In this study, we report on the ability of resorbable poly(L-lactic acid) (PLLA) nonwoven scaffolds to support the attachment, growth, and differentiation of marrow stromal cells (MSCs) under fluid flow. Rat MSCs were isolated from young male Wistar rats and expanded using established methods. The cells were then seeded on PLLA nonwoven fiber meshes. The PLLA nonwoven fiber meshes had 99% porosity, 17 microm fiber diameter, 10 mm scaffold diameter, and 1.7-mm thickness. The nonwoven PLLA meshes were seeded with a cell suspension of 5 x 10(5) cells in 300 microl, and cultured in a flow perfusion bioreactor and under static conditions. Cell/polymer nonwoven scaffolds cultured under flow perfusion had significantly higher amounts of calcified matrix deposited on them after 16 days of culture. Microcomputed tomography revealed that the in vitro generated extracellular matrix in the scaffolds cultured under static conditions was denser at the periphery of the scaffold while in the scaffolds cultured in the perfusion bioreactor the extracellular matrix demonstrated a more homogeneous distribution. These results show that flow perfusion accelerates the proliferation and differentiation of MSCs, seeded on nonwoven PLLA scaffolds, toward the osteoblastic phenotype, and improves the distribution of the in vitro generated calcified extracellular matrix.
The resorbable polymers polyglycolic acid (PGA) and polylactic acid (PLA) are gaining increasing importance in tissue engineering and cell transplantation. The present investigation was focused on the biocompatibility and cell retaining behavior of PGA/poly-L-lactide (PLLA) (90/10) and PLLA nonwoven structures for the in vitro development of chondrocyte-polymer constructs. The effect of the relevant monomers to chondrocytes was analyzed. Type II collagen and poly-L-lysine were compared to improve loading of PGA/PLLA and PLLA polymer nonwovens with chondrocytes. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetra-zoliumbrom ide (MTT) test was applied for quantification. At concentrations above 2 mg/mL, glycolic acid was more cytotoxic than lactic acid. As shown by pH equilibration, the cytotoxic effect is not due merely to the acidity of the alpha-hydroxy acids. Regarding the degradation products, glycolic acid, and L(+) lactic acid, nonwovens of PLLA are more biocompatible with chondrocytes than nonwovens of polyglycolide. Collagen type II and poly-L-lysine generally improved cell seeding on resorbable polymers in tissue engineering; however, their efficiency varies depending on the type of fiber structure.
Textiles have a very long history, but they are far from becoming outdated. They gain new importance in technical applications, and man-made fibers are at the center of this ongoing innovation. The development of high-tech textiles relies on enhancements of fiber raw materials and processing techniques. Today, melt spinning of polymers is the most commonly used method for manufacturing commercial fibers, due to the simplicity of the production line, high spinning velocities, low production cost and environmental friendliness. Topics covered in this review are established and novel polymers, additives and processes used in melt spinning. In addition, fundamental questions regarding fiber morphologies, structure-property relationships, as well as flow and draw instabilities are addressed. Multicomponent melt-spinning, where several functionalities can be combined in one fiber, is also discussed. Finally, textile applications and melt-spun fiber specialties are presented, which emphasize how ongoing research efforts keep the high value of fibers and textiles alive.
Resorbable augmentation devices for cruciate ligament surgery have been developed to temporarily protect healing tendon grafts or sutured ligaments against high tensile loads during the postoperative healing period. Materials available at present [e.g., polydioxanone (PDS)] show a half-life tensile strength of only 4-6 weeks, whereas the process of revitalization and recovering of the transplanted tendon graft can take up to 12 months. Therefore, a device that provides gradually decreasing mechanical properties with a half-time strength of at least 6 months would be desirable. In order to obtain a suitable material, we investigated the degradation kinetics of a variety of different resorbable fibers made of poly(L-lactide) and poly(L-lactide-co-glycolide). The fiber materials differed in processing and treatment parameters like thermal posttreatment, irradiation, and fiber diameter. The fibers were degraded in vitro and were tested for mechanical properties and molecular weight at various time points up to 72 weeks. The half-time strength of the materials ranged between 5 and 64 weeks, depending on their treatment parameters. In contrast, the stiffness did not decrease adequately. However, an augmentation stiffness that does not change much versus time could not provide a gradual increase in graft load, which is important to stimulate the orientation of the collagenous tissue. Therefore, design of an augmentation construct braided out of more than one quickly degrading fiber materials is suggested. After the breakdown of the faster-degrading fiber components the stiffness would automatically decrease by the diminution of the load-carrying fiber volume.
Adult adipose-derived stem cells (ASCs) are considered to be an alternative cell source for cell-based cartilage repair because of their multiple differentiation potentials. This article addresses the chondrogenic differentiation of ASCs seeded into poly-lactide-co-glycolide (PLGA) scaffolds after implantation in a subcutaneous pocket of nude mice. Human ASCs were seeded into PLGA (polylactic acid:polyglycolic acid = 90:10) scaffolds and cultured in transforming growth factor beta 1 (TGF-beta1)-containing medium for 3 weeks in vitro. Then specimens were implanted into a subcutaneous pocket of severe combined immunodeficiency mice and harvested after 8 weeks. Chondrospecific messenger RNA (mRNA) expression was analyzed using reverse transcriptase polymerase chain reaction. Corresponding extracellular matrix (ECM) synthesis was demonstrated using immunohistochemical staining. Chondrospecific marker molecules such as collagen type II and type X, cartilage oligomeric matrix protein, and aggrecan subsequently increased during the 3 weeks period in vitro. After a further 8 weeks, in vivo samples pretreated with TGF-beta1 continued expressing collagen type II and aggrecan mRNA, and collagen type II was found within the ECM using immunohistochemistry. Chondrospecific mRNA was not detected in control samples. ASC-seeded PLGA scaffolds express a stable chondrogenic phenotype in a heterotopic model of cartilage transplantation and represent a suitable tool for tissue engineering of cartilage.
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