Gold and Hydroxyapatite Nano-Composite Scaffolds for Anterior Cruciate Ligament Reconstruction: In Vitro Characterization

Abstract

Current anterior cruciate ligament (ACL) graft replacement materials often fail due to the lack of biological integration. While many newly developed extracellular matrix based scaffolds show good biocompatibility they often do not entice cellular remodeling and the rebuilding of a functional ligament. We have proposed the conjugation of gold nanoparticles (AuNP) and hydroxyapatite nanoparticles (nano-HAp) to acellular tissue to enhance cell attachment and proliferation while maintaining an improved degradation resistance and open microstructure. We are the first to investigate the double conjugation of AuNP and nano-HAp onto decellularized tissue to improve the tissue remodeling response. Decellularized porcine diaphragm was crosslinked with two types of nano-HAp and amine-functionalized AuNP with 1-ethyl-3-(3-dimethlaminopropyl) carbodiimide (EDC) crosslinker. Scaffolds were characterized using electron microscopy, differential scanning calorimetry, and fibroblast assays. Results demonstrated that scaffolds with nano-HAp have increased thermal stability at low levels of crosslinking. The open microstructure of the scaffold was not compromised allowing for cell migration while still providing increased degradation resistance. The addition of < 200 nm nano-HAp decreased cell viability compared to scaffolds without nanoparticles, but the addition of AuNP to scaffolds showed enhanced cell viability in the presence of < 200 nm nano-HAp. The addition of < 40 nm nano-HAp showed an increase in cell viability compared to scaffolds crosslinked without nanoparticles. It is concluded that attaching AuNP and < 40nm nano-HAp to extracellular matrices may improve overall properties.

Preclinical Development of Bioengineered Allografts Derived from Decellularized Human Diaphragm

“…In this work, human diaphragm harvested from cadavers was considered as an ideal, but still little investigated, source for the development of decellularized biological scaffolds for VML treatment. To the best of our knowledge, only 13 works in the literature have reported the decellularization of the diaphragmatic muscle [27][28][29][30][31][32]34,[39][40][41][42][43][44], plus 5 research articles that considered the diaphragm together with other tissues for methodological studies about decellularization procedures or analysis techniques validating acellular ECM quality [33,[35][36][37][38] (Table S1).…”

“…In particular, human diaphragmatic skeletal muscle may represent a favourable tissue source to obtain biological scaffolds with myogenic potential. The decellularization of mouse [27][28][29][30], rat [31][32][33][34][35][36][37][38], bovine [39,40], and pig [41][42][43] diaphragm was previously reported to fabricate biological prosthesis mainly intended for orthotopic implants in the regeneration of diaphragmatic defects [27,28,32], including neonatal congenital hernia [29][30][31]34,43]. Regarding different clinical targets, the development of decellularized diaphragmatic muscle was also conceptualised for chronic wounds treatment [41], abdominal wall repair [39,40], and anterior cruciate ligament reconstruction [42].…”

“…The decellularization of mouse [27][28][29][30], rat [31][32][33][34][35][36][37][38], bovine [39,40], and pig [41][42][43] diaphragm was previously reported to fabricate biological prosthesis mainly intended for orthotopic implants in the regeneration of diaphragmatic defects [27,28,32], including neonatal congenital hernia [29][30][31]34,43]. Regarding different clinical targets, the development of decellularized diaphragmatic muscle was also conceptualised for chronic wounds treatment [41], abdominal wall repair [39,40], and anterior cruciate ligament reconstruction [42]. Interestingly, Davari and collaborators firstly reported the preclinical investigation of decellularized human hemidiaphragm for large defect reconstruction of the muscle [44].…”

Transforming Wound Management: Nanomaterials and Their Clinical Impact

“…Scaffolds crosslinked with varying concentrations of AgNPs and two different types of nano-HAp were characterized for surface microstructure, thermal properties, cell proliferation, and cell viability. Adding gold nanoparticles and nano-HAp to scaffolds was hypothesized to increase cell proliferation and viability compared to non-nanoparticulated scaffolds [ 114 ].…”

Enhancement of in vitro proliferation and bioactivity of human anterior cruciate ligament fibroblasts using an in situ tissue isolation method and basic fibroblast growth factor culture conditions

“…The ability of the human ACL to self-repair is weaker than other soft tissues due to the poor tissue-blood supply. [1] Three lakhs (300,000) patients have newly acquired ACL injuries, and the overwhelming majority of these patients will require ACL reconstruction surgery. [2–4]…”