Tissue engineering techniques using novel scaffolding materials offer potential alternatives for managing tendon disorders. An ideal tendon tissue engineered scaffold should mimic the three-dimensional (3D) structure of the natural extracellular matrix (ECM) of the native tendon. Here, we propose a novel electrospun nanoyarn network that is morphologically and structurally similar to the ECM of native tendon tissues. The nanoyarn, random nanofiber, and aligned nanofiber scaffolds of a synthetic biodegradable polymer, poly(l-lactide-co-ecaprolactone) [P(LLA-CL)], and natural collagen I complex were fabricated using electrospinning. These scaffolds were characterized in terms of fiber morphology, pore size, porosity, and chemical and mechanical properties for the purpose of culturing tendon cells (TCs) for tendon tissue engineering. The results indicated a fiber diameter of 632 -81 nm for the random nanofiber scaffold, 643 -97 nm for the aligned nanofiber scaffold, and 641 -68 nm for the nanoyarn scaffold. The yarn in the nanoyarn scaffold was twisted by many nanofibers similar to the structure and inherent nanoscale organization of tendons, indicating an increase in the diameter of 9.51 -3.62 mm. The nanoyarn scaffold also contained 3D aligned microstructures with large interconnected pores and high porosity. Fourier transform infrared analyses revealed the presence of collagen in the three scaffolds. The mechanical properties of the sample scaffolds indicated that the scaffolds had desirable mechanical properties for tissue regeneration. Further, the results revealed that TC proliferation and infiltration, and the expression of tendon-related ECM genes, were significantly enhanced on the nanoyarn scaffold compared with that on the random nanofiber and aligned nanofiber scaffolds. This study demonstrates that electrospun P(LLA-CL)/collagen nanoyarn is a novel, 3D, macroporous, aligned scaffold that has potential application in tendon tissue engineering.

Background: Previous cell culture and animal in vivo studies indicate the obvious effects of mechanical compression on disc cell biology. However, the effects of dynamic compression magnitude, frequency and duration on the immature nucleus pulposus (NP) from an organ-cultured disc are not well understood.Objective: To investigate the effects of a relatively wide range of compressive magnitudes, frequencies and durations on cell apoptosis and matrix composition within the immature NP using an intelligent and mechanically active bioreactor.Methods: Discs from the immature porcine were cultured in a mechanically active bioreactor for 7 days. The discs in various compressive magnitude groups (0.1, 0.2, 0.4, 0.8 and 1.3 MPa at a frequency of 1.0 Hz for 2 hours), frequency groups (0.1, 0.5, 1.0, 3.0 and 5.0 Hz at a magnitude of 0.4 MPa for 2 hours) and duration groups (1, 2, 4 and 8 hours at a magnitude of 0.4 MPa and frequency of 1.0 Hz) experienced dynamic compression once per day. Discs cultured without compression were used as controls. Immature NP samples were analyzed using the TUNEL assay, histological staining, glycosaminoglycan (GAG) content measurement, real-time PCR and collagen II immunohistochemical staining.Results: In the 1.3 MPa, 5.0 Hz and 8 hour groups, the immature NP showed a significantly increase in apoptotic cells, a catabolic gene expression profile with down-regulated matrix molecules and up-regulated matrix degradation enzymes, and decreased GAG content and collagen II deposition. In the other compressive magnitude, frequency and duration groups, the immature NP showed a healthier status regarding NP cell apoptosis, gene expression profile and matrix production.Conclusion: Cell apoptosis and matrix composition within the immature NP were compressive magnitude-, frequency- and duration-dependent. The relatively high compressive magnitude or frequency and long compressive duration are not helpful for maintaining the healthy status of an immature NP.

To provide applied anatomical evidence of the preoperative assessment of oblique lumbar interbody fusion (OLIF), the anatomical parameters of the OLIF operative window were observed through computed tomography angiography (CTA). We selected imaging data from 60 adults (30 males, 30 females) who underwent abdominal CTA and T12-S1 vertebral computed tomography (CT) with three-dimensional reconstruction. The OLIF operative windows at the L1-2, L2-3, L3-4, L4-5 and L5-S1 levels were as follows: the vascular window, bare window, psoas major window, ideal operative window, and actual operative window. Each level's actual operative window was statistically analyzed based on an actual operative window of <1 cm and ≥1 cm. The vascular window was largest at L4-5 (1.72 ± 0.58 cm). The bare window was largest at L5-S1 (1.59 ± 0.93 cm) and smallest at L3-4 (1.37 ± 0.51 cm). The psoas major window was largest at L3-4 (1.14 ± 0.35 cm) and smallest at L1-2 (0.41 ± 0.34 cm). The ideal operative window was largest at L4-5 (3.74 ± 0.36 cm) and smallest at L1-2 (3.23 ± 0.30 cm). The actual operative window was largest at L3-4, followed by L2-3, L4-5, L1-2, and L5-S1, which were 2.51 ± 0.56 cm, 2.28 ± 0.54 cm, 2.01 ± 0.74 cm, 1.80 ± 0.45 cm and 1.59 ± 0.93 cm, respectively (P = 0.000), and the percentages of the actual surgical window were 69%, 66%, 53%, 56% and 43%, respectively. The actual surgical window was <1 cm in 2 cases at L1-2 (3.3%), 4 cases at L4-5 (6.7%), and 17 cases at L5-S1 (28.3%) (11 males and 6 females). The regional anatomy of each level related to OLIF has its own peculiarities, and not all levels are suitable for OLIF. Before OLIF surgery, surgeons should analyze the imaging anatomy and select the appropriate surgical procedures.

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Mechanical overloading-induced nucleus pulposus (NP) cells senescence plays an important role in the pathogenesis of intervertebral disc degeneration (IVDD). The silent mating type information regulator 2 homolog-1 (SIRT1)-mediated pathway preserves the normal NP cell phenotype and mitochondrial homeostasis under multiple stresses. We aimed to investigate the role of SIRT1 in IVDD by assessing the effects of SIRT1 overexpression on high-magnitude compression-induced senescence in NP cells. High-magnitude compression induced cellular senescence and mitochondrial dysfunction in human NP cells. Moreover, SIRT1 overexpression tended to alleviate NP cell senescence and mitochondrial dysfunction under compressive stress. Given the mitophagy-inducing property of SIRT1, activity of mitophagy was evaluated in NP cells to further demonstrate the underlying mechanism. The results showed that SIRT1-overexpression attenuated senescence and mitochondrial injury in NP cells subjected to high-magnitude compression. However, depletion of PINK1, a key mitophagic regulator, impaired mitophagy and blocked the protective role of SIRT1 against compression induced senescence in NP cells. In summary, these results suggest that SIRT1 plays a protective role in alleviating NP cell senescence and mitochondrial dysfunction under high-magnitude compression, the mechanism of which is associated with the regulation of PINK1-dependent mitophagy. Our findings may provide a potential therapeutic approach for IVDD treatment.

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Osmolarity fluctuations are inevitable within the nucleus pulposus (NP). However, the effects of osmolarity on NP cell apoptosis within the organ-cultured disc remain unclear. The objective of this study was to investigate effects of different osmolarity levels (hypo-, iso-, and hyper-) and osmolarity modes (constant and cyclic) on NP cell apoptosis in a disc perfusion culture and to study the role of the ERK1/2 pathway in this regulatory process. Porcine discs were cultured for 7 days in different osmotic medium, including constant hypo-, iso-, and hyper-osmolarity (330, 430, and 550 mOsm/L, respectively) and cyclic-osmolarity (430 mOsm/L for 8 h, followed by 550 mOsm/L for 16 h). The role of the ERK1/2 pathway was investigated by using the pharmacological inhibitor U0126. NP cell apoptosis was analyzed by TUNEL staining, caspase-3 activity, gene expression of Bcl-2, Bax and caspase-3 and protein expression of cleaved caspase-3, and cleaved PARP. Our results showed that NP cell apoptosis was increased in hypo-and hyperosmolarity cultures compared to iso-or cyclic-osmolarity culture, whereas the level of apoptosis in the iso-osmolarity culture was lower than that in the cyclic-osmolarity culture. When the ERK1/2 pathway was inhibited in the iso-and cyclic-osmolarity cultures, the level of NP cell apoptosis was significantly increased. In conclusion, the effects of osmolarity on NP cell apoptosis depend on the osmolarity level (hypo-, iso-, or hyper-) and osmolarity mode (constant or cyclic). Futhermore, inhibition of the ERK1/2 pathway promotes NP cell apoptosis in this process. Keywords: intervertebral disc; osmolarity; apoptosis; nucleus pulposus; ERK1/2Intervertebral disc degeneration (IDD) affects approximately 80% of adult humans and is the primary cause of lower back pain which leads to personal disability and a heavy socioeconomic burden. 1 Although many research programs and clinical studies have been performed, the accurate etiopathogenesis of disc degeneration remains unclear.The intervertebral disc (IVD) consists of the surrounding annulus fibrosus (AF), the central nucleus pulposus (NP) and the upper and lower cartilage endplates (CEP). 2 With these three structurally distinct parts, the IVD has an outstanding mechanical property of supporting spine motion and stability. During disc degeneration, the decrease of viable NP cells can lead to decreased NP matrix production and ultimately changes in the disc structure and/or disc function. 3,4 Moreover, apoptosis of NP cells is a typical cellular phenomenon, which is closely associated with disc degeneration. 5 Therefore, NP cell apoptosis during disc degeneration has become a new area of research focus.Apoptosis is a process of programmed cell death executed through the activation of caspases, such as caspase-3, 6, and 7) 6 and may be a critical cause of the loss of disc cellularity during degeneration. 3,7 It is universally accepted that un-physiologic loading is implicated in the initiation and progression of disc degeneration. 8 Previous st...