffold for tissue engineering and regenerative therapy. This scaffold is designed for the in vitro culture of high-quality cartilage tissue to be then transplanted in vivo to enable recovery from congenital malformations in the maxillofacial area or crippling jaw disease. The three-dimensional scaffold for in-plate culture is designed with interface chemistry capable of stimulating cartilage formation and maintaining its structure through counteracting the dedifferentiation of mesenchymal stem cells (MSCs) during the formation of cartilage tissue. The developed interface chemistry enabled high efficiency in both growth rate and tissue quality, thus satisfying the requirements of large volumes, high matrix quality, and superior mechanical properties needed in cartilage transplants. We characterized the cartilage tissue in vitro grown on a NanoCliP-FD gel scaffold by human periodontal ligament-derived stem cells (a type of MSC) with cartilage grown by the same cells and under the same conditions on a conventional (porous) atelocollagen scaffold. The cartilage tissues produced by the MSCs on different scaffolds were comparatively evaluated by immunohistochemical and spectroscopic analyses. Cartilage differentiation occurred at a higher rate when MSCs were cultured on the NanoCliP-FD gel scaffold compared to the atelocollagen scaffold, and produced a tissue richer in cartilage matrix. In situ spectroscopic analyses revealed the cell/scaffold interactive mechanisms by which the NanoCliP-FD gel scaffold stimulated such increased efficiency in cartilage matrix formation. In addition to demonstrating the high potential of human periodontal ligament-derived stem cell cultures on NanoCliP-FD gel scaffolds in regenerative cartilage therapy, the present study also highlights the novelty of Raman spectroscopy as a non-destructive method for the concurrent evaluation of matrix quality and cell metabolic response. In situ Raman analyses on living cells unveiled for the first time the underlying physiological mechanisms behind such improved chondrocyte performance.

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Section: Introductionmentioning

confidence: 53%

Section: Methodsmentioning

confidence: 99%

Three-Dimensional Culture of Cartilage Tissue on Nanogel-Cross-Linked Porous Freeze-Dried Gel Scaffold for Regenerative Cartilage Therapy: A Vibrational Spectroscopy Evaluation

Adachi

Miyamoto

Imamura

et al. 2022

IJMS

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“…Recently, Du et al [ 41 ] adopted Raman spectroscopy to explore the microstructure and chemical components of the new bone tissue in an animal model of bone defect. Moreover, Atachi et al [ 42 ] evaluated the in vivo performance of a 3D scaffold specifically developed for bone tissue engineering also with the support of Raman spectroscopy.…”

Section: Discussionmentioning

confidence: 99%

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

Sartore

Borsani

et al. 2021

Materials

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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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“…However, the porous structure of materials is also beneficial for wound healing—the dressing may be a breathable protective barrier that maintains the optimal microenvironment for the wound healing process, as well may constitute a micro-skeleton for the migrating cells involved in granulation and epithelialization [ 5 , 6 , 7 , 8 ]. Various biopolymers may be employed in the preparation of porous materials, such as collagen [ 9 , 10 ], gelatin [ 11 , 12 ], chitosan [ 13 ], fibroin [ 14 ], sodium alginate [ 15 ], pullulan [ 16 ] and cellulose [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

Lyophilized Emulsions in the Form of 3D Porous Matrices as a Novel Material for Topical Application

Prus-Walendziak

Kozłowska

2021

Materials

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Researchers are constantly searching for innovations that can be applied to the cosmetic industry. Production of porous materials stored in a lyophilized form and swollen directly before use may be beneficial considering their facilitated packaging, transport and storage. In this study, we propose porous materials based on sodium alginate, gelatin, glycerol and lipids (cottonseed oil and beeswax) obtained by freeze-drying and cross-linking. Material composition with the most promising properties was modified by the addition of PLA microparticles with Calendula officinalis flower extract. The structure and properties of obtained porous materials were analyzed. ATR-FTIR, mechanical properties, residual moisture content, porosity and density were assessed, as well as swelling properties and degradation after their cross-linking. The loading capacity and in vitro release of Calendula officinalis flower extract were performed for samples with incorporated PLA microparticles containing plant extract. The modification of the composition and fabrication method of materials significantly influenced their physicochemical properties. The selected plant extract was successfully incorporated into polymeric microparticles that were subsequently added into developed materials. Prepared materials may be considered during designing new cosmetic formulations.

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In Vivo Regeneration of Large Bone Defects by Cross-Linked Porous Hydrogel: A Pilot Study in Mice Combining Micro Tomography, Histological Analyses, Raman Spectroscopy and Synchrotron Infrared Imaging

Cited by 14 publications

References 43 publications

Three-Dimensional Culture of Cartilage Tissue on Nanogel-Cross-Linked Porous Freeze-Dried Gel Scaffold for Regenerative Cartilage Therapy: A Vibrational Spectroscopy Evaluation

Three-Dimensional Culture of Cartilage Tissue on Nanogel-Cross-Linked Porous Freeze-Dried Gel Scaffold for Regenerative Cartilage Therapy: A Vibrational Spectroscopy Evaluation

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

Lyophilized Emulsions in the Form of 3D Porous Matrices as a Novel Material for Topical Application

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