Adipose tissue regeneration in a 3D-printed poly(lactic acid) frame-supported space in the inguinal region of rats.

BACKGROUND: Adipose tissue engineering has been studied as an alternative to current options for breast reconstruction, such as lipofilling, flap reconstruction, and silicone implants. Previously, we demonstrated that a poly(L-lactic acid) mesh containing a collagen sponge, containing neither cells nor growth factors, could be filled with the regenerated adipose tissues when implanted in rodent models. However, the main factor contributing to adipogenesis remained unclear. OBJECTIVE: We aimed to clarify whether adipogenesis can be achieved by the space provided by the mesh or by the bioactivity of collagen. METHODS: A three-dimensional (3D) poly(lactic acid) (PLA) frame, which was stiff enough to maintain its shape, was fabricated by 3D printing. The frame with (PLA+ColI) or without (PLA only) a type I collagen hydrogel was implanted in the inguinal region of rats for up to 12 months. Adipose tissue regeneration in the PLA only and PLA+ColI groups was evaluated histologically. RESULTS: The 3D PLA frame maintained its structure for 12 months in vivo and oil red O (ORO)-positive adipose tissues were regenerated in the frame. No significant difference in the ORO-positive area was detected between the PLA only and PLA+ColI groups. CONCLUSION: The space supported by the frame was a key factor in adipogenesis in vivo.

Anti-platelet adhesion and in situ capture of circulating endothelial progenitor cells on ePTFE surface modified with poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) and hemocompatible peptide 1 (HCP-1).

We recently reported in vitro suppression of platelet adhesion on expanded polytetrafluoroethylene (ePTFE) by surface grafting of poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC). However, this may be inadequate for long-term hemocompatibility of blood-contacting biomaterials, and it has led us to develop a strategy of circulating mononuclear cell-capture. ePTFE was treated with argon (Ar) plasma, and grafted with 2-methacryloyloxyethyl phosphorylcholine (MPC) and methacrylic acid (MAA), by glycidyl methacrylate (GMA)-anchored graft polymerization. Next, it was immobilized with integrin α4ß1-positive circulating blood cell-specific peptides, i.e., the traditional arginine-glutamic acid-aspartic acid-valine (REDV), and our original hemocompatible peptide-1 (HCP-1). Both the surfaces retained the anti-platelet property just like the PMPC-grafted surface, and revealed considerable affinity to human umbilical vein endothelial cells (HUVEC), which is a well-known in vitro integrin α4ß1-positive model. Better HUVEC spreading and proliferation was also confirmed, in terms of the cell extension property. Since coagulation and endothelialization on the materials compete in the body, they cannot be properly evaluated separately, in vitro. They were assessed by using an in situ porcine closed-circuit system for 18 h in the present study. Our findings suggest that poly(MPC-co-MAA) is a great ePTFE surface modifier, exhibiting good hemocompatibility in association with REDV/HCP-1 immobilization, which suppresses anti-platelet adhesion and enhances circulating cell capture simultaneously.

De novo adipogenesis using a bioabsorbable implant without additional cells or growth factors.

Adipose tissue regeneration in breast cancer patients without additional growth factors or adipose-tissue-derived stromal cells is desirable because of the possibility of recurrence and metastasis. We report that a poly-L-lactic acid (PLLA) mesh implant containing a collagen sponge (CS) maintained the internal space in vivo for up to 12 months and substituted for adipose tissue. We developed a PLLA capsule that maintained the internal space longer than that of PLLA mesh and compared adipose tissue formation at 12 and 24 months after implantation between the PLLA mesh with CS implant and the PLLA capsule implant with or without CS in a rabbit model. After 12 months, all implants maintained the internal space, and the adipose tissue that formed in all implant groups was larger than that in the control group. At 24 months, PLLA mesh maintained the internal space just as well as that at 12 months, while the PLLA capsule collapsed and accumulated a large number of macrophages. The formed adipose tissue in the PLLA mesh group was maintained up to 24 months; however, those in two PLLA capsule groups decreased and showed no difference from the control group. In conclusion, the internal space of the PLLA mesh implant with CS was substituted for adipose tissue at 12 months and sustained the formed adipose tissue after 24 months. The PLLA mesh implant containing CS is a desirable bioabsorbable implant that can be replaced by autologous adipose tissue after implantation in vivo without using any growth factors or cells.

Short-term evaluation of thromboresistance of a poly(ether ether ketone) (PEEK) mechanical heart valve with poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC)-grafted surface in a porcine aortic valve replacement model.

Improved thromboresistance of mechanical valves is desired to decrease the risk of thromboembolism and thrombosis and reduce the dosage of anticoagulation with a vitamin K antagonist (e.g., warfarin). For several mechanical valves, design-related features are responsible for their improved thromboresistance. However, it remains unclear whether material-related features provide a practical level of thromboresistance to mechanical valves. Here, we studied the effect of a bileaflet valve made of poly(ether ether ketone) (PEEK) with a poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC)-grafted surface (PEEK-g-PMPC). PMPC is a well-known thromboresistant polymeric material. A short-term (<26 h) porcine aortic valve replacement model using neither an anticoagulant nor an antiplatelet agent showed that the PEEK-g-PMPC valve opened and closed normally with an allowable transvalvular gradient. Unlike an untreated PEEK valve, no thrombus formed on the PEEK-g-PMPC valves on gross anatomy examination in addition to the absence of traveled thrombi in the kidney and lung tissues. Material (PEEK-g-PMPC)-related thromboresistance appeared to decrease the risk of thromboembolism and thrombosis for patients with mechanical valves. However, thromboresistance of the PEEK-g-PMPC valve requires improvement because fibrous fouling was still observed on the leaflet. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1052-1063, 2019.

A surface graft polymerization process on chemically stable medical ePTFE for suppressing platelet adhesion and activation.

An effective surface grafting method for chemically inert and elaborately porous medical expanded-polytetrafluoroethylene (ePTFE) was developed. Although surface graft polymerization onto basic polymeric biomaterials has been widely studied, successful modification of the ePTFE surface has been lacking due to its high chemical resistance. Herein, we succeeded in surface graft polymerization onto ePTFE through glycidyl methacrylate (GMA) as a bridge linkage following argon (Ar) plasma treatment. The epoxy group of GMA was expected to react with the peroxide groups produced on ePTFE by Ar plasma exposure, and its methacrylic groups can copolymerize with various monomers. In the present study, we selected 2-methacryloyloxyethyl phosphorylcholine (MPC) as a model monomer and the blood compatibility of modified ePTFE was evaluated. Two sequences of surface grafting were compared. In a two-step graft polymerization, GMA was first immobilized onto Ar plasma treated ePTFE, and then MPC was polymerized. In a one-step graft copolymerization, MPC and GMA were mixed and copolymerized simultaneously onto Ar plasma treated ePTFE, resulting in a poly(MPC-co-GMA) (PMG) graft surface. The roughness of the node-and-fibril structure of ePTFE was reduced by the uniform polymer layer, and the modified ePTFE had a good hydrophilic nature even after being stored in an aqueous environment for 30 days. The indispensable GMA in graft polymerization improved the surface grafting on ePTFE. The one-step and two-step graft polymerization methods could decrease the number of adhered platelets, and almost inhibit platelet activation. We concluded that graft polymerization with the GMA linker provides a novel strategy to modify the chemically inert ePTFE surfaces for functionalizing as new medical devices.

Recombinant production and film properties of full-length hornet silk proteins.

Full-length versions of the four main components of silk cocoons of Vespa simillima hornets, Vssilk1-4, were produced as recombinant proteins in Escherichia coli. In shake flasks, the recombinant Vssilk proteins yielded 160-330mg recombinant proteinl(-1). Films generated from solutions of single Vssilk proteins had a secondary structure similar to that of films generated from native hornet silk. The films made from individual recombinant hornet silk proteins had similar or enhanced mechanical performance compared with films generated from native hornet silk, possibly reflecting the homogeneity of the recombinant proteins. The pH-dependent changes in zeta (ζ) potential of each Vssilk film were measured, and isoelectric points (pI) of Vssilk1-4 were determined as 8.9, 9.1, 5.0 and 4.2, respectively. The pI of native hornet silk, a combination of the four Vssilk proteins, was 4.7, a value similar to that of Bombyx mori silkworm silk. Films generated from Vssilk1 and 2 had net positive charge under physiological conditions and showed significantly higher cell adhesion activity. It is proposed that recombinant hornet silk is a valuable new material with potential for cell culture applications.

Effect of RGDS-expressing fibroin dose on initial adhesive force of a single chondrocyte.

Initial chondrocyte-material interactions are important for cell behaviors such as proliferation, phenotypic expression and matrix synthesis. Previously, we showed that chondrocytes cultured in/on silk fibroin scaffolds proliferate without dedifferentiating into fibroblast-like cells and that RGDS sequences genetically interfused in the fibroin light chain protein enhance cartilage tissue formation. In the present study, the adhesive force of chondrocytes was measured on fibroin substrates containing RGDS-expressing fibroin molecules produced by transgenic silkworms at the different densities of 0, 0.6, 1.5 and 3.0 mol%. The degree of chondrocyte attachment to fibroin substrates increased with the number of RGDS-expressing fibroin molecules. Moreover, the adhesive force per unit spreading area of a single cultured chondrocyte exhibited a peak that was higher with increased RGDS concentrations. The results of this study indicate that the RGDS sequences genetically interfused in the fibroin light chain protein exert effects on chondrocytes' adhesive behavior and can enhance cartilage tissue organization.