Repair of segmental ulna defects using a β-TCP implant in combination with a heparan sulfate glycosaminoglycan variant

Rai, Bina; Chatterjea, Anindita; Lim, Zophia X.H.; Tan, Tuan Chun; Sawyer, Andrew J.; Hosaka, Yoshinao Z.; Murali, Sadasivam; Lee, J.J.L.; Fenwick, Steven; Hui, James; Nurcombe, Victor; Cool, Simon M.

doi:10.1016/j.actbio.2015.09.008

Cited by 25 publications

(25 citation statements)

References 36 publications

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“…Post hoc testing for GDF5 indicated this effect was only significant at doses ≥25 nM ( p < 0.0001), while for BMP2, significance was seen at the lowest dose tested of ≥5 nM ( p < 0.0001). We then proceeded to look at the dose-dependent effect of heparin, HS, and HS3 (a purified HS fraction known to increase BMP2 bioactivity 68,69 ) on GDF5-/BMP2-induced ALP production, using the lowest growth factor concentration that gave a significant increase in ALP (Fig. 6C, D).…”

Section: Resultsmentioning

confidence: 99%

“…Our previous work and ongoing investigations indicate that the isolation or synthesis of HS variants, for example those based on their affinity for particular growth factors, enables the selection of saccharides markedly more potent at promoting the bioactivity of growth factors compared to using unfractionated HS starting material, with its intrinsically high level of compositional heterogeneity. 26,59,68,69 Thus, there is the possibility to potentiate the activity of specific growth factors, to the same or greater extent than heparin, while avoiding the adverse and off-target effects of heparin caused by its pleiotropic nature. Indeed, in this study, we have shown that the HS3 variant (selected through its affinity for BMP2 and shown to be effective in promoting BMP2 bioactivity 68,69 ) could not inhibit the GDF5-induced ALP activity in the same manner as heparin.…”

Section: Discussionmentioning

confidence: 99%

“…26,59,68,69 Thus, there is the possibility to potentiate the activity of specific growth factors, to the same or greater extent than heparin, while avoiding the adverse and off-target effects of heparin caused by its pleiotropic nature. Indeed, in this study, we have shown that the HS3 variant (selected through its affinity for BMP2 and shown to be effective in promoting BMP2 bioactivity 68,69 ) could not inhibit the GDF5-induced ALP activity in the same manner as heparin. Therefore, by using this particular HS3 variant, there is the possibility of promoting BMP2 activity (as with heparin) without affecting the activity of GDF5, which would also be at the injury site and important for the repair process.…”

Section: Discussionmentioning

confidence: 99%

“…Cells were then stimulated with the stated doses of GDF5/GAG for 3 days. GAGs tested were heparin, HS, and BMP2 affinity-selected HS (termed HS3; see Murali et al , 68 Rai et al 69 ). The ALP activity was measured in triplicate by lysing cells in the ALP buffer 1 (0.1 M glycine, 1% [v/v] Nonidet P-40, 1 mM MgCl 2 , and 1 mM ZnCl 2 , pH 9.6) for 1 h at room temperature with gentle shaking, followed by the addition of one volume of ALP buffer 2 (5 mM p -nitrophenyl phosphate [ p -NPP], 0.1 M glycine, 1 mM MgCl 2 , and 1 mM ZnCl 2 , pH 9.6).…”

Section: Methodsmentioning

confidence: 99%

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Growth Differentiation Factor 5-Mediated Enhancement of Chondrocyte Phenotype Is Inhibited by Heparin: Implications for the Use of Heparin in the Clinic and in Tissue Engineering Applications

Ayerst

Smith

Nurcombe

et al. 2017

Tissue Engineering Part A

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The highly sulfated glycosaminoglycan (GAG) heparin is widely used in the clinic as an anticoagulant, and researchers are now using it to enhance stem cell expansion/differentiation protocols, as well as to improve the delivery of growth factors for tissue engineering (TE) strategies. Growth differentiation factor 5 (GDF5) belongs to the bone morphogenetic protein family of proteins and is vital for skeletal formation; however, its interaction with heparin and heparan sulfate (HS) has not been studied. We identify GDF5 as a novel heparin/HS binding protein and show that HS proteoglycans are vital in localizing GDF5 to the cell surface. Clinically relevant doses of heparin (≥10 nM), but not equivalent concentrations of HS, were found to inhibit GDF5's biological activity in both human mesenchymal stem/stromal cell-derived chondrocyte pellet cultures and the skeletal cell line ATDC5. We also found that heparin inhibited both GDF5 binding to cell surface HS and GDF5-induced induction of Smad 1/5/8 signaling. Furthermore, GDF5 significantly increased aggrecan gene expression in chondrocyte pellet cultures, without affecting collagen type X expression, making it a promising target for the TE of articular cartilage. Importantly, this study may explain the variable (and disappointing) results seen with heparin-loaded biomaterials for skeletal TE and the adverse skeletal effects reported in the clinic following long-term heparin treatment. Our results caution the use of heparin in the clinic and in TE applications, and prompt the transition to using more specific GAGs (e.g., HS derivatives), with better-defined structures and fewer off-target effects.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Growth Differentiation Factor 5-Mediated Enhancement of Chondrocyte Phenotype Is Inhibited by Heparin: Implications for the Use of Heparin in the Clinic and in Tissue Engineering Applications

Ayerst

Smith

Nurcombe

et al. 2017

Tissue Engineering Part A

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“…The use of carriers that can reduce the quantity of exogenous growth factors required, through their localisation, protection or enhancement, or indeed that could harness endogenously produced growth factors, would therefore be ideal. Research into the use of GAGs for these approaches is proving particularly promising, and presents an attractive therapeutic opportunity for TE strategies [163,195,292,293,294,295]. …”

Section: Glycosaminoglycansmentioning

confidence: 99%

The Good the Bad and the Ugly of Glycosaminoglycans in Tissue Engineering Applications

2017

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High sulfation, low cost, and the status of heparin as an already FDA- and EMA- approved product, mean that its inclusion in tissue engineering (TE) strategies is becoming increasingly popular. However, the use of heparin may represent a naïve approach. This is because tissue formation is a highly orchestrated process, involving the temporal expression of numerous growth factors and complex signaling networks. While heparin may enhance the retention and activity of certain growth factors under particular conditions, its binding ‘promiscuity’ means that it may also inhibit other factors that, for example, play an important role in tissue maintenance and repair. Within this review we focus on articular cartilage, highlighting the complexities and highly regulated processes that are involved in its formation, and the challenges that exist in trying to effectively engineer this tissue. Here we discuss the opportunities that glycosaminoglycans (GAGs) may provide in advancing this important area of regenerative medicine, placing emphasis on the need to move away from the common use of heparin, and instead focus research towards the utility of specific GAG preparations that are able to modulate the activity of growth factors in a more controlled and defined manner, with less off-target effects.

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Reconstruction of critical‐size segmental defects in rat femurs using carbonate apatite honeycomb scaffolds

Sakemi

Hayashi

Tsuchiya

et al. 2021

J Biomedical Materials Res

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Critical‐size segmental defects are formidable challenges in orthopedic surgery. Various scaffolds have been developed to facilitate bone reconstruction within such defects. Many previously studied scaffolds achieved effective outcomes with a combination of high cost, high‐risk growth factors or stem cells. Herein, we developed honeycomb scaffolds (HCSs) comprising carbonate apatite (CO3Ap) containing 8% carbonate, identical to human bone composition. The CO3Ap HCSs were white‐columned blocks harboring regularly arranged macropore channels of a size and wall thickness of 156 ± 5 μm and 102 ± 10 μm, respectively. The compressive strengths of the HCSs parallel and perpendicular to the macropore channel direction were 51.0 ± 11.8 and 15.6 ± 2.2 MPa, respectively. The HCSs were grafted into critical‐sized segmental defects in rat femurs. The HCSs bore high‐load stresses without any observed breakage. Two‐weeks post‐implantation, calluses formed around the HCSs and immature bone formed in the HCS interior. The calluses and immature bone matured until 8 weeks via endochondral ossification. At 12 weeks post‐implantation, large parts of the HCSs were gradually replaced by newly formed bone. The bone reconstruction efficacy of the CO3Ap HCSs alone was comparable to that of protein and cell scaffolds, while achieving a lower cost and increased safety.

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Repair of segmental ulna defects using a β-TCP implant in combination with a heparan sulfate glycosaminoglycan variant

Cited by 25 publications

References 36 publications

Growth Differentiation Factor 5-Mediated Enhancement of Chondrocyte Phenotype Is Inhibited by Heparin: Implications for the Use of Heparin in the Clinic and in Tissue Engineering Applications

Growth Differentiation Factor 5-Mediated Enhancement of Chondrocyte Phenotype Is Inhibited by Heparin: Implications for the Use of Heparin in the Clinic and in Tissue Engineering Applications

The Good the Bad and the Ugly of Glycosaminoglycans in Tissue Engineering Applications

Reconstruction of critical‐size segmental defects in rat femurs using carbonate apatite honeycomb scaffolds

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