Insulin in Tissue Engineering of Cartilage: A Potential Model System for Growth Factor Application

Kellner, Karin; Schulz, Michaela B.; Göpferich, Achim; Blunk, Torsten

doi:10.3109/10611860108998778

Cited by 31 publications

(39 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Well in agreement with the obtained results, similar effects of insulin on chondrocyte-based tissue engineering products have previously been observed [9]. Insulin is homologous to parts of the IGF-1 molecule [8]; IGF-1 is a potent stimulator of extracellular matrix production in chondrocytes [2][3][4][5] and has also anabolic effects on mesenchymal progenitor cells undergoing chondrogenesis at a concentration of 100 ng/ml in the medium [12,14,20].…”

Section: Discussionsupporting

confidence: 76%

See 1 more Smart Citation

Insulin is essential for in vitro chondrogenesis of mesenchymal progenitor cells and influences chondrogenesis in a dose-dependent manner

Mueller

Blunk

Appel

et al. 2012

International Orthopaedics (SICOT)

View full text Add to dashboard Cite

Purpose Insulin is a commonly used additive in chondrogenic media for differentiating mesenchymal stem cells (MSCs). The indispensability of other bioactive factors like TGF-β or dexamethasone in these medium formulations has been shown, but the role of insulin is unclear. The purpose of this study was to investigate whether insulin is essential for MSC chondrogenesis and if there is a dose-dependent effect of insulin on MSC chondrogenesis. Methods We cultivated human MSCs in pellet culture in serum-free chondrogenic medium with insulin concentrations between 0 and 50 μg/ml and assessed the grade of chondrogenic differentiation by histological evaluation and determination of glycosaminoglycan (GAG), total collagen and DNA content. We further tested whether insulin can be delivered in an amount sufficient for MSC chondrogenesis via a drug delivery system in insulin-free medium. Results Chondrogenesis was not induced by standard chondrogenic medium without insulin and the expression of cartilage differentiation markers was dose-dependent at insulin concentrations between 0 and 10 μg/ml. An insulin concentration of 50 μg/ml had no additional effect compared with 10 μg/ml. Insulin was delivered by a release system into the cell culture under insulin-free conditions in an amount sufficient to induce chondrogenesis. Conclusions Insulin is essential for MSC chondrogenesis in this system and chondrogenic differentiation is influenced by insulin in a dose-dependent manner. Insulin can be provided in a sufficient amount by a drug delivery system. Therefore, insulin is a suitable and inexpensive indicator substance for testing drug release systems in vitro.

show abstract

Section: Discussionsupporting

confidence: 76%

“…Insulin is structurally similar to IGF-1 and can activate the IGF-1 receptor, and insulin has been shown to be a potent stimulator of ECM production in chondrocyte-based tissue engineering products [8,9].…”

Section: Introductionmentioning

confidence: 99%

Insulin is essential for in vitro chondrogenesis of mesenchymal progenitor cells and influences chondrogenesis in a dose-dependent manner

Mueller

Blunk

Appel

et al. 2012

International Orthopaedics (SICOT)

View full text Add to dashboard Cite

show abstract

“…The angular velocity of the vessel was set in order to maintain constructs in a continual freefall condition [21] and was increased during culture time to account for construct growth. For each experimental condition (i.e., static, mixed flask or RCCS culture), cell-scaffold constructs were cultured for 4 weeks in 10 ml of complete medium further supplemented with 10ng/ml TGF-3, 0.1 mM ascorbic acid 2-phosphate and 10 g/ml human insulin, in order to enhance chondrogenesis [22,23], with medium changes twice a week.…”

Section: Cell Isolation and Expansionmentioning

confidence: 99%

Use of hydrodynamic forces to engineer cartilaginous tissues resembling the non-uniform structure and function of meniscus

et al. 2006

View full text Add to dashboard Cite

The measurement and calculation of tensile modulus could be more clearly explained, in particular the dimensions that were recorded during each test (with reference to Fig. 1b, or diagram) and how the measurements at each strain were processed to obtain the data presented in Fig. 6. Also, why were the same tests not performed on native meniscus for direct comparison?Data presented in Fig. 6 were obtained using standard calculations to derive the elastic modulus of materials based on stress-strain curves. In order to better characterize the dimensions recorded, the description has been revised as follows:"Changes in dimensions during loading were obtained with a digital image analysis system (Adapted QuickCam® Pro 3000, Logitech Europe S.A., Switzerland and the GNU Image Manipulation Program, www.gimp.org), and were always lower than 10% of the gauge length considered." (Methods, page 6,1 st paragraph). The same tensile tests could not be performed on the inner or outer regions of native meniscus: in fact, out of those regions, having a semilunar shape, it would not be possible to punch ring-shaped specimens. This is now discussed in the following sentence: We now clarify that "Since the mechanical properties of the non-woven mesh scaffold used are negligible, the mechanical functionality of the bi-zonal engineered constructs was mainly due to the differential deposition of extracellular matrix by chondrocytes." (Discussion, page 9, last paragraph) and discuss that "The HYAFF ® -11 non-woven meshes used in this study were supportive of the formation of bi-zonal tissues. The composition and open structure of the meshes may have contributed respectively to the deposition of GAG in the inner region and to the outgrowth of cells towards the outer capsule. " (Discussion, page 8, last paragraph)To what extent might these differences also be accounted for by the increase in cell number and viability when cultured under dynamic as opposed to static conditions? Likewise, is the preferential orientation of the collagen fibers not due simply to the increased rate of growth of the fibrous capsule when cultured under conditions of improved mass transport rather than hydrodynamic shear per se (N.B. the RCCS is considered a low shear environment)?We now describe that "In the outer capsule, cells were elongated and at a higher density than in the central region" (Results, Page 6, last paragraph) and discuss that "More fundamental studies would be required to determine the role of increased mass transport and/or increased cell density following application of hydrodynamic flow in the development of the bi-zonal tissues." (Page 9, 1 st paragraph) The term has been deleted.Methods p3, line 13: replace 'due to' with 'to account for ' The change has been made. Yes, the volume of medium was in each case equivalent to 10 ml per construct p4, line 9: The constructs were examined blind.The change has been made. The change has been made. The sentence now reads "SEM analysis indicated that collagen fibers were randomly oriented in the ...

show abstract

“…Insulin is essential for maintenance of glucose homeostasis and for proper maintenance of the energetic balance, but also contributes to maintain anabolic processes in several tissues, including cartilage, as it was shown to induce anabolic and inhibit catabolic responses in adult chondrocytes and cartilage explant cultures from various species [12,26]. In DM2 and the metabolic syndrome, hyperinsulinemia occurs as an attempt at maintaining glucose homeostasis, at least while pancreatic β cells remain functional.…”

Section: Discussionmentioning

confidence: 99%

Hyperglycemia and Hyperinsulinemia-Like Conditions Independently Induce Inflammatory Responses in Human Chondrocytes

Rufino

Ribeiro

Ferreira

et al. 2017

JFMK

View full text Add to dashboard Cite

To elucidate the mechanisms by which type 2 Diabetes Mellitus (DM2) constitutes a risk factor for the development and progression of osteoarthritis (OA), this work determined whether high glucose and/or high insulin, the hallmarks of DM2, are capable of activating the transcription factor, Nuclear Factor-κB (NF-κB), which plays a critical role in OA by inducing the expression of pro-inflammatory and catabolic genes. For this, we analyzed NF-κB activation by measuring the nuclear levels of p65 by western blot. As readouts of NF-κB activity, Interleukin-1β, Tumor Necrosis Factor-α, and inducible nitric oxide synthase (iNOS) expression were analyzed by real time RT-PCR and western blot. Culture of the human chondrocytic cell line, C28-I2, in high glucose (30 mM) increased nuclear NF-κB p65 levels in a time-dependent manner, relative to cells cultured in medium containing 10 mM glucose (regular culture medium). High glucose-induced NF-κB activation was inhibited by co-treatment with its specific inhibitor, Bay 11-7082, 5 µM. Culture of primary human chondrocytes under high glucose for 24 h increased IL-1β and TNF-α mRNA levels by 97% (p = 0.0066) and 85% (p = 0.0045), respectively, while iNOS mRNA and protein levels and NO production increased by 61% (p = 0.0017), 148% (p = 0.0089), and 70% (p = 0.049), respectively, relative to chondrocytes maintained in 10 mM glucose. Treatment of chondrocytic cells with 100 nM insulin was also sufficient to increase nuclear NF-κB p65 levels, independently of the glucose concentration in the culture medium. This study shows that hyperglycemia and hyperinsulinemia are independently sufficient to induce inflammatory responses in human chondrocytes, namely by activating NF-κB. This can be a relevant mechanism by which DM type 2 and other conditions associated with impaired glucose and insulin homeostasis, like obesity and the metabolic syndrome, contribute to the development and progression of OA.

show abstract

Insulin in Tissue Engineering of Cartilage: A Potential Model System for Growth Factor Application

Cited by 31 publications

References 30 publications

Insulin is essential for in vitro chondrogenesis of mesenchymal progenitor cells and influences chondrogenesis in a dose-dependent manner

Insulin is essential for in vitro chondrogenesis of mesenchymal progenitor cells and influences chondrogenesis in a dose-dependent manner

Use of hydrodynamic forces to engineer cartilaginous tissues resembling the non-uniform structure and function of meniscus

Hyperglycemia and Hyperinsulinemia-Like Conditions Independently Induce Inflammatory Responses in Human Chondrocytes

Contact Info

Product

Resources

About