Abstract
The capacity to study the deposition of mineral within a hydrogel structure is of significant interest to a range of therapies that seek to replace the hard tissues and the hard-soft tissue interface. Here, a method is presented that utilises Confocal Raman microscopy as a tool for monitoring mineralisation within hydrogels. Synthetic hard-soft material interfaces were fabricated by apposing brushite (a sparingly soluble calcium phosphate) and biopolymer gel monoliths. The resulting structures were matured over a period of 28 days in phosphate buffered saline. Confocal Raman microscopy of the interfacial region showed the appearance of calcium phosphate salt deposits away from the original interface within the biopolymeric structures. Furthermore, the appearance of octacalcium phosphate and carbonated hydroxyapatite was observed in the region of the brushite cement opposing the biopolymer gel. This study describes not only a method for analysing these composite structures, but also suggests a method for recapitulating the graduated tissue structures that are often found in vivo.
Highlights
Interfacial tissue engineering aims to regenerate the intricate structure of musculoskeletal interfaces.[1]
The TruFit® Bone Graft Substitute (BGS) has a chondral region manufactured from polyglycolic acid and a bone region consisting of porous poly-D,L-lactide-co-glycolide (PGLA) and calcium sulphate whereas the Osseofit® consists of a type I collagen chondral region and a porous tricalcium phosphate–polylactic acid mix underneath.[18]
We have reported the creation of a whole bone-ligament-bone construct from a cell-seeded hydrogel and artificial bone blocks manufactured from a bioresorbable calcium phosphate cement.[21,22,23]
Summary
Interfacial tissue engineering aims to regenerate the intricate structure of musculoskeletal interfaces.[1]. At the junction between the soft tissue sinews tendon/ligament and bone (the osteotendinous junction), the interface is comprised of four interconnected regions, namely the tendon/ ligament proper, unmineralised fibrocartilage, mineralised fibrocartilage and bone.[3,4,5] the junction between bone and cartilage (the osteochondral junction) has a multiphasic structure, with 5 different regions; bone, mineralised cartilage, and deep, middle and superficial layers of cartilage.[6,7] In both tissue transitions, the gradient of mineralisation, along with changes in cell type, matrix composition and collagen fibre alignment, affects the mechanical properties. Despite their use in the repair of damaged cartilage, it is unknown whether mineral deposition across the
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