Combined Biomaterials: Amniotic Membrane and Adipose Tissue to Restore Injured Bone as Promoter of Calcification in Bone Regeneration: Preclinical Model

Abstract

Discarded tissues, like human amniotic membranes and adipose tissue, were investigated for the application of Decellularized Human Amniotic Membrane (DAM) as a viable scaffold for transplantation of Adipose-derived stromal cells (ASCs) in bone regeneration of non-healing calvarial defects in rats. Amniotic membrane was decellularized to provide a scaffold for male Wistar rats ASCs expansion and transplantation. ASCs osteoinduction in vitro promoted the deposition of a mineralized bone-like matrix by ASCs, as calcified globular accretions associated with the cells on the DAM surface and inside the collagenous matrix. Non-healing calvarial defects on male Wistar rats were randomly divided in control without treatment, treatment with four layers of DAM, or four layers of DAM associated with ASCs. After 12 weeks, tissue blocks were examined by micro-computed tomography and histology. DAM promoted osteoconduction by increasing the collagenous matrix on both DAM treatments. DAM with ASCs stimulated bone deposition, demonstrated by a higher percentage of bone volume and trabecular bone number, compared to control. Besides the osteogenic capacity in vitro, ASCs stimulated the healing of calvarial defects with significant DAM graft incorporation concomitant with higher host bone deposition. The enhanced in vivo bone regeneration by undifferentiated ASCs loaded onto DAM confirmed the potential of an easily collected autologous cell source associated with a broadly available collagenous matrix in tissue engineering.

Adipose-Derived Stromal Cells and Mineralized Extracellular Matrix Delivery by a Human Decellularized Amniotic Membrane in Periodontal Tissue Engineering

“…As an interfacial matrix between old and new bone, the presence of the reversal line demarcated the pattern of bone deposition in the furcation defect and demonstrated the osteoconduction of the amniotic membrane collagenous matrix. The collagenous niches for osteogenesis were confirmed between the DAM folds and between loose collagenous fibers of the DAM, as observed previously in multilayered DAM [32].…”

“…DAM has been investigated in vivo for guided bone regeneration [ 28 ], assessing different amniotic membrane preservation methods [ 29 ]. The association of DAM with different cell sources has been examined in bone defects with periodontal ligament cells [ 30 ], osteoblasts [ 31 ], and ASCs [ 32 , 33 ].…”

Use of Amniotic Membrane and Its Derived Products for Bone Regeneration: A Systematic Review

“…No statistical difference between AM and seeded AM was evidenced in this study. Dziedzic et al showed a significant higher bone formation using seeded AM compared the defect left empty (Dziedzic et al, 2021 ). However, they also failed to evidence significant difference between AM and seeded AM in contrast with the study of Semyari et al which observed an earlier bone regeneration when rabbit ADSCs-seeded AM was implanted in a rabbit calvarial bone defect (Semyari et al, 2016 ).…”

“…Most of the included studies pointed that AM is mainly applied as a single layer. Few studies suggested to create a multilayered AM (Li et al, 2015 ; Dziedzic et al, 2021 ). This might enhance its thickness that initially ranges from 0.02 to 0.5 mm (Bourne, 1962 ).…”

Periosteal Tissue Engineering: Current Developments and Perspectives

“…Discarded tissues like amniotic membrane are easier to obtain than periosteum ECM. [ 67 ] Amniotic membrane is the innermost layer of the placenta, and it contains angiogenic growth factors, cytokines, and other active substances. Ghanmi et al.…”