A New Osteogenic Membrane to Enhance Bone Healing: At the Crossroads between the Periosteum, the Induced Membrane, and the Diamond Concept

Abstract

The lack of viability of massive bone allografts for critical-size bone defect treatment remains a challenge in orthopedic surgery. The literature has reviewed the advantages of a multi-combined treatment with the synergy of an osteoconductive extracellular matrix (ECM), osteogenic stem cells, and growth factors (GFs). Questions are still open about the need for ECM components, the influence of the decellularization process on the latter, the related potential loss of function, and the necessity of using pre-differentiated cells. In order to fill in this gap, a bone allograft surrounded by an osteogenic membrane made of a decellularized collagen matrix from human fascia lata and seeded with periosteal mesenchymal stem cells (PMSCs) was analyzed in terms of de-/recellularization, osteogenic properties, PMSC self-differentiation, and angiogenic potential. While the decellularization processes altered the ECM content differently, the main GF content was decreased in soft tissues but relatively increased in hard bone tissues. The spontaneous osteogenic differentiation was necessarily obtained through contact with a mineralized bone matrix. Trying to deepen the knowledge on the complex matrix–cell interplay could further propel these tissue engineering concepts and lead us to provide the biological elements that allow bone integration in vivo.

Collagen molecular organization preservation in human fascia lata and periosteum after tissue engineering

“…We did not investigate irradiated periosteum since the priority, for the creation of the new regenerative membrane, was the use of irradiated decellularized HFL ( Manon et al, 2023 ).…”

“…Each protocol ended with a 12 h DNase treatment. D5 protocol is the one routinely used in the institutional tissue bank ( van Steenberghe et al, 2017 ; Manon et al, 2023 ). In brief, it includes sequential immersion in pure acetone to degrease, ether to neutralize the previous compounds and 70° ethanol to further degrease.…”

“…The DNA content was measured using the Quant-iT PicoGreen DNA assay kit (ThermoFisher, Waltham, MA, United States) just after being extracted using the DNeasy ® Blood & Tissue kit (Qiagen, Hilden, Germany). The very precise steps are previously described ( Manon et al, 2022 ; Manon et al, 2023 ). Samples were then stored at −80°C.…”

“…Bone repair therefore remains a major concern for orthopedic surgeons ( Evans, Chang, and Knothe Tate, 2013 ; Delloye et al, 2014 ; Dalisson et al, 2021 ) which has led to the emergence of a series of alternative tissue engineering approaches in recent decades ( Chen et al, 2015 ; Lou et al, 2021 ; Manon et al, 2023 ). Among these, Manon et al recently showed the suitability of using human fascia lata matrices as a periosteal-like scaffold for bone regeneration, mainly through their collagen type I composition and appropriate mechanical properties ( Manon et al, 2022 ; Manon et al, 2023 ). The human fascia lata (HFL) is a dense connective tissue made up of three layers of collagen fiber bundles with little cellularity ( Pancheri et al, 2014 ; Blottner et al, 2019 ).…”

“…In the present study harvested human periosteum (HP) and HFL samples were treated with several protocols of decellularization ( Manon et al, 2023 ) as well as by irradiation. Whereas the general microstructure, composition, and biosafety of the ECM-derived scaffold had received considerable attention, a special analysis of collagen per se is required as the main component contributing to matrix-cell crosstalks and, hence, to the global integrity of the periosteum.…”