Supermacroporous poly(vinyl alcohol)-carboxylmethyl chitosan-poly(ethylene glycol) scaffold: an in vitro and in vivo pre-assessments for cartilage tissue engineering

Lee, Si-Yuen; Wee, Ai-Sze; Lim, Chin-Keong; Abbas, Azlina Amir; Selvaratnam, Lakshmi; Merican, Azhar M.; Ahmad, Tunku Sara; Kamarul, Tunku

doi:10.1007/s10856-013-4907-4

Cited by 17 publications

(8 citation statements)

References 24 publications

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“…Good understanding of the cartilage structure, physiology, and the molecular basis of chondrogenesis is key to in vitro cartilage production, either for use in tissue engineering or clinics (Bhat, Tripathi & Kumar, 2011; Lee et al, 2013; Li et al, 2012). The state-of-the-art concept of in vitro cartilage tissue development combines the use of biocompatible and biodegradable carrier materials, the application of growth factors, the use of different cell types (stem or already differentiated) and different approaches to simulate the native mechanical stimulation (Gardner et al, 2013; Hildner et al, 2011; Khan et al, 2013; Naranda et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

Naranda

Gradišnik

Gorenjak

et al. 2017

PeerJ

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BackgroundCartilage tissue engineering is a fast-evolving field of biomedical engineering, in which the chondrocytes represent the most commonly used cell type. Since research in tissue engineering always consumes a lot of cells, simple and cheap isolation methods could form a powerful basis to boost such studies and enable their faster progress to the clinics. Isolated chondrocytes can be used for autologous chondrocyte implantation in cartilage repair, and are the base for valuable models to investigate cartilage phenotype preservation, as well as enable studies of molecular features, nature and scales of cellular responses to alterations in the cartilage tissue.MethodsIsolation and consequent cultivation of primary human adult articular chondrocytes from the surgical waste obtained during total knee arthroplasty (TKA) was performed. To evaluate the chondrogenic potential of the isolated cells, gene expression of collagen type 2 (COL2), collagen 1 (COL1) and aggrecan (ACAN) was evaluated. Immunocytochemical staining of all mentioned proteins was performed to evaluate chondrocyte specific production.ResultsCartilage specific gene expression of COL2 and ACAN has been shown that the proposed protocol leads to isolation of cells with a high chondrogenic potential, possibly even specific phenotype preservation up to the second passage. COL1 expression has confirmed the tendency of the isolated cells dedifferentiation into a fibroblast-like phenotype already in the second passage, which confirms previous findings that higher passages should be used with care in cartilage tissue engineering. To evaluate the effectiveness of our approach, immunocytochemical staining of the evaluated chondrocyte specific products was performed as well.DiscussionIn this study, we developed a protocol for isolation and consequent cultivation of primary human adult articular chondrocytes with the desired phenotype from the surgical waste obtained during TKA. TKA is a common and very frequently performed orthopaedic surgery during which both femoral condyles are removed. The latter present the ideal source for a simple and relatively cheap isolation of chondrocytes as was confirmed in our study.

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Section: Introductionmentioning

confidence: 99%

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

Naranda

Gradišnik

Gorenjak

et al. 2017

PeerJ

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“…In our previous study, we have found CMCS could promote proliferation and inhibit apoptosis in cultured Schwann cells [10][11][12][13], we also found CMCS could inhibit the apoptosis of cultured nucleus pulposus cells [14]. CMCS has been widely studied in OA related diseases in recent years in vivo and in vitro in recent days [15][16][17]. We have also used CMCS in cultured chondrocytes and found CMCS also has the inhibitory effect on NO-induced apoptosis in cultured chondrocytes [18,19].…”

Section: Introductionmentioning

confidence: 86%

Mitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide-induced apoptosis in chondrocytes

et al. 2020

BMC Complement Med Ther

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Background: Chondrocyte apoptosis activated by the mitochondrial dependent pathway serves a crucial role in cartilage degeneration of osteoarthritis (OA). In the present study, the protective effects of CMCS against sodium nitroprusside (SNP)-induced chondrocyte apoptosis were evaluated and the underlying molecular mechanisms were elucidated. Methods: Chondrocytes were isolated from articular cartilage of SD rats and identified by type II collagen immunohistochemistry. The chondrocytes stimulated with or without SNP to induce apoptosis, were treated by CMCS for various concentrations. The cell viability were determined by MTT and LDH assays. Cell apoptotic ratio was determined by Annexin V-FITC/PI staining. Mitochondrial membrane potential (ΔΨm) was detected by using Rhodamine123 (Rho123) staining. To understand the mechanism, the mRNA expression levels of Bcl-2, Bax, cytochrome c (Cyt c) and cleaved caspase-3 were detected by real-time PCR and western blot analysis, respectively. Results: It was shown using the MTT and LDH assays that CMCS protected the viability of chondrocyte against SNP damage. Annexin V-FITC/PI and Rho123 staining showed that CMCS not only inhibited the cell apoptosis but also restored the reduction of the ΔΨm in chondrocytes. In SNP-induced chondrocytes, CMCS down-regulated the expression of Bax, Cyt c and cleaved caspase-3 but upregulated the expression of Bcl-2, as shown by real-time PCR and western blot. Conclusions: Taken together, these results indicated that CMCS has the protective effect on chondrocytes against SNPinduced apoptosis, at least partly, via inhibiting the mitochondrial dependent apoptotic pathway. Thus, CMCS may be potentially used as a biological agent for prevention and treatment of OA.

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“…[1][2][3][4] Multiple techniques are used to adjust the physicomechanical properties of PVA, many of which depend on a thermal transition wherein noncovalent intermolecular bonds form during the PVA crystallization process upon cooling to below the freezing temperature of water, resulting in a physically cross-linked polymer network with a porous structure. [5][6][7][8][9][10] PVA hydrogels can be further tuned via chemical modification of the PVA backbone or by modification of the hydroxyl groups. [11][12][13][14][15][16] One limitation of direct PVA chemical modification is the potential for adverse functional groups to induce toxicity of the hydrogel; therefore, nonchemical modifications can be preferred, despite the decreased thermal stability.…”

Section: Introductionmentioning

confidence: 99%

Effects of cryo-processing on the mechanical and biological properties of poly(vinyl alcohol)-gelatin theta-gels

et al. 2020

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Poly(vinyl alcohol) (PVA), a synthetic, nontoxic polymer, is widely studied for use as a biomedical hydrogel due to its structural and physicomechanical properties. Depending on the synthesis method, PVA hydrogels can exhibit a range of selected characteristics-strength, creep resistance, energy dissipation, degree of crystallinity, and porosity. While the structural integrity and behavior of the hydrogel can be finetuned, common processing techniques result in a brittle, linear elastic material. In addition, PVA lacks functionality to engage and participate in cell adhesion, which can be a limitation for integrating PVA materials with tissue in situ. Thus, there is a need to further engineer PVA hydrogels to optimize its physicomechanical properties while enhancing cell adhesion and bioactivity. While the inclusion of gelatin into PVA hydrogels has been shown to impart cell-adhesive properties, the optimization of the mechanical properties of PVA-gelatin blends has not been studied in the context of traditional PVA hydrogel processing techniques. The incorporation of poly(ethylene glycol) with PVA prior to solidification forms an organized, cell instructive hydrogel with improved stiffness. The effect of cryo-processing, i.e., freezethaw (FT) cycling was elucidated by comparing 1 FT and 8 FT theta-cryo-gels and cryo-gels. To confirm the viability of the gels, human mesenchymal stem cell (hMSC) protein and sulfated glycosaminoglycan assays were performed to verify the nontoxicity and influence on hMSC differentiation. We have devised an elastic PVA-gelatin hydrogel utilizing the theta-gel and cryo-gel processing techniques, resulting in a stronger, more elastic material with greater potential as a scaffold for complex tissues.

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Supermacroporous poly(vinyl alcohol)-carboxylmethyl chitosan-poly(ethylene glycol) scaffold: an in vitro and in vivo pre-assessments for cartilage tissue engineering

Cited by 17 publications

References 24 publications

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

Mitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide-induced apoptosis in chondrocytes

Effects of cryo-processing on the mechanical and biological properties of poly(vinyl alcohol)-gelatin theta-gels

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