Kaarlo Paakinaho

Effects of lactide monomer on the hydrolytic degradation of poly(lactide-co-glycolide) 85L/15G

The hydrolytic degradation of oriented poly(L-lactide-co-glycolide) 85 L/15 G (PLGA 85/15) sample materials with various amounts of lactide monomer was monitored in vitro at 37 °C. The materials were manufactured from medical grade PLGA 85/15 by a two-step melt extrusion-die drawing process. Results showed that the hydrolytic degradation rate depended highly on the lactide monomer content, which in turn influenced the retention of mechanical properties, mass loss, crystallinity, and dimensional stability. Even small quantities of lactide monomer (0.05-0.20 wt%) affected especially the retention of mechanical properties, which started to decline rapidly upon the inherent viscosity reaching 0.6-0.8 dl/g due to hydrolytic degradation. Based on our hydrolytic degradation data, we constructed a simplified mathematical model of degradation-related strength retention and recommend it as a functional quality control tool for melt-processed biodegradable medical devices manufactured from poly(L-lactide-co-glycolide) 85 L/15 G.

Human Adipose Stem Cells Differentiated on Braided Polylactide Scaffolds Is a Potential Approach for Tendon Tissue Engineering

Growing number of musculoskeletal defects increases the demand for engineered tendon. Our aim was to find an efficient strategy to produce tendon-like matrix in vitro. To allow efficient differentiation of human adipose stem cells (hASCs) toward tendon tissue, we tested different medium compositions, biomaterials, and scaffold structures in preliminary tests. This is the first study to report that medium supplementation with 50 ng/mL of growth and differentiation factor-5 (GDF-5) and 280 μM l-ascorbic acid are essential for tenogenic differentiation of hASCs. Tenogenic medium (TM) was shown to significantly enhance tendon-like matrix production of hASCs compared to other tested media groups. Cell adhesion, proliferation, and tenogenic differentiation of hASCs were supported on braided poly(l/d)lactide (PLA) 96l/4d copolymer filament scaffolds in TM condition compared to foamed poly(l-lactide-co-ɛ-caprolactone) (PLCL) 70L/30CL scaffolds. A uniform cell layer formed on braided PLA 96/4 scaffolds when hASCs were cultured in TM compared to maintenance medium (MM) condition after 14 days of culture. Furthermore, total collagen content and gene expression of tenogenic marker genes were significantly higher in TM condition after 2 weeks of culture. The elastic modulus of PLA 96/4 scaffold was more similar to the elastic modulus reported for native Achilles tendon. Our study showed that the optimized TM is needed for efficient and rapid in vitro tenogenic extracellular matrix production of hASCs. PLA 96/4 scaffolds together with TM significantly stimulated hASCs, thus demonstrating the potential clinical relevance of this novel and emerging approach to tendon injury treatments in the future.

Programmed water-induced shape-memory of bioabsorbable poly(d,l-lactide): activation and properties in physiological temperature

This study reports of the novel water-induced shape-memory of bioabsorbable poly(d,l-lactide). We have developed an orientation-based programming process that generates an ability for poly(d,l-lactide) to transform its shape at 37°C in an aqueous environment without external energy and to adapt to a predefined stress level by stress generation or relaxation. In this orientation-programming process, polymer material is deformed and oriented at an elevated temperature and subsequently cooled down while retaining its deformed shape, tension, and polymer chain entanglements. At body temperature and in an aqueous environment, the shape-memory is activated by the plasticizing effect of water molecules diffused into the polymer matrix causing an entropy-driven directed relaxation of oriented and preloaded polymer chains. This plasticizing effect is clearly seen as a decrease of the onset glass transition temperature by 10–13°C. We found that γ-irradiation used for sterilizing the orientation-programmed materials strongly affected the shape-recovery rate, but not the recovery ratio. Both non-γ-irradiated and γ-irradiated sample materials showed excellent shape-recovery ratios during a ten-week test period: 94 and 97%, respectively. The orientation-programmed materials generated a predefined load in a 37°C aqueous environment when their shape-recovery was restricted, but when external tension was applied to them, they adapted to the predefined level by stress relaxation. Our results show that functionality in terms of shape-memory can be generated in bioabsorbable polymers without tailoring the polymer chain structure thus shortening the time from development of technology to its utilization in medical devices.

In Vitro Degradation of Borosilicate Bioactive Glass and Poly(l-lactide-co-ε-caprolactone) Composite Scaffolds

Composite scaffolds were obtained by mixing various amounts (10, 30 and 50 weight % [wt %]) of borosilicate bioactive glass and poly(l-lactide-co-ε-caprolactone) (PLCL) copolymer. The composites were foamed using supercritical CO2. An increase in the glass content led to a decrease in the pore size and density. In vitro dissolution/reaction test was performed in simulated body fluid. As a function of immersion time, the solution pH increased due to the glass dissolution. This was further supported by the increasing amount of Ca in the immersing solution with increasing immersion time and glass content. Furthermore, the change in scaffold mass was significantly greater with increasing the glass content in the scaffold. However, only the scaffolds containing 30 and 50 wt % of glasses exhibited significant hydroxyapatite (HA) formation at 72 h of immersion. The compression strength of the samples was also measured. The Young’s modulus was similar for the 10 and 30 wt % glass-containing scaffolds whereas it increased to 90 MPa for the 50 wt % glass containing scaffold. Upon immersion up to 72 h, the Young’s modulus increased and then remained constant for longer immersion times. The scaffold prepared could have great potential for bone and cartilage regeneration.

µCT Based Characterization of Biomaterial Scaffold Microstructure Under Compression

Scaffolds are often designed with progressive degradation to make way for cell proliferation of seeded cells for native tissue. The viability of the scaffold has been shown to depend on, among other things, the microstructure. Common parameters, that are used to describe microstructure, are porosity, material thickness, pore size and surface area. These properties quantify the suitability of the scaffold as a substrate for cell adhesion, fluid exchange and nutrient transfer. Bone and cartilage scaffolds are often placed or operated under loads (predominantly compression). This can alter the structural parameters depending on the stiffness of the scaffold and applied deformation. It is important to know, how scaffolds’ parameters change under deformation. In this study, two scaffolds (PLCL-TCP and collagen-PLA) intended for use in bone and cartilage applications, were studied through micro computed tomography based imaging and in situ mechanical testing. The scaffolds were subjected to uniaxial compressive deformation up to 50% of the original size. The corresponding changes in the individual scaffold bulk characteristics were analyzed. Our results show an expected decrease in porosity with increasing deformation (with PLCL-TCP scaffold 52% deformation resulted in 56% decrease in porosity). Especially in the sandwich constructs of collagen-PLA, but also in PLCL-TCP composites, it was evident that different materials are affected differently which may be of significance in applications with mechanical loading. Our results are a step towards understanding the changes in the structure of these scaffolds under loading.

Porous poly-l-lactide-co-ɛ-caprolactone scaffold: a novel biomaterial for vaginal tissue engineering

The surgical reconstruction of functional neovagina is challenging and susceptible to complications. Therefore, developing tissue engineering-based treatment methods for vaginal defects is important. Our aim was to develop and test a novel supercritical carbon dioxide foamed poly-l-lactide-co-ɛ-caprolactone (scPLCL) scaffold for vaginal reconstruction. The scaffolds were manufactured and characterized for porosity (65 ± 4%), pore size (350 ± 150 µm) and elastic modulus (2.8 ± 0.4 MPa). Vaginal epithelial (EC) and stromal cells (SC) were isolated, expanded and characterized with flow cytometry. Finally, cells were cultured with scPLCL scaffolds in separate and/or co-cultures. Their attachment, viability, proliferation and phenotype were analysed. Both cell types strongly expressed cell surface markers CD44, CD73 and CD166. Strong expression of CD326 was detected with ECs and CD90 and CD105 with SCs. Both ECs and SCs attached and maintained viability on scPLCL. Further, scPLCL supported the proliferation of especially ECs, which also maintained epithelial phenotype (cytokeratin expression) during 14-day assessment period. Interestingly, ECs expressed uroplakin (UP) Ia, UPIb and UPIII markers; further, UPIa and UPIII expression was significantly higher on ECs cultured on scPLCL than on cell culture plastic. In conclusion, the scPLCL is potential scaffold for vaginal tissue engineering and the results of this study further illustrate the excellent biocompatibility of PLCL.

Novel osteoconductive β-tricalcium phosphate/poly(L-lactide-co-e-caprolactone) scaffold for bone regeneration: a study in a rabbit calvarial defect

AbstractThe advantages of synthetic bone graft substitutes over autogenous bone grafts include abundant graft volume, lack of complications related to the graft harvesting, and shorter operation and recovery times for the patient. We studied a new synthetic supercritical CO2 –processed porous composite scaffold of β-tricalcium phosphate and poly(L-lactide-co-caprolactone) copolymer as a bone graft substitute in a rabbit calvarial defect. Bilateral 12 mm diameter critical size calvarial defects were successfully created in 18 rabbits. The right defect was filled with a scaffold moistened with bone marrow aspirate, and the other was an empty control. The material was assessed for applicability during surgery. The follow-up times were 4, 12, and 24 weeks. Radiographic and micro-CT studies and histopathological analysis were used to evaluate new bone formation, tissue ingrowth, and biocompatibility. The scaffold was easy to shape and handle during the surgery, and the bone-scaffold contact was tight when visually evaluated after the implantation. The material showed good biocompatibility and its porosity enabled rapid invasion of vasculature and full thickness mesenchymal tissue ingrowth already at four weeks. By 24 weeks, full thickness bone ingrowth within the scaffold and along the dura was generally seen. In contrast, the empty defect had only a thin layer of new bone at 24 weeks. The radiodensity of the material was similar to the density of the intact bone. In conclusion, the new porous scaffold material, composed of microgranular β-TCP bound into the polymer matrix, proved to be a promising osteoconductive bone graft substitute with excellent handling properties.