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The tissue engineering of tendon was studied using highly elastic poly(L-lactide-co-ε-caprolactone) (PLCL) scaffolds and focusing on the effect of dynamic tensile stimulation. Tenocytes from rabbit Achilles tendon were seeded (1.0 × 106 cells/scaffold) onto porous PLCL scaffolds and cultured for periods of 2 weeks and 4 weeks. This was performed in a static system and also in a bioreactor equipped with tensile modulation which mimicked the environmental surroundings of tendons with respect to tensile extension. The degradation of the polymeric scaffolds during the culture was relatively slow. However, there was an indication that cells accelerated the degradation of PLCL scaffolds. The scaffold/cell adducts from the static culture exhibited inferior strength (at 2 weeks 350 kPa, 4 weeks 300 kPa) compared to the control without cells (at 2 weeks 460 kPa, 4 weeks 340 kPa), indicating that the cells contributed to the enhanced degradation. On the contrary, the corresponding values of the adducts from the dynamic culture (at 2 weeks 430 kPa, 4 weeks 370 kPa) were similar to, or higher than, those from the control. This could be explained by the increased quantity of cells and neo-tissues in the case of dynamic culture compensating for the loss in tensile strength. Compared with static and dynamic culture conditions, mechanical stimulation played a crucial role in the regeneration of tendon tissue. In the case of the dynamic culture system, cell proliferation was enhanced and secretion of collagen type I was increased, as evidenced by DNA assay and histological and immunofluorescence analysis. Thus, tendon regeneration, indicated by improved mechanical and biological properties, was demonstrated, confirming the effect of mechanical stimulation. It could be concluded that the dynamic tensile stimulation appeared to be an essential factor in tendon/ligament tissue engineering, and that elastic PLCL co-polymers could be very beneficial in this process. [ABSTRACT FROM AUTHOR]

For repairing cartilage defects by cartilage tissue engineering, it is important that engineered cartilage that is fabricated with scaffolds and cells can maintain the biological and physiological functions of cartilage, and also can induce three-dimensional spatial organization of chondrocytes. In this sense, hydrogels such as fibrin gels (FG) and hyaluronan (HA) are widely used for application in cartilage treatment. However, the use of hydrogels alone as a scaffold has a physical weakness; the mechanical properties of hydrogels are too weak to endure complex loading in the body. In this study, for mimicking a native cartilage microenvironment, we made cell–hybrid scaffold constructs with poly(L-lactide-co-ε-caprolactone) (PLCL) scaffolds and hydrogels to guide three-dimensional spatial organization of cells and extracellular matrix. A highly elastic scaffold was fabricated from PLCL with 85% porosity and 300–500 μm pore size using a gel-pressing method. The mixture of rabbit chondrocytes and hydrogels was seeded on PLCL scaffolds, and was subcutaneously implanted into nude mice for up to eight weeks. The cell seeding efficiency of the hybrid scaffolds with FG or HA was higher than that of the PLCL scaffolds. From in vivo studies, the accumulation of cartilaginous extracellular matrices of constructs, which was increased by hybridization of hydrogels and PLCL scaffolds, showed that the cell–hybrid scaffold constructs formed mature and well-developed cartilaginous tissue. In conclusion, the hybridization of hydrogels and PLCL scaffold for three-dimensional spatial organization of cells would provide a biomimetic environment where cartilage tissue growth is enhanced and facilitated. It can enhance the production of cartilaginous extracellular matrices and, consequently, improve the quality of the cartilaginous tissue formed. [ABSTRACT FROM AUTHOR]

Tissue-engineered vascular grafts have been investigated as a substitute for prosthetic vascular grafts. The current scaffolds have several limitations due to weak mechanical properties in withstanding the pressure of blood vessel. A gel-spinning molding device including three-separate drivers that make a cylindrical shaft turn on its axis, orbit, and concurrently move up and down was developed for preparing seamless fibrous tubular scaffolds for vascular grafts. A seamless double-layered tubular scaffold, which was composed of an outer fibrous network and inner porous layer, was fabricated by using the device for the spinning of poly(L-lactide-co-caprolactone) (PLCL, 50:50) solution as a gel state on a rotating cylindrical shaft that had been dip-coated with the mixture of PLCL solution and NaCl particles. A scaffold that had an inner layer fabricated with 30% salts, below 20 μm in salt size, and more than 100 μm in thickness, was found to be optimal from a blood leakage test. The burst pressures of the scaffolds were more than 900 mmHg. The scaffolds exhibited 550–670% elongation-at-break. The measured circumferential and longitudinal tensile strengths of the scaffolds were 3.62 ± 0.68 and 2.64 ± 0.41 MPa, respectively. The suture retention strength of the scaffold was measured to be 7.68 ± 0.75 N. These mechanically strong and elastic properties of the newly developed scaffolds provide an important basis for blood vessel tissue engineering. [ABSTRACT FROM AUTHOR]

In cartilage tissue regeneration, it is important that an implant inserted into a defect site can maintain its mechanical integrity and endure stress loads from the body, in addition to being biocompatible and able to induce tissue growth. These factors are crucial in the design of scaffolds for cartilage tissue engineering. We developed an elastic biodegradable scaffold from poly(L-lactideco-ε-caprolactone) (PLCL) for application in cartilage treatment. Biodegradable PLCL co-polymer was synthesized from L-lactide and ε-caprolactone in the presence of stannous octoate as a catalyst. A highly elastic PLCL scaffold was fabricated by a gel-pressing method with 80% porosity and 300–500 μm pore size. The tensile mechanical and recovery tests were performed in order to examine mechanical and elastic properties of the PLCL scaffold. They could be easily twisted and bent and exhibited almost complete (over 94%) recoverable extension up to breaking point. For examining cartilaginous tissue formation, rabbit chondrocytes were seeded on scaffolds. They were then cultured in vitro for 5 weeks or implanted in nude mice subcutaneously. From in vitro and in vivo tests, the accumulation of extracellular matrix on the constructs showed that chondrogenic differentiation was sustained onto PLCL scaffolds. Histological analysis showed that cells onto PLCL scaffolds formed mature and well-developed cartilaginous tissue, as evidenced by chondrocytes within lacunae. From these results, we are confident that elastic PLCL scaffolds exhibit biocompatibility and as such would provide an environment where cartilage tissue growth is enhanced and facilitated. [ABSTRACT FROM AUTHOR]

It is known that complex loading is involved in the development and maintenance of articular cartilage in the body. It means the compressive mechanical stimulation is a very important factor for formation of articular cartilage using a tissue-engineering technique. The objective of this study is to engineer cartilaginous constructs with mechano-active scaffolds and to evaluate the effect of dynamic compression for regeneration of cartilage. The mechano-active scaffolds were prepared from a very elastic poly(L-lactide-co-ε-caprolactone) (PLCL) with 85% porosity and 300–500 μm pore size using a gel-pressing method. The scaffold was seeded with 2 × 106 chondrocytes and the continuous compressive deformation of 5% strain was applied with 0.1 Hz for 10 days and 24 days, respectively. Then, the chondrocytes-seeded constructs were implanted subcutaneously into nude mice. Mechano-active scaffolds with complete rubber-like elasticity showed almost complete (over 97%) recovery at an applied strain of up to 500%. The amount of chondral extracellular matrix was increased significantly by mechanical stimulation on the highly elastic mechano-active scaffolds. Histological analysis showed the mechanically stimulated implants formed mature and well-developed cartilaginous tissue, as evidenced by the chondrocytes within lacunae and the abundant accumulation of sulfated GAGs. However, unhealthy lacunae shapes and hypertrophy forms were observed in the implants stimulated mechanically for 24 days, compared with those stimulated for 10 days. In conclusion, the proper periodical application of dynamic compression can encourage chondrocytes to maintain their phenotypes and enhance the production of GAGs, which would improve the quality of cartilaginous tissue formed both in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Biodegradable macroporous scaffolds have been developed for tissue-engineering applications. We fabricated and characterized a new tubular, macroporous, fibrous scaffold using a very elastic biodegradable co-polymer, poly(L-lactide-co-caprolactone) (PLCL, 5:5) in a gel-spinning process. A viscous PLCL solution was spun as a gel-phase under swirl-flow conditions and was subsequently fabricated to produce a tubular fibrous scaffold on a rotating cylindrical shaft in a methanol solution. The porosity and median pore size of the fibrous PLCL scaffolds were 55–75% and 120–150 μm, respectively, using a 5–10% PLCL solution. The use of a 7.5% (w/v) solution resulted in scaffolds with tensile strength and elastic modulus of 3.39 MPa and 1.22 MPa, respectively. The scaffolds exhibited 500–600% elongation-at-break. The tensile strength and modulus of fibrous PLCL scaffolds were proven to decrease on lowering the concentration of the PLCL spinning solution; however, the tensile strength and modulus of fibrous PLCL scaffolds, produced from 5% solutions, are approximately 4- and 5-times higher than those of extruded PLCL scaffolds. These properties indicated that the fibrous PLCL scaffolds were very elastic and mechanically strong. The scaffolds appeared to be well interconnected between the pores as determined by SEM imaging analysis. In addition, the cell-seeding efficiency was 2-fold higher using gel-spun scaffolds than using extruded scaffolds. These results suggest that the gel-spun fibrous PLCL scaffold is an excellent matrix for vascular tissue-engineering applications. [ABSTRACT FROM AUTHOR]

A soft and very elastic poly(lactide-co-ε-caprolactone) (PLCL)(50:50, Mn 185 × 103) was synthesized. Tubular scaffolds were prepared by an extrusion-particulate leaching method for mechano-active vascular tissue engineering. The copolymer was very flexible but completely rubber-like elastic. Even the high porous PLCL scaffolds (90% salt wt) exhibited 200% elongation, but recovery over 85% in a tensile test. Moreover, the PLCL scaffolds maintained their high elasticity also in culture media under cyclic mechanical strain conditions. The highly porous scaffold (90% salt wt) withstood for an initial 1 week without any deformation and sustained for 2 weeks in culture media under cyclic stress of 10% amplitude and at 1 Hz frequency which are similar to the natural vascular conditions. Vascular smooth muscle cells (VSMCs) were seeded on to the PLCL scaffolds. The cell adhesion and proliferation on the scaffolds of various pore-size were increased with increasing pore size. For the pore sizes of 50-100 μm, 100-150 μm, 150-200 μm and 200-250 μm, the ratios of cell numbers were about 1:1.2:1.9:2.2, respectively, at both 12 h and 5 days. Similarly, the higher porous scaffolds exhibited more cell adhesion and proliferation compared to lower porous one, where the effect was more pronounced in the longer proliferation period. SMC-seeded scaffolds were implanted subcutaneously in athymic nude mice to confirm the biocompatibility. Such a high elastic property and proper biocompatibility to SMCs of PLCL scaffolds prepared in this study will be very useful to engineer SM-containing tissues such as blood vessels under mechanically dynamic environments (mechano-active tissue engineering). [ABSTRACT FROM AUTHOR]