Novel biocompatible polymeric blends for bone regeneration: Material and matrix design and development.

摘要

The first part of the work presented in this dissertation is focused on the design and development of novel miscible and biocompatible polyphosphazene-polyester blends as candidate materials for scaffold-based bone tissue engineering applications. Biodegradable polyesters such as poly(lactide-co-glycolide) (PLAGA) are among the most widely used polymeric materials for bone tissue engineering. However, acidic degradation products resulting from the bulk degradation mechanism often lead to catastrophic failure of the structure integrity, and adversely affect biocompatibility both in vitro and in vivo. One promising approach to circumvent these limitations is to blend PLAGA with other macromolecules that can buffer the acidic degradation products with a controlled degradation rate. Biodegradable polyphosphazenes (PPHOS), a new class of biomedical materials, have proved to be superior candidate materials to achieve this objective due to their unique buffering degradation products. A highly practical blending approach was adopted to develop novel biocompatible, miscible blends of these two polymers. In order to achieve this miscibility, a series of amino acid ester, alkoxy, aryloxy, and dipeptide substituted PPHOS were synthesized to promote hydrogen bonding interactions with PLAGA. Five mixed-substituent PPHOS compositions were designed and blended with PLAGA at different weight ratios producing candidate blends via a mutual solvent method. Preliminary characterization identified two specific side groups namely glycylglycine dipeptide and phenylphenoxy that resulted in improved blend miscibility and enhanced in vitro osteocompatibility. These findings led to the synthesis of a mixed-substituent polyphosphazene poly[(glycine ethyl glycinato)1(phenylphenoxy)1phosphazene] (PNGEGPhPh) for blending with PLAGA. Two dipeptide-based blends having weight ratios of PNGEGPhPh to PLAGA namely 25:75 (Matrix1) and 50:50 (Matrix2) were fabricated. Both of the blends were characterized for miscibility, mechanical properties, degradation kinetics, and in vitro osteocompatibility. Primary rat osteoblasts (PRO) isolated from rat calvaria were used to evaluate their in vitro osteocompatibility. The blends were also characterized for in vivo biodegradability and biocompatibility using a rat subcutaneous implantation model.;Successful in vivo scaffold-based tissue regeneration greatly depends on the scaffold material biocompatibility, mechanical stability, and scaffold architecture to promote tissue in-growth. The other part of the work in the dissertation is focused on the development of mechanically competent bioresorbable nano-structured three-dimensional (3D) hiomimetic scaffolds for bone tissue engineering applications. Scaffold material selection was based on achieving improved mechanical stability, in vitro osteoblast performance, and in vivo biocompatibility. A miscible PNGEGPhPh-PLAGA blend system developed and characterized in the first part of the thesis work was chosen to fabricate a nanofiber-based mechanically competent biomimetic scaffold via electrospinning. Due to its versatility, controllability and reproducibility, the technique of electrospinning was adopted to produce blend nanofibers. The polymer solution concentration and electrospinning parameters were optimized to produce blend fibers in the range of 50-500 nm to mimic dimensions of collagen fibrils present in the natural extracellular matrix of native bone. These blend nanofiber matrices supported PRO adhesion, proliferation and showed an elevated phenotype expression compared to PLAGA nanofibers. Orienting electrospun nanofibers in a concentric manner with an open central cavity created a mechanically competent 3D scaffold mimicking the bone marrow cavity, as well as, the lamellar structure of bone. The 3D biomimetic scaffold exhibited a similar characteristic mechanical behavior to that of native bone. Compressive modulus of the scaffold was found to be within the range of human trabecular bone. To our knowledge this is the first mechanically competent 3D electrospun nanofiber scaffold with mechanical properties in the middle range of human trabecular bone. The potential of this scaffold for bone repair was further investigated by monitoring the cellular activity and mechanical performance over time using in vitro culture. This biomimetic scaffold supported the robust PRO growth throughout the scaffold architecture and maintained osteoblast phenotype expression in vitro, which resulted in a similar cell-matrix organization to that of native bone and maintenance of structure integrity. (Abstract shortened by UMI.).