Transforming Growth Factor Beta 3-Loaded Decellularized Equine Tendon Matrix for Orthopedic Tissue Engineering

Abstract

Transforming growth factor beta 3 (TGFβ3) promotes tenogenic differentiation and may enhance tendon regeneration in vivo. This study aimed to apply TGFβ3 absorbed in decellularized equine superficial digital flexor tendon scaffolds, and to investigate the bioactivity of scaffold-associated TGFβ3 in an in vitro model. TGFβ3 could effectively be loaded onto tendon scaffolds so that at least 88% of the applied TGFβ3 were not detected in the rinsing fluid of the TGFβ3-loaded scaffolds. Equine adipose tissue-derived multipotent mesenchymal stromal cells (MSC) were then seeded on scaffolds loaded with 300 ng TGFβ3 to assess its bioactivity. Both scaffold-associated TGFβ3 and TGFβ3 dissolved in the cell culture medium, the latter serving as control group, promoted elongation of cell shapes and scaffold contraction (p < 0.05). Furthermore, scaffold-associated and dissolved TGFβ3 affected MSC musculoskeletal gene expression in a similar manner, with an upregulation of tenascin c and downregulation of other matrix molecules, most markedly decorin (p < 0.05). These results demonstrate that the bioactivity of scaffold-associated TGFβ3 is preserved, thus TGFβ3 application via absorption in decellularized tendon scaffolds is a feasible approach.

TGF–β3 Loaded Electrospun Polycaprolacton Fibre Scaffolds for Rotator Cuff Tear Repair: An in Vivo Study in Rats

“…The modified fibre scaffolds (PCL modified with CS-g-PCL plus alginate) were incubated in this solution for 15 min. Therewith a TGF-β3 loading capacity of 250 ng/scaffold was reached, which is in line with different in vitro studies (e.g., [46,68]). Before further use, the fibre scaffolds were washed twice for one minute with Millipore water.…”

“…Other authors could observe enhanced chondrogenesis after subcutaneous implantation of PDLLA-PEG hydrogel combined with mesenchymal stem cells and TGF-β3 [40] when compared to hydrogels without TGF-β3. In an in vitro study, mesenchymal stromal cells seeded onto decellularised equine superficial digital flexor tendon scaffolds with absorbed TGF-β3 showed elongated cell shapes, scaffold contraction and a musculoskeletal gene expression with an up-regulation of tenascin c and down-regulation of matrix molecules [46]. However, there are also studies which could not find any positive effect of TGF-β3 although TGF-β 3 was delivered with an osmotic pump throughout the complete observation period of four weeks [47].…”

Comparative Analysis of Tenogenic Gene Expression in Tenocyte-Derived Induced Pluripotent Stem Cells and Bone Marrow-Derived Mesenchymal Stem Cells in Response to Biochemical and Biomechanical Stimuli

“…In our study, all the tested stimuli enhanced the expression of TNC in teno-iPSCs, and similar results were obtained from BMSCs treated with all stimuli except BMP12. Although these data did not agree with the studies showing decreased TNC in mechanical stressed C3H10T1/2 cells [ 57 ], increased TNC in BMP12-treated rat BMSCs [ 46 ], and variable expression of TNC in TGF- β 3-treated equine ESCs and iPSCs [ 18 ], they are in agreement with other studies reporting increased TNC in mechanical strained human BMSCs [ 68 ] and in TGF- β 3-treated equine ADMSCs [ 69 ].…”

Sterilization and disinfection methods for decellularized matrix materials: Review, consideration and proposal

“…Applications : Alcohol is commonly used in disinfection of skin and precision instruments, etc. There are many reports on the use of alcohol to disinfect dECM, which are mainly derived from pericardium [ 11 ], valve [ 123 , 124 ], blood vessel [ 12 , 121 , 125 , 126 ], liver [ [127] , [128] , [129] ], pancreas [ 130 ], trachea [ 131 ], bladder [ 130 , [132] , [133] , [134] , [135] ], kidney [ 48 , 136 ], bone [ 137 , 138 ], tendon [ [139] , [140] , [141] ] and nerve [ 142 ].…”

“…Tendons mainly come from bovine, horse and pig, and a small number of tendons are from human, rabbit, sheep, dog. The sterilization and disinfection methods of decellularized tendon mainly include ethanol [ [139] , [140] , [141] ], EO [ [78] , [79] , [80] ], PAA [ 95 , 96 ], irradiation [ 27 , [54] , [55] , [56] ], antibiotics [ 153 , 179 ] and ScCO 2 [ 55 ], etc.…”