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

Dilcele Silva Moreira Dziedzic,Júlio César Francisco,Priscila Elias Ferreira Stricker,Bassam Felipe Mogharbel,Juliana Floriano,Ana Carolina Irioda,Katherine Athayde Teixeira De Carvalho,Lúcia De Noronha,Célia Regina Cavichiolo Franco,Eltyeb Abdelwahid

doi:10.1007/s00223-020-00793-1

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.

Highlights

Bone presents continuous remodeling and a substantial regenerative capacity compared to other tissues
The decellularization process with 0.1% SDS was effective in removing the cells and the innermost epithelial layer, which lies adjacent to the fetus (Fig. 1a)
The Decellularized Human Amniotic Membrane (DAM) preserved its structural integrity during the decellularization, and throughout the cell culture with regular medium, and both osteoinduction media

Summary

Introduction

Bone presents continuous remodeling and a substantial regenerative capacity compared to other tissues. Lifelong bone remodeling is responsible for skeletal development, responses to mechanical stimuli, and maintaining mineral homeostasis. Intrinsic regeneration capacity guarantees bone integrity as an injury repair process. Moderate-sized bone defects repair without the need for a graft. Complex clinical conditions require a considerable amount of bone, in which natural bone regeneration capacity is not sufficient to establish functional tissue recovery. Bone engineering strategies may be necessary to guide or accelerate

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Conclusion